Antimicrobial

Physiol Behav, 2002 Nov, 77(2-3), 291 - 9
Acute stress decreases inflammation at the site of infection . A role for nitric oxide; Campisi J et al.; Exposure to acute stress modulates immune function . Most research regarding stress and immunity has described the deleterious effects of stress . Recent studies, however, indicate that acute stress enhances many features of innate immunity . For example, exposure to acute stress reduced the time required to resolve inflammation produced by subcutaneous injection of streptomycin-killed, benign bacteria . It is unclear if this change in inflammation would be advantageous to the organism if challenged with living, infectious bacteria . Thus, the current experiments examined the effect of acute stressor exposure on inflammation development and resolution after a naturalistic, live bacterial challenge . In addition, nitric oxide (NO), an important bactericidal mediator, was measured at the inflammatory site . Rats (F344) were exposed to acute stress (100, 5-s, 1.6 mA tailshocks) and subcutaneously injected with live Escherichia coli ( approximately 2.5 x 10(9) colony forming units {CFU}) . Stressed rats attained their peak inflammatory size quicker, resolved their inflammation 10-14 days faster, experienced less bacterial-induced weight loss and released 300% greater NO at the inflammatory site than nonstressed controls . Thus, acute stress improved recovery from bacterially induced inflammation possibly due to local elevations in NO.

J Hosp Infect, 2002 Nov, 52(3), 219 - 24
Intra-laboratory reproducibility of the hand hygiene reference procedures of EN 1499 (hygienic handwash) and EN 1500 (hygienic hand disinfection); Kampf G et al.; The bactericidal efficacy of hand antiseptic products is determined in Europe using two norms--EN1499 (hygienic handwash), and EN 1500 (hygienic hand disinfection) based on reducing the counts of bacteria on artificially contaminated hands . Each requires 12-15 data sets per test and comparison with a reference procedure . Recent research using EN 1500 suggested that most alcohol-based hand gels are significantly less effective than the reference alcohol (2-propanol 60%), whereas liquid alcohol-based rubs are not . However concerns about the accuracy and reproducibility of the norm reference procedures have been raised . We therefore analysed 23 experiments carried out using EN 1500 representing 342 hand disinfection procedures, and 12 experiments using EN 1499 representing 178 handwashes, all performed in the same laboratory for reproducibility of the reference procedures . The reference alcohol gave a mean log(10) reduction factor (RF) of 4.64 +/- 0.93; only one data set gave a significantly higher result (5.14, P = 0.034), and one significantly lower (4.05; P = 0.034) . Analysis of all 23 means revealed no significant difference (P = 0.188; ANOVA model) . The reference soap gave a mean reduction of 2.82 +/- 0.49 . Two data sets were significantly higher than this (3.35,P < 0.001; 3.12, P < 0.001) and two significantly lower (2.55, P = 0.031; 2.47,P = 0.004) . Analysis of all the means did reveal a significant difference (P < 0.001, ANOVA model), which is probably explained by the smaller standard deviations of these results . Pre-values (bacteria recovered from fingers before a reference procedure) correlated significantly with RFs for both hand disinfection (correlation coefficient: 0.291;P = 0.01) and handwash (correlation coefficient: 0.372, P = 0.01) . Overall both procedures gave accurate and reproducible results .

Mar Environ Res, 2002 Sep-Dec, 54(3-5), 547 - 51
Bacterial killing by Mytilus hemocyte monolayers as a model for investigating the signaling pathways involved in mussel immune defence; Canesi L et al.; The signaling pathways involved in mussel immune defence were investigated utilizing a model of killing of Escherichia coli by Mytilus galloprovincialis hemocytes in a co-culture setting . In particular, the role played by different mitogen activated protein kinases (MAPKs) and by the production of eicosanoids were investigated utilising specific cell permeant, pharmacological enzyme inhibitors . Hemocyte pretreatment with the p38 MAPK inhibitor SB203580 significantly reduced bacterial killing, whereas PD98059 (an inhibitor of ERK--extracellularly regulated kinase--MAPK activation) had no significant effect . Wortmannin also inhibited bacterial killing, indicating a crucial role for PI3-kinase activation in the immune response . Killing of E . coli was also reduced by inhibitors of both PLA2 and cyclooxygenase activities, indicating that eicosanoid production is involved in mediating the response to bacterial challenge . The results demonstrate that bacterial killing by mussel hemocytes is particularly sensitive to inhibitors of the key steps involved in the transduction of bacterial signals into the host cell . Moreover, these data indicate that the hemocyte bactericidal activity can be suitably utilized not only for identifying the signaling pathways involved in the response to bacterial infection, but also as a potential investigative-toxicology model to test drugs and contaminants for their effect on the overall mussel immune defence.

J Lab Clin Med, 2002 Oct, 140(4), 228 - 35
In vivo interaction of endotoxin and recombinant bactericidal/permeability-increasing protein (rBPI23): hemodynamic effects in a human endotoxemia model; Jellema WT et al.; The cardiovascular derangement that results from the administration of endotoxin in healthy subjects is qualitatively similar to what is observed in patients in septic shock . The biological response to endotoxin is attributed in part to cytokine release . In experimental endotoxemia, recombinant bactericidal/permeability increasing protein (rBPI(23)) has shown a protective effect by binding endotoxin with the subsequent inhibition of the endotoxin-induced cytokine release and of neutrophil activation . In a controlled, blinded crossover study the early cardiovascular effects of rBPI(23) were investigated in an experimental endotoxemia model in humans . The beat-to-beat changes in arterial pressure and cardiac output following infusion of endotoxin (40 EU/kg body weight) and rBPI(23) (1 mg/kg) or placebo (human serum albumin, 0.2 mg/kg) were studied for 2 hours in 8 healthy male adults . Endotoxin or rBPI(23) alone did not induce significant cardiovascular changes . Endotoxin following rBPI(23) infusion elicited a fall in total peripheral resistance with its nadir after 4 minutes to 40% (range 16-53; P <.001) of control level . Mean arterial pressure showed little change, and the fall in total peripheral resistance was associated with a reflex increase in heart rate and cardiac output (32%; range 43-106) . Changes in cardiovascular variables in the subsequent 2 hours were not significant . In vitro activation of the contact system by, respectively, rBPI(23), LPS, and LPS-rBPI(23) complexes was assessed . Following incubation with rBPI(23), LPS, and LPS-rBPI(23) complexes, complex levels were generated at levels comparable to those observed in the buffer control . The rapid vasodilatation by endotoxin administered concomitantly with rBPI(23) is not mediated by complement or contact system activation . The early vasodilatation is compensated by an increase in cardiac output, which therefore does not result in arterial hypotension . The monitoring of continuous cardiac output allows for the detection of rapid effects on systemic flow and conductance that go unnoticed in a recording of arterial pressure.

Antimicrob Agents Chemother, 2002 Nov, 46(11), 3637 - 40
Erythromycin resistance in Borrelia burgdorferi; Terekhova D et al.; Susceptibility testing of laboratory strains and clinical isolates of Borrelia burgdorferi indicates that resistance to erythromycin is present in them . Evaluation of the MICs, minimal bactericidal concentrations, and kinetics of bacterial killing of erythromycin suggests that this resistance is increased by preexposure to the antibiotic, is dependent on inoculum size, and may be the result of selection of subpopulations of bacterial cells with increased resistance.

Otolaryngol Pol, 2002, 56(4), 409 - 13
A newly discovered function of palatine tonsils in immune defence: the expression of defensins; Weise JB et al.; The palatine tonsils have an undoubted role in the immune defence system . After antigen contact an effective adaptive immune response by B- and T-cell lymphocytes will be released . In addition the palatine tonsils seem to exert influence to the defence by the innate immune system . Therefore, we studied the ability of palatine tonsils to express different alpha and beta defensins and to find out any distinctions in chronic inflamed tonsils . Total RNA of 49 specimens of hyperplastic tonsils and chronic tonsillitis with pathological provided evidence of Actinomyces israelii was isolated using TRIzol protocol, reverse transcribed and the HNP-1, HNP-4, HBD-1 and HBD-2 gene expression densitometric determined, standardised in relation to glycerinaldehyd-3-phosphatdehydrogenase gene expression, after a semiquantitative polymerase chain reaction was performed . mRNA of HNP-1, HNP-4, HBD-1 and HBD-2 was detected in tissue samples, but their amount differed within the two defensin families and tissue of origins . HBD-1 was detected in all 49 tissues of hyperplastic tonsils and chronic tonsillitis . Only in chronic inflamed tonsils the amount of HBD-2 mRNA expression was significant increased . In these specimens also mean relative expression rate of all defensins was observed to be manifestly increased . Palatine tonsils express mRNA for different alpha and beta defensins and this expression suggest a newly supposed function in immune defence: the participation in the innate, non-adaptive immune system . Thus, palatine tonsils have a potentially influence in the growth and control of the physiological mouth bacteria by their bactericidal activity.

Dev Comp Immunol, 2002 Nov, 26(9), 817 - 30
Differential effects of age on chicken heterophil functional activation by recombinant chicken interleukin-2; Kogut M et al.; Interleukin-2 (IL-2) exercises an array of biological effects on many cells including the functional activation of cells of the innate immune response . Heterophils, the avian equivalent of the neutrophil, function as professional phagocytes to aid in regulation of innate host defenses . The objective of the present studies was to examine the effects of recombinant chicken IL-2 (rChIL-2) on functional activities of heterophils from chickens during the first 3 weeks after hatch . Peripheral blood heterophils were isolated and incubated with either COS cell-derived rChIL-2 or supernatants from mock-transfected COS cells . rChIL-2 had no effect on the functional activities of heterophils from day-of-hatch chickens, but significantly increased the phagocytosis and bactericidal activity of heterophils from 7- and 14-day-old chickens . rChIL-2 induced no direct stimulation of the respiratory burst by heterophils, but primed heterophils from 7- and 14-day-old birds for an enhanced respiratory burst in response to phorbol ester stimulation . Lastly, rChIL-2 had neither direct nor priming effects on heterophil degranulation . The enhancing effects on heterophil functional activity by rChIL-2 were abated by a neutralizing anti-chicken IL-2 mAb and were therefore specific for this cytokine . These results show that rChIL-2 can directly activate chicken heterophils to exert effector functions, and that heterophil activation by rChIL-2 is also an age-dependent event.

Bull Exp Biol Med, 2002 Mar, 133(3), 308 - 10
Evaluation of antibiotic sensitivity of Chlamydia trachomatis using RT-PCR; Misyurina OY et al.; The sensitivity of 11 clinical strains of Chlamydia trachomatis to azithromycin, ofloxacin, doxycycline, and erythromycin was evaluated . The minimum inhibiting concentrations of all antibiotics for 90% strains, determined by PCR with reverse transcription of omp3B gene RNA (GenBank U68443) corresponded to, and those with reverse transcription of 16S rRNA gene RNA (GenBank X54451) far surpassed the minimum bactericidal concentrations for 90% strains determined by direct immunofluorescence with monoclonal antibodies to the major outer membrane protein.

NPN Med, 1983 May 2, 3(50), 591 - 6
{Vaginal contraception: mechanical or chemical?}; Monard S; PIP: Vaginal contraception is the oldest method of protection against pregnancy . Current methods operate on the same principles as those mentioned in ancient Egyptian papyrus: stopping the migration of sperm with mechanical barriers or destroying them with spermicidal substances . The combination of diaphragm and spermicide was among the most widely used methods before 1960, and recently has been regaining popularity because of fear of side effects of oral contraceptives and IUDs . Diaphragms and cervical caps are vaginal mechanical methods used by the female . Diaphragms must be fitted individually and the woman must be instructed to use them properly . Both devices must be used each time intercourse takes place, always with a spermicide, and must be left in place for 3-8 hours afterwards . Diaphragms and caps are difficult to prescribe because they are time consuming to fit and require a high degree of motivation . Local chemical methods are composed of 1 of a number of active ingredients which have antiinfectious effects, sperm immobilizing action, and lytic power . Spermicides cause the permeability of the cell membrane to increase until rupture occurs . Spermicides may be used with a diaphragm or, less surely, by themselves . They are available in foam, gel, cream, or vaginal tablets, and should be applied 3-5 minutes before intercourse . A vaginal tampon containing spermicide which could be left in place for 3 days and is as easy to use as a menstrual tampon is under study . Side effects of spermicides are minor and no contraindications have been identified . Spermicides have a bactericidal action against some sexually transmitted diseases, and 2 studies have indicated lower rates of cervical cancer in diaphragm users . A Pearl index of 1.5-3 is possible if vaginal contraception is used correctly and consistently . The acceptability of vaginal methods is limited by psychological resistance on the part of both partners . The diaphragm in particular must be inserted each time and the spermicide may cause an exaggerated lubrication in some women .

Contracept Fertil Sex (Paris), 1982 Nov, 10(11), 759 - 69
{Vaginal contraception in 1982 . Realities and perspectives (author's transl)}; Limouzin-lamothe MA; PIP: In recent years, there has been a definite renewal of interest in vaginal contraception . Locally applied suppositories and creams contain surfactants such as benzalkonium chloride or phenyl-mercury nitrate which are powerful spermicidal drugs capable of achieving spermatolysis of previously immobilized spermatozoa . They are highly efficient spermicides as indicated by a Pearl's index close to 1.5, provided they are employed exactly as prescribed . Sponge-soaked spermicides remain active for 2 or 3 days which is a good palliative to the short-lived efficacy of other galenical preparations . Locally applied spermicides have the added advantage of possessing bactericidal properties and preventing M.S.T . Vaginal rings inhibit ovulation through a continuous release of progestogen with a strong antigonadotropic action . A number of different rings have been tested with varying degrees of success . When anovulation is actually obtained, it is frequently accompanied by hyperestrogeny, spotting, and amenorrhea . Furthermore, metabolic and in particular lipid disorders induced by progestogens cannot be avoided . (author's modified)

Contracept Fertil Sex (Paris), 1985 Jan, 13(1 Suppl), 431 - 6
{Disease and contraception . Recent aspects}; Rozenbaum H; PIP: This article reviews several different articles which have contributed to an understanding of the harmful or beneficial effects of oral contraceptives (OCs) on various diseases . The Royal College of General Practitioners study found that current OC users compared to women who had never used OCs had relative risks of .52 for menorrhagia, .37 for dysmenorrhea, .65 for irregular cycles, .72 for intermenstrual bleeding, and .71 for premenstrual syndrome . Several studies found combined OCs to offer protection against ovarian cysts . Microdose progestin only pills did not ameliorate most menstrual problems and aggravated ovarian cysts . Despite some theoretical grounds for suspecting an association between pituitary prolactinomas and OC use, recent studies have failed to find an increased relative risk for prolactinomas in women using OCs for contraceptive purposes, although 1 study found an increased risk in women using OCs for cycle control . 1 study reported 11 pregnancies in 30 diabetic women in 15 months of IUD use; the high rate was attributed to abnormal patterns of mineral deposit on the IUD surface . The 11 pregnancies occurred with 5 Gravigardes, 5 Saf-T-Coils, and 1 Dalkon Shield . Other studies on the contrary have noted no difference in pregnancy rates among 103 diabetic women using Copper Ts or 118 diabetic women using Lippes loops . Combined OCs appear to reduce the incidence of rheumatoid arthritis by 1/2 among current OC users and to protect former users as well . Combined OCs aggravate lupus erythmatous but synthetic progestins alone are effective without aggravating the condition . It has recently been argued that low dose OCs are not contraindicated in cases of sickle cell disease and may even offer protection against thromboembolic vascular accidents for women with sickle cell anemia . Estimates of relative risk of pelvic infection among IUD users vary from 1.5 to 6.5, with the risk apparently greatest for women under 25 . Recent studies have indicated that copper IUDs do not have the bactericidal power formerly attributed to them . Numerous in vitro studies and statistical comparisons of the effect of spermicides in vivo have demonstrated that local methods provide protection against sexually transmitted diseases . OCs may favor vaginal infection, but some recent studies have indicated that they offer protection against pelvic infections . The protective effect of the condom against sexually transmitted diseases is well known . It has been estimated that, relative to non-users of OCs, each 100,000 users will have 235 fewer cases of benign breast disease, 35 fewer of ovarian cysts, 320 fewer of iron deficiency anemia, 600 fewer of pelivc infection, 117 fewer of extrauterine pregnancy, 32 fewer of rheumatoid arthritis, 1 fewer of endometrial cancer, and 3 fewer of ovarian cancer .

Contracept Deliv Syst, 1981 Jul, 2(3), 225 - 9
Germicidal effect of pure electrolytic copper on the gonococcus; Mandouvalos H et al.; PIP: Asymptomatic gonorrhea is now considered to be the main cause of the high current incidence of gonorrhea, especially among women . Both in vivo and in vitro studies were carried out to assess the germicidal effect of copper on gonococcus strains . Of cultures taken from 32 men who exhibited suspicious symptomatology of gonorrhea and 27 women known to have had intercourse with them, 32 of the male and 20 of the female cultures were positive . 28 of the male and 16 of the female cultures exhibited a sensitivity to copper, i.e., the development of the gonococcus cultures was restrained by the introduction of copper to the culture medium . Of the 27 gonorrhea-exposed women who were tested, 2 had multiple locations of the disease and 20 had infections which were localized in the cervical mucus . For the last 20, culture and sensitivity tests of the gonococcus strains showed 14 to be sensitive to copper . These 14 women were fitted with a copper IUD in the endometrium; no other treatment was followed . The results were promising, giving completely negative culture results in every case where the disease had been localized in the cervical mucus . Therefore, although the bactericidal activity of copper and its salts is known to be of low degree, copper and its ions do seem to suppress or limit the spread of gonorrhea . Only large amounts would be effective in actually killing the gonococci in relatively short time periods .

IPPF Med Bull, 1984 Aug, 18(4), 1 - 2
Role of copper in IUDs; Chantler EN; PIP: This report analyzes the contraceptive potential of copper IUDs . The antifertility action of copper in IUDs is considered to involve 1) inhibition of zinc-containing metalloenzymes in the uterus, 2) reduced activity in the endometrial steroid receptor, 3) production of low levels of human chorionic gonadotropin in the luteal phase, 4) depression of ovum transport through the uterine tubes and inhibition of the penetration of cervical mucus by sperm, 5) elevation of the fibrinoloytic activity of the endometrium, and 6) depression of the synthesis of prostaglandins . These biochemical and physiological changes in response to copper released from the IUD are thought to cause only a local effect and systemic accumulation of copper from the device is considered unlikely . The rate of release of copper varies during the time the device remains in the uterus, declining exponentially during the 1st 2 years of use and then increasing as a result of destabilization and fragmentation of the accumulated layer of copper corrosion products . Accelerated copper loss is associated with menorrhagis, higher parity, and the presence of a cervical lesion at the time of IUD insertion . However, accelerated loss of the copper coil does not seem to cause either copper toxicity or decreased contraceptive efficiency . Women with insulin-dependent diabetes may manifest a different copper corrosion process and possibly a higher incidence of contraceptive failure . Since copper IUDs must be replaced more often than inert devices, there is a risk of increased incidence of pelvic inflammatory disease, reported to occur more frequently after replacement of an IUD . The longterm retention of copper IUDs probably has the same risk of infection as that associated with inert devices, and there is no evidence to link the presence of copper with any bacteriostatic or bactericidal action in vivo . The probability of fragmentation is reduced by new types of devices in which the copper wire is replaced by copper bands or a core of an inert metal is included inside the copper wire . It is concluded that copper IUDs, especially the newer devices with a more stable copper component, provide a successful fertility control method .

Pediatr Med Chir, 2002 May-Jun, 24(3), 188 - 93
{Oral antibiotic therapy: problems and perspectives}; Caramia G; As all therapies, oral antibiotic therapy has its own scientific rationale . Infact, for an optimal oral therapy the drug must be absorbed completely, reach the infection site and diffuse in the tissues . It should also remain active for as long as possible at the minimal bactericidal level of concentration . It is therefore necessary to use drugs with a specific bactericidal action for the pathogens in question and that these drugs be administered at right doses for the necessary duration so that the infection is eliminated . It is an inappropriate therapy that which not only fails to eliminate the infection, but leads to the development of resistance, which is an increasingly serious and diffused problem . In hospital and at home it is possible to switch from an injected antibiotic form to the more practical and less expensive oral form, therefore creating a sequential therapy which is better accepted by patients and their families . The acceptance of prescribed therapies is an aspect that should never be underestimated, especially during the development age . Infact, despite the general understanding that unsatisfactory clinical results are a consequence of unadequate compliance with therapy, the desire to recover is rarely the same as the will to follow such plans, even if the patient expects a quick and full recovery . It is therefore necessary to use those therapies which are best accepted but always effective in order to avoid legal implications.

Infect Immun, 2002 Oct, 70(10), 5865 - 9
A uropathogenicity island contributes to the pathogenicity of Escherichia coli strains that cause neonatal meningitis; Houdouin V et al.; We report that the archetypal Escherichia coli strain C5 causing neonatal meningitis harbors a pathogenicity island (PAI) designated PAI I(C5) that is similar to the PAI II(J96) of uropathogenic E . coli J96 inserted in the leuX-tRNA gene . PAI-negative C5 mutants had a lower capacity than C5 to induce high-level bacteremia in a neonatal rat model . However, no change in their resistance to the bactericidal effect of serum and their capacity to cross the blood-brain barrier was observed.

Expert Opin Ther Targets, 2002 Apr, 6(2), 135 - 46
Potential of fumarate reductase as a novel therapeutic target in Helicobacter pylori infection; Ge Z; Approximately 50% of the world's population carries Helicobacter pylori, a gastric bacterial pathogen linked to diseases including gastritis, ulcers and gastric cancer . Chemotherapies are being routinely used to treat systemic H . pylori infection . The common regimens consist of proton pump inhibitors (PPIs) or ranitidine bismuth citrate (RBC) and two antibiotics . Although these regimens efficiently eradicate H . pylori, the emergence of antibiotic-resistant H . pylori strains, their severe side effects and high costs are major drawbacks of these treatments . More efficient, economic and friendly drugs need to be developed . Fumarate reductase (FRD) catalyses the reduction of fumarate to succinate in the Krebs cycle and is also a key enzyme in anaerobic respiration with fumarate as the terminal electron acceptor for many facultative bacteria . H . pylori FRD contains three subunits, FrdA, FrdB and FrdC . Genome analysis and experimental evidence indicate that this enzyme appears to play an important role in the energy metabolism of H . pylori . In addition, FRD is essential for the colonisation of H . pylori in the acidic stomach as demonstrated in the mouse model of infection . Furthermore, three FRD inhibitors used to cure helminthic infection in animals and humans have both inhibitory and bactericidal effects on H . pylori . These lines of evidence indicate that FRD may be a promising chemotherapeutic target . Given that FrdA is strongly immunogenic in the sera from H . pylori-positive patients, this protein may also be used as a candidate for the development of an anti-H . pylori vaccine.

Hunan Yi Ke Da Xue Xue Bao, 2000 Aug 28, 25(4), 371 - 2
{Effect of human serum on the growth of Helicobacter pylori}; Fan XG et al.; To investigate whether sheep blood could be replaced with human serum or fetal calf serum for Helicobacter pylori(Hp) culture, Hp was inoculated onto the culture plates containing 7% of sheep blood, human serum, or fetal calf serum respectively . The colonies were counted after 5 days of culture . Similar culture results were obtained in the plates containing sheep blood, human serum, or fetal calf serum respectively . No effect of anti-Hp-IgG antibody in human serum on the growth of Hp was found (P > 0.05) . However, heat-inactivated human serum significantly suppressed the growth of Hp (P < 0.01) . Human serum or fetal calf serum may be used for Hp culture where sheep blood is not available . Bactericidal activity against Hp in human serum is mediated by complements.

Biomol Eng, 2002 Aug, 19(2-6), 263 - 8
Surface implantation treatments to prevent infection complications in short term devices; Davenas J et al.; Surface treatments of short term devices are actually evaluated to reduce the risk of infections, which in particular are one of the main causes of complications following catheter insertion . We have investigated the efficacy of ion beam techniques to reduce bacterial adhesion-or to induce bactericidal activity of different polymer materials: PVC, silicone rubber, poly(urethane) and poly(ethylene) . Two routes have been evaluated, based on the production of non fouling surfaces, through the production of diamond-like surfaces upon irradiation with rare gases, or the implantation of silver, known for its bactericidal action . In this contribution we discuss more specifically the treatment of poly(ethylene), where a broad range of surface characterisation techniques could show that the biological activity resulted from the formation of metallic colloidal silver near the surface of the polymer, associated to the formation of a dense surface acting as a diffusion barrier . Reduction of the implantation energy to 10 keV, led to activity enhancement resulting from the easier accessibility of surface colloids evidenced by AFM microscopy . This study emphasises the specific processes induced by the formation of silver nano-particles at low energy implantation, which differs basically from Ion Beam Assisted Deposition (IBAD technique) leading to the formation of a continuous silver coating (Artif . Organs 18 (1994) 266; International Patent (PCT) WO 95/18637 (1995)).

Vox Sang, 2002 Aug, 83(2), 119 - 24
Prestorage leucocyte reduction of red cell components prevents release of bactericidal permeability increasing protein and defensins; Fransen EJ et al.; BACKGROUND AND OBJECTIVES: In this study we examined whether prestorage leucocyte reduction prevents the accumulation of bioreactive substances in red cell units . MATERIALS AND METHODS: Measurements were performed in the supernatants of buffy-coat-depleted (standard red cells) and leucocyte-reduced (filtered red cells) red cell units . The effect of storage was evaluated by taking repetitive samples up to 35 days after donation . We determined the concentrations of polymorphonuclear neutrophil (PMN)-derived bactericidal permeability increasing protein (BPI), defensins and annexin A5 . In addition, leucocyte counts (using nageotte chamber) were performed on days 0 and 35 . RESULTS: During storage, the concentrations of BPI, defensins and annexin A5 in standard red cells gradually increased . However, in the filtered red cells BPI and defensins were found in only a few samples, whereas the annexin A5 concentration in these units did not change during storage . Haemolysis data in both types of red cell components were similar at all time-points, except prestorage . Significant correlations were found between the release of BPI, defensins and annexin A5 into red cell units and the loss of leucocytes during storage . CONCLUSIONS: PMNs lose their membrane integrity during cold storage and release their contents into red cell components . Prestorage leucocyte reduction of red cell components prevents the accumulation of BPI, defensins and annexin A5.

Vet Res, 2002 Jul-Aug, 33(4), 413 - 9
Bovine milk fat globules do not inhibit C5a chemotactic activity; Rainard P; The C5a complement fragment is a potent inflammatory molecule but its contribution to the inflammatory response of the mammary gland remains uncertain . One of the unresolved questions is the possible interference of whole milk with C5a . In this study, the chemotactic activity ofpurifled bovine C5a was tested in the presence of whole or skimmed milk . Milk from healthy glands acted as a chemoattractant, which could mask any inhibitory effect on C5a activity . To circumvent milk activity, washed milk fat globules were incubated with optimal (1 nM) or suboptimal (0.1 nM) concentrations of C5a, and the eventual chemoattractant activity was measured . There was no reduction in C5a-induced migration through a polycarbonate filter or shape-change of neutrophils . The concentrations of C5a determined by ELISA in the fluid phase were not reduced after incubation with the fat globules . It can be concluded that bovine milk fat globules do not trap or degrade C5a . Although these results do not explain the inhibitory effect of whole milk in the C5a-induced recruitment of neutrophils in milk, they suggest that milk does not inhibit the second major activity of C5a (apart from chemotaxis), i.e . the stimulation of phagocytic and bactericidal activities of neutrophils.

Angiogenesis, 2001, 4(4), 289 - 98
cDNA microarray analysis of the gene expression profile of VEGF-activated human umbilical vein endothelial cells; Abe M et al.; Vascular endothelial growth factor (VEGF) is one of the most important factors that stimulate angiogenesis and vascular permeability . To clarify the role of VEGF, we analysed a human cDNA chip containing 7267 human genes to identify genes induced by VEGF in human umbilical vein endothelial cells (HUVECs) . One hundred thirty-nine cDNAs, including ninety-nine previously known and forty poorly characterized or novel sequences, were increased more than two-fold by VEGF within twenty-four hours of stimulation . Among them, only five are known to regulate angiogenesis: cyclooxygenase 2 (COX2), heparin-binding epidermal growth factor-like growth factor, early growth response 1 (EGR 1), CYR61, and angiopoietin 2 . Fifty-three genes induced within the first two hours were thought to be directly induced by VEGF . Of these, Down syndrome candidate region 1 (maximum induction = 6.1-fold) was the most profoundly induced, followed by Mifl (KIAA0025; 5.5-fold), COX2 (4.7-fold), EGR 3 (3.7-fold), EGR 2 (3.2-fold), bactericidal/permeability-increasing protein (3.1-fold), and CD1B antigen, b polypeptide (3.1-fold) . In addition to the genes mentioned above, there were many poorly characterized or novel genes induced by VEGF . Further analysis of these genes may aid in the elucidation of the molecular mechanisms of angiogenesis or vascular permeability stimulated by VEGF.

Infect Immun, 2002 Sep, 70(9), 4826 - 32
virB-Mediated survival of Brucella abortus in mice and macrophages is independent of a functional inducible nitric oxide synthase or NADPH oxidase in macrophages; Sun YH et al.; The Brucella abortus virB locus is required for establishing chronic infection in the mouse . Using in vitro and in vivo models, we investigated whether virB is involved in evasion of the bactericidal activity of NADPH oxidase and the inducible nitric oxide synthase (iNOS) in macrophages . Elimination of NADPH oxidase or iNOS activity in macrophages in vitro increased recovery of wild-type B . abortus but not recovery of a virB mutant . In mice lacking either NADPH oxidase or iNOS, however, B . abortus infected and persisted to the same extent as it did in congenic C57BL/6 mice up until 60 days postinfection, suggesting that these host defense mechanisms are not critical for limiting bacterial growth in the mouse . A virB mutant did not exhibit increased survival in either of the knockout mouse strains, indicating that this locus does not contribute to evasion of nitrosative or oxidative killing mechanisms in vivo.

Bull Exp Biol Med, 2002 Jan, 133(1), 62 - 4
Lysosomal-cationic test and NBT reduction test just partially reflect the completeness of phagocytic process in human granulocytes; Belokrylov GA et al.; The results of NBT test correlated with the index of phagocytosis completeness in only 43% of 21 clinically healthy volunteers . The level of the lysosomal-cationic test was significantly reduced only if the phagocytosis completeness index was markedly decreased . The latter is an integral value reflecting the bactericidal activity of granulocytes.

J Leukoc Biol, 2002 Aug, 72(2), 239 - 48
Neutrophil-Kupffer cell interaction: a critical component of host defenses to systemic bacterial infections; Gregory SH et al.; Most bacteria that enter the bloodstream are taken up and eliminated within the liver . The specific mechanisms that underlie the role of the liver in the resolution of systemic bacterial infections remain to be determined . The vast majority of studies undertaken to date have focused on the function of resident tissue macrophages (Kupffer cells) that line the liver sinusoids . Indeed, it is often reported that Kupffer cells ingest and kill the bulk of organisms taken up by the liver . Recent studies indicate, however, that phagocytosis by Kupffer cells is not the principal mechanism by which organisms are eliminated . Rather, elimination depends on the complex interaction of Kupffer cells and bactericidal neutrophils that immigrate rapidly to the liver in response to infection . We discuss the critical role of neutrophil-Kupffer cell interaction in innate host defenses and, conceivably, the development and expression of adaptive immunity in the liver.

J Dairy Sci, 2002 Jun, 85(6), 1454 - 9
Phagocytosis and serum susceptibility of Escherichia coil cultured in iron-deplete and iron-replete media; Wise AJ et al.; The susceptibility of Escherichia coli cultured in either iron-deplete or iron-replete media to phagocytosis by bovine neutrophils and the bactericidal activity of bovine serum was tested in vitro . Fourteen E . coli isolates from naturally occurring intramammary infections (IMI) were cultured overnight at 37 degrees C in iron-replete media and iron-deplete media . The iron-replete media were trypticase soy broth or a chemically defined medium . The iron-deplete media were either trypticase soy broth plus 0.2 mM alpha, alpha' dipyridyl and 1mM citrate, or the chemically defined medium plus 0.2 mM alpha, alpha' dipyridyl, and 1 mM citrate . Iron-replenished medium was the chemically defined iron-deplete medium plus 40 mM ferric citrate . Bacteria grown in iron-deplete media were less susceptible to phagocytosis compared with bacteria grown in iron-replete media . Replenishing the chemically defined iron-deplete medium with ferric citrate obliterated the decreased susceptibility to phagocytosis observed in iron-deplete media . The iron availability in media used to culture E . coli before assay did not affect the bactericidal action of either the classical pathway of complement or the antibody independent alternative pathway of complement in serum . The growth of bacteria in iron-deplete medium did not alter the expression of capsule compared with growth in iron-replete medium . Iron availability during culture of E . coli altered the susceptibility of isolates to phagocytosis by neutrophils, but had no effect on the susceptibility of isolates to the bactericidal activity of serum.

Int J Med Microbiol, 2002 Jun, 291 Suppl 33, 141 - 6
Immune evasion of Borrelia burgdorferi: insufficient killing of the pathogens by complement and antibody; Kraiczy P et al.; The innate immune system and, in particular, the complement system play a key role in the elimination of micro-organisms after entrance in the human host . Like other pathogens, borreliae must develop strategies to inactivate host defence mechanisms . By investigating serum (NHS)-susceptibility of borreliae, we found that mainly B . afzelii isolates are serum-resistant, whereas the majority of B . burgdorferi s . s . isolates display an intermediate serum-sensitive phenotype . In contrast, B . garinii isolates are killed effectively by complement and therefore are classified as serum-sensitive . Up to now, we have identified two distinct proteins of 27.5 kDa and 20.7 kDa expressed on the outer surface of borreliae, which interact directly with FHL-1/reconectin and factor H, the two major regulators of the alternative complement pathway . These borrelial proteins are termed CRASPs (complement regulator-acquiring surface proteins) . CRASPs are detectable only on serum-resistant borreliae and, accordingly, binding of FHL-1/reconectin and factor H only occur with serum-resistant borrelial isolates . We conclude from these results that the control of complement activation on the borrelial surface is due to the interaction of borrelial CRASPs with host complement regulatory proteins . Thus, CRASPs represent an important mechanism of immune evasion on the part of borrelial isolates belonging mostly to the genospecies B . afzelii . By analysing the humoral adaptive immune response of patients, we detected sera that killed NHS-resistant borreliae . Borreliacidal activity is observed most frequently with sera of patients at stage III of the disease . The killing of NHS-resistant isolates by these immune sera always requires the combination of antibodies and complement . Bactericidal activity, however, is not detected in all immune sera at the different disease stages, although specific anti-Borrelia antibodies are present according to serological test results . This observation suggests that not all borrelial antigens are able to induce a borreliacidal immune response . In an extensive analysis of 24 immune sera, we identified up to 12 borrelial antigens, including OspC, which possess the greatest potential for the induction of borreliacidal antibody . The borreliacidal potential of anti-OspC antibodies was tested directly on an OspC-expressing borrelial wild-type isolate and a corresponding variant lacking OspC . In these studies, only the wild-type isolate expressing OspC on its surface proved positive for the lytic complement complex, thereby indicating the great importance of this antigen for the control of the infection . Additional studies are required to identify further "protective" antigens among these 12 proteins, all of which are candidates for infection control according to our studies involving patient immune sera . These antigens may include the recently detected CRASPs.

Microb Pathog, 2002 Jun, 32(6), 279 - 85
Effect of Shiga toxins on granulocyte function; Aoki Y et al.; We already showed that injection of Shiga toxin (Stx) 2 into mice caused severe granulocytosis in the peripheral blood . In this study we further clarified changes of granulocyte function by Stx 2 . The activity of medullasin, a neutral serine protease in granulocytes that injures endothelial cells in vessels, significantly increased when Stx 2 was injected into mice intraperitoneally . Since granulocyte count in the peripheral blood of mice was markedly increased after intraperitoneal injection of Stx 2, medullasin activity in the peripheral blood was remarkably elevated . In contrast to Stx 2, injection of Stx 1 into mice caused no elevation of medullasin activity in granulocytes nor increase in granulocyte count in the peripheral blood . Cathepsin G levels in granulocytes increased only slightly after Stx 2 injection . Granulocytes obtained from mice injected with Stx 2 showed reduced superoxide-producing activity compared with those from controls . Addition of Stx 2 or Stx 1 to human mature granulocytes in vitro decreased their superoxide-producing activity when stimulated with agonists . Therefore, these toxins produced from Escherichia coli augment toxic effect of the bacteria by reducing bactericidal activity of granulocytes . Tissue injury in organisms infected with Shiga toxin-producing E . coli is mainly derived from elevated neutral proteases, such as medullasin, in granulocytes rather than direct toxic effect of superoxide from granulocytes . Hemolytic uremic syndrome caused by Shiga toxin-producing E . coli infection is due, at least in part, to the elevation of medullasin levels produced by granulocytes.

Zhonghua Nei Ke Za Zhi, 2002 Jun, 41(6), 404 - 7
{Clinicopathological characteristics of propylthiouracil-induced antineutrophil cytoplasmic antibodies-positive vasculitis and their target antigens: a report of 4 cases and literature review}; Xu X et al.; OBJECTIVE: To study the clinicopathological manifestations and target antigens of propylthiouracil (PTU)-induced antineutrophil cytoplasmic antibody (ANCA) positive vasculitis . METHODS: Four hospitalized patients with PTU-induced ANCA positive vasculitis in recent two years were studied . Target antigens and antibody titers were investigated with ELISA using seven highly purified known ANCA antigens as solid phase ligands . The known antigens included proteinase 3 (PR3), myeloperoxidase (MPO), human leukocyte elastase (HLE), lactoferrin (LF), cathepsinG (CG), bactericidal/permeability-increasing protein (BPI) and azurocidin (AZU) . RESULTS: Four patients with Grave's disease, 2 female and 2 male with a mean age of 30 (11 approximately 57) years who had been treated with PTU for 7 approximately 60 months suffered from ANCA positive vasculitis . All the 4 patients had renal, lung, skin, joint, muscle and hematological involvement . Sera from all the 4 patients were ANCA positive and recognized MPO, LF and CG . Sera from 3 patients recognized HLE and AZU and 2 recognized PR3 . None of the sera recognized BPI . The majority of the autoantibodies had high titers >/= 1:25 600 . All the sera from 30 patients with Grave's disease and PTU treatment but without vasculitis were ANCA negative . All the 4 patients had pauci- immune glomerular lesions in renal biopsy; 2 had crescentic glomerulonephritis and the remaining 2 had minor glomerular abnormalities . All the 4 patients responded to withdrawal of propylthiouracil; 3 patients were treated with immunosuppressive agents . All the patients achieved clinical remission . However, one patient with crescentic glomerulonephritis was dialysis dependent . After withdrawal of PTU and administration of immunosuppressants, ANCA titres declined, but did not turn to negative in 1-6 months in most of the patients . CONCLUSIONS: PTU can induce ANCA positive vasculitis . The autoantibodies were polyclonal and recognized multiple target antigens of neutrophil cytoplasm . Early withdrawal of PTU and administration of immunosuppressive agents might improve the prognosis.

Lasers Surg Med, 2002, 30(5), 360 - 4
The Er:YAG laser in endodontics: results of an in vitro study; Schoop U et al.; BACKGROUND AND OBJECTIVE: Until recently, the main field of Er:YAG laser application was the removal of dental hard substances within the scope of cavity preparation . Nowadays, several new delivery-systems are available, permitting the application of the Er:YAG laser in endodontics . The aim of the present study was to assess the effects of Er:YAG laser irradiation on root canals in vitro . STUDY DESIGN/MATERIALS AND METHODS: For this purpose, 220 extracted human teeth were endodontically processed and subsequently irradiated at different settings using an Er:YAG laser imitating in vivo irradiation procedures . The teeth were then subdivided into three groups and subjected to bacteriological evaluations, scanning electron microscopy, and temperature measurements . RESULTS: The bacteriological evaluation revealed a decisive bactericidal effect of the Er:YAG laser in the root canal . The bactericidal effect was dependent on the applied output power and specific for the different species of bacteria investigated . Scanning electron microscopy showed discrete removal of dentine from the root canal walls . The temperature rise during irradiation was moderate when standardized power settings were used . CONCLUSION: The investigations indicate that the Er:YAG laser is a suitable tool for the elimination of bacteria in root canals under in vitro conditions .

Curr Drug Targets, 2002 Aug, 3(4), 335 - 44
Streptogramin antibiotics: mode of action and resistance; Johnston NJ et al.; The streptogramin antibiotics were discovered over 40 years ago but are only now emerging as important therapeutic agents for the treatment of infection caused by a variety of bacteria . The streptogramins consist of mixtures of two structurally distinct compounds, type A and type B, which are separately bacteriostatic, but bactericidal in appropriate ratios . These antibiotics act at the level of inhibition of translation through binding to the bacterial ribosome . Resistance to streptogramins occurs through a number of mechanisms including target modification, efflux, and enzyme catalyzed antibiotic modification . This review describes the current understanding of streptogramin function and resistance with emphasis on molecular mechanism and epidemiology.

Int Microbiol, 2002 Mar, 5(1), 25 - 31
An in vitro study of the susceptibility of mobile and cystic forms of Borrelia burgdorferi to hydroxychloroquine; Brorson O et al.; In this work the susceptibility of mobile and cystic forms of Borrelia burgdorferi to hydroxychloroquine (HCQ) was studied . The minimal bactericidal concentration (MBC) of HCQ against the mobile spirochetes was > 32 microg/ml at 37 degrees C, and > 128 microg/ml at 30 degrees C . Incubation with HCQ significantly reduced the conversion of mobile spirochetes to cystic forms . When incubated at 37 degrees C, the MBC for young biologically active cysts (1-day old) was > 8 microg/ml, but it was > 32 microg/ml for old cysts (1-week old) . Acridine orange staining, dark-field microscopy and transmission electron microscopy revealed that the contents of the cysts were partly degraded when the concentration of HCQ was > or = MBC . At high concentrations of HCQ (256 microg/ml) about 95% of the cysts were ruptured . When the concentration of HCQ was > or = MBC, core structures did not develop inside the cysts, and the amount of RNA in these cysts decreased significantly . Spirochetal structures inside the cysts dissolved in the presence of high concentrations of HCQ . When the concentration of HCQ was > or = MBC, the core structures inside the cysts were eliminated . These observations may be valuable in the treatment of resistant infections caused by B . burgdorferi, and suggest that a combination of HCQ and a macrolide antibiotic could eradicate both cystic and mobile forms of B . burgdorferi.

Clin Diagn Lab Immunol, 2002 Jul, 9(4), 901 - 7
Bactericidal activity in whole blood as a potential surrogate marker of immunity after vaccination against tuberculosis; Cheon SH et al.; The development of new tuberculosis (TB) vaccines will require the identification of correlates of human protection . This study examined the balance between immunity and virulence in a whole blood infection model in which intracellular mycobacterial survival was measured using BACTEC . In the blood of tuberculin-negative donors, counts of Mycobacterium tuberculosis H(37)Ra organisms fell by 0.14 log(10) CFU during 96 h of whole blood culture, whereas counts of Mycobacterium bovis BCG, M . tuberculosis H(37)Rv, and a clinical TB isolate's organisms increased by 0.13, 0.43, and 1.04 log(10) CFU, respectively (P < 0.001), consistent with their relative virulence . Inhibition of tumor necrosis factor alpha by the addition of methylprednisolone or pentoxifylline or removal of CD4(+) or CD8(+) T cells by magnetic beads had deleterious effects on immune control of intracellular growth only in the blood of tuberculin-positive donors . Repeated vaccination of eight tuberculin-negative volunteers with M . bovis BCG resulted in a 0.3 log (50%) reduction in BCG CFU counts in the model compared to baseline values (P < 0.05) . Three of the volunteers responded only after the second vaccination . These experiments indicate that whole blood culture may be used to measure immunity to M . tuberculosis and that further studies of repeated BCG vaccination are warranted.

Front Biosci, 2002 Jul 1, 7, d1634 - 41 Epub 2002 Jul 01.
The effect of phospholipid transfer protein on lipoprotein metabolism and atherosclerosis; Jiang XC; Phospholipid transfer protein (PLTP) is a member of the lipid transfer/lipopolysaccharide binding protein gene family . Recently, the crystal structure of one of the members of the gene family, bactericidal permeability increasing protein, was solved, providing potential insights into the mechanisms of action of PLTP . These molecules contain intrinsic lipid binding sites and appear to act as carrier proteins that shuttle between lipoproteins to redistribute lipids . The phenotype of PLTP transgenic and gene knock out mice indicates that PLTP plays a major role in the metabolism of high-density lipoprotein (HDL) and apoB-containing lipoproteins and thereby influences the concentration, apolipoprotein content, and size of lipoprotein particles in plasma . Recent data indicate that PLTP deficiency in mice is associated with a decrease of atherosclerosis, despite decreased HDL levels . At lease two underlined mechanisms are involved in the reduction of atherosclerosis in PLTP deficient status, 1) reduction of apoB-containing lipoprotein production and levels; and 2) increase of anti-oxidation potential . Human studies indicated that PLTP activity positively correlated with aging, obesity, diabetes and coronary artery disease . A challenge for the future will be the inhibition of PLTP for therapeutic benefit.

J Cell Sci, 2002 Jul 15, 115(Pt 14), 2975 - 83
Granule-specific ATP requirements for Ca2+-induced exocytosis in human neutrophils . Evidence for substantial ATP-independent release; Theander S et al.; Ca2+-induced exocytosis in neuronal and neuroendocrine cells involves ATP-dependent steps believed to 'prime' vesicles for exocytosis . Primed, docked vesicles are released in response to Ca2+ influx through voltage-gated Ca2+ channels . Neutrophils, however, do not possess voltage-gated Ca2+ channels and appear to have no docked vesicles . Furthermore, neutrophils have several types of granules with markedly different Ca2+ requirements for exocytosis . These differential Ca2+ dependencies were used as a tool to investigate the ATP dependence of different granule populations . Here we demonstrate distinct ATP requirements for release of neutrophil granule populations, with respect to rate as well as amplitude . Intracellular ATP was depleted to various levels, and exocytosis was stimulated with different Ca2+ concentrations and measured with the patch-clamp capacitance technique or by detecting enzyme release . Primary granule exocytosis displayed a distinct ATP dependence with an apparent K(m) of approximately 80 microM ATP and no ATP-independent exocytosis . Release of secondary and tertiary granules displayed a more shallow ATP dependence (K(m) approximately 330 microM), and more than 50% of secondary and tertiary granules appeared not to need ATP at all for their release . Individual granules in human neutrophils have distinct ATP requirements for exocytosis, suggesting that the ATP-sensitive elements are localised to the granules . Primary granule exocytosis has a very low affinity for ATP . Furthermore, substantial ATP-independent exocytosis of secondary and tertiary granule occurs despite the absence of docked granules . These characteristics should help neutrophils to fulfil their bactericidal functions at poorly irrigated sites of infection with low glucose supply.

Antimicrob Agents Chemother, 2002 Jul, 46(7), 2116 - 23
Enzymes associated with reductive activation and action of nitazoxanide, nitrofurans, and metronidazole in Helicobacter pylori; Sisson G et al.; Nitazoxanide (NTZ) is a redox-active nitrothiazolyl-salicylamide prodrug that kills Helicobacter pylori and also many anaerobic bacterial, protozoan, and helminthic species . Here we describe development and use of a spectrophotometric assay, based on nitroreduction of NTZ at 412 nm, to identify H . pylori enzymes responsible for its activation and mode of action . Three enzymes that reduce NTZ were identified: two related NADPH nitroreductases, which also mediate susceptibility to metronidazole (MTZ) (RdxA and FrxA), and pyruvate oxidoreductase (POR) . Recombinant His-tagged RdxA, FrxA, and POR, overexpressed in nitroreductase-deficient Escherichia coli, each rapidly reduced NTZ, whereas only FrxA and to a lesser extent POR reduced nitrofuran substrates (furazolidone, nitrofurantoin, and nitrofurazone) . POR exhibited no MTZ reductase activity either in extracts of H . pylori or following overexpression in E . coli; RdxA exhibited no nitrofuran reductase activity, and FrxA exhibited no MTZ reductase activity . Analysis of mutation to rifampin resistance (Rif(r)) indicated that NTZ was not mutagenic and that nitrofurans were only weakly mutagenic . Alkaline gel DNA electrophoresis indicated that none of these prodrugs caused DNA breakage . In contrast, MTZ caused DNA damage and was strongly mutagenic . We conclude that POR, an essential enzyme, is responsible for most or all of the bactericidal effects of NTZ against H . pylori . While loss-of-function mutations in rdxA and frxA produce a Mtz(r) phenotype, they do not contribute much to the innate susceptibility of H . pylori to NTZ or nitrofurans.

Biofizika, 2002 May-Jun, 47(3), 570 - 4
{A negative result of the experimental verification of distant effect of bactericidal agents on the growing bacterial culture}; Dergacheva IP; The incontact action of penicillin and ethidium bromide on dividing bacterial culture Escherichia coli through ampoule glass or by means of antennae was studied . No statistically significant differences in cell division between samples exposed to distant action and control were found, though contact action of both substances reduced division considerably.

Probl Tuberk, 2002, (3), 38 - 42
{Some metabolic characteristics of circulating phagocytes in patients with different types of pulmonary tuberculosis}; Kaminskaia GO et al.; Before and 2 and 3 months after chemotherapy, spontaneous and BCG-stimulated nitroblue tetrazolium (NBT) tests, superoxide dismutase (SOD) and catalase activities, the levels of malonic dialdehyde (MDA) and platelet activation factor (PAF) in isolated mononuclear cells and neutrophils were estimated in 169 patients with different forms of pulmonary tuberculosis . Before treatment, in the patients the monocytic bactericidal potential assessed by NBT test parameters was found to increase, but the neutrophilic one reduced due to their basal overirritation . All other test parameters were higher in both types of cells . The magnitude and balance of changes were interrelated to the onset and nature of the process . Following 3 months of treatment, the bactericidal potential increased much more in the monocytes and returned to normal values if the changes in the processes were favourable . In both types of the cells, SOD activity became normal to normal values, MDA levels were maintained in the normal range due to the a preserved enhancement in catalase activity . PAF remained in the upper normal range . Both types of the cells showed drastically increased levels of MDA and drastically decreased PAF levels with progression due to therapy.

Probl Tuberk, 2002, (3), 21 - 5
{The use of immunomodulator likopid in the combined treatment pulmonary tuberculosis}; Svistunova AS et al.; Licopid is a synthetic analogue of a cell wall component of all bacteria . The monocytic macrophageal system is the main target of licopid's action . Addition of the immunomodulator to combined therapy for patients with pulmonary tuberculosis in order to enhance phagocytic activity exerted a marked clinical effect appeared as ceased bacterial isolation in 80% of the patients, a fall in the amount of purulent sputum, no symptoms of intoxication following 2-3 weeks, accelerated resolution of infiltrative changes . The positive clinical effect coincided with the immunological one manifested as increases in the absolute counts of CD3+CD4+ and CD3+CD8+ cells, in the absorptive and bactericidal functions of phagocytes . Such effects were not observed in patients receiving routine treatment . Licopid is recommended for supplementation to the combined treatment regimen for pulmonary tuberculosis.

Crit Rev Food Sci Nutr, 2002, 42(3 Suppl), 267 - 72
Fighting infectious diseases with inhibitors of microbial adhesion to host tissues; Sharon N et al.; The majority of infectious diseases are initiated by the adhesion of pathogenic organisms to the tissues of the host . In many cases this adhesion is mediated by lectins present on the surface of the infectious organism that bind to complementary carbohydrates on the surface of the host tissues . Soluble carbohydrates recognized by the bacterial lectins block the adhesion of the bacteria to animal cells in vitro . Moreover, such carbohydrates have been shown to protect against experimental infection by lectin-carrying bacteria of different mammals, such as mice, rabbits, calves, and monkeys . Agents other than carbohydrates also block adhesion, as demonstrated with cranberry juice as well as with low and high molecular weight preparations isolated from the juice . Both kinds of preparation inhibited the adhesion in vitro of Escherichia coli to different animal cells . In addition, the high molecular weight material acted similarly on the adhesion of Helicobacter pylori to human gasrointenstinal cells, and on the coaggregation of oral bacteria . Furthermore, in limited clinical studies regular drinking of cranberry juice had a significant preventive effect on bacteriuria, and the high molecular weight constituent of the juices was also effective in decreasing the salivary level of Streptococus mutans, the major cause of tooth decay . Because the inhibitors of adhesion examined are not bactericidal, the selection of resistant inhibitor strains is unlikely to occur . Together, these findings may lead to new therapeutic strategies that are in dire need because of the world-wide increase in antibiotic resistant bacteria.

Inflamm Res, 2002 Apr, 51(4), 201 - 5
Effect of enrofloxacin treatment on plasma endotoxin during bovine Escherichia coli mastitis; Dosogne H et al.; OBJECTIVE AND DESIGN: To investigate the effect of enrofloxacin on endotoxin resorption during bovine Escherichia coli mastitis . ANIMALS: 12 healthy early post partum Holstein cows . TREATMENT: Mastitis was induced by intramammary infusion of 10(4) cfu E . coli P4:032 . Six cows were treated twice according to the usual enrofloxacin therapy: 5 mg/kg enrofloxacin 1) intravenously at 10 h and 2) subcutaneously at 30 h after challenge . The other 6 cows served as non-treated controls . METHODS: Blood and milk samples were collected at several time points after challenge . LPS in plasma was quantified using the limulus amoebocyte lysate (LAL) assay . The somatic cell count (SCC) and cfu of milk samples were also analysed . RESULTS: Occasional LPS peaks were detected in the plasma of 2 control cows at 6 h post-challenge and of 1 enrofloxacin-treated cow at 10 h post-challenge (P < 0.01 and P < 0.05, respectively, in comparison with time 0), just before enrofloxacin treatment . After enrofloxacin treatment, no significant LPS amounts were detected in the plasma of treated cows, but neither in the control cows . CONCLUSION: During induced coliform mastitis, LPS resorption in plasma occured only sporadically and within 10 h post-challenge . Whereas enrofloxacin treatment clearly limited bacterial growth in milk, significant effects on LPS resorption could not be detected . This suggests that enrofloxacin treatment of E . coli mastitis is predominantly beneficial by its bactericidal activity and is not associated with enhanced resorption of endotoxins.

J Immunol, 2002 Jun 15, 168(12), 6358 - 65
Mycobacterium tuberculosis promotes apoptosis in human neutrophils by activating caspase-3 and altering expression of Bax/Bcl-xL via an oxygen-dependent pathway; Perskvist N et al.; In addition to direct bactericidal activities, such as phagocytosis and generation of reactive oxygen species (ROS), neutrophils can regulate the inflammatory response by undergoing apoptosis . We found that infection of human neutrophils with Mycobacterium tuberculosis (Mtb) induced rapid cell death displaying the characteristic features of apoptosis such as morphologic changes, phosphatidylserine exposure, and DNA fragmentation . Both a virulent (H37Rv) and an attenuated (H37Ra) strain of Mtb were equally effective in inducing apoptosis . Pretreatment of neutrophils with antioxidants or an inhibitor of NADPH oxidase markedly blocked Mtb-induced apoptosis but did not affect spontaneous apoptosis . Activation of caspase-3 was evident in neutrophils undergoing spontaneous apoptosis, but it was markedly augmented and accelerated during Mtb-induced apoptosis . The Mtb-induced apoptosis was associated with a speedy and transient increase in expression of Bax protein, a proapoptotic member of the Bcl-2 family, and a more prominent reduction in expression of the antiapoptotic protein Bcl-x(L) . Pretreatment with an inhibitor of NADPH oxidase distinctly suppressed the Mtb-stimulated activation of caspase-3 and alteration of Bax/Bcl-x(L) expression in neutrophils . These results indicate that infection with Mtb causes ROS-dependent alteration of Bax/Bcl-x(L) expression and activation of caspase-3, and thereby induces apoptosis in human neutrophils . Moreover, we found that phagocytosis of Mtb-induced apoptotic neutrophils markedly increased the production of proinflammatory cytokine TNF-alpha by human macrophages . Therefore, the ROS-dependent apoptosis in Mtb-stimulated neutrophils may represent an important host defense mechanism aimed at selective removal of infected cells at the inflamed site, which in turn aids the functional activities of local macrophages.

Biochem Biophys Res Commun, 2002 May 24, 293(5), 1571 - 8
Relationship between p38 mitogen-activated protein kinase and small GTPase Rac for the activation of NADPH oxidase in bovine neutrophils; Yamamori T et al.; Superoxide production by NADPH oxidase is essential for bactericidal properties of neutrophils . However, molecular mechanisms underlying the activation of this enzyme remain largely unknown . Here, using bovine neutrophils we examined the role of p38 mitogen-activated protein kinase (p38 MAPK) in the signaling pathways of the NADPH oxidase activation . Superoxide production was induced by stimulation with serum-opsonized zymosan (OZ) and attenuated by p38 MAPK inhibitor, SB203580 . OZ stimulation induced the translocation of p47(phox) and Rac to the plasma membrane and SB203580 completely blocked the translocation of Rac, but only partially blocked that of p47(phox) . Furthermore, SB203580 abolished the OZ-elicited activation of Rac, which was assessed by detecting the GTP-bound form of this protein . Phosphatidylinositol 3-kinase (PI3K) inhibitors, wortmannin and LY294002, blocked not only p38 MAPK activation but also Rac activation . However, SB203580 showed no effect on the PI3K activity . These results suggested that PI3K/p38 MAPK/Rac pathway was present in the activation of NADPH oxidase in bovine neutrophils.

Int Microbiol, 2001 Dec, 4(4), 209 - 15
Susceptibility of motile and cystic forms of Borrelia burgdorferi to ranitidine bismuth citrate; Brorson O et al.; Gastrointestinal symptoms accompanying Lyme disease have not been considered in the treatment of Lyme patients yet . Here we examine the effect of ranitidine bismuth citrate (RBC) on motile and cystic forms of Borrelia burgdorferi in vitro, to determine whether it could cure this bacterial infection in the gastrointestinal tract . When motile forms of B . burgdorferi were exposed to RBC for 1 week at 37 degrees C, the minimal bactericidal concentration (MBC) was > 64 mg/ml . At 30 degrees C, the MBC was > 256 mg/ml . When the incubation lasted for 2 weeks at 37 degrees C, the MBC dropped to > 2 mg/ml . Bismuth aggregates were present on the surface of B . burgdorferi when RBC > or = MBC, as shown by transmission electron microscopy (TEM) . Cystic forms of B . burgdorferi, exposed to RBC for 2 weeks at 37 degrees C, were examined by cultivation in BSK-H medium (Sigma B3528) . They were stained with acridine orange (pH 6.4, pH 7.4) and studied by TEM . The MBC for RBC for young cystic forms (1 day old) and old cysts (8 months old) was estimated to be > 0.125 mg/ml and > 2 mg/ml, respectively . Bismuth aggregates were attached to the cysts and, in some, the pin-shaped aggregates penetrated the cyst wall . The bismuth aggregates also bound strongly to blebs and granules of B . burgdorferi when RBC > or = MBC . When B . burgdorferi is responsible for gastrointestinal symptoms, bismuth compounds may be candidates for eradication of the bacterium from the gastrointestinal tract.

J Chemother, 1991 Jan, 3 Suppl 1, 28 - 9
In-vitro activity of roxithromycin against Chlamydia trachomatis; Moroni A et al.; Ten Chlamydia trachomatis isolates were tested for their sensitivity to roxithromycin, in comparison with erythromycin and tetracycline . The minimum inhibitory and minimum bactericidal concentrations of roxithromycin ranged from 0.03-0.12 and 0.25-1 microg/ml, respectively.

J Gen Physiol, 2002 Jun, 119(6), 571 - 80
Absence of proton channels in COS-7 cells expressing functional NADPH oxidase components; Morgan D et al.; Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is an enzyme of phagocytes that produces bactericidal superoxide anion (O(2)(-)) via an electrogenic process . Proton efflux compensates for the charge movement across the cell membrane . The proton channel responsible for the H(+) efflux was thought to be contained within the gp91(phox) subunit of NADPH oxidase, but recent data do not support this idea (DeCoursey, T.E., V.V . Cherny, D . Morgan, B.Z . Katz, and M.C . Dinauer . 2001 . J . Biol . Chem . 276:36063-36066) . In this study, we investigated electrophysiological properties and superoxide production of COS-7 cells transfected with all NADPH oxidase components required for enzyme function (COS(phox)) . The 7D5 antibody, which detects an extracellular epitope of the gp91(phox) protein, labeled 96-98% of COS(phox) cells . NADPH oxidase was functional because COS(phox) (but not COS(WT)) cells stimulated by phorbol myristate acetate (PMA) or arachidonic acid (AA) produced superoxide anion . No proton currents were detected in either wild-type COS-7 cells (COS(WT)) or COS(phox) cells studied at pH(o) 7.0 and pH(i) 5.5 or 7.0 . Anion currents that decayed at voltages positive to 40 mV were the only currents observed . PMA or AA did not elicit detectable H(+) current in COS(WT) or COS(phox) cells . Therefore, gp91(phox) does not function as a proton channel in unstimulated cells or in activated cells with a demonstrably functional oxidase.

Biol Chem, 2002 Mar-Apr, 383(3-4), 709 - 13
Efficacy of HOCl scavenging by sulfur-containing compounds: antioxidant activity of glutathione disulfide?
den Hartog GJ, Haenen GR, Vegt E, van der Vijgh WJ, Bast A.
Hypochlorous acid (HOCl) is a bactericidal compound formed by activated neutrophils during inflammation . Overproduction of HOCl causes damage to tissues at the site of neutrophil accumulation . The deleterious effects of excessive HOCl formation can be attenuated using antioxidants . Thiols and thioethers are known to be very effective HOCl scavengers . In the present study, the potency of several sulfur-containing compounds to protect acetylcholinesterase, glutathione S-transferase P1-1 (GST P1-1) and alpha1-antiproteinease against inactivation by HOCl was determined . Surprisingly, glutathione disulfide was an effective protector of acetylcholinesterase against hypochlorous acid . Glutathione disulfide did not provide protection for GST P1-1 and alpha1-antiproteinease against oxidative inactivation by HOCl . The implications of this finding are discussed.

J Immunol, 2002 Jun 1, 168(11), 5638 - 44
Cloning and characterization of the homolog of mammalian lipopolysaccharide-binding protein and bactericidal permeability-increasing protein in rainbow trout Oncorhynchus mykiss; Inagawa H et al.; We cloned two cDNAs denoted as RT-LBP/BPI-1 and RT-LBP/BPI-2, respectively, which were derived from the mRNA of head kidney from rainbow trout . They showed structural homology with LPS-binding protein (LBP) and bactericidal/permeability-increasing protein (BPI) in mammals . The full-length cDNA of RT-LBP/BPI-1 and RT-LBP/BPI-2 is 1666 and 1741 bp, respectively . Both cDNAs encoded 473 aa residues, including the amino acids conserved in mammalian LBP and BPI proteins that were assumed to be involved in LPS binding . The overall coding sequence of RT-LBP/BPI-1 has 33% amino acid homology to human LBP and 34% to human BPI, and RT-LBP/BPI-2 has 32% amino acid homology to human LBP and 33% to human BPI . Three-dimensional structure analysis by three-dimensional/one-dimensional (3D-1D) methods also demonstrated that RT-LBP/BPI-1 and RT-LBP/BPI-2 proteins showed significant similarity to human BPI, having a boomerang shape with N-terminal and C-terminal barrels . Phylogenetic analysis showed that the LBP and BPI genes seemed to be established after the divergence of mammals from teleosts . These results suggested that RT-LBP/BPI-1 and RT-LBP/BPI-2 may be a putative ortholog for mammalian LBP and/or BPI genes . This is the first study to identify the LBP family genes from nonmammalian vertebrates.

Antimicrob Agents Chemother, 2002 Jun, 46(6), 1875 - 9
Bactericidal activity of increasing daily and weekly doses of moxifloxacin in murine tuberculosis; Yoshimatsu T et al.; Moxifloxacin (MXF) is a new 8-methoxyquinolone with potent activity against Mycobacterium tuberculosis and a half-life of 9 to 12 h in humans . Previous in vivo studies using daily doses of 100 mg/kg of body weight have demonstrated bactericidal activity comparable to that of isoniazid (INH) in a murine model of tuberculosis (TB) . Recent pharmacokinetic data suggest that MXF may have been underadministered in these studies and that a 400-mg/kg dose in mice better approximates the area under the concentration-time curve obtained in humans after a 400-mg oral dose . Therefore, the bactericidal activity of MXF in doses up to 400 mg/kg given daily or weekly for 28 days was assessed in mice infected intravenously with 5 x 10(6) CFU of M . tuberculosis . INH was used as a positive control . After 3 days of daily therapy, the CFU counts from splenic homogenates for mice treated with MXF in doses of 100 to 400 mg/kg/day were lower than those from pretreatment controls . No significant differences in CFU counts were seen when mice receiving INH or MXF at 50 mg/kg/day were compared to pretreatment controls . After 28 days of therapy, dose-dependent reductions in CFU counts in splenic homogenates were seen for daily MXF therapy . The maximum bactericidal effect was seen with daily doses of 400 mg/kg, which resulted in a reduction in CFU counts of 1 log(10) greater than that with INH treatment, although the difference was not statistically significant . CFU counts from lung homogenates after 28 days of therapy were significantly lower in all treatment groups than in untreated controls . The weekly administration of MXF in doses ranging from 50 to 400 mg/kg resulted in no significant bactericidal activity . Mice receiving daily MXF doses of 200 and 400 mg/kg/day failed to gain weight and appeared ill after 28 days of therapy, findings suggestive of drug toxicity . In conclusion, MXF has dose-dependent bactericidal activity against M . tuberculosis in the mouse when given in doses up to 400 mg/kg, where its pharmacokinetic profile better approximates that of standard human dosages . Combination regimens which take advantage of the enhanced pharmacodynamic profile of MXF at these doses have the potential to shorten the course of antituberculous therapy or allow more intermittent (i.e., once-weekly) therapy and should be evaluated in the mouse model of TB.

Compend Contin Educ Dent Suppl, 1997, 18(21), S2 - 7; quiz S45
The use of sodium bicarbonate in oral hygiene products and practice; Newbrun E; Early dentifrices contained natural ingredients, mostly in coarse particle form, and were quite abrasive . Salts, either sodium chloride, sodium bicarbonate, or a mixture of both, have also been used for tooth cleaning because of their ready availability and low cost . Because of both their relatively low intrinsic hardness and their high solubility, another advantage is low abrasivity . Their biggest disadvantage is a salty, unpalatable taste . Many modern dentifrices that contain sodium bicarbonate, either as the sole abrasive or one of several, disguise the saltiness with flavoring and sweetening agents . An almost inverse relationship exists between the percentage of baking soda in a dentifrice and its abrasivity . Sodium bicarbonate has no anticaries activity per se but is compatible with fluoride . In high concentrations, sodium bicarbonate is bactericidal against most periodontal pathogens . Most clinical studies have not found significant differences in periodontal response to baking soda as compared with other commercial dentifrices, probably because of its rapid clearance from the gingival sulcus . Sodium bicarbonate may not be the "magic bullet" for curing dental diseases, but its safety (if ingested), low abrasivity, low cost, and compatibility with fluoride make it a consummate dentifrice ingredient.

Infect Immun, 2002 Jun, 70(6), 2908 - 14
Lipopolysaccharide-induced granule mobilization and priming of the neutrophil response to Helicobacter pylori peptide Hp(2-20), which activates formyl peptide receptor-like 1; Bylund J et al.; The cecropin-like bactericidal peptide Hp(2-20) from Helicobacter pylori induces activation of the NADPH oxidase in human neutrophils via formyl peptide receptor-like 1 (FPRL1) (J . Bylund, T . Christophe, F . Boulay, T . Nystrom, A . Karlsson, and C . Dahlgren, Antimicrob . Agents Chemother . 45:1700-1704, 2001) . Here we investigated the ability of bacterial lipopolysaccharide (LPS) to prime this response . Neutrophils treated with LPS for 30 min at 37 degrees C produced substantially more superoxide anion than control cells upon stimulation with Hp(2-20) . Hence, LPS primed the cells for subsequent stimulation through FPRL1 . To study the molecular background of this priming phenomenon, we measured the degrees of granule mobilization and concomitant receptor upregulation to the cell surface in LPS-treated cells . Exposure of complement receptors 1 and 3 as well as the formyl peptide receptor (FPR) was markedly increased after LPS treatment . Since approximately 60% of the gelatinase granules were mobilized while the specific granules were retained, we hypothesized that the gelatinase granules were potential stores of FPRL1 . The presence of FPRL1 mainly in the gelatinase granules was confirmed by Western blotting of subcellular fractions of resting neutrophils . These results suggest that the mechanism behind the LPS-induced priming of FPRL1-mediated responses lies at the level of granule (receptor) mobilization.

Crit Care Med, 2002 May, 30(5), 1118 - 22
Omeprazole treatment diminishes intra- and extracellular neutrophil reactive oxygen production and bactericidal activity; Zedtwitz-Liebenstein K et al.; OBJECTIVE: Neutrophils play a crucial role in host defense against infectious disease . The objective was to analyze the effect of omeprazole treatment on indexes of neutrophil function in healthy subjects . DESIGN: Open . SETTING: University hospital . SUBJECTS: Ten healthy subjects . INTERVENTION: Analysis of blood samples before and after omeprazole administration . MEASUREMENTS AND MAIN RESULTS: Neutrophil Escherichia coli phagocytosis was assessed by microscopy and flow cytometry . Intracellular production of reactive oxygen intermediates was measured by flow cytometry . Extracellular reactive oxygen intermediate production was assessed with a cytochrome c reduction assay . Neutrophil bactericidal capacity and intracellular concentrations of Ca2+ were determined by fluorometry . Four hours after a single 40-mg dose of omeprazole, intra- and extracellular reactive oxygen intermediate production by neutrophils was significantly reduced compared with pretreatment values: -30% (24% to 42%) (median and range) and -22% (21% to 68%; p <.05 for both) . The intracellular Ca2+ concentrations in resting neutrophils were significantly increased (+33%, 21% to 39%, compared with pretreatment concentrations, p <.001) and neutrophilic bactericidal activity was decreased (-30%, 19% to 47%, compared with pretreatment concentrations, p <.0001) . Intracellular Ca2+ concentrations correlated with intracellular reactive oxygen intermediate production and neutrophilic bactericidal capacity (r =.730 and r =.618, p <.05 for both, respectively) . In contrast, phagocytosis rates were not impaired by omeprazole . CONCLUSIONS: Our results imply that omeprazole impairs production of reactive oxygen intermediates by neutrophils . Whether specific impairments of neutrophil host defenses occur in vivo remains uncertain . Reduced bactericidal activity is associated with an increase of intracellular Ca2+ concentrations in resting neutrophils.

Amyloid, 2002 Mar, 9(1), 13 - 23
Channel formation by serum amyloid A: a potential mechanism for amyloid pathogenesis and host defense; Hirakura Y et al.; Serum amyloid A (SAA) is a family of closely related apolipoproteins associated with high density lipoprotein (HDL) . Subclasses of SAA isoforms are differentially expressed constitutively and during inflammation . During states of infection or inflammation, levels of HDL bound, acute phase isoforms of SAA rise as much as 1000-fold in the serum, suggesting that it might play a role in host defense . Following recurrent or chronic inflammation, an N-terminal peptide fragment of SAA known as amyloid A (AA) assembles into fibrils causing extensive damage to spleen, liver, and kidney, and rapidly progressing to death . In the present paper, we report the novel finding that a recombinant acute phase isoform variant of human SAA 1.1 (SAAp) readily forms ion-channels in planar lipid bilayer membranes at physiologic concentrations . These channels are voltage-independent, poorly selective, and are relatively long-lived This type of channel would place a severe metabolic strain on various kinds of cells . Expression of human SAA 1.1 in bacteria induces lysis of bacterial cells, while expression of the constitutive isoform (human SAA4) does not . Secondary structural analysis of the SAA isoforms in dicates a strong hydrophobicity of the N-terminal of the acute phase isoform relative to the constitutive SAA4 isoform, which may be responsible for the bactericidal activity of the former, in keeping with the notion that SAA 1 targets cell membranes and forms channels in them . Channel formation may thus be related to a host defense role of acute phase SAA isoforms and may also be the mechanism of end organ damage in AA and other amyloidoses.

Biochim Biophys Acta, 2002 Apr 12, 1561(2), 159 - 70
Conformational changes of colicin Ia channel-forming domain upon membrane binding: a solid-state NMR study; Huster D et al.; Channel-forming colicins are bactericidal proteins that spontaneously insert into hydrophobic lipid bilayers . We have used magic-angle spinning solid-state nuclear magnetic resonance spectroscopy to examine the conformational differences between the water-soluble and the membrane-bound states of colicin Ia channel domain, and to study the effect of bound colicin on lipid bilayer structure and dynamics . We detected (13)C and (15)N isotropic chemical shift differences between the two forms of the protein, which indicate structural changes of the protein due to membrane binding . The Val C(alpha) signal, unambiguously assigned by double-quantum experiments, gave a 0.6 ppm downfield shift in the isotropic position and a 4 ppm reduction in the anisotropic chemical shift span after membrane binding . These suggest that the alpha-helices in the membrane-bound colicin adopt more ideal helical torsion angles as they spread onto the membrane . Colicin binding significantly reduced the lipid chain order, as manifested by (2)H quadrupolar couplings . These results are consistent with the model that colicin Ia channel domain forms an extended helical array at the membrane-water interface upon membrane binding.

Crit Care Med, 2002 Mar, 30(3), 591 - 7
Tumor necrosis factor-alpha-stimulated polymorphonuclear leukocytes suppress migration and bactericidal activity of polymorphonuclear leukocytes in a paracrine manner; Grutkoski PS et al.; OBJECTIVE: Polymorphonuclear leukocytes (PMN) and tumor necrosis factor-alpha (TNF-alpha) play prominent roles in acute respiratory distress syndrome, ischemia-reperfusion injury, trauma, and sepsis . Whereas direct effects of TNF-alpha on PMN function and viability are well documented, little data are available addressing the ability of PMN to communicate with each other in response to cytokine stimulation . Therefore, the aim of this study was to determine whether TNF-alpha can modulate PMN function by inducing PMN to secrete products upon stimulation, which would affect other PMN in vitro in a manner independent of cell contact . METHODS: PMN were purified daily from blood obtained from a pool of 22 healthy volunteers . Conditioned media (CM-TNF) was prepared by incubating PMN in Hanks' balanced salt solution plus TNF-alpha for 1-4 hrs . Freshly isolated PMN were resuspended in CM-TNF and analyzed for 1) phagocytosis of opsonized Escherichia coli, 2) oxidative metabolism as measured as an index of DCF-DA activation, and 3) migration to chemoattractants through Transwell inserts . RESULTS: CM-TNF decreased PMN phagocytotic activity by 8% to 15% and completely suppressed oxidative metabolism but did not modulate the expression of receptors associated with these functions . CM-TNF suppressed the migration of PMN to two biologically relevant agents, N-formyl-methionyl-leucyl-phenylalanine and leukotriene B4, by approximately 65%, but had no effect on PMN migration to interleukin-8 . This suppression was observed for migration across plastic filters as well as extracellular matrix proteins . CONCLUSION: Our data demonstrate that PMN stimulated with TNF-alpha suppress the immunologic function and migration of other PMN independent of cell-cell contact and suggest that TNF-alpha may participate in a negative feedback loop by inducing a PMN-derived factor that counteracts its activity.

Hum Mol Genet, 2002 Apr 15, 11(8), 937 - 43
PLUNC: a novel family of candidate host defence proteins expressed in the upper airways and nasopharynx; Bingle CD et al.; The upper respiratory tract, including the nasal and oral cavities, is the major route of entry of pathogens into the body, and early recognition of bacterial products in this region is critical for host defence . A well-established family of four proteins involved in this process are bactericidal/permeability-increasing protein (BPI) and lipopolysaccharide-binding protein (LBP), which are central to the host defence against bacteria, and cholesteryl ester transfer protein (CETP) and phospholipid transfer protein (PLTP), which have also been implicated in this response . In this paper, we demonstrate the existence of a related family of seven human proteins, which we designate PLUNC proteins . The PLUNC proteins are encoded by adjacent genes found within a 300 kb region of chromosome 20, suggesting that they may be under transcriptional control of shared genomic elements, and expression data shows that these proteins are found in overlapping regions of the pulmonary, nasopharyngeal and oral epithelium, sites where the previously described BPI family members are not expressed . Whereas the BPI family are predicted to share very closely similar three-dimensional structures, the PLUNC family is predicted to have much greater variability in the N-terminal domain, corresponding to the active domain of BPI, thus creating the notion of a BPI/PLUNC structural superfamily . We suggest that members of the PLUNC family may function in the innate immune response in regions of the mouth, nose and lungs, which are sites of significant bacterial exposure.

Antioxid Redox Signal, 2002 Feb, 4(1), 141 - 59
Reactive nitrogen and oxygen intermediates and bacterial defenses: unusual adaptations in Mycobacterium tuberculosis; Zahrt TC et al.; The production of reactive oxygen and reactive nitrogen intermediates is an important host defense mechanism mediated in response to infection by bacterial pathogens . Not surprisingly, intracellular pathogens have evolved numerous defense strategies to protect themselves against the damaging effects of these agents . In enteric bacteria, exposure to oxidative or nitrosative stress induces expression of numerous pathways that allow the bacterium to resist the toxic effects of these compounds during growth in the host . In contrast, members of pathogenic mycobacterial species, including the frank human pathogens Mycobacterium tuberculosis and Mycobacterium leprae, are dysfunctional in aspects of the oxidative and nitrosative stress response, yet they remain able to establish and maintain productive acute and persistent infections in the host . This article reviews the current knowledge regarding reactive oxygen and nitrogen intermediates, and compares the adaptative mechanisms utilized by enteric organisms and mycobacterial species to resist the bactericidal and bacteriostatic effects resulting from exposure to these compounds.

Antioxid Redox Signal, 2002 Feb, 4(1), 61 - 8
Modulation of the cytosolic and phagosomal pH by the NADPH oxidase; Jankowski A et al.; Proton (equivalents) are primary participants in the control and potency of the NADPH oxidase . Both the cytosolic and intraphagosomal pH are influenced during oxidase activation, and maintenance of the optimal environment requires the coordinated action of a series of sophisticated, highly regulated H(+) transporters, including the Na(+)/H(+) exchange, vacuolar H(+)-ATPase, and H(+)-conductive pathway(s) . In addition, protons that are produced during some of the NADPH oxidase reactions then are substrates for the dismutation of superoxide, which precedes production of additional bactericidal agents . In this review, pH homeostasis is shown in conjunction with the NADPH oxidase to present an integrated picture of leukocyte physiology during the phagocytic response.

J Appl Microbiol, 2002, 92(4), 784 - 9
A rapid method for assessing the suitability of quenching agents for individual biocides as well as combinations; Johnston MD et al.; AIMS: To develop a novel, rapid method for testing the ability of quenching agents to neutralize disinfectants . METHODS AND RESULTS: Tests were performed to determine the suitability of different neutralizers for a range of disinfectants, using a new method based on the Bioscreen optical density analyser . Results showed that during disinfection tests, efficacy could be over-estimated due to poor, or no, neutralization of the disinfectant after a specified time of exposure to the bacteria . The failure to distinguish adequately between bacteriostatic and bactericidal effects can lead to false results during disinfectant testing . Experiments also showed that dilution of the disinfectant, following exposure to the bacteria, was not always sufficient to stop the activity of the disinfectant for chemicals with low dilution coefficients . CONCLUSIONS: The quench test proved to be very quick and easy to perform, with results being available within 18 h . Using the Bioscreen, the test is automated and determines whether dilution into a particular neutralizer is able to inactivate a disinfectant within 30 s . SIGNIFICANCE AND IMPACT OF THE STUDY: This new approach allows the efficacy of quenching agents to be determined, prior to undertaking each disinfection study, and can help in the development of more suitable quenching solutions . The test has also been used to find suitable neutralizers for mixtures of disinfectants which might be used during studies on synergistic biocide combinations.

Shock, 2002 Apr, 17(4), 329 - 33
The significance of changes in high mobility group-1 protein mRNA expression in rats after thermal injury; Fang WH et al.; There has been a widespread impression that tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) mediate the toxicity of high doses of lipopolysaccharide (LPS, endotoxin) and are key factors in septic shock . However, the clinical efficacy of treatment with antagonists of TNF-alpha and IL-1beta is still controversial, suggesting that mediators other than TNF-alpha and IL-1beta might contribute causally to endotoxin-induced death . Recent studies implicated high mobility group-1 (HMG-1) protein as a late mediator of endotoxin lethality in mice . However, the role of HMG-1 in mediating multiple organ damage-associating trauma has not been studied . This study was designed to investigate changes in HMG-1 gene expression in vital organs, and its potential role in mediating multiple organ damage following major burns . Wistar rats were subjected to a 35 percent full-thickness thermal injury, and randomly divided into three groups as follows: normal controls (n = 7), thermal injury (n = 24), and recombinant bactericidal/permeability-increasing protein (rBPI21) treatment (n = 12) . Tissue samples from liver and lungs were collected to measure tissue endotoxin levels and HMG-1 mRNA expression . In addition, blood samples were obtained for measurement of organ function parameters . Our data demonstrated a significant increase in HMG-1 gene expression in tissues at 24 h postburn, which remained markedly elevated up to 72 h after thermal injury (P< 0.05-0.01) . Treatment with rBPI21 could significantly decrease tissue HMG-1 mRNA expression in the liver and lung (P < 0.01) . In addition, there were high positive correlations between hepatic HMG-1 mRNA and serum aminoleucine transferase (ALT) and aspartate aminotransferase (AST) levels, and also between pulmonary HMG-1 mRNA and myeloperoxidase activities (P < 0.05-0.01) . Taken together, these findings indicate that thermal injury per se can markedly enhance HMG-1 gene expression in various organs . Up-regulation of HMG-1 expression may be involved in the pathogenesis of endogenous endotoxin-mediated multiple organ damage secondary to major burns.

J Comp Pathol, 2002 Feb-Apr, 126(2-3), 171 - 82
Capsular polysaccharide conjugate vaccines against contagious bovine pleuropneumonia: Immune responses and protection in mice; Waite ER et al.; The immunogenicity of Mycoplasma mycoides subsp . mycoides small colony biotype (MmmSC) vaccines was investigated in BALB/c mice . Groups of mice were vaccinated with either (1) unconjugated capsular polysaccharide (CPS), (2) CPS covalently conjugated to ovalbumin via a carbodiimide reaction, (3) CPS non-covalently bound to latex microspheres, (4) CPS non-covalently complexed with rabbit anti-CPS IgG, and (5) whole inactivated, ultrasonically disrupted (WID) MmmSC . Only mice immunized with the CPS-ovalbumin conjugate exhibited a significant (P<0 small middle dot001) antibody response against CPS . Mice immunized with WID vaccine exhibited a high ELISA antibody titre against non-CPS (protein) antigens only . Mice given WID vaccine were immune against challenge with live MmmSC, and exhibited a significantly reduced degree of mycoplasmaemia (both in incidence and duration) as compared with non-vaccinated controls (P<0 small middle dot001) . Mice immunized with the CPS-ovalbumin conjugate did not exhibit a reduction in mycoplasmaemia . The bactericidal activity of rabbit MmmSC-antiserum in an in-vitro growth inhibition test was related to the CPS antibody titre . This was not observed with antisera from the vaccinated mice . None of the mouse antisera exhibited growth inhibiting activity, irrespective of a high CPS or protein antibody titre (CPS-ovalbumin or WID vaccine groups, respectively) . Thus, it would seem that protection against an MmmSC-induced mycoplasmaemia in the mouse is based upon cell-mediated rather than humoral immunity . The results suggest that conjugation to ovalbumin significantly increases the antibody response to CPS in the mouse; the lack of bactericidal activity of mouse anti-CPS as compared with rabbit anti-CPS in vitro suggests either that the titre of growth inhibiting antibodies is lower in the mouse or that the mechanism of growth inhibition differs between antibodies of the two species . Copyright Harcourt Publishers Ltd.

Anal Bioanal Chem, 2002 Feb, 372(4), 601 - 4 Epub 2001 Dec 21.
A novel chemiluminescent method for the determination of salicylic acid in bactericidal solutions; Cui H et al.; A new flow-injection procedure has been developed for the determination of salicylic acid based on the enhancement of the chemiluminescence from the cerium(IV)-Tween 20 reaction by salicylic acid in acidic medium . The method is simple, selective and sensitive with a detection limit of 2.5x10(-9) g mL(-1) . It is applicable to the determination of salicylic acid in the concentration range of 4.0x10(-9)-1.1x10(-6) g mL(-1) . The relative standard deviation (RSD) is 0.85% for 4.0x10(-7) g mL(-1) salicylic acid (n=11) . The method has been successfully applied to the determination of salicylic acid in bactericidal solutions . Furthermore, it is suggested that light emission from cerium(IV)-Tween 20 reaction is probably because of the formation of singlet oxygen 1O2* and the emitter is excited oxygen molecular pairs O2(1delta(g))O2(1sigma(g)-).

Vet Hum Toxicol, 2002 Apr, 44(2), 83 - 6
Suppressive effect of zearalenone, an estrogenic mycotoxin, on bovine neutrophil chemiluminescence; Murata H et al.; The effects of zearalenone (ZEA), an estrogenic mycotoxin produced by Fusarium fungi, on bovine neutrophils were investigated in vitro using chemiluninescence, a bactericidal parameter . ZEA suppressed luminol-dependent, phorbol 12-myristate 13-acetate (PMA)-elicited chemiluminescence in a dose-dependent manner at concentrations of 10(-4) M and 10(-5) M . No significant suppression was observed at concentrations lower than 10(-6) M . The possible mode ofaction of 10(-4) M ZEA on the cell activity was investigated with special reference to intracellular Ca2+ ({Ca2+}i) release and estrogen receptors . The 10(-4) M ZEA treatment significantly impaired {Ca2+}i release . When pretreated with a low dose (10(-6) M) of PMA, the cells resisted the ZEA-induced chemiluminescence suppression . However, pretreatment of the cells with the estrogen receptor blockers Tamoxifen and ICl 182,780 (both at 10(-6) M) did not annul the suppressive ZEA action . Considering that PMA is an activator of protein kinase C (PKC), a signal transducing enzyme, and in association with a rise in {Ca2+}i causes cytosolic PKC to shift to the plasma membrane where the activated PKC triggers a varied array of cellular responses, the pharmacological dose of ZEA might have suppressed chemiluminescence by hindering the release of {Ca2+}i and the PKC shift . The results of pretreatment with estrogen receptor blockers, however, did not support the suggestion that the ZEA treatment affected the cells via estrogen receptor pathways.

J Mol Biol, 2002 Mar 29, 317(3), 327 - 35
In vivo dissection of the Tat translocation pathway in Escherichia coli; Ize B et al.; The bacterial Tat pathway is capable of exporting folded proteins carrying a special twin arginine (RR) signal peptide . By using two in vivo reporter proteins, we assessed factors that affect Tat pathway transport . We observed that, like the intact RR signal peptide, those with a KR or RK substitution were still capable of mediating the translocation of the folded green fluorescent protein (GFP) . However, the translocation efficiency decreased in the order of RR>KR>RK . The KK motif was unable to mediate GFP translocation . The translocation of the RR-GFP fusion required TatA, TatB and TatC proteins . By exploiting the periplasmic bactericidal property of colicin V (ColV), we constructed a translocation-suicide probe, RR-ColV . The translocation of RR-ColV fully inhibited the growth of wild-type Escherichia coli and those of the DeltatatD and DeltatatE mutants . In contrast, the deletion of the tatC gene blocked RR-ColV in the cytoplasm and this strain exhibited a normal growth phenotype . Interestingly, the growth of DeltatatA and tatB mutants was inhibited partially by RR-ColV . Moreover, KR, RK and KK motifs were capable of mediating the ColV translocation with a decreasing RR=KR>RK>KK efficiency . In addition to TatE and TatC proteins, either TatA or TatB was sufficient for the translocation of RR-ColV or KR-ColV . In contrast, TatA plus the conserved N-terminal domain of TatB were required to mediate the killing effect of ColV fused to the less-efficient RK signal peptide . Taken together, these results suggest that a fully efficient Tat pathway transport is determined by the sequence of the signal peptide, the composition of the Tat apparatus, and the intrinsic characteristics of exported proteins .

Biochem Cell Biol, 2002, 80(1), 113 - 7
Effect of lactoferrin on Helicobacter felis induced gastritis; Dial EJ et al.; Lactoferrin possesses antibiotic, antiinflammatory, and immune-modulating properties that may be active against the gastritis-, ulcer- and cancer-inducing bacterium Helicobacter pylori . In vitro testing of bovine and human lactoferrin by several laboratories has shown significant bacteriostatic and bactericidal activity . Subsequent in vivo testing of bovine lactoferrin in animal models of H . pylori infection has shown beneficial effects of this agent . Our laboratory has utilized a mouse model that is infected with the feline strain of this bacterium, H . felis . The resulting gastritis that develops in this model and the effects of bovine lactoferrin and recombinant human lactoferrin (from Aspergillus niger var . awamori, Agennix Inc., Houston, Tex.) treatment were assessed by various measures . Infected animals treated with orally administered lactoferrin showed reversals in all parameters . In addition, when recombinant human lactoferrin was used in combination with low doses of amoxicillin or tetracycline, there was an enhancement in gastritis-reducing activity . Possible mechanisms for these effects of lactoferrin are discussed . Lactoferrin has significant, orally active in vivo actions and should be further investigated for clinical situations involving Helicobacter infections where it may have utility when administered alone and also when given in combination with established antibiotic agents.

Vaccine, 2002 Mar 15, 20(13-14), 1754 - 60
The outer membrane proteins UspA1 and UspA2 of Moraxella catarrhalis are highly conserved in nasopharyngeal isolates from young children; Meier PS et al.; UspA1 and UspA2 of Moraxella catarrhalis are vaccine candidates . The aims of this study were to determine: (1) the frequencies of occurrence and (2) the degrees of conservation of two surface-exposed epitopes of the uspA1 and uspA2 genes and their respective gene products in 108 nasopharyngeal isolates from young children . The uspA1 and uspA2 genes were detected in 107 (99%) and 108 (100%) isolates, respectively . Twenty-three of 108 uspA2 genes (21%) were identified as the variant gene uspA2H . One-hundred and five isolates (97%) expressed the mAb17C7-reactive epitope shared by UspA1 and UspA2, and 103 isolates (95%) reacted with the UspA1-specific mAb24B5 . The only isolate which lacked a uspA1 gene demonstrated reduced adherence to HEp-2 cells and complement sensitivity . The data indicate that both uspA genes and the expression of at least two surface-exposed epitopes are virtually ubiquitous in isolates from a population at risk for otitis media . A vaccine capable of inducing a bactericidal immune response against the mAb17C7- and/or mAb24B5-reactive epitopes appears promising.

Chin J Traumatol, 1998 Nov 15, 1(1), 21 - 24
Protective effect of bactericidal/permeability-increasing protein in mice with E . coli sepsis; Jiang J et al.; OBJECTIVE: To investigate the effect of bactericidal/permeability-increasing protein(BPI) on the outcome of sepsis in mice and its possible mechanism . METHODS: Sepsis was induced by injection of 2x10(6) colony-formed unit E . coli J5 via the tail vein . BPI of 5 mg/kg or equal volume of normal saline(NS) were injected intravenously at the same time . Endotoxin and TNFalpha levels in serum were assayed using a chromogenic limulus amebocyte lysate test and ELISA respectively . RESULTS: Seventy-two hour survival rate of septic mice was significantly higher in the BPI group (15/18) than in the NS group(8/18, P<0.01) . Serum endotoxin levels in the BPI group (1.3+/-0.3 and 0.7+/-0.4 &mgr;g/L) were significantly lower than those in the NS group (3.9+/-0.8 and 2.5+/-0.9 &mgr; g/L, P<0.01) 0.5 and 1 hour following injection of bacteria respectively . The peak levels of serum tumor necrosis factor-alpha(TNFalpha)in the BPI group (1.9+/-0.6 &mgr;g/L) were also markedly lower than those in the NS group (3.8+/-0.8 &mgr;g/L, P<0.01) 1.5 hours following bacterial injection . But there was no significant difference in blood bacterial count between the BPI and NS groups 0.5, 1.5 and 3.0 hours after injection of bacteria . CONCLUSIONS: BPI has a marked protective effect on E . coli sepsis, which might be related to its action against bacterial endotoxin and its inhibition of TNFalpha production in sepsis.

Int J Urol, 2001 Nov, 8(11), 604 - 8
Changes in cellular immunity during chemotherapy for testicular cancer; Kubota Y et al.; BACKGROUND: The changes in vivo in immunocyte functions during chemotherapy that is administered in combination with granulocyte colony-stimulating factor (G-CSF) in humans have not been fully investigated . This study was designed to examine neutrophil functions and the activities of natural killer (NK) cells, during the administration of chemotherapy and G-CSF for the treatment of testicular cancer . METHODS: Seven patients with germ cell tumors at stage IIA, IIB or IIIB, who were treated with bleomycin, etoposide and cisplatin (BEP), were enrolled in the study . Numbers and activities of neutrophils and NK cells were measured at various times during and after the first course of chemotherapy . Neutrophil phagocytosis was quantitated by flow cytometry with fluorescent latex beads . Bactericidal activity was measured in terms of colony-forming units . The activity of NK cells was measured by monitoring the release of 51Cr . RESULTS: After BEP chemotherapy, CD16+ and CD56+ cell counts, and neutrophil granulocyte counts decreased while there were no significant changes in the number of lymphocytes . Phagocytosis by neutrophils was enhanced after administration of G-CSF . The activity of NK cells was severely impaired after chemotherapy and did not change after administration of G-CSF . CONCLUSIONS: After BEP chemotherapy for testicular cancer with G-CSF, neutrophil function was not at all inferior to those before treatment . Natural killer cell activity was suppressed by the BEP chemotherapy and did not change after administration of G-CSF.

Eur J Clin Invest, 2001 Dec, 31(12), 1064 - 9
In vitro effect of advanced glycation end-products on human polymorphonuclear superoxide production; Bernheim J et al.; BACKGROUND: Advanced glycation end-products (AGEs) are elevated in the sera of diabetic patients . The latter are prone to severe bacterial infections . Advanced glycation end-products have been shown to modulate immune competent cell activities . In this study we examined the in vitro effect of advanced glycation end-products on superoxide anion generation by human polymorphonuclear leukocytes . MATERIALS AND METHODS: Advanced glycation end-products were prepared by incubation of bovine serum albumin (BSA) with glucose for 90 days . Superoxide production was measured as the superoxide dismutase-inhibitable reduction of ferricytochrome c . The effect of advanced glycation end-products on superoxide production was evaluated in both baseline (nonstimulated) and stimulated (by either formyl-methionyl-leucyl-phenylalanine, or phorbol-myristate-acetate) polymorphonuclear leukocytes . RESULTS: The baseline superoxide production of polymorphonuclear leukocytes was significantly increased by advanced glycation end-products in a dose-dependent manner . In contrast, in stimulated polymorphonuclear leukocytes advanced glycation end-products significantly inhibited superoxide production, again in a dose-dependent manner . This inhibitory effect of advanced glycation end-products was observed after dialyzing AGE-BSA, thereby eliminating the possible influence of reactive carbohydrates . No modification of superoxide production was seen with BSA and only a mild inhibitory effect of glucose at high concentrations . CONCLUSIONS: Advanced glycation end-products depress superoxide production by stimulated polymorphonuclear leukocytes . As superoxide plays an essential role in bactericidal activity, this polymorphonuclear leukocyte dysfunction may be a contributory factor to the increased prevalence and severity of bacterial infection seen in diabetic patients.

Vestn Khir Im I I Grek, 2001, 160(6), 51 - 3
{Prophylaxis of recurrent multiple echinococcosis of the liver in children}; Allabergenov AT et al.; The article presents an experience with treatment of 47 children aged from 3 to 14 with multiple echinococcosis of the liver . The authors have made a comparative analysis of the antiparasitic remedies and of a wide spectrum bactericidal solution of Betadin . The results of microscopic, histological and electron microscopic investigations have shown Betadin to be highly effective and harmless for the intraoperative prophylactics of recurrent echinococcosis . It is expedient to use Betadin for multiple echinococcal lesion of the liver in children.

Chin J Traumatol, 1999 Nov 15, 2(2), 84 - 86
Effect of bactericidal/permeability-increasing protein on sepsis induced by intra-abdominal infection in rats; Jiang J et al.; OBJECTIVE: To investigate the effect and mechanism of bactericidal/permeability-increasing protein (BPI) on sepsis induced by intra-abdominal infection in rats . METHODS: Cecal ligation and puncture (CLP) was made on 20 rats with sepsis induced by intra-abdominal infection . BPI or equal volume of physiological saline (PS) was intra-abdominally given immediately and 12 h after CLP, respectively (2.5 mg/kg of BPI each time) . Plasma endotoxin levels were determined with limulus amebocyte chromogenic assay . RESULTS: (1) The survival time in BPI group was significantly higher than that in PS group . (2) The values of the mean arterial pressure (MAP), the left ventricular systolic pressure (LVSP), the isovolumic ventricular pressure (IP), and the maximal change of left intraventricular pressure (+/-dp/dtmax) in BPI group, although decreasing, were markedly higher than those in PS group . (3) Plasma glutamic-pyruvic transaminase (GPT) and urea nitrogen levels in BPI group, though increasing, were obviously lower than those in PS group . (4) There was no significant change of plasma endotoxin levels in BPI group, while plasma endotoxin levels markedly increased in PS group . CONCLUSIONS: BPI has obvious protective effect on sepsis induced by intra-abdominal infection, which might be related to its neutralization of endotoxin.

Curr Mol Med, 2001 Dec, 1(6), 739 - 54
Mammalian secreted phospholipases A2 and their pathophysiological significance in inflammatory diseases; Touqui L et al.; Phospholipases A2 (PLA2s) represent a growing family of enzymes that catalyze the hydrolysis of phospholipids at the sn-2 position leading to the generation of free fatty acids and lysophospholipids . Mammalian PLA2s are divided into two major classes according to their molecular mass and location: intracellular PLA2 and secreted PLA2 (sPLA2) . Type-IIA sPLA2 (sPLA2-IIA), the best studied enzyme of sPLA2, plays a role in the pathogenesis of various inflammatory diseases . Conversely, sPLA2-IIA can exert beneficial action in the context of infectious diseases since recent studies have shown that this enzyme exhibits potent bactericidal effects . Induction of the synthesis of sPLA2-IIA is generally initiated by endotoxin and a limited number of cytokines via paracrine and/or autocrine processes . If the mechanisms involved in the regulation of sPLA2-IIA gene expression have been relatively clarified, little is known on the mechanisms that regulate the expression of other sPLA2 . There have been substantial progresses in understanding the transcriptional regulation of sPLA2-IIA expression . Recently, transcription factors including NF-kappaB, PPAR, C/EBP have been identified to play a prominent role in the regulation of sPLA2-IIA gene expression . The activation of these transcription factors is under the control of distinct signaling pathways (PKC, cAMP ...) . Accumulating evidences in the literature suggest that cytosolic PLA2 together with two sPLA2 isozymes (sPLA2-IIA and sPLA2-V) are functionally coupled with cyclooxygenase-1 and 2 pathways, respectively, for immediate and delayed PG biosynthesis . This spatio-temporal coupling of cyclooxygenase enzymes with PLA2s may represent a key mechanism in the propagation of inflammatory reaction . Unraveling the mechanisms involved in the regulation of the expression of sPLA2s is important for understanding their pathophysiological roles in inflammatory diseases.

J Food Prot, 2002 Mar, 65(3), 476 - 83
Addition of fumaric acid and sodium benzoate as an alternative method to achieve a 5-log reduction of Escherichia coli O157:H7 populations in apple cider; Comes JE et al.; A study was conducted to develop a preservative treatment capable of the Food and Drug Administration-mandated 5-log reduction of Escherichia coli O157:H7 populations in apple cider . Unpreserved apple cider was treated with generally recognized as safe acidulants and preservatives before inoculation with E . coli O157:H7 in test tubes and subjected to mild heat treatments (25, 35, and 45 degrees C) followed by refrigerated storage (4 degrees C) . Fumaric acid had significant (P < 0.05) bactericidal effect when added to cider at 0.10% (wt/vol) and adjusted to pH 3.3, but citric and malic acid had no effect . Strong linear correlation (R2 = 0.96) between increasing undissociated fumaric acid concentrations and increasing log reductions of E . coli O157:H7 in apple cider indicated the undissociated acid to be the bactericidal form . The treatment that achieved the 5-log reduction in three commercial ciders was the addition of fumaric acid (0.15%, wt/vol) and sodium benzoate (0.05%, wt/vol) followed by holding at 25 degrees C for 6 h before 24 h of refrigeration at 4 degrees C . Subsequent experiments revealed that the same preservatives added to cider in flasks resulted in a more than 5-log reduction in less than 5 and 2 h when held at 25 and 35 degrees C, respectively . The treatment also significantly (P < 0.05) reduced total aerobic counts in commercial ciders to populations less than those of pasteurized and raw ciders from the same source (after 5 and 21 days of refrigerated storage at 4 degrees C, respectively) . Sensory evaluation of the same ciders revealed that consumers found the preservative-treated cider to be acceptable.

Comp Biochem Physiol A Mol Integr Physiol, 2002 Apr, 131(4), 917 - 22
The effect of vitamin C on the non-specific immune response of the juvenile soft-shelled turtle (Trionyx sinensis); Zhou X et al.; The study was conducted to determine the effect of supplementation vitamin C in dietary non-specific immunity in juvenile soft-shelled turtles . The soft-shelled turtles were fed with vitamin C supplementation at dosages of 0, 250, 500, 2500, 5000 and 10000 mg/kg diets, respectively, for 4 weeks . The results showed that there were no differences in the phagocytosis of blood cells, serum bacteriolytic activity and bactericidal activity among soft-shelled turtles fed with vitamin C supplementation in 0-500 mg/kg diets . However, firm indicators were significantly enhanced in soft-shelled turtles fed with vitamin C supplementation at 2500 mg/kg diets compared with those fed at 0 and 250 mg/kg diets . The soft-shelled turtles fed with 5000 mg/kg diets had only significantly higher bactericidal activity than those fed vitamin C-deficient diets . The vitamin C supplementation in 10000 mg/kg diets had no notable effects on the phagocytosis, bacteriolytic activity and bactericidal activity . These results suggest that vitamin C seems have an upper and lower threshold for improving non-specific immune function, and the optimum dose was 2500 mg/kg.

J Chemother, 2002 Feb, 14(1), 3 - 12
Immunomodulating activity of quinolones: review; Riesbeck K; Fluorinated quinolones exert their bactericidal activity by inhibiting bacterial type II topoisomerases . At therapeutic concentrations, quinolones superinduce interleukin-2 (IL-2) and interferon-gamma production by mitogen-activated human peripheral blood T lymphocytes . At the molecular level, a stronger activation of the nuclear factor AP-1 ('activator protein-1') is observed in cells incubated with ciprofloxacin, resulting in enhanced cytokine gene transcription . Several cytokine and immediate early (e.g., c-fos and c-jun) mRNAs are upregulated by ciprofloxacin, possibly reflecting a mammalian stress response . In cultures with murine splenocytes, quinolones enhance IL-3 and granulocyte-macrophage colony stimulating factor (GM-CSF) synthesis . The stimulation of these hematopoietic growth factors prolongs survival of mice with depressed bone marrow and prevents experimental antiphospholipid syndrome (APS) . In contrast, quinolones inhibit both human and mouse monocytic IL-1 and TNF-alpha synthesis, an effect that is beneficial in rat experimental type II collagen induced arthritis and LPS-induced septic chock in mice . The intriguing immunomodulatory activities of fluoroquinolones warrant future investigations with new tailored derivatives.

Stomatologiia (Mosk), 2002, 81(1), 44 - 7
{Validation of the optimal concentration of Metrogil-denta preparation in the treatment of periodontal inflammations}; Grudianov AI et al.; Fifty-two patients (22-68 years) with chronic generalized periodontitis of different severity and 223 with fulminant severe periodontitis were observed . The bacteriostatic and bactericidal effects of Metrogil-denta depended on the severity of periodontal involvement and metronidasole concentration in the drug . The maximum bacteriostatic effect was observed at 20-25% and the maximum bactericidal effect at 25% concentration of metronidasole in the preparation irrespective of periodontitis severity.

Crit Care Med, 2002 Feb, 30(2), 428 - 34
The ability of endotoxin-stimulated enterocytes to produce bactericidal factors; Ogle CK et al.; OBJECTIVE: Bactericidal peptides, specifically defensins, are produced by polymorphonuclear cells . Intestinal epithelial cells also produce bactericidal peptides, perhaps as part of their barrier function, to the greatest load of endogenous bacteria present in the body . We sought to determine whether and under what conditions intestinal cell lines could produce bactericidal compounds . DESIGN: Laboratory investigation . SETTING: Children's burn hospital . SUBJECTS: Caco-2, IEC-6, and HT-29 cell lines . INTERVENTIONS: Three different enterocyte lines were cultured for 1 day +/- lipopolysaccharide (1 or 10 microg/mL), and their supernatants were tested for bactericidal activity . Also, reverse transcription-polymerase chain reaction of Caco-2 cells was performed to assess the expression of defensin-6 mRNA . MEASUREMENTS AND MAIN RESULTS: After culture, enterocytes all were found to release one or more soluble factors with bactericidal activity (as measured fluorometrically by using a metabolizable dye) when stimulated by lipopolysaccharide (1 microg/mL) . The bactericidal activity of these culture supernatants was saturated by increased bacterial load, additive to the effects of normal human peripheral blood polymorphonuclear cells, and was reduced by serial supernatant dilution . Enterocyte stimulation with larger amounts of lipopolysaccharide (10 microg/mL) resulted in greater bactericidal activity . After supernatant fractionation based on molecular weight, the bactericidal effect was best retained in the <10-kDa fraction . In addition, the expression of mRNA for defensin-6, a bactericidal peptide produced by neutrophils, was seen in Caco-2 cells . CONCLUSION: Enterocytes are shown to produce a soluble, low molecular weight, bactericidal compound in response to endotoxin stimulation . The expression of defensin-6 mRNA in Caco-2 cells suggests that intestinal cells may release defensins as bactericidal peptides . This experimental system provides an in vitro model to study the activity and production of bactericidal factors by enterocytes.

J Immunol, 2002 Mar 15, 168(6), 2914 - 21
Macrophage effector functions controlled by Bruton's tyrosine kinase are more crucial than the cytokine balance of T cell responses for microfilarial clearance; Mukhopadhyay S et al.; Macrophages from X-linked immunodeficient (xid) mice lacking functional Bruton's tyrosine kinase (Btk) show poor NO induction and enhanced IL-12 induction, and contribute to delayed clearance of injected microfilaria (mf) in vivo . We now show that Btk is involved in other macrophage effector functions, such as bactericidal activity and secretion of proinflammatory cytokines (TNF-alpha, IL-1beta), but not the T cell-directed cytokine IL-12 . Induction of some transcriptional regulators of the NF-kappaB family, crucial for the expression of proinflammatory cytokines, is also poor in Btk-deficient macrophages . Thus, Btk appears to be involved in signaling for inducible effector functions, but not APC functions, in macrophages . Furthermore, adoptive transfer of T cells from mf-infected xid or wild-type mice did not alter the course of mf clearance in recipients, mf clearance was unaltered in IFN-gamma-deficient mice, and improved mf clearance was seen only if greater inducibility of IL-12 was accompanied by greater NO secretion from macrophages, as seen in Ity(r) C.D2 mice as compared with congenic Ity(s) BALB/c mice . Thus, delayed mf clearance in xid mice was correlated not with the high IL-12/Th1 phenotype but with low NO induction levels . Also, xid macrophages showed poor toxicity to mf in vitro as compared with wild-type macrophages . Inhibition of NO production blocked this mf cytotoxicity, and an NF-kappaB inhibitor blocked both NO induction and mf cytotoxicity . Thus, Btk is involved in inducing many macrophage effector functions, and delayed mf clearance seen in Btk-deficient xid mice is due to poor NO induction in macrophages, resulting in compromised microfilarial toxicity.

Zhonghua Shao Shang Za Zhi, 2000 Jun, 16(3), 157 - 60
{The significance of the expressions of lipopolysaccharide binding protein mRNA and lipopolysaccharide receptor CD14 mRNA in the liver of burned rat}; Fang W et al.; OBJECTIVE: To explore the significance and the correlation between enteric endotoxin translocation and hepatic endotoxin -- sensitivity -- enhancing system--lipopolysaccharide -- binding protein (LBP)/lipopolysaccharide receptor CD14 (LBP/CD14) after burn . METHODS: Wistar rats subjected to 35% III degree burn were employed as the model . The rats were randomly divided into three groups: i.e . normal control (C, n = 8) group, thermal injury (T, n = 10) group and recombinant bactericidal/permeability -- increasing protein (rBPI(21)) treatment (R, n = 6) group . The rats in T and R groups were sacrificed at 12 postburn hour (PBH) and the liver tissue was collected for the detection of the mRNA expressions of LBP, CD14 and tumor necrosis factor (TNFalpha) and blood samples collected for hepatic functional indices . RESULTS: The hepatic endotoxin content increased significantly postburn (P < 0.01), and the expressions of LBP/CD14 and TNFalpha mRNA in liver tissue increased obviously . However the use of rBPI(21) could evidently lower hepatic endotoxin content and inhibit the expression of tissue LBP/CD14 and TNFalpha . In addition, rBPI(21) could also significantly decrease serum level of glutamic -- pyruvic transaminase (ALT) (P < 0.01) . CONCLUSION: Aggregation of endotoxin in liver due to postburn translocation might obviously stimulate the expression of LBP/CD14 mRNA locally . The up -- regulation of the expression of LBP/CD14 might be the principle molecular basis enhandced activity of translocated endotoxin on inflammatory cells.

Clin Sci (Lond), 2002 Mar, 102(3), 287 - 95
Effects of ornithine 2-oxoglutarate on neutrophils in stressed rats: evidence for the involvement of nitric oxide and polyamines; Moinard C et al.; Diets enriched in ornithine 2-oxoglutarate (ornithine alpha-ketoglutarate; OKG) improve immune status during stress . We described previously the ability of OKG to increase the respiratory burst in polymorphonuclear neutrophils (PMNs), but the underlying mechanisms remain unclear . OKG is usually recognized as generating glutamine, arginine and polyamines . The aim of the present study was first to determine the effects of OKG on PMN bactericidal functions (chemotaxis and respiratory burst) in stressed rats, and whether these effects could be reproduced by glutamine- or arginine-enriched diets . Secondly, we investigated the metabolic pathway involved in these actions, using three metabolic inhibitors: methionine sulphoximine (an inhibitor of glutamine synthetase), S-methylthiourea (an inhibitor of inducible nitric oxide synthase) and difluoromethylornithine (an inhibitor of ornithine decarboxylase) . OKG, arginine and glutamine all increased the production of reactive oxygen species (evaluated by chemiluminescence, ferricytochrome c reduction and flow cytometry) . Only OKG markedly enhanced the chemotaxis index (5-fold) . Inhibition of glutamine synthetase showed that glutamine production was not involved in the action of OKG . The use of S-methylthiourea and difluoromethylornithine demonstrated that OKG modulated the respiratory burst via nitric oxide (NO*) and polyamine generation . Moreover, OKG stimulated PMN migration via NO*, but arginine administration failed to reproduce this effect . These data suggest that OKG (or its metabolites) and arginine are channelled differently in PMNs . This hypothesis deserves further study.

Probl Tuberk, 2001, (9), 56 - 8
{Surgery on tuberculosis lymphatic system and abdominal organs}; Barinov VS et al.; The paper presents data on surgery for tuberculosis of the lymphatics and abdominal organs . Removal of tuberculosis-affected peripheral lymph nodes during etiotropic therapy yields positive therapeutical results in 84.6% of patients . The efficiency of early operations has been first evidenced by a dynamic study of the systemic immunity . Complicated abdominal tuberculosis is encountered in 44.2% and forced emergency and planned operations on the abdomen to be performed; good late results should be consolidated by combined bactericidal therapy.

Probl Tuberk, 2001, (9), 42 - 5
{Atlantoaxial dislocation in spondylitis}; Lavrov VN et al.; The paper summarizes the outcomes of surgical treatment in 6 patients with nonspecific and tuberculous spondylitis of the craniovertebral area . These patients underwent decompressively stabilizing operations that consists of two steps: at first occipitospondylodesis was performed with a wire and protacryl then via transpharyngeal access with sanitation of the abscess cavity, by removing necrotic tissues--a saving resection of bony tissue sites within the healthy tissues, anterior stabilization of an affected part with an osseous autograft . The above procedure of surgical treatment along with bactericidal therapy yielded positive results by recovering spinal cord function, by forming a bony unit at the site of spinal inflammatory lesion.

Probl Tuberk, 2001, (9), 20 - 1
{Results of surgical intervention depending on duration of preoperative treatment of pulmonary tuberculosis patients}; Grigorian VA et al.; The case histories of 428 patients operated on for tuberculosis were analyzed . Three groups were identified . They were as follows: 1) 121 patients untreated with bactericidal drugs before surgery; 2) 247 patients treated less than 6 months before it; 3) 160 patients treated more than 6 months before surgery . Various complications due to resection of the lung were observed in 30 (7%) patients undergone surgery . They were 6.6, 6.8, and 7.5% in Groups 1, 2, and 3, respectively . A clinical effect was achieved in 99.8% of cases . The late outcomes of surgical intervention were studied within 1 to 10 years in 354 patients, including 102, 119, and 133 patients in Groups 1, 2, and 3, respectively . Progressive and recurrent tuberculosis was revealed in 7 (6.8%), 11 (9.2%), and 18 (13.5%), respectively . Thus, immediate and late outcomes of surgical treatment were not worse in patients with tuberculomas untreated with bactericidal drugs before surgery than in those who receive long-term therapy that substantially reduces the duration of therapy, which is a most important task of modern phthisiology . So patients should be operated on when they are found to have pulmonary tuberculomas without signs of a progressive tuberculous process.

Probl Tuberk, 2001, (9), 10 - 1
{Lung resection in drug resistant tuberculosis patients}; Golovchenko RN et al.; Three and seventy-one case histories of patients operated on for different forms of pulmonary tuberculosis in whom their Mycobacteria tuberculosis (MT) were resistant to bactericidal drugs (BD) were analysed . Fibrocavenous or cavernous pulmonary tuberculosis was detected in most patients (73.3%) . Various postoperative complications occurred in 20% of the patients operated on, in them empyema with bronchial fistula was stated in 6.5% of cases . The immediate outcomes of lung resections were satisfactory in 90% of patients and poor (progressive tuberculosis, uncorrected empyema) in 6%, mortality was 4% . Comparing these data with the outcomes of 6033 resections whose efficiency was 96.4% and mortality 1.4% leads to the conclusion that MT resistance to BD negatively affect the outcomes of resections.

Clin Microbiol Infect, 1999 Jun, 5(6), 331 - 338
Impacts of interleukin-12 on multiplication of Mycobacterium tuberculosis and Mycobacterium avium complex in mice; Lounis N et al.; OBJECTIVE: To evaluate the potential impacts of exogenous administration of murine recombinant interleukin-12 (IL-12) on multiplication of Mycobacterium tuberculosis and M . avium complex (MAC) in murine models . METHODS: Swiss or beige mice were infected intravenously with M . tuberculosis H37Rv or MAC respectively, and were treated by subcutaneous injection with various doses of IL-12, either alone or in combination with chemotherapy . Effectiveness of treatment was assessed by the enumeration of CFUs in the spleens and lungs, together with other indicators . RESULTS: Multiplication of M . tuberculosis was reduced by IL-12 in a dose-dependent manner if the treatment began at day 1, whereas no statistically significant suppression was observed if the treatment began at day 14 . Combination with IL-12 did not enhance the bactericidal activity of antituberculosis chemotherapy . The growth curves of MAC in IL-12-treated mice were almost identical to those of untreated controls, indicating that IL-12 did not affect the multiplication of MAC in beige mice . In both experiments, the dosing of IL-12 approached levels of severe toxicity for the mouse strains used . CONCLUSIONS: IL-12 had a positive affect on early multiplication of M . tuberculosis . It had no effect on early multiplication of M . avium complex.

J Bacteriol, 2002 Mar, 184(5), 1474 - 80
The flavoenzyme ferredoxin (flavodoxin)-NADP(H) reductase modulates NADP(H) homeostasis during the soxRS response of Escherichia coli; Krapp AR et al.; Escherichia coli cells from strain fpr, deficient in the soxRS-induced ferredoxin (flavodoxin)-NADP(H) reductase (FPR), display abnormal sensitivity to the bactericidal effects of the superoxide-generating reagent methyl viologen (MV) . Neither bacteriostatic effects nor inactivation of oxidant-sensitive hydrolyases could be detected in fpr cells exposed to MV . FPR inactivation did not affect the MV-driven soxRS response, whereas FPR overexpression led to enhanced stimulation of the regulon, with concomitant oxidation of the NADPH pool . Accumulation of a site-directed FPR mutant that uses NAD(H) instead of NADP(H) had no effect on soxRS induction and failed to protect fpr cells from MV toxicity, suggesting that FPR contributes to NADP(H) homeostasis in stressed bacteria.

Zhonghua Wai Ke Za Zhi, 1999 May, 37(5), 267 - 70
{Biopterin synthesis and GTP cyclohydrolase I mRNA expression in rats following endotoxic shock}; Yao Y et al.; OBJECTIVE: To investigate the potential role of changes in biopterin (tetrahydrobiopterin and more oxidized species) synthesis and GTP cyclohydrolase I (GTP-CHI) mRNA expression in endotoxic shock . METHODS: SD rats were subjected to endotoxic shock by a bolus injection of lipopolysaccharide (15 mg/kg, i.p.), biopterin and GTP-CHI mRNA levels in liver, kidney, and intestine were determined at 4, 8 hours following endotoxin challenge . RESULTS: Tissue biopterin levels significantly elevated in liver, kidney, and intestine, with high levels sustaining for 8 hours after endotoxin challenge (P < 0.05, P < 0.01) . Similarly, GTP-CHI mRNA induction in various tissues substantially increased within 4 hours compared to normal controls (P < 0.05, P < 0.01) . It was also shown that hepatic and renal biopterin levels were positively correlated with glutamic pyruvic transaminase (GPT) and also blood urea nitrogen (BUN) values (both P < 0.01) . In addition, treatment with recombinant bactericidal/permeability increasing protein (rBPI(21)) markedly reduced tissue biopterin and GTP-CHI mRNA levels, concomitant with significant decreases in GPT, BUN as well as D-lactate values . CONCLUSIONS: Biopterin synthesis and release may be involved in the development of endotoxin-induced multiple organ dysfunction, and early use of rBPI(21) is effective to inhibit marked GTP-CHI mRNA expression and biopterin formation following septic shock.

Zhonghua Wai Ke Za Zhi, 1999 Feb, 37(2), 120 - 2
{Protective effects of bactericidal/permeability-increasing protein on the vital organ function after endotoxic shock in rats}; Jiang J et al.; OBJECTIVE: To investigate the protective effects of BPI on the vital organ function after endotoxic shock in rats, and to provide experimental evidence for further clinical studies . METHODS: 12.5 mg/kg of E . coli 026:B6 LPS were bolus injected via i.a . to reproduce endotoxic shock . Immediately after injection of LPS, 5 mg/kg of BPI or equal volume of physiological saline were bolus injected via i.a . RESULTS: The survival time of shocked animals was prolonged in the BPI group, 24 h survival rate was higher than that in the saline group . The values of mean arterial pressure, left intraventricular systolic pressure, isovolemic ventricular pressure, and dp/dtmax in the BPI group were markedly higher than those in the saline group . The plasma glutamic-pyruvic transaminase and urea nitrogen levels were obviously lower than those in the saline group . The plasma endotoxin and TNF levels in the BPI group were significantly lower than those in the saline group . CONCLUSIONS: BPI has good protective effects on cardiac, hepatic, and renal function in endotoxic shock in rats, indicating that BPI might be useful in clinic treatment of sepsis/septic shock.

Alcohol Alcohol, 2002 Jan-Feb, 37(1), 81 - 6
Concentrations of lipopolysaccharide-binding protein, bactericidal/permeability-increasing protein, soluble CD14 and plasma lipids in relation to endotoxaemia in patients with alcoholic liver disease; Schafer C et al.; There is increasing evidence that gut leakage in persons with chronic alcohol misuse leads to endotoxaemia, which might contribute to the development of alcoholic hepatitis or cirrhosis . In addition, it was recently shown that the endotoxin-binding capacity of whole blood is reduced in these patients . To analyse this phenomenon, we measured the concentration of functionally important endotoxin-binding plasma components which modify the action of endotoxin . In patients with minimal (n = 10), intermediate (n = 9), and cirrhotic alcoholic liver disease (n = 11), and healthy controls (n = 11), plasma endotoxin was determined in a limulus assay . The concentration of lipoproteins was assessed by measuring apolipoproteins, the other factors were directly measured in immunoassays . In the entire group of alcoholics, endotoxin and the concentration of binding factors that are involved in the action of endotoxin on its target cells (LPS-binding protein and sCD14) were increased . Endotoxin antagonists, such as bactericidal/permeability-increasing protein and high-density lipoprotein, were increased in the pre-cirrhotic stages, whereas a significant reduction of the latter was observed in cirrhosis . Low-density lipoprotein remained unchanged . The elevation of binding factors in the pre-cirrhotic stages of alcoholic liver disease might attenuate the effects of endotoxaemia, whereas in cirrhosis the reduction of high density lipoprotein, to which large quantities of endotoxin bind, may contribute to its pro-inflammatory effects.

Angiogenesis, 2001, 4(1), 45 - 51
Designed beta-sheet peptides that inhibit proliferation and induce apoptosis in endothelial cells; Mayo KH et al.; Novel beta-sheet-forming peptide 33 mers, beta pep peptides, have been designed by using a combination approach employing basic folding principles and incorporating short sequences or proposed key residues from the beta-sheet domains of interleukin-8 (IL-8), platelet factor-4 (PF4) and bactericidal/permeability increasing protein (B/PI) . Since PF4 and B/PI are anti-angiogenic and IL-8 is angiogenic, the library of 30 beta pep peptides was assayed for the ability to affect the growth of endothelial cells . Results indicate that five beta pep peptides (beta pep-2, 7, 8, 21 and 25) demonstrate greater than 50% anti-proliferative activity at 30 micrograms/ml, and one of those (beta pep-25) is similarly active at 10 micrograms/ml . Insight into the mechanism of action was probed in an apoptosis assay . Anti-proliferative activity was found to be correlated with the induction of apoptosis . For example, at 100 micrograms/ml beta pep-25 induces 85% of endothelial cells to undergo apoptosis within 2 days . These effects from beta pep peptides appear to be selective for endothelial cell (EC) because normal cells (fibroblasts and leukocytes) and various tumor cells are not significantly affected at peptide concentrations used in this study . Comparison of active and inactive beta pep sequences allows structure-function relationships to be deduced . Five hydrophobic residues and two lysines appear to be crucial to activity . This research contributes to the development of novel anti-angiogenic peptides.

Hong Kong Med J, 1999 Jun, 5(2), 158 - 162
Helicobacter pylori infection: the reduced need for ulcer surgery; Chu KM; The success of the eradication treatment for Helicobacter pylori in managing ulcer disease has further refined the indications for ulcer surgery . More patients are spared unnecessary surgery and its untoward sequelae . It is intriguing that surgery has been effective, given that Helicobacter pylori infection is the underlying culprit . Recent studies show that the prevalence of Helicobacter pylori infection remains high after a vagotomy but is more markedly reduced following a partial gastrectomy . Such a reduction may be due to the removal of distal stomach, which is the usual site of infection . In addition, bile appears to be bactericidal to Helicobacter pylori . The association between Helicobacter pylori infection and perforated duodenal ulcer has not been completely settled . A recent study demonstrates that male gender and positive Helicobacter pylori status are independent factors that are associated with recurrent duodenal ulcer after surgery for perforation . Hence, eradication therapy should be given to any patient who has a Helicobacter pylori infection, after surgery has been performed for perforated duodenal ulceration.

J Biol Chem, 2002 Apr 12, 277(15), 12762 - 9 Epub 2002 Jan 30.
Plunc, a member of the secretory gland protein family, is up-regulated in nasal respiratory epithelium after olfactory bulbectomy; Sung YK et al.; Subtraction suppression hybridization was used with high throughput screening to identify transcripts of genes that are differentially expressed in nasal epithelium following lesioning of the olfactory bulb, termed bulbectomy . We isolated the rat homologue of plunc, a murine gene highly expressed in lung and nasopharyngeal regions, by this method . Rat plunc encodes a 270-amino acid protein containing a putative signal peptide . plunc up-regulation in respiratory epithelium was confirmed by Northern blot and in situ hybridization . plunc mRNA was expressed in nasal epithelium, heart, lung, thymus, and salivary gland in adult rodent . plunc was expressed in nasal epithelium, thymus, and salivary gland during embryogenesis . Antibodies against Plunc detected a 31-kDa protein in lung, heart, and spleen . Rat nasal epithelium displayed robust immunoreactivity that was highly localized to the microvilli layer of respiratory epithelium . The expression of plunc was up-regulated after bulbectomy in respiratory epithelium . We also detected secreted plunc in rat and human mucus . Sequence and homology analyses suggest that Plunc is a member of the secretory gland protein family with putative bactericidal/bacteriostatic function . This is the first protein found in respiratory epithelium whose expression is regulated by olfactory neuronal injury and may provide protection against infection subsequent to injury.

Folia Histochem Cytobiol, 2001, 39 Suppl 2, 73 - 4
The coexistence of tuberculosis infection and lung cancer in patients treated in pulmonary department of Medical Academy in Lublin during last ten years (1990-2000); Rybacka-Chabros B et al.; The coexistance of tuberculous infection (TB-infection) and lung cancer in patients treated in Pulmonary Department of Medical Academy in Lublin during last ten years (1990-2000) has been evaluated . Inclusion criteria involved: aging from 50 to 80 years, tobacco smoking, tuberculous infection in present or in past, lung cancer . All analyzed patients (32 males, 13 females) were heavy smokers (from 10 to 70 cigarettes per day, during at least 5 years) . 27 patients were suffered from lung tuberculosis in past, the rest of them had active tuberculous infection . In 19 cases we detected carcinoma planoepitheliale, in 13 cases carcinoma macrocellulare, in 7 cases carcinoma microcellulare and in 6 cases adenocarcinoma . We concluded, that increased occurrence of lung cancer in TB reinfected patients may be connected with immunodepression caused by chronic TB infection . In patients with new active TB-infection in whom the clinical status and chest X-ray were getting worse in spite of antituberculotic treatment recommended procedures for cancer diagnosis were performed . We suggest that bad results of anti-tuberculotic treatment in TB-infected patients are not always caused by bactericidal resistance . In these cases, the proper diagnosis of lung cancer should be considered.

Wien Med Wochenschr, 2001, 151(24), 600 - 14
{Prevention and therapy of infections in tumor patients}; Burgmann H; The neutropenic patient with fever remains a great challenge . Because of the high risk of life-threatening bacterial infections, all febrile neutropenic patients should be treated with broad-spectrum bactericidal antibiotics promptly by the intravenous route and in maximal therapeutic dosages . Although the currently available diagnostic tests are not sufficiently rapid, sensitive, or specific for identifying or excluding the microbial cause of a febrile episode, a careful diagnostic work-up should be done . The concern about the problem of emerging drug resistance bacteria due to extensive antibiotic use, plus the fact that such prophylaxis has not been shown to reduce mortality rates, led to the recommendation that routine prophylaxis with these antibiotics in neutropenic patients be avoided . Vaccination should be emphasized in these patients.

J Leukoc Biol, 2002 Feb, 71(2), 255 - 61
Activation of phagocytic cell NADPH oxidase by norfloxacin: a potential mechanism to explain its bactericidal action; El Bekay R et al.; The mechanisms underlying the bactericidal power of fluoroquinolones against intracellular parasites in host macrophages remain poorly understood . We have analyzed the effect of norfloxacin, a fluoroquinolone antibiotic, on the production of reactive oxygen intermediates (O(2)(*-) and H(2)O(2)) and NADPH oxidase activity in mouse macrophages . The generation of anion superoxide (O(2)(*-)) was found to be significantly greater in macrophages incubated with norfloxacin than in untreated controls . This enhancing effect of norfloxacin was dose-dependent and reached maximal values within 10 min after its addition . The O(2)(*-) generated was mainly intracellular, as determined by the use of specific dyes, such as lucigenin and luminol, and able to diffuse freely through the cell membrane . Also, the production of H(2)O(2) was increased in macrophages in response to norfloxacin . The positive effect of norfloxacin was associated to an enhanced mobilization of NADPH oxidase subunits p47(phox) and p67(phox) from the cytosol to the plasma membrane in phagocytic cells . The effect of the antibiotic persisted in vivo for several hours . These data support the notion that norfloxacin inhibits mycobacterial growth within phagocytic cells by enhancing intracellular production of O(2)(*-) and other reactive oxygen species.

J Infect Chemother, 2001 Sep, 7(3), 195 - 7
Inhibition by epigallocatechin gallate (EGCg) of conjugative R plasmid transfer in Escherichia coli; Zhao WH et al.; Epigallocatechin gallate (EGCg) in tea catechins blocked or significantly diminished the transfer of conjugative R plasmid between Escherichia coli C600 with plasmid R-222 (donor) and E . coli K-12 RC85 (recipient) in a dose-dependent manner . The inhibition rates of R plasmid transfer by EGCg were 42%-67% at 50-200 microg/ml, and up to 99% at 800 microg/ml . Nevertheless, EGCg, even at the concentration of 1600 microg/ml, was not sufficient to kill E . coli cells in 1 h, as confirmed by determining viable cells after incubation with various concentrations of EGCg . The result indicates that the bactericidal activity of EGCg against the two strains was not the main factor responsible for the inhibition of R plasmid transfer . This observation suggests the possibility that tea consumption may be effective in preventing R plasmid transfer in enteric bacteria.

J Chemother, 2001 Dec, 13(6), 615 - 20
Desirable and undesirable immunotropic effects of antibiotics: immunomodulating properties of cefaclor; Dabrowski MP et al.; The effect exerted on complex immunoregulatory functions of the immune system is an important criterion when selecting an antibiotic . When assessing the effect of an antibiotic on the immune system, one should also take into consideration the existence of functional relationships between the immune system and the nervous and hormonal systems . Among these three systems, there are common factors that modify biological processes . This makes it possible for the antibiotic not only to interact directly with the elements of the immune system, but also to exert indirect influences on potential neurotropic and endocrinotropic effects of the drug . Besides highly effective bactericidal activity, cefaclor demonstrates the ability to exert a favorable effect on some of the specific and non-specific immune responses and immunoregulation mechanisms, which may be important from a clinical point of view . Cefaclor enhances phagocytosis and bactericidal activity of granulocytes and macrophages, and favorably modifies the cooperation of monocytes and T lymphocytes . In this way, it corrects, both in vivo and in vitro, the immunoregulatory disturbances induced and aggravated by an infection . This effect is reflected by an improvement in the impaired immunoregulating activity of T lymphocytes, and is manifest both as an increase in suppressive activity and a correction of the monokine level ratio in relation to the decrease of proinflammatory monokine IL-1 and a relative increase of antiinflammatory IL-1ra . By normalizing the disturbed immunoregulation mechanism, cefaclor enhances the protective potential of the immune reaction while it also reduces the risk of immunogenic clinical complications such as persistent inflammatory conditions and allergic and/or autoaggressive responses . Such immunomodulating properties of cefaclor may be useful in the clinical treatment of patients with immune disorders leading to chronic inflammation and secondary allergic or autoaggressive reactions.

Shock, 2002 Jan, 17(1), 23 - 9
Effect of in vivo infusion of granulocyte colony-stimulating factor on immune function; Valente JF et al.; As the applications of hematopoietic growth factors increase, their complex impact on host defense and immune responses continues to unfold . The effect of the administration of granulocyte colony-stimulating factor (G-CSF) on bacterial defense, proliferation of lymphocytes, and cytokine production by lymphocytes and peripheral blood mononuclear cells (PBMC) was studied . The effect of G-CSF administration on the phenotype of the cells in the major hematopoietic organs was studied as well . ACI rats were given 10 mg/kg/day G-CSF or vehicle daily for 4 days . Isolated bone marrow neutrophils and enterocytes from treated animals showed a greater bactericidal activity than controls . Proliferation of mitogen-stimulated lymphocytes and PBMC was reduced in G-CSF-treated animals . The production of proinflammatory cytokines, tumor necrosis factor (TNF), and interleukin 6 (IL-6) by lymphocytes and PBMC was reduced by G-CSF pretreatment . G-CSF administration caused an increase in IL-4 (Th2 cytokine) release and a decrease in interferon-gamma (IFNgamma, Th1 cytokine) release by mitogen-stimulated lymphocytes . Cytometric analysis of cells in the progenitor cell region indicated a large increase in immature cells in the bone marrow of G-CSF-treated animals compared with sham along with an increase in B cells and a decrease in polymorphonuclear leukocytes (PMNs) . In addition, cytometric analysis showed a large increase in PMNs in blood and splenocytes of the treated animals compared with sham . This study confirms and extends previous observations that G-CSF administration has a number of effects that might simultaneously enhance host defense while reducing the risk of developing uncontrolled systemic inflammation . This may also be efficacious in prolonging graft survival and reducing graft vs . host disease.

Chem Res Toxicol, 2002 Jan, 15(1), 83 - 92
Hypochlorite-induced damage to DNA, RNA, and polynucleotides: formation of chloramines and nitrogen-centered radicals; Hawkins CL et al.; Stimulated monocytes and neutrophils generate hypochlorite (HOCl) via the release of the enzyme myeloperoxidase and hydrogen peroxide . HOCl is a key bactericidal agent, but can also damage host tissue . As there is a strong link between chronic inflammation and some cancers, we have investigated HOCl damage to DNA, RNA, and polynucleotides . Reaction of HOCl with these materials is shown to yield multiple semistable chloramines (RNHCl/RR'NCl), which are the major initial products, and account for 50-95% of the added HOCl . These chloramines decay by thermal and metal-ion catalyzed processes, to give nucleoside-derived, nitrogen-centered, radicals . The latter have been characterized by EPR spin trapping . The propensity for radical formation with polynucleotides is cytidine > adenosine = guanosine > uridine = thymidine . The rates of decay, and yield of radicals formed, are dependent on the nature of the nucleobase on which they are formed, with chloramines formed from ring heterocyclic amine groups being less stable than those formed on exocyclic amines (RNH2 groups) . Evidence is presented for chlorine transfer from the former, kinetically favored, sites to the more thermodynamically favored exocyclic amines . EPR experiments have also provided evidence for the rapid addition of pyrimidine-derived nitrogen-centered radicals to other nucleobases to give dimers and the oxidation of DNA by radicals derived from preformed nucleoside chloramines . Direct reaction of HOCl with plasmid DNA gives rise to single- and double-strand breaks via chloramine-mediated reactions . Preformed nucleoside chloramines also induce plasmid cleavage, though this only occurs to a significant extent with unstable thymidine- and uridine-derived chloramines, where radical formation is rapid . Overall the data rationalize the preferential formation of chlorinated 2'-deoxycytidine and 2'-deoxyadenosine in DNA and suggest that DNA damage induced by HOCl, and preformed chloramines, occurs at sequence-specific sites.

J Clin Laser Med Surg, 1999 Dec, 17(6), 267 - 72
Er:YAG and alexandrite laser radiation propagation in root canal and its effect on bacteria; Jelinkova H et al.; OBJECTIVE: The objective of this study was to compare the bactericidal effect of the Er:YAG (wavelength 2.94 microm) and the Alexandrite (wavelength 0.75 microm) laser radiation . The spreading laser energy in the surrounding hard dental tissues round the root canal was evaluated and the bactericidal effect of both these different laser wavelengths was analyzed . SUMMARY BACKGROUND DATA: The use of a laser to clean and shape the root canal space is the latest method used for cleaning of root canals . The interest in laser endodontics was concentrated on the possibility to extirpate the contents of the root canal, to sterilize and to "melt" the walls of the root canals . The previous reports were performed with CO2, excimer, argon, Nd:YAG, Ho:YAG, and Er:YAG lasers . METHODS: Er:YAG laser system (2.94 microm, energy 100 mJ or 300 mJ, repetition rate 1 Hz, 30 pulses) and alexandrite laser system (0.75 microm, energy 250 mJ, repetition rate 1 Hz, 30 pulses) were prepared and three experimental arrangements were used during the measurements . First the energy transport through the tooth tissue was observed (frontal and side experimental setups) and then, the bactericidal effect was evaluated . RESULTS: It was demonstrated that due to the absorption in the hydroxyapatite and water content in the dentin, the Er:YAG laser radiation is fully absorbed in the root canal wall . This direct influence of the radiation could be expected only close to the sapphire tip . It was found that the tissue, which was not directly affected by the laser radiation, cannot be disinfected by the subablative effect of Er:YAG laser radiation . In the second part of the experiment the real bactericidal effect of Er:YAG ablative energy (300 mJ) could be observed . It was also shown that the alexandrite laser radiation with a wavelength of 0.75 microm spreads through the canal system space and leaks into the surrounding tooth tissues . Both lasers have bactericidal effect . CONCLUSIONS: The pulsed Er:YAG and alexandrite lasers can be efficiently used for killing dental bacteria but the spreading of their radiation in the tooth tissues are different.

Zhonghua Nei Ke Za Zhi, 1999 Jul, 38(7), 451 - 4
{The diagnostic significance of antineutrophil cytoplasmic antibodies in ulcerative colitis}; Liu X et al.; OBJECTIVE: To inquire into the diagnostic significance of antineutrophil cytoplasmic antibodies (ANCA) in ulcerative colitis (UC) . METHODS: Serum ANCA from 58 UC patients, 43 non-UC patients and 58 healthy donors were detected with indirect immunofluorescence (IIF), enzyme linked immunosorbent assay (ELISA)and Western blot . RESULTS: The sensitivity and specificity of the presence of ANCA in the diagnosis of UC were 37.93% and 100% respectively . In the group of UC, patients with mild, moderate and severe clinical manifestations had positive ANCA rates of 17.65%, 41.67% and 52.94% respectively . The occurrence rates of mucosal vasculitis and grade III-V mucosal inflammation were 78.95% and 78.95% respectively in ANCA-positive group, but 37.04% and 44.44% in ANCA-negative group . The levels of the binding of the five kinds of ANCA antigens i.e . myeloperoxidase (MPO), bactericidal/permeability increasing protein (BPI), lactoferrin (LF), cathepsin G (CG) and proteinase-3 (PR-3) with the serum of UC patients were 13.99%, 13.79%, 10.34% 10.34% and 8.62% respectively . Specific protein strips were observed in 48.28% of the UC patients with Western blot technique, 47,000 polypeptide being the most common and making up 22.41% of them . CONCLUSION: The detection of ANCA was an auxiliary method for the diagnosis of UC . At present, UC-associated target antigens were not yet clarified, 47,000 polypeptide may be one of them . ANCA may participate in the pathogenesis of UC.

Rom J Physiol, 1999 Jul-Dec, 36(3-4), 205 - 18
Research concerning the radioprotective and immunostimulating effects of deuterium-depleted water; Bild W et al.; Mice fed for 15 days with Deuterium-Depleted Water (30 ppm deuterium) had a statistically significant increased survival rate compared with control groups fed with normal distilled water (150 ppm deuterium), after 8.5 Gy irradiation (61% survival in the test group versus 25% in the control group) . The hematological picture showed that normal WBC, RBC and platelet counts were maintained in the test groups . Immunological parameters (serum opsonic and bactericidal capacity, bactericidal capacity of the peritoneal macrophages) showed a marked increase in the test groups compared to a severe decrease in the control groups . Auxiliary tests using chemical radiomimetics (hydrochloric embihine) and immunosuppressors (cyclophosphamide) showed a strong protective effect of deuterium-depleted water against the decrease of the leukocyte counts and other immunologic parameters . In conditions of experimental inflammation induced with subcutaneous-implanted pellets, deuterium-depleted water feeding resulted in a statistically significant increase of the inflammatory response, demonstrated by increased percentages of PMN and lymphocytes in the peripheral blood and the increased phagocytic capacity of the peripheral blood PMN . Experimental infections induced with K . pneumoniae 506 and S . pneumoniae 558 in mice irradiated or treated with cyclophosphamide showed increased, non-specific immunity parameters . All results show a marked intensification of the immune defenses and increased proliferation of the peripheral blood cells, probably accounting for the radioprotective effects.

Sheng Wu Gong Cheng Xue Bao, 2001 Sep, 17(5), 587 - 9
{Expression of recombinant BPI23-Fc gamma 1 fusion protein in CHO cells}; Xu JJ et al.; The fusion gene of BPI23 and human Fc gamma 1 was obtained by PCR method, and the expression plasmid was constructed to express recombinant BPI23-Fc gamma 1 fusion protein in CHO cells . After transfection with the plasmid and selection by methotrexate, the cell lines expressing the fusion protein were obtained . The recombinant protein was purified using cation-exchange chromatography and its bioactivity was proved with bactericidal assays.

Eur J Clin Pharmacol, 2001 Nov, 57(9), 631 - 6
In vitro and in vivo inhibition of myeloperoxidase with 5-fluorouracil; Akbiyik F et al.; OBJECTIVE: Myeloperoxidase (MPO) exists in neutrophils and has an important bactericidal role . During phagocytosis, MPO catalyzes a peroxidative reaction using chloride ion and hydrogen peroxide (H2O2) as substrate . The aim of the present study was to investigate whether 5-fluorouracil (5-FU), a chemotherapeutic agent, has a direct inhibitory effect on MPO and to evaluate some properties of this inhibition . METHODS: The inhibitory effect of 5-FU on MPO was studied in rat tissue, human leukocytes, and leukocytes from cancer patients under 5-FU therapy . MPO was solubilized in a detergent-containing buffer . MPO activity was measured spectrophotometrically through the oxidation of a synthetic substrate tetramethyl benzidine in the presence of H2O2 . RESULTS: 5-FU inhibited tissue-associated MPO activity in a dose-dependent but not time-dependent manner with an IC50 value of 0.6 mg/ml . 5-FU also inhibited MPO activity in isolated human leukocytes in a dose-dependent manner, and the IC50 value was 0.75 mg/ml . Using this 5-FU concentration, the inhibitory effect was monitored at different substrate concentrations . Leukocyte MPO activities of patients receiving 5-FU therapy were compared before treatment and after the first, second, and third administration cycles . 5-FU treatment resulted in a significant decrease in leukocyte MPO activity, and repeated 5-FU treatment caused additional decrease . CONCLUSION: Our data showed that 5-FU directly inhibited the MPO activity of human leukocytes in vitro and in vivo . We concluded that, the patients treated with 5-FU should be intensively followed for the risk of infections.

Int Immunopharmacol, 2002 Jan, 2(1), 1 - 13
Receptors for chemotactic formyl peptides as pharmacological targets; Le Y et al.; Leukocytes accumulate at sites of inflammation and immunological reaction in response to locally existing chemotactic mediators . N-formyl peptides, such as fMet-Leu-Phe (fMLF), are some of the first identified and most potent chemoattractants for phagocytic leukocytes . In addition to the bacterial peptide fMLF and the putative endogenously produced formylated peptides, a number of novel peptide agonists have recently been identified that selectively activate the high-affinity fMLF receptor FPR and/or its low-affinity variant FPRL1, both of which belong to the seven-transmembrane (STM), G protein-coupled receptor (GPCR) superfamily . These agonists include peptide domains derived from the envelope proteins of human immunodeficiency virus type 1 (HIV-1) and at least three amyloidogenic polypeptides, the human acute phase protein serum amyloid A, the 42 amino acid form of beta amyloid peptide and a 21 amino acid fragment of human prion . Furthermore, a cleavage fragment of neutrophil granule-derived bactericidal cathelicidin, LL-37, is also a chemotactic agonist for FPRL1 . Activation of formyl peptide receptors results in increased cell migration, phagocytosis, release of proinflammatory mediators, and the signaling cascade culminates in heterologous desensitization of other STM receptors including chemokine receptors CCR5 and CXCR4, two coreceptors for HIV-1 . Thus, by interacting with a variety of exogenous and host-derived agonists, formyl peptide receptors may play important roles in proinflammatory and immunological diseases and constitute a novel group of pharmacological targets.

J Biol Chem, 2002 Mar 8, 277(10), 7970 - 8 Epub 2001 Dec 31.
The carboxyl-terminal domain of closely related endotoxin-binding proteins determines the target of protein-lipopolysaccharide complexes; Iovine N et al.; The bactericidal/permeability increasing (BPI) and lipopolysaccharide (LPS)-binding (LBP) proteins are closely related two-domain proteins in which LPS binding is mediated by the NH(2)-terminal domain . To further define the role of the COOH-terminal domain of these proteins in delivery of LPS to specific host acceptors, we have compared interactions of LBP, BPI, LBP(N)-BPI(C) (NH(2)-terminal domain of LBP, COOH-terminal domain of BPI), and BPI(N)-LBP(C) with purified (3)H-LPS and, subsequently, with purified leukocytes and soluble (s)CD14 . The COOH-terminal domain of LBP promotes delivery of LPS to CD14 on both polymorphonuclear leukocytes and monocytes resulting in cell activation . In the presence of Ca(2+) and Mg(2+), LBP and BPI each promote aggregation of LPS to protein-LPS aggregates of increased size (apparent M(r) > 20 x 10(6) Da), but only LPS associated with LBP and BPI(N)-LBP(C) is disaggregated in the presence of CD14 . BPI and LBP(N)-BPI(C) promote apparently CD14-independent LPS association to monocytes without cell activation . These findings demonstrate that the carboxyl-terminal domain of these closely related endotoxin-binding proteins dictates the route and host responses to complexes they form with endotoxin.

Vet Q, 2001 Nov, 23(4), 153 - 62
How does Mycobacterium avium subsp . paratuberculosis resist intracellular degradation?
Tessema MZ, Koets AP, Rutten VP, Gruys E.
Paratuberculosis is a chronic, progressive disease of mainly ruminants caused by the facultative intracellular bacterium, Mycobacterium avium subsp . paratuberculosis . Infection usually occurs in young animals through oral uptake of food contaminated with the organisms . The ingested bacteria are transcytosed through M-cells overlying the Peyer's patches and are released in the stroma, where they are taken up by macrophages . Inside the macrophage, the mycobacteria resist enzymatic and toxic degradation and multiply until the infected macrophage ruptures . The thick, lipid-rich cell envelope is mainly responsible for micobacterial resistance . In addition to its barrier effect, which provides protections, the mycobacterial cell wall also contains several biologically active components that down-regulate the bactericidal function of macrophages . The basic survival strategy of pathogenic mycobacteria can be viewed at three levels: selective use of relatively safe entry pathways that do not trigger oxidative attack, modification of the intracellular trafficking of mycobacteria-containing phagosomes, and modulation of the cooperation between the innate and specific immunity . In doing so, pathogenic mycobacteria are successful intracellular organisms that survive and multiply inside macrophages . Current understanding about the survival strategies of M . a . paratuberculosis and its implications in the epidemiology, diagnosis, and control of the disease are discussed.

Curr Opin Hematol, 2002 Jan, 9(1), 36 - 42
Neutrophil specific granule deficiency and mutations in the gene encoding transcription factor C/EBP(epsilon); Gombart AF et al.; Neutrophil specific granule deficiency (SGD) is a rare congenital disorder . The neutrophils of these patients display atypical bilobed nuclei; lack expression of at least one primary and all secondary and tertiary granule proteins; and possess defects in chemotaxis, disaggregation, receptor upregulation, and bactericidal activity . SGD patients suffer frequent and severe bacterial infections . Although the first of five patients worldwide was reported in the early 1970s, the molecular basis for the defect was discovered only recently . This review presents data implicating the functional loss of the myeloid transcription factor CCAAT/enhancer binding protein (C/EBP(epsilon)) as a causative agent in the development of SGD . The murine model for SGD provides evidence for defects in eosinophil granule gene expression and indicates abnormalities in macrophage maturation and function . Deficiencies in multiple myeloid lineages, in addition to neutrophils, indicate the importance of C/EBP(epsilon) in regulating important innate immune and inflammatory responses critical for host defense.

Curr Opin Hematol, 2002 Jan, 9(1), 11 - 7
Nox/Duox family of nicotinamide adenine dinucleotide (phosphate) oxidases; Lambeth JD; Reactive oxygen species are classically described as occurring as an accidental byproduct of respiration, and are generally thought to be deleterious to biologic systems . The phagocyte nicotinamide adenine dinucleotide phosphate oxidase provides an example of deliberate reactive oxygen species generation, but the function of this enzyme is to oxidatively modify bacteria as part of bactericidal mechanisms . The discovery of a family of nicotinamide adenine dinucleotide (phosphate) oxidases related to the phagocyte oxidase, the Nox/Duox family, provides additional examples of deliberate generation of reactive oxygen species . This article describes this new family of enzymes and considers hypotheses for their function . Potential roles of Nox/Duox in generation of reactive oxygen species that function in cell signaling (related to growth and angiogenesis), immune function, hypoxic response, and oxidative modification of extracellular matrix proteins are discussed.

Am J Respir Cell Mol Biol, 2002 Jan, 26(1), 105 - 13
Acid stimulation reduces bactericidal activity of surface liquid in cultured human airway epithelial cells; Nakayama K et al.; To examine the effects of acid exposure with moderate acidity (pH 3.0-5.0) on bactericidal activity of airway surface liquid (ASL), ASL was collected by washing the surface of primary cultures of human tracheal epithelial cells 24 h after treatment with phosphate-buffered saline (PBS) adjusted to a pH of 3.0, 4.0, or 5.0 . In all ASL, bactericidal activity was sensitive to sodium concentration . Escherichia coli (500 colony forming units {CFU}) was incubated in ASL, and the number of surviving bacteria was examined . The number of surviving bacteria in ASL from cultured cells with acid exposure at pH 3.0-5.0 was significantly higher than that in control ASL . The minimum inhibitory dilution ratio of ASL against 500 CFU of E . coli was also examined by microdilution assays . According to this assay, the bactericidal activity in ASL with acid challenge at a pH of 3.0 was less than half of that in control ASL . Reverse transcription-polymerase chain reaction and Western blot analysis showed that the production of mRNA and protein of human beta-defensin (HBD)-1 were significantly decreased by acid exposure at pH 3.0-5.0 . In contrast, acid exposure did not change the production of mRNA and protein of HBD-2 and beta-actin mRNA . These results indicate that acid exposure, even with moderate acidity, may inhibit the production of bactericidal molecules, including HBD-1, in airway epithelial cells . Acid exposure may reduce bactericidal activity of ASL in human airway epithelial cells and may increase susceptibility of the airway to bacterial infection.

Infect Immun, 2002 Jan, 70(1), 403 - 6
Growth phase influences complement resistance of Bordetella pertussis; Barnes MG et al.; The BrkA protein of Bordetella pertussis inhibits killing by the antibody-dependent classical pathway of complement; however, susceptibility to complement can be highly variable . Log-phase bacteria grown in Stainer-Scholte (SS) broth plated on Bordet-Gengou (BG) agar were about 500 times more sensitive to killing by complement than stationary-phase SS-BG cultures . While always more susceptible to complement than the wild-type strain, a BrkA mutant displayed a similar growth phase variation in susceptibility to complement . Growth phase susceptibility to complement was also observed for a mutant constitutive for Bvg activation of BrkA, suggesting that modulation of virulence factor expression was not responsible for sensitivity to complement . Susceptibility was not due to differential antigenic expression, since serum adsorbed with complement-resistant, stationary-phase SS-BG cultures lacked bactericidal activity against B . pertussis harvested at all times during the growth cycle . These results suggest that log-phase susceptibility to complement is not due to variable expression of BrkA or antigenic differences and may be an inherent property of rapidly growing cultures . Implications for vaccine development are discussed.

J Leukoc Biol, 2001 Dec, 70(6), 881 - 6
Senescence in innate immune responses: reduced neutrophil phagocytic capacity and CD16 expression in elderly humans; Butcher SK et al.; Elderly humans are more susceptible to bacterial infections because of declining immune status . We have investigated the effect of aging on neutrophil bactericidal responses, comparing neutrophil function in healthy, young (23-35 years) and elderly (>65 years) volunteers . Superoxide generation in response to fMLP was slightly increased in neutrophils from elderly donors, and serum from the elderly was able to opsonize E . coli efficiently . In contrast, phagocytic index was significantly lower in neutrophils from the elderly, compared with young donors (P<0.005) . CD11a and CD11b expression was not affected by age, but CD16 was significantly reduced in neutrophils from elderly donors (P<0.0001) . CD16 expression and phagocytic index were measured in the same neutrophils using FITC-labeled E . coli, PE-conjugated anti-CD16 antibody, and CD16 expression correlated with phagocytic index (r=0.83; P<0.05) . In elderly patients with bacterial infection, CD16 expression remained low . We propose that reduced neutrophil CD16 expression and phagocytosis contribute to human immunesenescence.

J Clin Periodontol, 2001 Dec, 28(12), 1106 - 14
Bacterial survival rate on tooth- and interdental brushes in relation to the use of toothpaste; Quirynen M et al.; BACKGROUND, AIMS: Previous studies indicated that oral hygiene aids can play a role in the intra-oral translocation of pathogens . The survival rate of cariogenic and periodontopathogenic species on toothbrushes, with and without toothpaste, and interdental brushes was presently investigated . MATERIAL AND METHODS: 12 periodontitis patients had their interdental spaces professionally cleaned with interdental brushes and their teeth with new toothbrushes with or without different dentifrices . Each time brushes were rinsed with tap water and stored dry at room temperature . At different time intervals an interdental brush or 4 tufts from a toothbrush were processed for vitality staining and selective and non-selective culturing procedures . RESULTS: Immediately after rinsing, a toothbrush without toothpaste harboured 10(7), 10(8) and 10(7) colony forming units (CFU) of respectively aerobic, anaerobic and black pigmented species . An insignificant decrease occurred the first 24 hours and after 48 hours still 10(4) CFU of aerobic and anaerobic species could be cultured . No periodontopathogen remained detectable at 8 hours, except for Fusobacterium nucleatum . The proportion of vital bacteria decreased in 48 hours from 50% to 30% . Comparable results were obtained for interdental brushes . The bacterial survival rate on toothbrushes was significantly reduced by the use of a detergent containing toothpaste by 2 log at baseline, another 2 log at 4 hours and an extra log more at 8 hours for aerobic and anaerobic species . A toothpaste without detergent only had an insignificant bactericidal effect . CONCLUSION: Toothpaste detergents decrease the survival rate of pathogenic species on a toothbrush and can thus limit the risk for bacterial translocation.

Crit Rev Biomed Eng, 2001, 29(3), 502 - 19
Design and application of low-frequency ultrasound and its combination with laser radiation in surgery and therapy; Zharov VP et al.; Further development of combined laser-ultrasonic medical technologies is presented . An analysis is made of the main effects produced by low-frequency ultrasound on biological tissues . These effects include cavitation, acoustic streamings, acoustic pressure, and mechanical influence . Possible application of low-frequency ultrasound is considered in regard to tumor therapy, bactericidal treatment of infectious tissues, and spray treatment of head and neck wounds . Some limitations of ultrasound application in medicine are discussed, and the feasibility of combining laser radiation with ultrasound is studied . A number of new laser-ultrasonic combinations are put forward and developed . They embrace the combination of photodynamic therapy with ultrasonic tumor therapy (sonophotodynamic therapy), laser-ultrasonic wound treatment (spray treatment), and laser-ultrasonic drug delivery (photophonophoresis) . Some of the above-mentioned technologies were tested experimentally, and the results obtained are considered.

J Biol Chem, 2002 Feb 1, 277(5), 3793 - 800 Epub 2001 Nov 27.
Modulation of mouse Paneth cell alpha-defensin secretion by mIKCa1, a Ca2+-activated, intermediate conductance potassium channel; Ayabe T et al.; Paneth cells in small intestinal crypts secrete microbicidal alpha-defensins in response to bacteria and bacterial antigens (Ayabe, T., Satchell, D . P., Wilson, C . L., Parks, W . C., Selsted, M . E., and Ouellette, A . J . (2000) Nat . Immunol . 1, 113- 38) . We now report that the Ca(2+)-activated K(+) channel mIKCa1 modulates mouse Paneth cell secretion . mIKCa1 cDNA clones identified in a mouse small intestinal crypt library by hybridization to human IKCa1 cDNA probes were isolated, and DNA sequence analysis showed that they were identical to mIKCa1 cDNAs isolated from erythroid cells and liver . The genomic organization was found to be conserved between mouse and human IKCa1 as shown by comparisons of the respective cDNA and genomic sequences . Reverse transcriptase-PCR experiments using nested primers amplified mIKCa1 from the lower half of bisected crypts and from single Paneth cells, but not from the upper half of bisected crypts, villus epithelium, or undifferentiated crypt epithelial cells, suggesting a lineage-specific role for mIKCa1 in mouse small bowel epithelium . The cloned mIKCa1 channel was calcium-activated and was blocked by ten structurally diverse peptide and nonpeptide inhibitors with potencies spanning 9 orders of magnitude and indistinguishable from that of the human homologue . Consistent with channel blockade, charybdotoxin, clotrimazole, and the highly selective IKCa1 inhibitors, TRAM-34 and TRAM-39, inhibited (approximately 50%) Paneth cell secretion stimulated by bacteria or bacterial lipopolysaccharide, measured both as bactericidal activity and secreted cryptdin protein, but the inactive analog, TRAM-7, did not block secretion . These results demonstrate that mIKCa1 is modulator of Paneth cell alpha-defensin secretion and disclose an involvement in mucosal defense of the intestinal epithelium against ingested bacterial pathogens.

J Soc Biol, 2001, 195(2), 143 - 50
{Elastase inhibitors}; Reboud-Ravaux M; The serine proteinase elastase is located in the azurophil granules of mature circulating polymorphonuclear neutrophils . This neutrophil elastase or NE is a potent non specific serine protease which plays a role as bactericidal agent and in the degradation of immune complexes by intraphagosomal processes . It promotes inflammation when the granule contents are secreted in the extracellular environment . In certain pathological circumstances, an imbalance between NE and its major plasmatic inhibitor alpha 1-PI (formerly, alpha 1-antitrypsin) leads to abnormal tissue destruction and disease development . Genetic or acquired alpha 1-PI deficiency is thought to be involved in the pathogenesis of pulmonary emphysema . A variety of degenerative and degradative disorders are also associated to uncontrolled proteolysis by NE (rheumatoid arthritis, glomerulonephritis, adult respiratory distress symptom, psoriasis, cancer) . Numerous inhibitors of NE have been reported . Various molecules are currently undergoing clinical trials for emphysema and other pulmonary diseases.

Chin Med J (Engl), 1999 Nov, 112(11), 998 - 1000
Induction of gastritis and gastric ulcer in Mongolian gerbils infected with Helicobacter pylori; Chi J et al.; OBJECTIVE: To induce the model of Mongolian gerbil infected by Helicobacter pylori (HP) and to observe the changes of the infiltration and the proliferation of inflammatory cells before and after the bactericidal treatment . METHODS: The animal model of HP infection was induced by inoculating the HP strain (American Type Culture Collection) to male Mongolian gerbil MGS/Sea (SPF) . Half a month, 1 month and 3 months after inoculation, kill the animals and make HE staining and immunohistochemistry staining of the stomach to observe the pathological changes . RESULTS: The infiltration of a great number of inflammatory cells was observed with the center of the mucosa of pylorus at the second week of HP infection . At the 3rd month of infection, ulcer appeared near the lesser curvature side of the pylorus . On the early stage of infection, the lesion was acute inflammation characterized by the neutrophilic infiltration, and then transformed into the chronic inflammation characterized by the lymphatic infiltration and the wide formation of lymphatic follicles . CONCLUSION: Mongolian gerbil is an ideal and standard animal model of HP infection.

Placenta, 2001 Nov, 22(10), 882 - 5
Enzyme-cytochemically detectable glucose-6-phosphate dehydrogenase in human villous macrophages (Hofbauer cells); Matsubara S et al.; Though various tissue macrophages possess high glucose-6-phosphate dehydrogenase (G6PD) activity, which plays an important role in their phagocytosis/bactericidal function, the presence of this enzyme in human placental villous macrophages (Hofbauer cells) has not been determined . We examined the ultrastructural localization of glucose-6-phosphate dehydrogenase (G6PD) in Hofbauer cells in first and second trimester placental villi, using a newly developed enzyme-cytochemistry (copper-ferrocyanide) method . Electron-dense deposits indicative of G6PD activity were clearly visible in the cytoplasm and on the cytosolic side of the endoplasmic reticulum of Hofbauer cells . Positive and negative cytochemical controls ensured specific detection of enzyme activity . These observations indicated that Hofbauer cells abundantly possessed enzyme-cytochemically detectable G6PD activity . Hofbauer cell G6PD may play a role in placental defense, by supplying NADPH-dependent enzymes (i.e . nitric oxide synthase or NADPH oxidase) with NADPH . This enzyme may also fuel Hofbauer cells with ribose 5-phosphate during their cell proliferation and cell division .

Am J Dent, 2001 Aug, 14(4), 227 - 32
Pulp capping with a modified bioglass formula (#A68-modified); Stanley HR et al.; PURPOSE: To evaluate a modified Bioglass (Formula #68 (MBF68) when used as a pulp capping agent . MATERIALS AND METHODS: After exposure, the pulps were contaminated with saliva for 60s, rinsed, dried and treated with Consepsis, acting as a hemostatic and bactericidal agent . After drying the Consepsis, the MFB68, having been pre-mixed in sterile saline, was carefully deposited over the pulp exposure, covered with polycarboxylate cement and restored with the ProBond bonding resin system . A calcium hydroxide group was used as control . RESULTS: There was no evidence of mummification, the incidence of properly positioned dentin bridge formation was higher and the incidence of extruded dentin bridge formation was reduced.

J Bacteriol, 2001 Dec, 183(23), 6794 - 800
Genetic evidence for parallel pathways of chaperone activity in the periplasm of Escherichia coli; Rizzitello AE et al.; The periplasm of Escherichia coli contains many proteins proposed to have redundant functions in protein folding . Using depletion analysis, we directly demonstrated that null mutations in skp and surA, as well as in degP and surA, result in synthetic phenotypes, suggesting that Skp, SurA, and DegP are functionally redundant . The Deltaskp surA::kan combination has a bacteriostatic effect and leads to filamentation, while the degP::Tn10 surA::kan combination is bactericidal . The steady-state levels of several envelope proteins are greatly reduced upon depletion of a wild-type copy of surA in both instances . We suggest that the functional redundancy of Skp, SurA, and DegP lies in the periplasmic chaperone activity . Taken together, our data support a model in which the periplasm of E . coli contains parallel pathways for chaperone activity . In particular, we propose that Skp and DegP are components of the same pathway and that SurA is a component of a separate pathway . The loss of either pathway has minimal effects on the cell, while the loss of both pathways results in the synthetic phenotypes observed.

J Org Chem, 1999 Feb 5, 64(3), 731 - 735
Axinellamines A-D, Novel Imidazo-Azolo-Imidazole Alkaloids from the Australian Marine Sponge Axinella sp .
Urban S, de Almeida Leone P, Carroll AR, Fechner GA, Smith J, Hooper JN, Quinn RJ.
Four imidazo-azolo-imidazole alkaloids, axinellamines A-D, have been isolated from an Australian marine sponge, Axinella sp . (order: Halichondrida: family: Axinellidae) . These compounds contain a unique perhydrocyclopenta-imidazo-azolo-imidazole carbon skeleton . Three of these compounds had bactericidal activity against Helicobacter pylori at 1000 &mgr;M.

Vaccine, 2001 Nov 12, 20(3-4), 352 - 8
Serum bactericidal activity and isotype distribution of antibodies in toddlers and schoolchildren after vaccination with RIVM hexavalent PorA vesicle vaccine; de Kleijn E et al.; A clinical phase II trial with the RIVM hexavalent OMV vaccine containing six different PorAs was carried out in toddlers (2-3 years) and schoolchildren (7-8 years) in The Netherlands . Children were vaccinated three times (0, 2, 8 months) . Sera after two and three vaccinations were analysed for serum bactericidal activity (SBA) and isotype distribution in whole cell enzyme linked immunosorbent assay (ELISA) . The SBA after vaccination against the six PorAs was significantly different . We investigated whether the age specific and PorA specific differences in SBA titers correlated with differences in PorA specific IgG isotype distribution . The SBA titers were higher in toddlers compared with schoolchildren . After vaccination, IgG1 antibodies dominated the response followed by IgG3 antibodies . IgG2 levels were low, whereas IgG4 was not detected . Irrespective of PorA, IgG total and isotype specific titers after two and three vaccinations were significantly higher in toddlers than in schoolchildren . A weak correlation was found between IgG total or IgG1 and SBA . Although the immunogenicity of the six PorAs is very different, the isotype distribution was similar for all six tested PorAs.We conclude that the RIVM hexavalent PorA vesicle vaccine induces bactericidal antibodies mainly of the IgG1 and IgG3 isotypes that are considered to be most important for protection against disease . The isotype distribution of the response is not age-dependent.

Gig Sanit, 2001 Sep-Oct, (5), 16 - 21
{Molecular and cellular mechanisms for protecting the respiratory organs from hazardous effects}; Velichkovskii BT; Knowledge of the mechanisms of the body's defense from poor exposures underlies the pathogenetical approach to examining the influence of environmental factors on human health . The respiratory organs are protected from poor exposures by several interrelated systems and mechanisms . The most important ones are an inspired air conditioning system, a respiratory tract self-cleansing system, a pulmonary non-specific bactericidal protection system, an alveolar respiratory surface self-cleansing system, a toxic substance metabolic and neutralizing system, and a specific immune defense system . Free radicals also play a great role in the physiology and pathology.

Int J Tuberc Lung Dis, 2001 Oct, 5(10), 952 - 7
Characterisation of the pncA gene in Mycobacterium tuberculosis isolates from Gauteng, South Africa; Bishop KS et al.; SETTING: The use of pyrazinamide (PZA) is important for the treatment of Mycobacterium tuberculosis as it is bactericidal to semi-dormant mycobacteria that are not affected by other drugs . The incidence of resistance to PZA and other drugs used in the treatment of M . tuberculosis is increasing in South Africa . OBJECTIVE: To characterise the pncA gene of M . tuberculosis isolates from Gauteng, South Africa, and to develop a rapid diagnostic method . DESIGN: The pncA gene and the putative regulatory gene were characterised by sequence analysis in a total of six PZA susceptible and 15 resistant isolates . The association with classical PZA susceptibility testing and PZase activity was determined . RESULTS: All PZA-resistant isolates were PZase negative as well as resistant to at least one other anti-tuberculosis drugs . Mutations were identified throughout the length of the pncA gene in 10/15 PZA-resistant isolates . Five lacked PZase activity, but the wild type pncA sequence was present . In all six PZase-positive strains, a PZA-susceptible pattern was obtained on BACTEC and the wild type pncA sequence was present . CONCLUSION: Sequencing is an effective means to identify mutations in the pncA gene in M . tuberculosis and therefore resistance to PZA . The fact that some PZA-resistant M . tuberculosis isolates lack mutations in the pncA gene suggests that alternative mechanisms for drug resistance exist . In PZase negative strains with no genetic changes which are resistant to 100 microg/ml and susceptible to 300 microg/ml, 300 microg/ml may be a more reliable breakpoint.

Presse Med, 2001 Sep 8, 30(25 Pt 2), XV - XVIII
{Experimental models: interaction of Synercid with other anti-Gram positive agents}; Fantin B; EXPERIMENTAL DATA: In case of resistance to quinupristin, the bacteriostatic synergism is preserved but the in vivo bactericidal effect of Synercid declines . On the other hand, no selection of resistant mutants has been observed . In case of isolated resistance to dafopristin, there is no reduction in the bactericidal effect of Synercid; there is however a possible risk of selecting resistant mutants . To become resistant to Synercid, S . aureus strains have to become resistant to both quinupristin and dafopristin, a highly unlikely situation . POTENTIAL COMBINATIONS: Among the combinations of Synercid with other antibiotics, the combination with vancomycin would have particular interest for clinical applications, increasing bactericidal activity . This would be the case for severe S . aureus infections with a large inoculum and even more so for meti-R resistant strains with a C-MLSB phenotype . Combination with rifampicin would be another possibility, but only for strains not resistant to quinupristin.

Zhonghua Yi Xue Za Zhi, 1999 Apr, 79(4), 289 - 91
{The effect of bactericidal/permeability-increasing protein on lipopolysaccharide-binding protein and lipopolysaccharide receptor CD14 mRNA expression in rats after thermal injury}; Fang W et al.; OBJECTIVE: To elucidate the possible mechanisms underlying beneficial effect of recombinant bactericidal/permeability-increasing protein (rBPI) on multiple organ damage secondary to major burns . METHODS: Wistar rats were subjected to a 35% full-thickness scald injury, and randomly divided into 3 groups: normal controls (n = 8), thermal injury (n = 21), and rBPI21 treatment (n = 12) . In thermal injury and rBPI21 treatment groups, animals were sacrificed at 12 and 24 hours after burn, respectively . Tissue samples from liver, kidneys, lungs and intestine were collected to measure lipopolysaccharide-binding protein (LBP)/lipopolysaccharide receptor CD14 and tumor necrosis factor-alpha (TNF-alpha) mRNA expression . In addition, blood samples were obtained for measurement of organ function parameters . RESULTS: Treatment with rBPI21 could significantly decrease tissue LBP and CD14 mRNA expression in various organs (P < 0.05-0.01) . Also, TNF-alpha mRNA expression in liver, kidneys and lungs were markedly inhibited by rBPI21 secondary to acute insults (P < 0.05-0.01) . Moreover, in the rBPI21 treatment group, significant reduction in serum GPT levels and elevation in intestinal DAO activities were found compared to those in the thermal injury group (P < 0.05-0.01) . CONCLUSIONS: Treatment with rBPI21 is effective in attenuating multiple organ damage resulted from gut origin endotoxin translocation after thermal injury, which might be associated with down-regulation effect of tissue LBP and CD14 gene expression by use of rBPI21.

Zhonghua Yi Xue Za Zhi, 1999 Apr, 79(4), 250 - 2
{Condition and mechanism of host antiinfect capacity induced by bypass-activated complement: experimental study}; Wang Z et al.; OBJECTIVE: To verify host antiinfect capacity induced by bypass-activated complement . METHODS: A monolayer culture of human PMNs and rats kupffer cells (KCs) were added with zymosan-activated human serum (ZAHS) and 3 parameters were measured . ZAHS had been neutralized in tube with antiserum against human C3 and C5 . RESULTS: In 0.07-0.09 ml ZAHS and 1-3 h, intracellular bactericidal activity (ICBA) showed a peak but significant top value at 2nd hour . The value dropped markedly with time and significantly in 4-6 h . Superoxide ions (O2-) dynamics was similar to ICBA . Acid phosphatase (ACP) curve distinguished from the former's with monotonous decrease (1-6 h) and more deep . With 0.11 ml ZAHS after 4 h, about 80% of KCs were fallen off . Three parameters of PMNs were decreased and reached the lowest point in the 6th hour after the adding 0.05-0.09 ml of ZAHS . After blocking test with antihuman C3 and C5 serum, the values of ICBA, O2- and ACP surpassed those in the experimental group . CONCLUSION: The possible mechanism of the origination of the harmful effects of ZAHS is suggested . Before KCs and PMNs undertake phagocytosis, they release the bactericidal and inflamagenic (O2- and ACP) so the intracellular bactericidal activity of KCs and PMNs are decreased . The accumulation of KCs and PMNs in tissues can damage themselves and the adjacent tissues barrier, which will lead to the occurrence of infection.

Antimicrob Agents Chemother, 2001 Nov, 45(11), 3276 - 8
In vitro susceptibility of Coxiella burnetii to linezolid in comparison with its susceptibilities to quinolones, doxycycline, and clarithromycin; Gikas A et al.; The in vitro susceptibility to linezolid shown by nine Greek isolates of Coxiella burnetii derived from patients with acute Q fever was investigated . MICs of linezolid were compared with those of pefloxacin, ciprofloxacin, ofloxacin, trovafloxacin, doxycycline, and clarithromycin using the shell vial assay . MICs of linezolid and clarithromycin ranged from 2 to 4 microg/ml; those of doxycycline, trovafloxacin, and ofloxacin ranged from 1 to 2 microg/ml; those of pefloxacin ranged from 1 to 4 microg/ml; and those of ciprofloxacin ranged from 4 to 8 microg/ml . Linezolid was effective in controlling intracellular parasites in cultures of Vero cells infected by C . burnetii . No bactericidal activity by linezolid was obtained against C . burnetii at 8 microg/ml.

Antimicrob Agents Chemother, 2001 Nov, 45(11), 3229 - 30
Efficacies of clarithromycin regimens against Mycobacterium xenopi in mice; Lounis N et al.; Mice were infected intravenously with 3.5 x 10(7) CFU of Mycobacterium xenopi and treated with various clarithromycin-containing regimens or left untreated for 4 weeks . All nine of the clarithromycin-containing regimens reduced the CFU counts to the levels below the pretreatment values, indicating that these regimens had a bactericidal effect on M . xenopi in mice . The rifampin-isoniazid-ethambutol regimen was significantly less bactericidal than clarithromycin alone or clarithromycin-containing combined regimens.

Antimicrob Agents Chemother, 2001 Nov, 45(11), 3092 - 7
Pharmacodynamics and bactericidal activity of moxifloxacin in experimental Escherichia coli meningitis; Rodriguez-Cerrato V et al.; Moxifloxacin, an 8-methoxyquinolone with broad-spectrum activity in vitro, was studied in the rabbit model of Escherichia coli meningitis . The purposes of this study were to evaluate the bactericidal effectiveness and the pharmacodynamic profile of moxifloxacin in cerebrospinal fluid (CSF) and to compare the bactericidal activity with that of ceftriaxone and meropenem therapy . After induction of meningitis, animals were given single doses of 10, 20, and 40 mg/kg or divided-dose regimens of 5, 10, and 20 mg/kg twice, separated by 6 h . After single doses, the penetration of moxifloxacin into purulent CSF, measured as percentage of the area under the concentration-time curve (AUC) in CSF relative to the AUC in plasma, was approximately 50% . After single doses of 10, 20, and 40 mg/kg, the maximum CSF concentration (C(max)) values were 1.8, 4.2, and 4.9 microg/ml, respectively; the AUC values (total drug) were 13.4, 25.4, and 27.1 microg/ml x h, respectively, and the half-life values (t(1/2)) were 6.7, 6.6, and 4.7 h, respectively . The bacterial killing in CSF for moxifloxacin, calculated as the Deltalog(10) CFU per milliliter per hour, at 3, 6, and 12 h after single doses of 10, 20, and 40 mg/kg were -5.70, -6.62, and -7.02; -7.37, -7.37, and -6.87; and -6.62, -6.62, and -6.62, respectively, whereas those of ceftriaxone and meropenem were -4.18, -5.24, and -4.43, and -3.64, -3.59, and -4.12, respectively . The CSF pharmacodynamic indices of AUC/MBC and C(max)/MBC were interrelated (r = 0.81); there was less correlation with T > MBC (r = 0.74) . In this model, therapy with moxifloxacin appears to be at least as effective as ceftriaxone and more effective than meropenem therapy in eradicating E . coli from CSF.

Nihon Hansenbyo Gakkai Zasshi, 2001 Aug, 70(3), 113 - 9
IL-12 and IL-18 synergistically induce the bactericidal activity of murine peritoneal cells against M . leprae; Nomaguchi H et al.; We examined the effect of IL-12 and IL-18 on bactericidal activities of mouse peritoneal cell (PC) against Mycobacterium leprae (M . leprae) . We demonstrated that IL-12 and IL-18 synergistically induced the NO-dependent bactericidal activity of PC by stimulating Natural Killer (NK) cells and T-cells through IFN-gamma production . IL-12 and IL-18 induced host cell death through NK-cells and T-cells . Therefore . IL-12 and IL-18 play an important role on direct killing of intracellular M . leprae and on indirect killing of them through inducing host cell death.

J Clin Laser Med Surg, 2000 Oct, 18(5), 259 - 62
350 ambulatory hemorrhoidectomies using a scanner coupled to a CO2 laser; Plapler H et al.; OBJECTIVE: The purpose of this study was to investigate how a scanner device would improve the outcome of laser hemorrhoidectomy regarding pain, healing, secretion, and hemostasis . BACKGROUND DATA: The technology of the laser has been applied since the 1960s in several clinical and surgical specialties . The inherent advantages, due to the biological characteristics of this instrument are: (1) it does not affect the neighborning tissues, (2) it is hemostatic, (3) it favors healing, (4) it is bactericidal, and (5) it presents less postoperative pain . Several authors have questioned these supposed advantages . After scanners were introduced, better results were accomplished, including in proctological surgery . METHODS: A total of 350 patients, consecutively, went through a laser hemorrhoidectomy using a CO2 laser coupled to a scanner device in a 3-year period . Patients were monitored for pain, healing, bleeding, and local secretion for up to 36 months . RESULTS: Almost 13% of the patients required bondage of arteries during the operation; 96% used analgesics for up to 3 days; and 5 patients (1.42%) complained of strong pain for several days . Only 2 patients (0.6%) reported intense pain for more than 7 days . All patients presented complete healing between the 30th and the 40th POD . In the 90th POD tissue regeneration appeared healthy . The immediate complications included 2 cases of nipple hemorrhage that demanded a review . Stenosis occurred in 4 patients . Residual "plicoma" was often present with the laser technique (20%) . Return to work, regardless of its nature, was an average, on the 3rd postoperative day . CONCLUSIONS: Our evaluation of the results led to the conclusion that the scanner coupled to the CO2 laser was fundamental for better postoperative outcome in our patients, at least compared to other laser techniques.

Berl Munch Tierarztl Wochenschr, 2001 Sep-Oct, 114(9-10), 375 - 7
Transport stress--consequences for bacterial translocation, endogenous contamination and bactericidal activity of serum of slaughter pigs; Seidler T et al.; On transport and at the abattoir animals are confronted with a lot of stressors, such as sound/noise, crowding/mixing, pollutants and infectious agents that act on the organism . After transport stress an endogenous contamination is often seen in slaughter carcasses and presents a hazard for the consumer . These events are often correlated with a rise in endotoxin level (Misawa et al., 1995; Morales et al., 1992) and a modified immune response . Previous own investigations confirm this hypothesis (Zucker and Kruger, 1998, Seidler et al., 2000) . The attempt was made to investigate the impact of selected stressors (short term transport (1 h), long term transport (7-8 hrs), high temperature, high humidity and intense handling/moving) on bacterial translocation, endogenous contamination, endotoxin levels and bactericidal activity of body fluids.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Jul-Aug, (4), 111 - 4
{Immunopathological responses in women with chronic inflammatory diseases of the uterus and appendages and their therapeutic correction}; Medvedev BI et al.; A total of 145 women with chronic inflammatory diseases of uterus and appendages (IDUA) were examined . Bacterioscopy of smears and culture of vaginal, cervical canal and uterine cavity contents were performed in all patients . Direct immunofluorescence and PCR were used for detection of chlamydiae in scrapes and smears and in biopsy specimens taken from endometrium . The number of T lymphocytes and their subpopulations, B lymphocytes as well as the level of plasmatic cells synthesizing IgA, IgM and IgG and secretory IgA (sIgA), immune complexes and C3 component of the complement were determined in biopsy specimens of the endometrium . Pathomorphological examination revealed structural changes characteristic of the immunocomplex disease (secondary deficiency of sIgA, pronounced suppression of IgA production by plasmocytes along with an essential increase in the number of IgG synthesizing cells and immune complexes fixed to the venule endothelium) were detected . Lectin-histochemical study revealed considerable changes in secretory activity of endometrial epitheliocytes manifested by severe suppression of bactericidal activity of the uterine mucus . The detected disturbances of local protective reactions in the endometrium of women IDUA gave grounds for including immunomodulators into the traditional treatment scheme . The latter made it possible to increase essentially the therapeutic effect.

Toxicol In Vitro, 2001 Aug-Oct, 15(4-5), 271 - 6
Mechanisms underlying chlorhexidine-induced cytotoxicity; Hidalgo E et al.; Chlorhexidine (CLX) is the most widely used antiseptic for wound and skin disinfection . Despite its potent bactericidal action, skin irritation is observed when it is used topically . This study aimed to evaluate the mechanisms underlying CLX-induced toxicity on human dermal fibroblasts with special emphasis on factors that may mediate or counteract its undesirable effects . Cells were exposed to CLX concentrations of 0.00005-0.025% for 3, 6, 8 or 24 h in the absence or presence of different concentrations of foetal calf serum (FCS) (2, 5 and 10%) . Depletion of cell ATP occurred, in a time- and concentration-dependent manner, in all experimental conditions at {CLX} >0.001% . At 24 h of CLX exposure time, the decrease in intracellular ATP was produced from a 10-times lower CLX concentration (0.0001%) . Concentrations > or =0.02% produced total loss of ATP . However, cell survival was maintained after CLX treatment for 3 and 8 h and CLX concentrations > or =0.005% were required to produce total cell death . CLX exerted an inhibitory concentration-dependent effect on DNA synthesis from concentrations as low as 0.0001% . Only FCS at 10% appeared to have a cytoprotective action against CLX-induced cytotoxicity.

Indian J Med Res, 2001 Apr, 113, 125 - 8
Use of vancomycin in the culture of Mycobacterium tuberculosis from gastric lavage; Mathew S et al.; BACKGROUND & OBJECTIVES: Earlier studies from the Tuberculosis Research Centre, Chennai, on culture of Mycobacterium tuberculosis from gastric lavage (GL) specimens in selective Kirchner's medium (SK) resulted in a loss of 60 per cent culture results due to contamination with aerobic spore bearers (ASB) . Addition of vancomycin to SK (SKV) effectively reduced the contamination rate to 20 per cent . The objective of the present study was to further reduce the contamination by collecting the specimens in bottles containing vancomycin, thus providing continuous exposure of the sample to the drug, which is bactericidal to ASB . METHODS: One thousand GL specimens collected from children in vancomycin containing bottles were decontaminated and cultured in SK medium, with and without vancomycin, subcultured on Lowenstein Jensen (LJ) medium and the culture results compared . RESULTS: The contamination of cultures in SK and SKV was 15 and 4 per cent respectively when the specimens were collected in bottles containing vancomycin compared to 60 and 20 per cent contamination reported in the earlier studies . INTERPRETATION & CONCLUSION: The reduced contamination in SK and SKV is most likely due to the collection of samples in vancomycin containing bottles . Although a concurrent comparison of samples processed in vancomycin free conditions would have been ideal, it could not be done due to practical difficulties . The study thus confirms the value of vancomycin as a major deterrent for contamination due to aerobic spores and better results can be obtained if vancomycin is used in sample collection bottles, transport media and liquid culture media used in mycobacteriology laboratories particularly in humid and tropical environment.

Natl Med J India, 2000 May-Jun, 13(3), 121 - 4
Impaired function of neutrophils in uraemic patients; Sharma A et al.; BACKGROUND: Immunodeficiency explains the very high frequency of bacterial infections in patients with chronic renal failure (CRF), which leads to high mortality and morbidity, despite improved therapeutic interventions . Among several factors, the decreased functional capacity of phagocytic leucocytes appears to be responsible for the defective host defence mechanisms against infection in CRF . We evaluated both oxygen-dependent and oxygen-independent microbicidal activity of neutrophils isolated from uraemic patients . METHODS: Forty patients with CRF (20 with mild-to-moderate CRF and 20 with advanced CRF) along with 20 age- and sex-matched healthy controls were studied . The assessment of phagocytic capability, ability to produce superoxide (O2.-) anion and H2O2, myeloperoxidase and granule-specific hydrolytic enzymes such as acid phosphatase, cathepsin D and lysozyme activity of the patient's neutrophils were performed to study their bactericidal activity . RESULTS: The phagocytic index (PI) in the control group was found to be 50.38 (4.58) . It was significantly reduced in both mild-to-moderate CRF and advanced CRF, as compared to controls . In mild-to-moderate and advanced CRF patients, O2.- production by resting polymorphonuclear neutrophils (PMN) was low . Also, on stimulation with PMA the O2.- production showed a relative reduction as compared to controls . H2O2 production by resting PMN from CRF patients was unaltered but on stimulation with PMA, the quantum of increase was significantly lower . A marked reduction in the level of intracellular myeloperoxidase activity in PMN was noted in CRF patients . Of the three intracellular lysosomal enzymes assayed, cathepsin D level was increased in the PMN of mild-to-moderate CRF patients; acid phosphatase level was elevated significantly in the PMN of both mild-to-moderate and advanced CRF patients . However, no change in lysozyme levels was observed . CONCLUSION: With increasing severity of uraemia, neutrophils from uraemic patients showed progressive impairment of phagocytic ability . Impairment of oxygen-dependent microbicidal mechanisms was indicated by a decrease in O2.- and H2O2 production . Increased activity of lysosomal enzymes such as cathepsin D and acid phosphatase suggest a state of neutrophil activation in uraemia . It is likely that the immunodeficiency state in uraemics is partly due to reduced bactericidal activity of the neutrophil cell population.

Am J Physiol Gastrointest Liver Physiol, 2001 Oct, 281(4), G957 - 63
Role of gut flora on intestinal group II phospholipase A2 activity and intestinal injury in shock; Rozenfeld RA et al.; We previously showed that group II phospholipase A2 (PLA2-II), a secretory, bactericidal, and proinflammatory protein in intestinal crypts, is upregulated after lipopolysaccharide (LPS) and platelet-activating factor (PAF) challenge . Here we examined whether germ-free environment (GF) or antibiotic treatment (ABX) affects the pathophysiological responses and intestinal PLA2-II activity after PAF (1.5 microg/kg) or LPS (8 mg/kg) injection . We found that LPS and PAF induced hypotension and mild intestinal injury in conventionally fed (CN) rats; these changes were milder in ABX rats, whereas GF rats showed no intestinal injury . PLA2-II enzyme activity was detected in normal rat small intestine; the basal level was not diminished in ABX or GF rats . PAF and LPS caused an increase in PLA2-II activity, which was abrogated in GF and ABX rats . Recolonization of GF rats by enteral contamination restituted their PLA2-II response to PAF and LPS and susceptibility to bowel injury . We conclude that PAF- and LPS-induced increases in PLA2-II activity are dependent on gut bacteria, and ABX and GF rats are less susceptible to LPS-induced injury than CN rats.

Fish Shellfish Immunol, 2001 Aug, 11(6), 523 - 35
Humoral immune parameters of cultured Atlantic halibut (Hippoglossus hippoglossus L.); Lange S et al.; Several humoral immune factors were studied in a group of cultured halibut (Hippoglossus hippoglossus L.) . The serum protein and IgM concentration was comparable to levels seen in other teleost species . A strong antibody activity against TNP-BSA was observed but not against other antigens tested . Lysozyme and anti-protease activity was detected and showed variable heat sensitivity . Unlike the anti-protease activity, the lysozyme activity of the sera was not sensitive to storage at -20 degrees C . No spontaneous haemolytic activity was observed and the sera had no bactericidal effect on any of the bacterial strains tested . Iron binding capacity of the sera was high . Individual variation was considerable in all the factors tested.

Infect Immun, 2001 Oct, 69(10), 5974 - 80
Interaction of Bartonella henselae with the murine macrophage cell line J774: infection and proinflammatory response; Musso T et al.; Bartonella henselae is the causative agent of cat scratch disease (CSD), a self-limiting condition characterized by a subacute regional lymphadenopathy that may develop into disseminated bartonellosis in immunocompromised subjects . Mice experimentally infected with B . henselae display typical liver and spleen granulomas rich in T cells and macrophages . So far there are no data on the interaction between bartonellae and macrophages . In order to clarify this topic, we investigated the interaction of B . henselae with J774, a mouse macrophage cell line . Analysis of bacterial uptake by functional assays and transmission electron microscopy indicates that bartonellae can enter and survive inside J774 . Entry occurred within 30 min postinfection and reached a plateau at 160 min . Infection of J774 was followed by a dose-dependent release of the proinflammatory cytokines tumor necrosis factor alpha, interleukin 1beta (IL-1beta), and IL-6 . Bartonellae persisted intracellularly without loss of viability for at least 8 h, and their number slightly decreased 24 h postinfection . Gamma interferon (IFN-gamma) treatment of J774 significantly decreased the number of recoverable bacteria at 8 and 24 h . This enhancement of macrophage bactericidal activity was associated with nitric oxide (NO) release and was prevented by the addition of the competitive inhibitor of NO synthesis N(G)-monomethyl L-arginine . These findings suggest that IFN-gamma-mediated activation of macrophages may be important for the clearing of B . henselae infection and that anti-B . henselae microbicidal activity of IFN-gamma-activated macrophages is mediated to a large extent by NO production.

Antibiot Khimioter, 2001, 46(4), 11 - 2
{Effect of pefloxacin on immune response}; Artsimovich NG et al.; The influence on cellular immune response of different doses of the pefloxacin was studied in vivo as well as in vitro experiments . The pefloxacin in super bactericidal concentrations (2.0 mg/ml and 0.4 mg/ml) possess pronounced supressing effect the T-lymphocyte proliferation in blast transformation reaction . While in concentration 0.08 mg/ml pefloxacin does not show such activity . The pefloxacin in maximal effective concentration (200 mg/kg) suppressed activity in delayed hypersensitivity skin reaction of intact mice towards sheep erythrocytes on 20.3 percent only.

J Immunol, 2001 Sep 15, 167(6), 3300 - 7
ATP-mediated killing of intracellular mycobacteria by macrophages is a P2X(7)-dependent process inducing bacterial death by phagosome-lysosome fusion; Fairbairn IP et al.; Mycobacterium tuberculosis survives within host macrophages by actively inhibiting phagosome fusion with lysosomes . Treatment of infected macrophages with ATP induces both cell apoptosis and rapid killing of intracellular mycobacteria . The following studies were undertaken to characterize the effector pathway(s) involved . Macrophages were obtained from p47(phox) and inducible NO synthase gene-disrupted mice (which are unable to produce reactive oxygen and nitrogen radicals, respectively) and P2X(7) gene-disrupted mice . RAW murine macrophages transfected with either the natural resistance-associated macrophage protein gene 1 (Nramp1)-resistant or Nramp1-susceptible gene were also used . The cells were infected with bacille Calmette-Guerin (BCG), and intracellular mycobacterial trafficking was analyzed using confocal and electron microscopy . P2X(7) receptor activation was essential for effective ATP-induced mycobacterial killing, as its bactericidal activity was radically diminished in P2X(7)(-/-) macrophages . ATP-mediated killing of BCG within p47(phox-/-), inducible NO synthase(-/-), and Nramp(s) cells was unaffected, demonstrating that none of these mechanisms have a role in the ATP/P2X(7) effector pathway . Following ATP stimulation, BCG-containing phagosomes rapidly coalesce and fuse with lysosomes . Blocking of macrophage phospholipase D activity with butan-1-ol blocked BCG killing, but not macrophage death . ATP stimulates phagosome-lysosome fusion with concomitant mycobacterial death via P2X(7) receptor activation . Macrophage death and mycobacterial killing induced by the ATP/P2X(7) signaling pathway can be uncoupled, and diverge proximal to phospholipase D activation.

J Immunol, 2001 Sep 15, 167(6), 3029 - 32
Cutting edge: the spirochetemia of murine relapsing fever is cleared by complement-independent bactericidal antibodies; Connolly SE et al.; Abs are the major effectors of host defense against infections with BORRELIA: Bactericidal murine mAbs and their Fabs destroy B . burgdorferi, the agent of Lyme disease, and relapsing fever Borrelia in the absence of complement . These in vitro observations led to the expansion of a search for functionally similar Abs in vivo . In this study, we demonstrate that functionally unique IgM Abs develop in vivo and are responsible for the elimination of spirochetemia in murine models of relapsing fever, without the assistance of complement . Mice deficient in the fifth or third component of complement can clear the spirochetemia, whereas B cell-deficient mice cannot . The B cell-deficient mice developed spirochetemia that was an order of magnitude higher and persisted for a longer period of time in comparison to the wild-type mice . Additionally, B cell-deficient mice passively immunized with immune IgM and with immune serum were protected from challenge.

Compend Contin Educ Dent Suppl, 1996, 17(19), S2 - 7
The use of sodium bicarbonate in oral hygiene products and practice; Newbrun E; Early dentifrices contained natural ingredients, mostly in coarse particle form, and were quite abrasive . Salts, either sodium chloride, sodium bicarbonate, or a mixture of both, have also been used for tooth cleaning because of their ready availability and low cost . Because of both their relatively low intrinsic hardness and their high solubility, another advantage is low abrasivity . Their biggest disadvantage is a salty, unpalatable taste . Many modern dentifrices that contain sodium bicarbonate, either as the sole abrasive or one of several, disguise the saltiness with flavoring and sweetening agents . An almost inverse relationship exists between the percentage of baking soda in a dentifrice and its abrasivity . Sodium bicarbonate has no anticaries activity per se but is compatible with fluoride . In high concentrations, sodium bicarbonate is bactericidal against most periodontal pathogens . Most clinical studies have not found significant differences in periodontal response to baking soda as compared with other commercial dentifrices, probably because of its rapid clearance from the gingival sulcus . Sodium bicarbonate may not be the "magic bullet" for curing dental diseases, but its safety (if ingested), low abrasivity, low cost, and compatibility with fluoride make it a consummate dentifrice ingredient.

Eur J Pharm Biopharm, 2001 Sep, 52(2), 151 - 8
In vivo-in vitro study of biodegradable and osteointegrable gentamicin bone implants; Sanchez E et al.; Three implants composed of phosphate (25% hydroxyapatite, 75% tricalcium phosphate), 20% poly(DL-lactide) (DL-PLA; weight-average molecular weight (Mw), 30 kD) and 3% gentamicin sulphate (GS) were assayed in vitro and in vivo to study their release profiles as potential drug delivery systems to prevent or treat osteomyelitis . To prolong GS release, some implants were coated with poly(lactide-co-glycolide) (PLGA; Mw, 100 kD; I-PLGA) or DL-PLA (Mw, 200 kD; I-PLA) . GS levels were measured in bone, kidney and blood after implantation into the femur of rats . The release profiles show a burst in the first few days, followed by a slower release rate . After I-PLA implantation, bone antibiotic concentrations higher than the minimum bactericidal concentration were maintained for 4 weeks . A linear correlation between in vitro and in vivo GS release was found to continue until complete drug release . Histological and radiological analysis showed that the implants were well tolerated and gradual new bone formation was observed.

Rheumatol Int, 2001 Jul, 20(5), 197 - 204
Antineutrophil cytoplasmic antibodies in patients with systemic lupus erythematosus: prevalence, antigen specificity, and clinical associations; Manolova I et al.; Fifty-five patients with systemic lupus erythematosus (SLE) were examined for antineutrophil cytoplasmic antibodies (ANCA) by indirect immunofluorescence (IIF) . Enzyme-linked immunosorbent assay (ELISA) for ANCA against myeloperoxidase (MPO), lactoferrin (LF), proteinase 3 (PR3), elastase (HLE), and bactericidal/permeability-increasing protein (BPI) was performed . The prevalence of ANCA by IIF was 29.1% (16/55 patients) . MPO-ANCA were found in 10.9% (6/55), LF-ANCA in 18.2% (10/55), PR3-ANCA in 12.7% (7/55), BPI-ANCA in 23.6% (13/55), and HLE-ANCA in 1.8% (1/55) . The levels of BPI-, LF-, and PR3-ANCA correlated with disease activity . A significant association between serositis and the presence of BPI-, LF-, and PR3-ANCA was observed, and PR3-ANCA were found to be associated with arthritis as well . Our results demonstrate that ANCA of various specificities occur in SLE, and BPI appears to be an important target antigen.

Chem Res Toxicol, 2001 Aug, 14(8), 1071 - 81
Hypochlorite-induced damage to nucleosides: formation of chloramines and nitrogen-centered radicals; Hawkins CL et al.; Stimulated monocytes and neutrophils generate hypochlorite (HOCl) via the release of the enzyme myeloperoxidase and hydrogen peroxide . HOCl is a key bactericidal agent, but can also damage host tissue . As there is a strong link between chronic inflammation and some cancers, we have investigated HOCl damage to DNA bases . We show that reaction of HOCl with the exocyclic -NH(2) groups of cytidine, adenosine, and guanosine, and the ring NH groups of all bases, yields chloramines (RNHCl/RR'NCl) . These are the major initial products . Chloramine decay can be accelerated by UV light and metal ions, and these reactions, together with thermal decomposition, give rise to nucleoside-derived nitrogen-centered radicals . Evidence is presented for the rapid addition of pyrimidine-derived nitrogen-centered radicals to another parent molecule to give dimers . Experiments with nucleoside mixtures show that the propensity for radical formation is cytidine > adenosine = guanosine > uridine = thymidine . These data are inconsistent with the selectivity of HOCl attack and the stability of the resulting chloramines, but can be rationalized if chlorine transfer between bases is rapid and yields the most stable chloramine, with such transfer preceding radical formation . Thus, though thymidine is the major initial site of chloramine formation, rapid chlorine atom transfer generates cytidine and adenosine chloramines . These reactions rationalize the preferential formation of chlorinated cytidine and adenosine in DNA.

Toxicol Appl Pharmacol, 2001 Aug 15, 175(1), 1 - 9
3-Methylcholanthrene increases phorbol 12-myristate 13-acetate-induced respiratory burst activity and intracellular calcium levels in common carp (Cyprinus carpio L) macrophages; Reynaud S et al.; Phagocytic cells play a key role in the fish immune system . They secrete reactive oxygen species (ROS) involved in their bactericidal activity . These cells are highly sensitive to pollution by polycyclic aromatic hydrocarbons and other organic pollutants . We have investigated the intracellular mechanisms by which 3-methylcholanthrene (3-MC) increased bactericidal activity of carp phagocytes . Macrophages isolated from head kidney (pronephros) and incubated 1 h with 3-MC enhanced their production of ROS when they were stimulated 1.25 h with phorbol 12-myristate 13-acetate (PMA), a direct activator of protein kinase C (PKC) . 3-MC also produced a rapid and a sustained increase in {Ca(2+)}(i) (2 h minimum) . However, the cytochrome P450 1A and Ah receptor inhibitor, alpha-naphtoflavone (alpha-NF), inhibited the potentiation of PMA-induced ROS production, suggesting 3-MC metabolic activation . Moreover, alpha-NF increased {Ca(2+)}(i) without macrophage ROS production, suggesting that some mechanism other than calcium release is playing a role in the stimulation of the macrophages by 3-MC . The rise in {Ca(2+)}(i) induced by 3-MC was potentiated by the inhibitor of the endoplasmic reticulum calcium ATPases, thapsigargin . And treating the cells with 3-MC decreased the calcium mobilization caused by thapsigargin . These results suggest that 3-MC acts on the endoplasmic reticulum, perhaps directly on calcium ATPases, to increase intracellular calcium levels in carp phagocytes .

Crit Care Med, 2001 Aug, 29(8), 1621 - 5
Activation of alveolar macrophages in acid-injured lung in rats: different effects of pentoxifylline on tumor necrosis factor-alpha and nitric oxide production; Kudoh I et al.; OBJECTIVE: To determine whether acid instillation augments tumor necrosis factor-alpha and nitric oxide production by alveolar macrophages in rats, and to study the effects of treatment with pentoxifylline before acid instillation on the production of these inflammatory mediators . DESIGN: Controlled laboratory investigation on tumor necrosis factor-alpha and nitric oxide production by alveolar macrophages of rats that had acid-induced lung injury . SETTING: University research laboratory . SUBJECT: Alveolar macrophages of rats . INTERVENTIONS: Alveolar macrophages were recovered by bronchoalveolar lavage at 4, 10, 16, 24, and 72 hrs after unilateral hydrochloric acid (pH, 1.0; volume, 0.1 mL) instillation into the lungs of rats . Alveolar macrophages then were cultured with or without lipopolysaccharide . One group of rats was pretreated with pentoxifylline before acid instillation . MEASUREMENTS AND MAIN RESULTS: Alveolar macrophages from both acid-instilled and contralateral lungs, which had recovered 24 hrs after acid instillation, produced significantly greater tumor necrosis factor-alpha and nitric oxide . Subsequent exposure to lipopolysaccharide, as a surrogate for bacterial infection, further promoted tumor necrosis factor-alpha and nitric oxide release . Alveolar macrophages from rats pretreated with pentoxifylline before acid instillation produced significantly less tumor necrosis factor-alpha and did not overproduce tumor necrosis factor-alpha when exposed to lipopolysaccharide . In contrast, pretreatment with pentoxifylline had no effect on nitric oxide production by alveolar macrophages . CONCLUSIONS: Acid instillation stimulates alveolar macrophages to produce tumor necrosis factor-alpha and nitric oxide . Pentoxifylline preserved innate production of tumor necrosis factor-alpha to lipopolysaccharide and did not inhibit the production of bactericidal nitric oxide . This may partly explain why pentoxifylline reduces acid aspiration-induced lung injury while maintaining the host's ability to combat bacterial infection after acid aspiration.

Ann Surg, 2001 Aug, 234(2), 233 - 44
Bactericidal/permeability-increasing protein attenuates systemic inflammation and acute lung injury in porcine lower limb ischemia-reperfusion injury; Harkin DW et al.; OBJECTIVE: To investigate the role of recombinant bactericidal/permeability-increasing protein (rBPI21) in the attenuation of the sepsis syndrome and acute lung injury associated with lower limb ischemia-reperfusion (I/R) injury . SUMMARY BACKGROUND DATA: Gut-derived endotoxin has been implicated in the conversion of the sterile inflammatory response to a lethal sepsis syndrome after lower torso I/R injury . rBPI21 is a novel antiendotoxin therapy with proven benefit in sepsis . METHODS: Anesthetized ventilated swine underwent midline laparotomy and bilateral external iliac artery occlusion for 2 hours followed by 2.5 hours of reperfusion . Two groups (n = 6 per group) were randomized to receive, by intravenous infusion over 30 minutes, at the start of reperfusion, either thaumatin, a control-protein preparation, at 2 mg/kg body weight, or rBPI21 at 2 mg/kg body weight . A control group (n = 6) underwent laparotomy without further treatment and was administered thaumatin at 2 mg/kg body weight after 2 hours of anesthesia . Blood from a carotid artery cannula was taken every half-hour for arterial blood gas analysis . Plasma was separated and stored at -70 degrees C for later determination of plasma tumor necrosis factor (TNF)-alpha, interleukin (IL)-6 by bioassay, and IL-8 by enzyme-linked immunosorbent assay (ELISA), as a markers of systemic inflammation . Plasma endotoxin concentration was measured using ELISA . Lung tissue wet-to-dry weight ratio and myeloperoxidase concentration were used as markers of edema and neutrophil sequestration, respectively . Bronchoalveolar lavage protein concentration was measured by the bicinclinoic acid method as a measure of capillary-alveolar protein leak . The alveolar-arterial gradient was measured; a large gradient indicated impaired oxygen transport and hence lung injury . RESULTS: Bilateral hind limb I/R injury increased significantly intestinal mucosal acidosis, intestinal permeability, portal endotoxemia, plasma IL-6 concentrations, circulating phagocytic cell priming and pulmonary leukosequestration, edema, capillary-alveolar protein leak, and impaired gas exchange . Conversely, pigs treated with rBPI21 2 mg/kg at the onset of reperfusion had significantly reduced intestinal mucosal acidosis, portal endotoxin concentrations, and circulating phagocytic cell priming and had significantly less pulmonary edema, leukosequestration, and respiratory failure . CONCLUSIONS: Endotoxin transmigration across a hyperpermeable gut barrier, phagocytic cell priming, and cytokinemia are key events of I/R injury, sepsis, and pulmonary dysfunction . This study shows that rBPI21 ameliorates these adverse effects and may provide a novel therapeutic approach for prevention of I/R-associated sepsis syndrome.

Auris Nasus Larynx, 2001 Aug, 28(3), 227 - 32
Fosfomycin nebulizer therapy to chronic sinusitis; Kamijyo A et al.; OBJECTIVE: effects of Fosfomycin (FOM) nebulizer therapy were studied in patients with chronic sinusitis . METHODS: about 28 patients with chronic sinusitis were administered 2 ml of FOM sodium (3% w/v) by nebulizer three times per week for 4 weeks . Levels of IL-1 beta, IL-6, IL-8, and TNF-alpha in nasal lavage were also measured before and at the end of treatment . RESULTS: the overall efficacy of this treatment on the basis of both subjective and objective symptoms, was 'excellent' for 28.6%, 'good' for 10.7%, 'fair' for 39.4%, and yield 'no change' for 21.4% of the patients . Both IL-1 beta and IL-6 concentrations were significantly decreased after treatment . Although the IL-8 level did not significantly decrease, it seems to be related to the overall efficacy . TNF-alpha was not detected in all of the samples . CONCLUSION: FOM nebulization therapy is highly effective in treatment for chronic sinusitis, and efficacy may be due to an immunomodulatory mechanism, as well as its bactericidal effect.

J Biomed Mater Res, 2001 Nov, 57(2), 313 - 20
Silica sol-gel for the controlled release of antibiotics . I . Synthesis, characterization, and in vitro release; Radin S et al.; Room temperature processed silica sol-gel (xerogel) was investigated as a novel controlled release carrier of antibiotics (vancomycin) . Xerogel characteristics, in vitro release properties, and bactericidal efficacy of the released antibiotic were determined . The xerogel/vancomycin composite showed a long-term sustained release (up to 6 weeks) . In addition, bactericidal efficacy of released vancomycin was retained . The kinetics of release and the amount released were dose dependent . The initial, first-order release was followed by a near-zero-order release . The time to transition from the first- to zero-order release increased with vancomycin load (from 2 to 3 weeks with load increase from 2.2 to 11.1 mg/g) . Regardless of the load, about 70% of the original vancomycin content was released by the transitional point, and the cumulative release after 6 weeks of immersion was about 90% . This study, combined with other reports documenting biocompatibility and controlled resorbability of the xerogel/drug composite in vivo, suggests that silica xerogel is a promising controlled release material for the treatment of bone infections .

Klin Khir, 2001 Feb, (2), 35 - 7
{Morphological substantiation of choice of the laser processing of residual cavity wall after hydatidectomy for pulmonary hydatid cyst}; Grubnik VV et al.; Morphological investigation of the pulmonary echinococcal cyst after applying the laser irradiation influence on it was done . The optimal way for performance of an adequate scolexo- and bactericidal treatment of fibrous capsule after the pulmonary hydatidectomy conduction is application of defocused to 5-10 mm irradiation of the CO2 and the Nd AIG-laser with 150-250 Wt/cm2 power density . In the cyst suppuration it is preferable to use Nd AIG-laser with increase of the exposition time in 3-4 times and of the energy absorbance up to 400-800 J/cm2 . Local coagulation using irradiation of the Nd AIG-laser by the contact method with the 20 Wt power output (250-400 Wt/cm2 power density) is applied if bronchi, which are confided into residual cavity, have diameter up to 3 mm.

Proc Natl Acad Sci U S A, 2001 Aug 14, 98(17), 9901 - 6 Epub 2001 Jul 31.
Peptide methionine sulfoxide reductase from Escherichia coli and Mycobacterium tuberculosis protects bacteria against oxidative damage from reactive nitrogen intermediates; St John G et al.; Inducible nitric oxide synthase (iNOS) plays an important role in host defense . Macrophages expressing iNOS release the reactive nitrogen intermediates (RNI) nitrite and S-nitrosoglutathione (GSNO), which are bactericidal in vitro at a pH characteristic of the phagosome of activated macrophages . We sought to characterize the active intrabacterial forms of these RNI and their molecular targets . Peptide methionine sulfoxide reductase (MsrA; EC ) catalyzes the reduction of methionine sulfoxide (Met-O) in proteins to methionine (Met) . E . coli lacking MsrA were hypersensitive to killing not only by hydrogen peroxide, but also by nitrite and GSNO . The wild-type phenotype was restored by transformation with plasmids encoding msrA from E . coli or M . tuberculosis, but not by an enzymatically inactive mutant msrA, indicating that Met oxidation was involved in the death of these cells . It seemed paradoxical that nitrite and GSNO kill bacteria by oxidizing Met residues when these RNI cannot themselves oxidize Met . However, under anaerobic conditions, neither nitrite nor GSNO was bactericidal . Nitrite and GSNO can both give rise to NO, which may react with superoxide produced by bacteria during aerobic metabolism, forming peroxynitrite, a known oxidant of Met to Met-O . Thus, the findings are consistent with the hypotheses that nitrite and GSNO kill E . coli by intracellular conversion to peroxynitrite, that intracellular Met residues in proteins constitute a critical target for peroxynitrite, and that MsrA can be essential for the repair of peroxynitrite-mediated intracellular damage.

Int J Biochem Cell Biol, 2001 Sep, 33(9), 877 - 88
Effects of chemical modifications of crotoxin B, the phospholipase A(2) subunit of crotoxin from Crotalus durissus terrificus snake venom, on its enzymatic and pharmacological activities; Soares AM et al.; Crotoxin B, the basic Asp49-PLA(2) subunit from crotoxin, the main component of Crotalus durissus terrificus venom, displays myotoxic, edema-inducing, bactericidal (upon Escherichia coli), liposomal-disrupting and anticoagulant activities . Chemical modifications of His (with 4-bromophenacyl bromide, BPB), Tyr (with 2-nitrobenzenesulphonyl fluoride, NBSF), Trp (with o-nitrophenylsulphenyl chloride, NPSC) and Lys (with acetic anhydride) residues of this protein, in addition to cleavage with cyanogen bromide (CNBr) and inhibition with ethylenediaminetetraacetic acid (EDTA), were carried out in order to study their effects on enzymatic and pharmacological activities . Lethality was reduced after modification of His or Lys residues, as well as after cleavage with CNBr, while enzymatic activity was completely abolished after modification of His or incubation with EDTA . Modification of Lys or Tyr, or cleavage with CNBr, partially reduced enzymatic activity . Anticoagulant activity was modified similarly to enzymatic activity, evidencing the dependency of this pharmacological effect on catalytic activity . Myotoxicity was reduced after modification of His or Lys, as well as after cleavage with CNBr, whereas EDTA reduced this effect to a lesser extent . Bactericidal effect was significantly reduced only after modification of Lys and after cleavage with CNBr . Edema-inducing activity was partially inhibited after treatment with EDTA and strongly reduced after acetylation of Lys residues and cleavage with CNBr, being only partially reduced after His alkylation . On the other hand, liposome disrupting activity was only partially reduced after modification of His and Tyr or after cleavage with CNBr . Modification of Trp residue partially reduced lethality and myotoxicity but did not affect enzymatic or anticoagulant activities . These data indicate that enzymatic activity is relevant for some pharmacological effects induced by crotoxin B (mainly lethal, myotoxic and anticoagulant activities), and also evidence that this subunit of crotoxin displays regions different from the active catalytic site which are involved in some of the toxic and pharmacological effects induced by this phospholipase A(2).

Vet Immunol Immunopathol, 2001 Aug 10, 80(3-4), 225 - 35
Inflammation-induced changes in serum modulate chicken macrophage function; Xie H et al.; Inflammation-induced changes in serum protein profiles and the effects of such serum on a chicken macrophage cell line HD11 were studied to find whether the changes in serum affect cellular immunity . Four-week-old male broiler chickens were injected subcutaneously with either olive oil or 50% croton oil mixed in olive oil to induce inflammation . The birds were bled at 48h after injection, and serum protein profiles were compared using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and densitometric evaluation . At 48h post-injection the serum from croton oil-injected birds showed distinct changes in protein profiles characterized by a selective increase or decrease in levels of several serum proteins . The protein bands which showed increases had relative molecular weights (Mr) corresponding to 65kilo Daltons (kD), 42kD, and two or more proteins with Mr> or =200kD . The levels of serum albumin (49kD), and a 56kD protein were reduced in croton oil-injected birds . The modulating effects of such serum on HD11 cells were studied using bacterial lipopolysaccharide (LPS) or phorbol myristate acetate (PMA) induced functional activation of these cells . The LPS-induced interleukin-6 (IL-6) production by HD11 cells was not affected by the presence of either olive oil-treated control or croton oil-treated inflammatory serum but nitrite production was enhanced by the inflammatory serum . Similarly, inflammatory serum also enhanced PMA-induced respiratory burst measured using dichlorofluorescein diacetate (DCF-DA) oxidation mediated by reactive oxygen intermediates . These results suggest that inflammatory serum can modulate macrophage function by influencing the production of reactive oxygen and nitrogen species which could affect their phagocytic and bactericidal activities.

Masui, 2001 Jun, 50(6), 672 - 5
{The effectiveness of ozonated water for hand washing before surgery}; Isosu T et al.; Using an ozonated water-dispensing machine for sterilization of hands (Mediaqua MA-III; Core Medical Co., Ltd, Tokyo, Japan), we investigated the effectiveness of ozonated water as a disinfectant for hand washing before surgery . The effectiveness of this new hand-washing method, using 4 ppm of ozonated water, which is expected to have a short-term bactericidal effect, and 0.2% benzalkonium chloride/83% ethanol solution (Welpas), which is expected to have a long-term bactericidal effect, was compared with that of the conventional hand-washing method (Furbringer's method using a scrubbing agent containing povidone-iodine) . The results showed no significant differences in the numbers of live bacteria and exponential reduction rates in live bacteria . Thus, this new method for hand washing using ozonated water is an effective method for sterilization of the hands before surgery.

Infect Immun, 2001 Aug, 69(8), 4799 - 807
Conformational nature of the Borrelia burgdorferi decorin binding protein A epitopes that elicit protective antibodies; Ulbrandt ND et al.; Decorin binding protein A (DbpA) has been shown by several laboratories to be a protective antigen for the prevention of experimental Borrelia burgdorferi infection in the mouse model of Lyme borreliosis . However, different recombinant forms of the antigen having either lipidated amino termini, approximating the natural secretion and posttranslational processing, or nonprocessed cytosolic forms have elicited disparate levels of protection in the mouse model . We have now used the unique functional properties of this molecule to investigate the structural requirements needed to elicit a protective immune response . Genetic and physicochemical alterations to DbpA showed that the ability to bind to the ligand decorin is indicative of a potent immunogen but is not conclusive . By mutating the two carboxy-terminal nonconserved cysteines of DbpA from B . burgdorferi strain N40, we have determined that the stability afforded by the putative disulfide bond is essential for the generation of protective antibodies . This mutated protein was more sensitive to thermal denaturation and proteolysis, suggesting that it is in a less ordered state . Immunization with DbpA that was thermally denatured and functionally inactivated stimulated an immune response that was not protective and lacked bactericidal antibodies . Antibodies against conformationally altered forms of DbpA also failed to kill heterologous B . garinii and B . afzelii strains . Additionally, nonsecreted recombinant forms of DbpA(N40) were found to be inferior to secreted lipoprotein DbpA(N40) in terms of functional activity and antigenic potency . These data suggest that elicitation of a bactericidal and protective immune response to DbpA requires a properly folded conformation for the production of functional antibodies.

J Pediatr, 2001 Jul, 139(1), 141 - 7
Clinical features of a human Rac2 mutation: a complex neutrophil dysfunction disease; Kurkchubasche AG et al.; The case of an infant with multiple, rapidly progressive, soft-tissue infections is presented . Despite features suggesting a neutrophil disorder, results of screening tests of phagocyte function were normal . A novel, multifaceted leukocyte disorder-distinguished by defects in shape change, chemotaxis, ingestion, degranulation, superoxide anion production, and bactericidal activity-was established secondary to a defect in Rac2.

Crit Care Med, 2001 Jul, 29(7), 1452 - 9
Effect of recombinant bactericidal/permeability-increasing protein on endotoxin translocation and lipopolysaccharide-binding protein/CD14 expression in rats after thermal injury; Fang WH et al.; OBJECTIVE: To investigate the potential mechanisms underlying the in vivo effect of recombinant bactericidal/permeability-increasing protein (rBPI21) on endogenous bacteria or endotoxin translocation and lipopolysaccharide-binding protein/CD14 expression secondary to thermal injury . DESIGN: Prospective, randomized, controlled animal study . SETTING: College hospital animal research laboratory . SUBJECTS: Thirty-six male Wistar rats weighing 250-300 g . INTERVENTIONS: The rats were anesthetized, and a 35% total body surface area full-thickness burn was created . Animals were randomized to receive treatment with either rBPI21 or the control protein (albumin) . rBPI21 (2 mg/kg body wt, BPI group) or a protein control preparation (burn group) in the same dose was administered in an intravenous bolus at 30 mins and 4 hrs after thermal injury . All animals were killed at 12 and 24 hrs postburn (six to ten rats for each interval) . In addition, eight rats were taken as normal controls . MEASUREMENT AND MAIN RESULTS: Our data showed that treatment with rBPI21 was effective in preventing endotoxin translocation secondary to severe burns . Meanwhile, tissue lipopolysaccharide-binding protein, CD14, and tumor necrosis factor-alpha mRNA expression in various organs were inhibited markedly by rBPI21 secondary to acute insults (p <.05-.01) . Furthermore, significant reduction in serum aminoleucine transferase concentrations and elevation in intestinal diamine oxidase activities in the rBPI21-treated group were found compared with controls (p <.05-.01) . CONCLUSIONS: These findings indicate that endotoxin accumulated in local sites after thermal injury can markedly up-regulate lipopolysaccharide-binding protein/CD14 and tumor necrosis factor-alpha mRNA expression in various organs . Meanwhile, up-regulation of lipopolysaccharide-binding protein/CD14 expression would be the major molecular mechanism of increasing sensitivity to endogenous endotoxin response after burns . Early treatment with rBPI21may be effective in attenuating multiple organ damage resulting from gut-origin endotoxin translocation . This might be associated with the down-regulation effects of tissue lipopolysaccharide-binding protein and CD14 gene expression by the use of rBPI21.

J Appl Microbiol, 2001 Jul, 91(1), 80 - 92
Possible mechanisms for the relative efficacies of ortho-phthalaldehyde and glutaraldehyde against glutaraldehyde-resistant Mycobacterium chelonae; Walsh SE et al.; AIMS: This investigation compared glutaraldehyde (GTA)-sensitive and -resistant strains of Mycobacterium chelonae and examined the effects of pretreatment of GTA-sensitive and -resistant strains of Myco . chelonae with chemical agents that interfere with cell wall synthesis . METHODS AND RESULTS: When exposed to 2% (v/v) GTA at 25 degrees C, GTA-resistant strains of Myco . chelonae dried on to glass carriers were not inactivated to any significant extent . By contrast, GTA-sensitive strains of Myco . chelonae and a strain of Myco . terrae suffered a > 6 log reduction in viability in 5 min . However, ortho-phthalaldehyde (OPA; 0.5% w/v) achieved a corresponding inactivation against two GTA-resistant strains within 5-10 and 10-20 min, respectively . Electron microscopy, using a non-aldehyde fixation process and also negative staining, failed to detect any extensive changes in GTA-sensitive and -resistant cultures exposed to GTA or OPA . Thin-layer chromatography was unsuccessful in detecting differences between GTA-resistant and -sensitive strains of Myco . chelonae . However, pretreatment of GTA-resistant cells with mycobacterial cell wall synthesis inhibitors increased their subsequent susceptibility further to OPA but not to GTA . CONCLUSION: Ortho-phthalaldehyde is an effective new biocidal agent that, at its in-use concentration, is rapidly bactericidal to non-sporulating bacteria, including GTA-sensitive and -resistant mycobacteria . SIGNIFICANCE AND IMPACT OF THE STUDY: Pretreatment of GTA-resistant cells with mycobacterial cell wall synthesis inhibitors increased their subsequent susceptibility to OPA but not to GTA.

Int J Med Microbiol, 2001 May, 291(2), 171 - 82
Regulation of virulence genes in Mycobacterium tuberculosis; Mehrotra J et al.; Mycobacterium tuberculosis has demonstrated remarkable ability to survive in diverse conditions encountered during the infection process . These involve surviving the bactericidal stresses within the macrophage, the anaerobic and nutritionally altered environment of the granuloma, and the metabolically inactive latent state . Understanding the molecular basis of this adaptive behavior lies in the identification of genes (or virulence determinants) specifically expressed under these varied conditions . Transcriptional control plays a key role in regulating gene expression in response to environmental signals . However, even after decades of investigation our knowledge about the function of these regulatory mechanisms in mycobacteria remains meagre . But the elucidation of the genome sequence and implementation of sophisticated molecular genetic approaches to this organism have made a revolutionary impact on the study of mycobacterial pathogenesis . Deletion and complementation of individual genes can be done at will facilitating the comparative analysis of mutants and wild-type strains . Novel and powerful technologies such as DNA microarrays, fluorescent beacons and proteomics have made possible the analysis of the expression levels of multiple genes in in vitro systems . More technically challenging uses of these techniques is being undertaken to explore pathogen gene expression within the host . This will lead to the identification of virulence factors and give definitive insight into their regulatory signals.

Cell Transplant, 2001, 10(3), 263 - 75
Poly-L-Lysine induces fibrosis on alginate microcapsules via the induction of cytokines; Strand BL et al.; Alginate-poly-L-lysine (PLL) microcapsules can be used for transplantation of insulin-producing cells for treatment of type I diabetes . In this work we wanted to study the inflammatory reactions against implanted microcapsules due to PLL . We have seen that by reducing the PLL layer, less overgrowth of the capsule is obtained . By incubating different cell types with PLL and afterwards measuring cell viability with MTT, we found massive cell death at concentrations of PLL higher than 10 microg/ml . Staining with annexin V and propidium iodide showed that PLL induced necrosis but not apoptosis . The proinflammatory cytokine, tumor necrosis factor (TNF), was detected in supernatants from monocytes stimulated with PLL . The TNF response was partly inhibited with antibodies against CD14, which is a well-known receptor for lipopolysaccharide (LPS) . Bactericidal permeability increasing protein (BPI) and a lipid A analogue (B-975), which both inhibit LPS, did not inhibit PLL from stimulating monocytes to TNF production . This indicates that PLL and LPS bind to different sites on monocytes, but because they both are inhibited by a p38 MAP kinase inhibitor, they seem to have a common element in the signal transducing pathway . These results suggest that PLL may provoke inflammatory responses either directly or indirectly through its necrosis-inducing abilities . By combining soluble PLL and alginate both the toxic and TNF-inducing effects of PLL were reduced . The implications of these data are to use alginate microcapsules with low amounts of PLL for transplantation purposes.

J Investig Allergol Clin Immunol, 2001, 11(1), 16 - 20
Are autoantibodies present in patients with subacute and chronic urticaria?
Ryhal B, DeMera RS, Shoenfeld Y, Peter JB, Gershwin ME.
Since several forms of autoimmunity have been associated with urticaria, we performed a detailed survey of autoantibodies in patients with idiopathic subacute and chronic urticaria . Sera from 25 consecutive patients referred for evaluation of urticaria were tested for the presence of autoantibodies and compared to sera from seventy-five control samples examined from individuals being treated for other allergic diseases . Study patients ranged in age from 15 to 73 years, with a mean of 48 . One patient had a diagnosis of inflammatory bowel disease and one had multiple myeloma, but otherwise there were no other diagnoses of disease specifically involving immunity other than atopy . No study patients had a concurrent diagnosis of autoimmune thyroid disease . The test sera were examined for autoantibodies and for antibodies to H . pylori . Antibodies to thyroid peroxidase (TPO) were found significantly (p < 0.01) more common in urticaria (20%} than in controls (0%) . Rheumatoid factor(RF) was also found in significantly (p < 0.05) increased in urticaria (16%) compared to controls {0%) . Neither H . pylori antibody nor other autoantibodies were present in significant numbers of urticaria patients when compared to controls . Tested autoantibodies included those to thyroglobulin, sDNA, SSA/SSB, ENA, cardiolipin, beta2-glycoprotein I, myeloperoxidase, proteinase-3, smooth muscle, ANA, human lysosomal-associated membrane protein, and bactericidal permeability increasing protein . Thus, patients with urticaria were somewhat more likely to have a thyroid autoantibody to TPO or to have RF . This survey demonstrates that while some markers of autoimmunity may be increased in urticaria patients, broad nonspecific autoimmunity is not found.

J Nat Prod, 2001 Jun, 64(6), 720 - 5
Trypanocidal withanolides and withanolide glycosides from Dunalia brachyacantha; Bravo B JA et al.; Two new withanolide glycosides, (20R,22R)-O-(3)-alpha-L-rhamnopyranosyl(1-->4)-beta-D-glucopyranosyl-1 alpha,12 beta-diacetoxy-20-hydroxywitha-5,24-dienolide (3) and (20R,22R)-O-(3)-beta-D-xylopyranosyl(1-->3)-{beta-D-xylopyranosyl(1-->4)}-beta-D-glucopyranosyl-1 alpha-acetoxy-12 beta,20-dihydroxywitha-5,24-dienolide (4), were isolated from the leaves and root of Dunalia brachyacantha . Their aglycones, (20R,22R)-1 alpha,12 beta-diacetoxy-3 beta,20-dihydroxywitha-5,24-dienolide (or 1 alpha,12 beta-diacetyldunawithagenine) and (20R,22R)-1 alpha-acetoxy-3 beta,12 beta,20-trihydroxywitha-5,24-dienolide (or 1 alpha-acetyl-12 beta-hydroxydunawithagenine), are novel . The known 18-acetoxywithanolide D (1) and 18-acetoxy-5,6-deoxy-5-withenolide D (2) were also isolated from the leaves . These last two compounds were shown to be responsible for the trypanocidal, leishmanicidal, and bactericidal activities manifested by the crude ethanolic extract . The structures were deduced from spectroscopic data and on the basis of chemical evidence.

Ter Arkh, 2001, 73(3), 15 - 9
{Evaluation of clinical effectiveness of intravenous laser irradiation of blood, plasmapheresis and their combination in patients with bronchial asthma}; Ishina TI et al.; AIM: To compare clinical response to intravenous laser radiation of blood (ILRB), plasmapheresis (PA) and ILRB + PA in patients with bronchial asthma (BA) . MATERIALS AND METHODS: 122 patients with endogenic BA of moderate severity were divided into four groups: group 1 was exposed to ILRB, group 2--to PA, group 3--to ILRB + PA, group 4 received only chemotherapy . The effect was assessed by body plethismography, peak flowmetry, NBT-test, LCP-test . RESULTS: Patients of group 1-3 vs group 4 demonstrated earlier disappearance of cough and normalization of lung auscultative picture, lower demand of oral glucocorticosteroids, 2 times longer remission, better external respiration function, earlier normalization of peak expiration flow . ILRB, PA and ILRB + PA activate function of oxygen-dependent bactericidal system of blood neutrophils and inhibited activity of non-oxygen-dependent system . CONCLUSION: Combined treatment of moderate severity BA with ILRB, PA alone and in combination is more effective than conventional drug therapy, the highest effect by remission terms being achieved in combination of ILRB with PA.

Ann Vasc Surg, 2001 May, 15(3), 326 - 31
Gut mucosal injury is attenuated by recombinant bactericidal/permeability-increasing protein in hind limb ischemia-reperfusion injury; Harkin DW et al.; Lower limb ischemia-reperfusion injury (IRI) is associated with increased gut permeability to endotoxin, which not only directly damages enterocytes but also stimulates a systemic inflammatory response syndrome (SIRS), compounding gut injury . Recombinant bactericidal/permeability-increasing protein (rBPI21) is a novel anti-endotoxin therapy with proven benefit in sepsis . Its potential role in modulating remote gut injury in hind limb IRI was studied . Male Wistar rats were chosen for a prospective randomized control trial (n = 10 per group) . The control group and two groups undergoing 3 hr bilateral hind limb ischemia with 2 hr reperfusion (I/R) were randomized to receive intravenously either control protein thaumatin at 2 mg/kg or rBPI21 at 2 mg/kg, respectively . Quantitative morphometric assessment of the small bowel was used as a measure of gut injury and, using an ex vivo everted gut sac model, translocation of 14C-labeled polyethylene glycol (PEG) was used as a measure of gut permeability . Our results indicate that hind limb IRI is associated with remote gut mucosal injury and increased permeability to macromolecules . rBPI21 anti-endotoxin therapy modulates remote gut injury associated with lower limb IRI in this model.

Antimicrob Agents Chemother, 2001 Jul, 45(7), 1972 - 6
Safety and bactericidal activity of rifalazil in patients with pulmonary tuberculosis; Dietze R et al.; Rifalazil, also known as KRM-1648 or benzoxazinorifamycin, is a new semisynthetic rifamycin with a long half-life of approximately 60 h . Rifalazil has potent bactericidal activity against Mycobacterium tuberculosis in vitro and in animal models of tuberculosis (TB) . Prior studies in healthy volunteers showed that once-weekly doses of 25 to 50 mg of rifalazil were well tolerated . In this randomized, open-label, active-controlled phase II clinical trial, 65 subjects with sputum smear-positive pulmonary TB received one of the following regimens for the first 2 weeks of therapy: 16 subjects received isoniazid (INH) (5 mg/kg of body weight) daily; 16 received INH (5 mg/kg) and rifampin (10 mg/kg) daily; 17 received INH (5 mg/kg) daily plus 10 mg of rifalazil once weekly; and 16 received INH (5 mg/kg) daily and 25 mg of rifalazil once weekly . All subjects were then put on 6 months of standard TB therapy . Pretreatment and day 15 sputum CFU of M . tuberculosis were measured to assess the bactericidal activity of each regimen . The number of drug-related adverse experiences was low and not significantly different among treatment arms . A transient decrease in absolute neutrophil count to less than 2,000 cells/mm(3) was detected in 10 to 20% of patients in the rifalazil- and rifampin-containing treatment arms without clinical consequences . Decreases in CFU counts were comparable among the four treatment arms; however, the CFU results were statistically inconclusive due to the variability in the control arms . Acquired drug resistance did not occur in any patient . Studies focused on determining a maximum tolerated dose will help elucidate the full anti-TB effect of rifalazil.

J Cataract Refract Surg, 2001 Jun, 27(6), 941 - 7
Corneal endothelial cytotoxicity of diluted povidone--iodine; Naor J et al.; PURPOSE: To assess corneal endothelial toxicity of diluted povidone-iodine (PI) in vivo and in vitro . SETTING: Cell Biology Laboratory and the Laboratory for Intraocular Microsurgery and Implants, Goldschleger Eye Research Institute, Sackler School of Medicine, Tel-Aviv University, Chaim Sheba Medical Center, Tel-Hashomer, Israel . METHODS: In an in vitro study, cultured bovine corneal endothelial cells were exposed to diluted PI . The degree of cell damage was determined by staining with trypan blue and by comparing the results to those in a control group . In an in vivo study, a single dose of diluted PI was injected into the anterior chamber of rabbit eyes, completely replacing the aqueous humor . The eyes were evaluated by clinical examination, specular microscopy, pachymetry, pneumotonometry, and histopathology and compared to a control group injected with a balanced salt solution . RESULTS: In vitro, PI concentrations of 0.05% or less did not induce endothelial cell damage . Significant damage was observed with a PI concentration of 0.1% . Calf serum concentrations of 1% and higher in the culture media protected the endothelial cell monolayer from cytotoxic damage by PI . Aqueous humor did not have a similar effect . In vivo, PI concentrations of 0.1% or less did not induce changes in corneal endothelium morphology or function as assessed by specular microscopy and pachymetry . A PI concentration of 1% served as a positive control, causing corneal edema and endothelial cell loss as demonstrated by pachymetry, histopathology, and elevated intraocular pressure . CONCLUSIONS: The concentrations of PI tolerated by animal endothelium in vitro and in vivo were higher than the reported bactericidal levels . These findings justify further investigation of the safety and efficacy of PI for intracameral prophylaxis during surgery.

J Infect Chemother, 2001 Mar, 7(1), 16 - 21
Comparison of antimycobacterial activity of grepafloxacin against Mycobacterium avium with that of levofloxacin: accumulation of grepafloxacin in human macrophages; Hirota M et al.; The bactericidal activity of two new quinolones, grepafloxacin and levofloxacin, against five strains of Mycobacterium avium was investigated in vitro . The minimum inhibitory concentrations (MICs) of these two quinolones, determined by the broth microdilution method, were comparable for all strains tested . In contrast, grepafloxacin suppressed the intracellular growth of all the strains in monocyte-derived macrophages more strongly than levofloxacin, when the cells infected with these strains were incubated for 7 days in the presence of various concentrations of the two new quinolones . To find the reason for the strengthened intracellular killing activity of grepafloxacin, we determined the ratio of the concentration of the new quinolones in the cells to that in the medium (C/M concentration ratio) . The C/M concentration ratio of grepafloxacin was increased to 34.7 by 7 days, whereas that of levofloxacin at 7 days was only 12.3 . These data suggested that a higher level of intraphagocytic accumulation of grepafloxacin endows it with greater mycobactericidal activity.

Anticancer Res, 2001 Mar-Apr, 21(2A), 965 - 70
Combination effect of lignin F and natural products; Jiang Y et al.; We investigated the effect of lignin F, isolated from the alkaline extract of the cone of Pinus parviflora Sieb . et Zucc, on the cytotoxic activity and radical intensity (measured by ESR spectroscopy) of various natural products . Lignin F slightly inhibited the proliferation of human oral tumor cell lines (human squamous cell carcinoma HSC-2, human salivary gland tumor HSG), but not that of human gingival fibroblast HGF, suggesting its tumor specific cytotoxic action . Lignin F enhanced the cytotoxic activity of vitamin K2, vitamin K3, sodium ascorbate (vitamin C), epigallocatechin gallate (EGCG) (a major component of green tea), gallic acid (structural unit of tannin), chlorogenic acid, and 6 tea extracts (Japanese green tea, Japanese barley tea, black tea, Chinese green tea, Chinese Jasmin tea, Chinese Oolong tea), to various extents . On the other hand, lignin inhibited the cytotoxic activity of curcumin and dopamine . ESR spectroscopy demonstrated that combination of lignin and vitamin K3, EGCG or gallic acid synergistically augmented the radical intensity . Lignin F enhanced the bactericidal activity of EGCG against E . coli . These data suggest the beneficial effect of the combination of lignin F and natural products.

Arch Biochem Biophys, 2001 Mar 15, 387(2), 188 - 96
Dissociation of enzymatic and pharmacological properties of piratoxins-I and -III, two myotoxic phospholipases A2 from Bothrops pirajai snake venom; Soares AM et al.; Piratoxins (PrTX) I and III are phospholipases A2 (PLA2s) or PLA2 homologue myotoxins isolated from Bothrops pirajai snake venom, which also induce myonecrosis, bactericidal activity against Escherichia coli, disruption of artificial membranes, and edema . PrTX-III is a catalytically active hemolytic and anticoagulant Asp49 PLA2, while PrTX-I is a Lys49 PLA2 homologue, which is catalytically inactive on artificial substrates, but promotes blockade of neuromuscular transmission . Chemical modifications of His, Lys, Tyr, and Trp residues of PrTX-I and PrTX-III were performed, together with cleavage of the N-terminal octapeptide by CNBr and inhibition by heparin and EDTA . The lethality, bactericidal activity, myotoxicity, neuromuscular effect, edema inducing effect, catalytic and anticoagulant activities, and the liposome-disruptive activity of the modified toxins were evaluated . A complex pattern of functional differences between the modified and native toxins was observed . However, in general, chemical modifications that significantly affected the diverse pharmacological effects of the toxins did not influence catalytic or membrane disrupting activities . Analysis of structural changes by circular dichroism spectroscopy demonstrated significant changes in the secondary structure only in the case of N-terminal octapeptide cleavage . These data indicate that PrTX-I and PrTX-III possess regions other than the catalytic site, which determine their toxic and pharmacological activities.

Crit Rev Oncog, 2000, 11(3-4), 255 - 305
Benzalkonium salts: effects on G protein-mediated processes and surface membranes; Patarca R et al.; Benzalkonium salts comprise a group of positively charged surface-active alkylamine biocides with the general formula alkyldimethylbenzylammonium chloride or bromide . They interact with guanine nucleotide triphosphate-binding proteins (G proteins), thereby affecting signal transduction in a variety of cell types and processes . This article reviews the known and potential basic science research and clinical applications and manifestations of benzalkonium salts . Benzalkonium salts have antiproliferative effects on a variety of cells (including T cells) through G protein-dependent pathways, affect cytokine gene expression (downregulate tumor necrosis factor expression), and are also effective bactericidal, fungicidal, and virucidal agents with multisite (direct and immunologically mediated) inhibitory activity against many pathogens, including the human immunodeficiency virus (HIV), papillomavirus, and herpesviruses . Therefore, benzalkonium salts not only appear to be effective as disinfectants and spermicides but may also prove useful in the prevention and treatment of neoplasias and other disease, particularly those linked to viruses and originating at the skin or mucosal surface.

Ther Apher, 2001 Apr, 5(2), 115 - 22
Pathophysiology of septic shock and multiple organ dysfunction syndrome and various therapeutic approaches with special emphasis on immunoglobulins; Werdan K; The pathophysiology of sepsis and septic shock is dominated by an imbalance of pro- and antiinflammatory mediators produced by toxin-activated inflammatory cells . Both the overshooting of proinflammatory mediators as well as the development of immune paralysis are deleterious to the patient . Available therapeutic approaches with monoclonal antibodies and antagonists targeted against toxins and mediators have focused mainly on inhibition of overshooting proinflammation: the results, however, have been disappointing . Due to these disappointing results of specific antiinflammatory regimens, adjuvant treatment of sepsis and septic shock with intravenous immunoglobulins (IVIgs) has regained interest although this indication has at best been validated in part . Likely beneficial mechanisms of action may include the improvement of serum bactericidal activity due to neutralizing and opsonizing IgG and IgM antibodies as well as stimulation of phagocytosis and neutralization of bacterial endo- and exotoxins; another attractive mode of action may represent Ig-mediated modification and specific suppression of proinflammatory cytokine release from endotoxin- and superantigen-activated blood cells . For the total group of patients with sepsis and septic shock, a reduction in mortality by IVIg could not be documented; however, in the SBITS study with 653 patients included, a moderate improvement in sepsis morbidity and multiple organ dysfunction syndrome was demonstrated . In defined sepsis sub-groups, a reduction in mortality by IVIg has been seen in each small, not yet confirmed trial . Thus, IVIg is not a magic bullet of sepsis treatment, but it may reduce morbidity and thereby may be useful in the therapeutic mosaic of sepsis treatment.

J Fr Ophtalmol, 2001 Apr, 24(4), 396 - 9
{Tuberculous uveitis}; El Bakkali M et al.; Ocular tuberculosis is relatively rare; however, the most common ocular lesion during ocular tuberculosis is uveitis . The recrudescence of this disease is probably caused by human immunodeficiency virus or long-term corticotherapy . The authors report a of chronic unilateral tuberculosis-related uveitis complicated by panophthalmia with subconjunctival abcesess and scleral fistula . A histopathological study revealed a granulomatous inflammation with caseous material . An extensive work-up revealed no extraocular lesion . A systemic bactericidal treatement associated with topical steroids decreased the local inflammation with phthisis bulbi, and with no spread of the infection . The authors discuss the etiopathogenic, clinical, and therapeutic aspects of tuberculosis-related uveitis.

J Appl Microbiol, 2001 May, 90(5), 741 - 8
Bactericidal effect of a whey protein concentrate with anti-Helicobacter pylori activity; Early EM et al.; AIMS: To investigate the effect of whey protein concentrate (WPC) enriched in anti-Helicobacter pylori antibodies on growth of the organism in vitro . METHODS AND RESULTS: A WPC rich in H . pylori-specific antibodies was produced by immunizing lactating cows against H . pylori and processing pooled bulk milk samples into whey powder . The antibodies bound several proteins within the bacterial homogenate and were active at pH 5 . In a complement-dependent reaction, the immune WPC was highly bactericidal against four H . pylori strains tested in vitro . CONCLUSION: WPC produced with milk from H . pylori-immunized cows contains antibodies that are active at the pH of the stomach, and bactericidal against H . pylori in vitro, via the classical complement pathway . SIGNIFICANCE AND IMPACT OF THE STUDY: This study has demonstrated the potential for use of WPC in the prevention/treatment of H . pylori infections.

J Immunol, 2001 May 15, 166(10), 6276 - 86
ATP-mediated killing of Mycobacterium bovis bacille Calmette-Guérin within human macrophages is calcium dependent and associated with the acidification of mycobacteria-containing phagosomes; Stober CB et al.; We previously demonstrated that extracellular ATP stimulated macrophage death and mycobacterial killing within Mycobacterium bovis Bacille Calmette-Guerin (BCG)-infected human macrophages . ATP increases the cytosolic Ca(2+) concentration in macrophages by mobilizing intracellular Ca(2+) via G protein-coupled P2Y receptors, or promoting the influx of extracellular Ca(2+) via P2X purinoceptors . The relative contribution of these receptors and Ca(2+) sources to ATP-stimulated macrophage death and mycobacterial killing was investigated . We demonstrate that 1) ATP mobilizes Ca(2+) in UTP-desensitized macrophages (in Ca(2+)-free medium) and 2) UTP but not ATP fails to deplete the intracellular Ca(2+) store, suggesting that the pharmacological properties of ATP and UTP differ, and that a Ca(2+)-mobilizing P2Y purinoceptor in addition to the P2Y(2) subtype is expressed on human macrophages . ATP and the Ca(2+) ionophore, ionomycin, promoted macrophage death and BCG killing, but ionomycin-mediated macrophage death was inhibited whereas BCG killing was largely retained in Ca(2+)-free medium . Pretreatment of cells with thapsigargin (which depletes inositol (1,4,5)-trisphosphate-mobilizable intracellular stores) or 1,2-bis-(2-aminophenoxy)ethane-N, N, N',N'-tetraacetic acid acetoxymethyl ester (an intracellular Ca(2+) chelator) failed to inhibit ATP-stimulated macrophage death but blocked mycobacterial killing . Using the acidotropic molecular probe, 3-(2,4-dinitroanilino)-3'-amino-N-methyl dipropylamine, it was revealed that ATP stimulation promoted the acidification of BCG-containing phagosomes within human macrophages, and this effect was similarly dependent upon Ca(2+) mobilization from intracellular stores . We conclude that the cytotoxic and bactericidal effects of ATP can be uncoupled and that BCG killing is not the inevitable consequence of death of the host macrophage.

J Endourol, 2001 Apr, 15(3), 303 - 6
In vitro bactericidal effect of a modified thermal Nitinol electrode; Liatsikos EN et al.; PURPOSE: A standard electrode surgical generator connected to a Nitinol coil was used in vitro to evaluate whether the generated electromagnetic energy had any bactericidal effect on Escherichia coli . MATERIALS AND METHODS: The ATCC 259222 E . coli strain was used . We mixed 135 mL of a 1.5% non-nutritive agar with 15 mL of a 10(6) CFU/mL inoculum and transferred it to gas-sterilized plastic containers lined with aluminium foil . A 22F cylindrical shape was cut from the center of the agar, and a Nitinol coil was placed in that space and connected to a standard electrode surgical generator . Electrical energy was then applied from 5 to 25 V at 5-V increments . Temperatures were measured with two thermocouples placed in the middle and periphery of each agar . The treatment was stopped when the temperature at the middle thermometer reached 50 degrees C . The control group was not treated and was embedded in a water bath at 45 degrees C . Three 3 x 7-mm pieces were sliced from the inner to the outer part of the agar and processed, and colony counts were performed . RESULTS: We observed statistically significant deleterious effects on E . coli in all three zones when the treatment voltage was 15 and 20 . When the potential was raised to 25 V, we observed a significant result only in the core zone . The treatment duration was 50 minutes for 5 and 10 V, 45 minutes for 15 V, 15 minutes for 20 V, and 10 minutes for 25 V . CONCLUSION: The bactericidal effect was mainly in the central area, decreasing linearly toward the periphery, and was related to the temperature reached during activation of the electrical generator . These results were disappointing with regard to the utility of Nitinol stents to treat bacterial prostatitis.

J Chromatogr B Biomed Sci Appl, 2001 Apr 5, 753(2), 269 - 78
Method for reducing endotoxin in Moraxella catarrhalis UspA2 protein preparations; Fiske MJ et al.; The UspA2 protein from the bacterium Moraxella catarrhalis is a potential vaccine candidate for preventing human diseases caused by this organism . Before a vaccine can be administered parentally, the level of endotoxin must be reduced as much as possible . However, in this case the endotoxin was very tightly complexed with the UspA2 protein and could not be dissociated with Triton X-100 . It was found that it dissociated from the protein with the zwitterionic detergents Zwittergent 3-12 and Zwittergent 3-14 . The endotoxin could then be separated from the protein by either ion-exchange or gel filtration chromatography . Using the limulus amoebocyte lysate assay for quantitation, the endotoxin was reduced approximately 20,000-fold . The removal of residual endotoxin from UspA2 preparations had no detrimental effect on the immunological properties of the protein . Mouse antisera raised against UspA2 prior to, and following endotoxin reduction exhibited comparable antibody and bactericidal titers against the tested strains . Further, mice immunized with both preparations, followed by pulmonary challenge with either a homologous or a heterologous isolate, exhibited comparable levels of clearance.

Avian Dis, 2001 Jan-Mar, 45(1), 52 - 60
The exacerbating effect of infectious bronchitis virus infection on the infectious bursal disease virus-induced suppression of opsonization by Escherichia coil antibody in chickens; Naqi S et al.; Chickens infected with infectious bronchitis virus (IBV) and infectious bursal disease virus (IBDV) commonly develop secondary infection of the respiratory tract with Escherichia coli, resulting in significant economic losses . To understand the host factors that may contribute to the E . coli infection, we investigated macrophage-mediated E . coli phagocytosis, intracellular bacterial killing, and development of opsonizing antibody in previously uninfected chickens and in those infected with IBV, IBDV, and IBDV plus IBV . Macrophages from the peripheral blood and the respiratory tracts of chickens infected with IBV or IBDV plus IBV efficiently performed in vitro phagocytosis of E . coli in the presence of positive-control serum (i.e., E . coli antiserum produced in normal chickens) . Those macrophages also had adequate bactericidal activity, indicating that IBV and IBDV infections had not affected their phagocytic activity or bactericidal function . The phagocytic activity of macrophages remained unaffected (P < 0.05) when the positive-control serum was replaced with E . coli antiserum produced in chickens infected with IBV alone . However, when E . coli antisera raised in IBDV-infected and, especially, that produced in IBDV plus IBV-infected chickens were supplemented, the percentage of phagocytosis and number of bacteria ingested per phagocyte were significantly (P < 0.05) less . These results indicate that although IBDV alone has the potential to markedly reduce opsonizing ability of antibody, this effect is significantly (P < 0.05) exacerbated by IBV infection.

Int J Food Microbiol, 2001 Apr 11, 65(1-2), 131 - 5
Effects of high-pressure carbon dioxide on Escherichia coli in nutrient broth and milk; Erkmen O; Bactericidal effects of high-pressure carbon dioxide against Escherichia coli were studied under 100, 75, 50 and 25 bar at 20 degrees C, 30 degrees C and 40 degrees C . E . coli suspended in nutrient broth (NB, pH = 6.75) was inactivated under 100, 75, 50, and 25 bar CO2 treatments for 50, 65, 100, and 140 min at 30 degrees C, respectively . Acidification of nutrient broth by dissolved CO2 alone might account for the bactericidal effect under pressure . E . coli was inactivated in NB with initial pH 5.50 and 4.5 at 100 bar for 80 and 95 min, respectively . Treatment at 100 bar CO2 pressure for 6 h caused a decrease of 6.42 and 7.24 log cycles in whole and skim milk, respectively.

Pharmacol Toxicol, 2001 Apr, 88(4), 209 - 12
Toxic effects of sarin in rats at three months following single or repeated low-level inhalation exposure; Kassa J et al.; Male albino Wistar rats were once or repeatedly exposed to three various low concentrations of sarin for 60 min . in the inhalation chamber . The clinical status of control as well as sarin-poisoned rats was tested 3 months after exposure to sarin using biochemical, haematological, neurophysiological, behavioural and immunotoxicological methods . While biochemical and haematological parameters, including the activities of cholinesterases in erythrocytes, plasma and various organs (brain, diaphragm), did not differ from the control values regardless of the sarin concentration used, few signs of sarin-induced neurotoxicity and immunotoxicity in sarin-poisoned rats were demonstrated . This was especially true when the single exposure of rats to non-convulsive symptomatic concentration and repeated exposure of rats to clinically asymptomatic concentration of sarin was used . In rats repeatedly poisoned with clinically asymptomatic concentrations of sarin, the alteration of the gait characterized by ataxia, the increase in the stereotyped behaviour, the increase in the excitability of the central nervous system following the administration of the convulsive drug pentamethylenetetrazol were observed . In rats poisoned with non-convulsive symptomatic concentration of sarin, the subtle supression of spontaneous, as well as lipopolysaccharides-stimulated, proliferation of spleen lymphocytes and the bactericidal activity of peritoneal macrophages was primarily observed besides the signs of neurotoxicity . Our findings confirm that both non-convulsive symptomatic and clinically asymptomatic concentrations of sarin can only cause very few, subtle long-term signs of neurotoxicity and immunotoxicity in sarin-poisoned rats when the rats were exposed to asymptomatic sarin concentrations repeatedly.

Crit Rev Biomed Eng, 2001, 29(1), 111 - 24
Methods for photoultrasonic treatment of festering wounds in oncological patients; Zharov VP et al.; This article presents further developments of photoultrasonic medical technology . It also analyzes its major effects and considers the most promising application of this combined technology in medicine, with particular emphasis given to the bactericidal treatment of infectious wounds . Moreover, this article describes the preliminary experimental results of the aforementioned technology.

Berl Munch Tierarztl Wochenschr, 2001 Mar-Apr, 114(3-4), 112 - 6
{Laboratory investigation of the chemical disinfection against different developmental stages of the housefly (Musca domestica Linnaeus, 1758).}; Mielke D et al.; Possibilities for disinfection of the developmental stages from the housefly (Musca domestica) were investigated under laboratory conditions . The developmental stages (eggs, larvae I, II, III, pupae and adults) were sprayed with solutions of disinfectants on the basis of p-chlorine-m-cresol and o-phenylphenol at different concentrations (0.025%-3%) . The effectiveness of both disinfectants was established by determining the emergence rate of eggs and pupae and of the live evidence of larvae and adults . Transmission electron microscopic investigations of the cover of the eggs revealed the bactericidal effect of both disinfectants . These results show the necessity of extending its use also to noxious arthropods.

J Leukoc Biol, 2001 Apr, 69(4), 522 - 30
The effects of extracellular pH on immune function; Lardner A; The effect of alterations in extracellular pH on cellular and humoral immune function is reviewed . Because acidic pH predominates at inflammatory loci and other sites of immune activity, most studies to date focus on the effect of acidic rather than alkaline pH . Investigations on polymorphonuclear leukocytes demonstrate mainly inhibition of chemotaxis, respiratory activity, and bactericidal capacity at reduced pH . Evidence of impaired lymphocyte cytotoxicity and proliferation at acidic pH is also beginning to emerge . Many of the clinical acidoses are accompanied similarly by immunodeficiency . Studies on macrophages and eosinophils are few and inconclusive . A small number of studies demonstrate acid-induced activation of complement proteins and the alternative complement pathway, plus increased antibody-binding to leukocytes at lowered pH . A differential effect of acidic pH on humoral and cellular immunity may, therefore, exist . Increasing recognition of the significance of extracellular pH in relation to immune function warrants further studies in this presently incomplete but rewarding field.

Am J Physiol Heart Circ Physiol, 2001 May, 280(5), H2357 - 63
Hydrogen peroxide stimulates macrophage vascular endothelial growth factor release; Cho M et al.; Neutrophils gather at the wound site shortly after trauma and release bactericidal reactive oxygen species (ROS) and H2O2 to kill bacteria and prevent infection . Macrophages arrive at the wound in response to environmental stimuli, phagocytose foreign particles, and release vascular endothelial growth factor (VEGF), an angiogenic factor crucial for wound healing . Because oxidants are released early in inflammation and have been found to regulate transcription factors, we investigated a possible role of H2O2 in VEGF stimulation . Human U937 macrophages exposed to H2O2 and allowed to recover in H2O2-free medium rapidly showed an increase in VEGF mRNA . The H2O2-mediated mRNA increase was dose dependent, blocked by catalase, and associated with elevated VEGF in conditioned media . The increase in VEGF was also found in primary rat peritoneal macrophages and the RAW 264.7 murine macrophage cell line . Transcriptional inhibition with actinomycin D revealed no significant difference in mRNA half-life . Transient transfections with a 1.6-kb VEGF promoter-luciferase construct (Shima DT, Kuroki M, Deutsch U, Ng YS, Adamis AP, and D'Amore PA . J Biol Chem 271: 3877-3883, 1996) showed a ninefold stimulation of VEGF gene promoter activity . We concluded that H2O2 increases macrophage VEGF through an oxidant induction of VEGF promoter . This oxidant stimulation can be mediated by activated neutrophils.

J Vasc Surg, 2001 Apr, 33(4), 840 - 6
Recombinant bactericidal/permeability-increasing protein attenuates the systemic inflammatory response syndrome in lower limb ischemia-reperfusion injury; Harkin DW et al.; OBJECTIVES: Hind limb ischemia-reperfusion (I/R) injury increases gut permeability, and resultant endotoxemia is associated with an amplified systemic inflammatory response syndrome leading to multiple organ dysfunction syndrome . We studied the potential role of recombinant bactericidal/permeability-increasing protein (rBPI(21) ), a novel antiendotoxin therapy, in modulating endotoxin-enhanced systemic inflammatory response syndrome in hind limb I/R injury . METHODS: In this prospective, randomized, controlled, experimental animal study, 48 male Wistar rats, weighing 300 to 350 g, were randomized to a control group (sham) and five groups undergoing 3 hours bilateral hind limb ischemia with 2 hours reperfusion (I/R) (n = 8 per group) . The control and untreated I/R groups received thaumatin, a control-protein preparation, at 2 mg/kg . Treatment groups were administered rBPI(21) intravenously at 1, 2, or 4 mg/kg body weight at the beginning of reperfusion; an additional group was administered rBPI(21) intravenously at 2 mg/kg after 1 hour of reperfusion . Plasma interleukin-6 concentration was estimated by bioassay as a measure of systemic inflammation . Plasma endotoxin concentration was determined by use of an amebocyte lysate chromogenic assay . Crossreactive immunoglobulin G and M antibodies to the highly conserved inner core region of endotoxin were measured by use of an enzyme-linked immunosorbent assay . The lung tissue wet-to-dry weight ratio and myeloperoxidase concentration were used as markers of edema and neutrophil sequestration, respectively . RESULTS: I/R provoked highly significant elevation in plasma interleukin-6 concentrations (1351.20 pg/mL {860.16 - 1886.40 pg/mL}) compared with controls (125.32 pg/mL {87.76-157.52 pg/mL; P <.0001}), but treatment with rBPI(21) 2 mg/kg at onset of reperfusion (715.89 pg/mL {573.36-847.76 pg/mL}) significantly decreased interleukin-6 response compared with the nontreatment group ( P <.016) . I/R increased plasma endotoxin concentrations significantly (21.52 pg/mL {6.20-48.23 pg/mL}), compared with control animals (0.90 pg/mL {0.00-2.30 pg/mL; P <.0001}), and treatment with rBPI(21) 4 mg/kg at reperfusion significantly decreased endotoxemia (1.30 pg/mL {1.20-2.20 pg/mL}), compared with the untreated group ( P <.001) . The lung tissue myeloperoxidase level was significantly increased in the untreated I/R group (208.18% {128.79%-221.81%}), compared with in controls (62.00% {40.45%-80.92%; P <.0001}), and attenuated in those treated with rBPI(21) 2 mg/kg (129.54% {90.49%-145.78%; P <.05}) . Data represent median and interquartile range, comparisons made with the nonparametric Mann-Whitney U test . CONCLUSIONS: These findings show that hind limb ischemia-reperfusion injury is associated with endotoxemia, elevations in plasma interleukin-6, and pulmonary leukosequestration . Treatment with rBPI(21) after ischemia reduces endotoxemia, the interleukin-6 response, and attenuates pulmonary leukosequestration in response to hind limb reperfusion injury.

Infect Immun, 2001 May, 69(5), 3067 - 72
BrkA protein of Bordetella pertussis inhibits the classical pathway of complement after C1 deposition; Barnes MG et al.; Bordetella pertussis produces a 73-kDa protein, BrkA (Bordetella resistance to killing), which inhibits the bactericidal activity of complement . In this study we characterized the step in the complement cascade where BrkA acts, using three strains: a wild-type strain, a strain containing an insertional disruption of brkA, and a strain containing two copies of the brkA locus . Following incubation with 10% human serum, killing was greatest for the BrkA mutant, followed by that for the wild-type strain, while the strain with two copies of brkA was the most resistant . Complement activation was monitored by enzyme-linked immunosorbent assay (ELISA) or Western blotting . ELISAs for SC5b-9, the soluble membrane attack complex, showed that production of SC5b-9 was greatest with the brkA mutant, less with the wild type, and least with the strain containing two copies of brkA . Deposition of complement proteins on the bacteria was monitored by Western blotting . A decrease in deposition on the bacteria of C4, C3, and C9 corresponded with decreased complement sensitivity . Deposition of C1, however, was not affected by the presence of BrkA . These studies show that BrkA inhibits the classical pathway of complement activation and prevents accumulation of deposited C4.

J Med Entomol, 2001 Jan, 38(1), 29 - 32
Meconial peritrophic membranes and the fate of midgut bacteria during mosquito (Diptera: Culicidae) metamorphosis; Moll RM et al.; The location of midgut bacteria relative to meconial peritrophic membranes (MPMs) and changes in bacterial numbers during midgut metamorphosis were studied in Anopheles punctipennis (Say), Culex pipiens (L.), and Aedes aegypti (L.) pupae and newly emerged adults . After adult emergence in Aedes, Anopheles, and most Culex, there were few to no bacteria in the midgut . In most newly emerged adult mosquitoes, few bacteria were found in either the lumen or within the MPMs/meconia . In a few Culex specimens, high numbers of bacteria were found in the MPMs/meconia and low numbers in the lumen . In all three species bacterial counts were high in fourth instars, decreased after final larval defecation, increased in young pupae, and increased further in old pupae . A very effective gut sterilization mechanism is operating during mosquito metamorphosis and adult emergence . This mechanism appears to involve the sequestration of remaining larval gut bacteria within the confines of the meconium and one or two MPMs and the possible bactericidal effect of the exuvial (molting) fluid, which is ingested during the process of adult emergence.

Eur J Clin Invest, 2001 Mar, 31(3), 258 - 63
Effect of vitamin C on neutrophil function after high-intensity exercise; Krause R et al.; High-intensity exercise leads to an increased risk of upper respiratory tract infections in athletes, which had been related to an exercise-induced impairment of neutrophil function . In this study, several indices of neutrophil function were analysed before and after a biathlon and the effect of oral vitamin C on neutrophil function was determined . Six athletes took 2 g vitamin C daily for 1 week prior to a biathlon and four athletes did not take any supplementation . Neutrophil phagocytosis was analysed by fluorescence microscopy and flow cytometry . Cytosolic calcium kinetics were assessed fluorometrically and neutrophil bactericidal ability was assessed by fluorescence microscopy . Reactive oxygen production was analysed by flow cytometry . Catecholamines were analysed by high-performance liquid chromatography . After high-intensity exercise there were significant reductions in the number of phagocytosed Escherichia coli per neutrophil and in neutrophil bactericidal ability . There was a significant exercise-dependent increase of catecholamines . There was no difference between the two groups of athletes . These results do not support the concept that vitamin C supplementation corrects neutrophil dysfunction after strenuous exercise.

J Leukoc Biol, 2001 Mar, 69(3), 414 - 8
Leptin: a potential regulator of polymorphonuclear neutrophil bactericidal action?
Caldefie-Chezet F, Poulin A, Tridon A, Sion B, Vasson MP.
It is well known that leptin, the ob gene product, is involved in the regulation of food intake and thermogenesis . Recent studies also demonstrate that leptin may be able to modulate functions of cells involved in nonspecific immune response such as phagocytosis and secretion of cytokines by macrophages . This and the prominent implication of polymorphonuclear neutrophils (PMNs) in infectious response suggested a possible role of leptin as a modulator of PMN functions . We detected a leptin receptor on the PMN membrane by immunocytochemistry with an anti-leptin receptor . Using chemiluminescence we then demonstrated that leptin enhances oxidative species production by stimulated PMNs . These results show for the first time that a functional leptin receptor is present on PMNs and that leptin may be able to influence their oxidative capacity.

Chem Res Toxicol, 2001 Mar, 14(3), 312 - 8
Reactive nitrogen oxygen species metabolize N-acetylbenzidine; Lakshmi VM et al.; A close association has been reported for certain types of cancers influenced by aromatic amines and infection/inflammation . Reactive nitric oxygen species (RNOS), components of the inflammatory response, are bactericidal and tumoricidal, and contribute to the deleterious effects attributed to inflammation on normal tissues . This study assessed the possible transformation of the aromatic amine N-acetylbenzidine (ABZ) by RNOS . RNOS were generated by various conditions to react with ABZ, and samples were evaluated by HPLC . Conditions which generate nitrogen dioxide radical (NO(2)(-) + myeloperoxidase + H(2)O(2), ONOO(-), and NO(2)(-) + HOCl) produced primarily a single new product termed 3'-nitro-ABZ . The myeloperoxidase-catalyzed reaction with 0.3 mM NO(2)(-) was completely inhibited by 1 mM cyanide, and not effected by 100 mM chloride with or without 1 mM taurine . In contrast, conditions which generate N(2)O(3), such as spermine NONOate, did not produce 3'-nitro-ABZ, but rather two compounds termed 4'-OH-AABP and AABP . (1)H NMR and mass spectrometry identified 3'-nitro-ABZ as 3'-nitro-N-acetylbenzidine, 4'-OH-AABP as 4'-OH-4-acetylaminobiphenyl, and AABP as 4-acetylaminobiphenyl . Human polymorphonuclear neutrophils incubated with {(3)H}ABZ and stimulated with beta-phorbol 12-myristate 13-acetate produced 3'-nitro-ABZ in the presence of NO(2)(-) (0.1-1 mM) . Neutrophil 3'-nitro-ABZ formation was verified by mass spectrometry and was consistent with myeloperoxidase oxidation of NO(2)(-) . The results demonstrate that ABZ forms unique products in the presence of nitrosating and nitrating RNOS, which could influence the carcinogenic process and serve as biomarkers for these reactive species.

J Vet Med B Infect Dis Vet Public Health, 2001 Feb, 48(1), 31 - 41
Variations among unbred heifers in the activities of polymorphonuclear leucocytes from the mammary gland and blood; Rysanek D et al.; The phenotypic characteristics are described for the activity of polymorphonuclear leucocytes NMN) obtained by either lavage of the cavity system of juvenile mammary glands stimulated with a synthetic muramyl dipeptide analogue or isolation from the peripheral blood . Attention was paid to the variability of characteristics and its sources, and to correlations among them . The following characteristics were investigated in 27 clinically healthy, unbred Bohemian Red Pied x Holstein heifers: migration activity in situ, number of phagocytosing PMN, phagocytotic index, bactericidal activity of PMN and unstimulated and zymosan-stimulated luminol-dependent chemiluminescence . Considerable individual variation was found in the characteristics . Significant differences between blood PMN and PMN from lavages after influx induction were found for bactericidal activity (P < 0.05) and chemiluminescence (P < 0.01) . A significant correlation between blood PMN and mammary gland PMN was found only for the number of phagocytosing cells (r = 0.329; P < 0.01) . Highly significant positive correlations (P < 0.01) were demonstrated between the number of phagocytosing PMN {a}, phagocytotic index {b}, and bactericidal activity {c} in both blood PMN (r(ab) = 0.602; r(ac) = 0.565; r(bc) = 0.529) and mammary gland PMN (r(ab) = 0.730, r(ac) = 0.618, r(bc) = 0.589) . No significant correlation was demonstrated for non-stimulated (NS), zymosan-stimulated (ZS), or opsonized zymosan-stimulated (OZS) chemiluminescence with any of the other characteristics of phagocytotic activity, in either blood PMN or mammary gland PMN (P > 0.05) . The animal was a highly significant source of variability for all the phagocytotic activity characteristics (P < 0.01) . Udder quarter was a non-significant source of variability for all the characteristics of phagocytotic activity except for NS chemiluminescence (P < 0.05) and ZS or OZS chemiluminescence (P < 0.01) . However, udder quarter was a non-significant source of variability of chemiluminescence indices ZS/NS and OZS/NS (P > 0.05) . It has been demonstrated that in situ migration activity, the number of phagocytosing PMN, phagocytotic index, bactericidal activity of PMN and chemiluminescence indices of PMN collected from juvenile mammary glands of unbred heifers after influx induction can be regarded as candidate early markers of resistance to mammary infections.

Mikrobiol Z, 2000 Nov-Dec, 62(6), 26 - 32
{Action of natural gamma-interferons on functional activity of phagocytes and antibody synthesis after vaccination}; Kishko IaG et al.; Natural swine and cattle gamma-IFNs were prepared for trials . One dose of gamma-suiferon contained 1000 IU, that of gamma-boviferon--2000 IU . Three series of researches were carried out to estimate the in vitro and in vivo absorbing activity of phagocytes (monocytes and neutrophiles), their bactericidal ability (on new born pigs and calves, 2 months old animals, sows and cows with calf) and antibodygenesis after immunization of animals by colibacteriosis vaccine . It has been shown in trials that gamma-IFN increased to significant degree (several times, as a rule) the absorbing activity of phagocytes (especially that of monocytes in new-born animals) . At the same time bactericidal activity of phagocytes sharply increased--their functional reserve in experimental animals was significantly higher (2-3-times), than in control . Immunization by colinebacteriosis vaccine with additional treatment by homologous gamma-IFN 3-4 times increased antibodygenesis in comparison with control.

Prog Neurobiol, 2001 Jun, 64(3), 277 - 305
Voltage-gated proton channels in microglia; Eder C et al.; Microglia, macrophages that reside in the brain, can express at least 12 different ion channels, including voltage-gated proton channels . The properties of H+ currents in microglia are similar to those in other phagocytes . Proton currents are elicited by depolarizing the membrane potential, but activation also depends strongly on both intracellular pH (pH(i)) and extracellular pH (pH(o)) . Increasing pH(o) or lowering pH(i) promotes H+ channel opening by shifting the activation threshold to more negative potentials . H+ channels in microglia open only when the pH gradient is outward, so they carry only outward current in the steady state . Time-dependent activation of H+ currents is slow, with a time constant roughly 1 s at room temperature . Microglial H+ currents are inhibited by inorganic polyvalent cations, which reduce H+ current amplitude and shift the voltage dependence of activation to more positive potentials . Cytoskeletal disruptive agents modulate H+ currents in microglia . Cytochalasin D and colchicine decrease the current density and slow the activation of H+ currents . Similar changes of H+ currents, possibly due to cytoskeletal reorganization, occur in microglia during the transformation from ameboid to ramified morphology . Phagocytes, including microglia, undergo a respiratory burst, in which NADPH oxidase releases bactericidal superoxide anions into the phagosome and stoichiometrically releases protons into the cell, tending to depolarize and acidify the cell . H+ currents may help regulate both the membrane potential and pH(i) during the respiratory burst . By compensating for the efflux of electrons and counteracting intracellular acidification, H+ channels help maintain superoxide anion production.

J Immunol, 2001 Mar 15, 166(6), 3873 - 81
Biphasic regulation of NF-kappa B activity underlies the pro- and anti-inflammatory actions of nitric oxide; Connelly L et al.; Expression of inducible NO synthase (iNOS) by macrophages is a prerequisite for the production of high output NO, which mediates many bactericidal and tumoricidal actions of these immune cells . The expression of iNOS in mammalian cells is governed predominantly by the transcription factor, NF-kappa B, which regulates the expression of many host defense proteins . In the present study, we characterize a novel, biphasic effect of NO on NF-kappa B activity in murine macrophages . This mechanism depends on the local concentration of NO and enables it both to up- and down-regulate the expression of host defense proteins including iNOS, cyclooxygenase-2, and IL-6 . This biphasic activity of NO appears to play a pivotal role in the time course of activation of these immune cells and, by inference, in facilitating the initiation of a defense response against pathogenic stimuli and in its termination to limit tissue damage . This mechanism may explain at least in part the reported ability of NO to act in both a pro- and anti-inflammatory manner.

Biofizika, 2001 Jan-Feb, 46(1), 69 - 76
{Membranotropic effects of electromagnetic radiation of extremely high frequency on Escherichia coli}; Trchunian A et al.; It was found that "sound" electromagnetic radiations of extremely high frequencies (53.5-68 GHz) or millimeter waves (wavelength range of 4.2-5.6 mm) of low intensity (power density 0.01 mW) have a bactericidal effect on Escherichia coli bacteria . It was shown that exposure to irradiation of extremely high frequencies increases the electrokinetic potential and surface change density of bacteria and decreases of membrane potential . The total secretion of hydrogen ions was suppressed, the H+ flux from the cytoplasm to medium decreased, and the flux of N,N'-dicyclohexylcarbodiimide-sensitive potassium ions increased, which was accompanied by changes in the stoichiometry of these fluxes and an increase in the sensitivity of H+ ions to N,N'-dicyclohexylcarbodiimide . The effects depended on duration of exposure: as the time of exposure increased, the bactericidal effect increased, whereas the membranotropic effects decreased . The effects also depended on growth phase of bacteria: the irradiation affected the cells in the stationary but not in the logarithmic phase . It is assumed that the H(+)-ATPase complex F0F1 is involved in membranotropic effects of electromagnetic radiation of extremely high frequencies . Presumably, there are some compensatory mechanisms that eliminate the membranotropic effects.

Surgery, 2001 Mar, 129(3), 282 - 91
The abnormal lipid spectrum in malignant obstructive jaundice in relation to endotoxin sensitivity and the result of preoperative biliary drainage; Kimmings N et al.; BACKGROUND: Biliary obstruction changes the spectrum of lipoproteins, which are now known to bind and neutralize endotoxin . Postoperative septic complications related to an increased susceptibility to endotoxin occur frequently in patients with obstructive jaundice . The effect of preoperative biliary drainage on changes in the lipoprotein spectrum and its relation to endotoxin sensitivity was studied . METHODS: Abnormalities in the lipoprotein spectrum were assessed in 15 patients with malignant obstructive jaundice before and 3 weeks after endoscopic biliary drainage . Changes in endotoxin responsiveness were assessed by using endotoxin-neutralizing reagents (anti-CD14 monoclonal antibody, polymyxin B, and recombinant bactericidal permeability increasing protein) to block cytokine production in whole blood cell cultures that were stimulated by cholestatic plasma taken before and after drainage . RESULTS: Drainage normalized very-low-density, low-density, and high-density lipoprotein cholesterol fractions from, respectively, 43% to 19%, 50% to 65%, and 6% to 16% (P <.01) . Ex vivo stimulation of whole blood with predrainage cholestatic plasma was 20-fold higher (P <.001) than with postdrainage plasma . Blocking the endotoxin response during the stimulation with predrainage cholestatic plasma with anti-CD14 monoclonal antibody, polymyxin B or recombinant bactericidal permeability increasing protein resulted in attenuation of the inflammatory response, reducing tumor necrosis factor-alpha levels at least 5-fold . CONCLUSIONS: Preoperative biliary drainage normalizes the changed lipid profile and the endotoxin-stimulating capacity of cholestatic plasma, and this signifies a change in sensitivity to endotoxin.

Front Biosci, 2001 Mar 01, 6, D366 - 87
Roles of plasma lipid transfer proteins in reverse cholesterol transport; Yamashita S et al.; Plasma lipid transfer proteins include plasma cholesteryl ester transfer protein (CETP) and phospholipid transfer protein (PLTP) . Plasma CETP facilitates the transfer of cholesteryl ester (CE) from high-density lipoprotein (HDL) to apolipoprotein (apo) B-containing lipoproteins, and is a key protein in reverse cholesterol transport which protects vessel walls from atherosclerosis . The importance of plasma CETP in lipoprotein metabolism was highlighted by the discovery of CETP-deficient subjects with a marked hyperalphalipoproteinemia (HALP) . The deficiency of CETP causes various abnormalities in the concentration, composition, and functions of both HDL and low-density lipoprotein (LDL) . Although the significance of CETP in terms of atherosclerosis has been controversial, the in vitro evidence showed that large CE-rich HDL particles in CETP deficiency are defective in cholesterol efflux . Recent epidemiological studies in Japanese-Americans and in Omagari area where HALP subjects with the intron 14 splicing defect of CETP gene are markedly frequent, have demonstrated an increased incidence of coronary atherosclerosis in CETP-deficient patients . Similarly, scavenger receptor BI (SR-BI) knockout mice show a marked increase in HDL-cholesterol but accelerated atherosclerosis in atherosclerosis-susceptible mice . Thus, CETP deficiency is a state of impaired reverse cholesterol transport which may possibly lead to the development of atherosclerosis . PLTP transfers phospholipids from triglyceride (TG)-rich lipoproteins to HDL during lipolysis . Human plasma PLTP has a 20% sequence homology to human CETP and human PLTP gene has a marked similarity in the exon-intron organization . Both CETP and PLTP belong to the lipid transfer/lipopolysaccharide binding protein (LBP) gene family, which also includes LBP and bactericidal/permeability-increasing protein (BPI) . Although these 4 proteins possess different physiological functions, they share marked biochemical similarities . The current review will also focus on the molecular genetics and function of plasma lipid transfer proteins, including CETP and PLTP.

Lepr Rev, 2000 Dec, 71 Suppl, S81 - 7
Combination of rifapentine-moxifloxacin-minocycline (PMM) for the treatment of leprosy; Ji B et al.; To further the development of a multidrug regimen for treatment of leprosy that is suitable for monthly administration and fully supervisable, the bactericidal activities against Mycobacterium leprae of HMR 3647 (HMR), moxifloxacin (MXFX) and rifapentine (RPT) were measured by the proportional bactericide technique in the mouse footpad system, and compared with those of the established antileprosy drugs clarithromycin (CLARI), ofloxacin (OFLO) and rifampicin (RMP) . Administered in five daily doses of 100 mg per kg body weight, HMR appeared slightly more bactericidal than CLARI, but the difference did not attain statistical significance . Administered as single doses, MXFX in a dosage of 150 mg per kg was more active than OFLO in the same dosage, and displayed the same level of activity as RMP in a dosage of 10 mg per kg; the combination MXFX-minocycline (MINO) (MM) was more bactericidal than the combination OFLO-MINO (OM); RPT in a dosage of 10 mg per kg was more bactericidal than RMP administered in the same dosage, and even more active than the combination RMP-OFLO-MINO (ROM); the combination RPT-MXFX-MINO (PMM) killed 99.9% of viable M . leprae, and was slightly more bactericidal than was RPT alone, indicating that the combination PMM showed an additive effect against M . leprae . These promising results justify a clinical trial among lepromatous patients, in which MM is being compared with OM, and PMM with ROM, in terms of efficacy and tolerance.

J Microencapsul, 2001 Jan-Feb, 18(1), 89 - 110
Controlled release of vancomycin from biodegradable microcapsules; Ozalp Y et al.; Poly D,L-lactic acid (PLA) and its copolymers with glycolide PLGA 90:10 and 70:30 were polymerized under various conditions to yield polymers in the molecular weight range 12000-40000 daltons, as determined by gel permeation chromatography . Vancomycin hydrochloride was the hydrophilic drug of choice for the treatment of methicillin resistant Staphyloccoccal infections . It was microencapsulated in the synthesized polymers using water-oil-water (w/o/w) double emulsion and solvent evaporation . The influence of microcapsule preparation medium on product properties was investigated . An increase in polymer-to-drug ratio from 1:1 to 3:1 caused an increase in the encapsulation efficiency (i.e . from 44-97% with PLGA) . An increase in the emulsifier (PVA) molecular weight from 14-72 kD caused an increase in encapsulation efficiency and microcapsule size . The in vitro release of vancomycin from microcapsules in phosphate buffer saline (pH 7.4) was found to be dependent on molecular weight and copolymer type . The kinetic behaviour was controlled by both diffusion and degradation . Sterilization with 60Co (2.5 Mrad) also affected the degradation rate and release profiles . Degradation of microcapsules could be seen by scanning electron microscopy, by the increase in the release rate from PLA and by the decrease in the Tg values of microcapsules . In vitro bactericidal effects of the microcapsule formulations on S . aureus were determined with a special diffusion cell after the preparations had been sterilized, and were found to have bactericidal effects lasting for 4 days.

J Parasitol, 2000 Dec, 86(6), 1223 - 8
A comparative study of mammalian and reptilian alternative pathway of complement-mediated killing of the Lyme disease spirochete (Borrelia burgdorferi); Kuo MM et al.; The potential bactericidal activity of the alternative complement pathway of mammalian and reptilian sera to Borrelia burgdorferi sensu stricto (s.s.) was evaluated in vitro . Complement-mediated killing was observed when cultured spirochetes were inoculated into sera from the western fence lizard (Sceloporus occidentalis) and from the southern alligator lizard (Elgaria multicarinata), but not when they were inoculated into serum from either the deer mouse (Peromyscus maniculatus) or from humans . Spirochetes were still alive after 4 hr in lizard serum that had been preheated at 56 C for 30 min to inactivate complement . Furthermore, when lizard serum was chelated with 10 mM ethylenediaminetetraacetic acid to block all complement activation, borreliacidal activity was arrested . When lizard serum was chelated with 10 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid plus 4 mM MgCl2 to block only classical complement pathway activation, >85% of spirochetes were immobilized within 1 hr . Differences in B . burgdorferi s.s . mortality were not observed when chelators with or without MgCl2 were added to serum from either deer mice or humans . Proteins comprising the alternative complement pathway are responsible for the borreliacidal activity observed in the blood of S . occidentalis and E . multicarinata.

Nephrol Dial Transplant, 2001 Jan, 16(1), 39 - 44
Analysis of anti-neutrophil cytoplasmic antibodies (ANCA): frequency and specificity in a sample of 191 homozygous (PiZZ) alpha1-antitrypsin-deficient subjects; Audrain MA et al.; BACKGROUND: ANCA are autoantibodies directed against polymorphonuclear cell antigens, mainly proteinase 3 (PR3) and myeloperoxidase (MPO), which are implicated in the pathogenesis of small-vessel necrotizing vasculitis . Alpha1-antitrypsin is the main inhibitor of neutral serine proteinase {i.e . human leukocyte elastase (HLE) and PR3} present in PMN alpha-granules (alphaGr) . An association first reported by us between PR3 ANCA and the deficient PiZZ phenotype in ANCA-positive systemic vasculitis, now widely confirmed by others, led us to study the incidence and specificity of ANCA among PiZZ subjects . METHODS: We tested a population of 191 PiZZ (273 sera) for ANCA activity versus 272 PiMM matched control subjects using alphaGr or antigen-specific ELISA {PR3, HLE, MPO, lactoferin (LF) and bactericidal/ permeability increasing protein (BPI)} . RESULTS: The incidence of antibodies directed against alphaGr and HLE but not PR3, MPO, LF or BPI was increased in the PiZZ as compared to the PiMM group (Fisher probability respectively P < 0.0001 and P < 0.05) . CONCLUSIONS: ANCA not directed against classical antigens (MPO and PR3) may be found in PiZZ patients . However, these patients do not develop systemic vasculitis features . Therefore, alpha1-antitrypsin deficiency is not sufficient to induce ANCA positive vasculitides, and may only act as a second hit amplifying factor.

J Surg Res, 2001 Mar, 96(1), 107 - 13
Bilirubin impairs bactericidal activity of neutrophils through an antioxidant mechanism in vitro; Arai T et al.; BACKGROUND: Liver failure accompanied by hyperbilirubinemia after major hepatic resection is profoundly associated with septic complications . Although the immune dysfunction in cholestasis has been intensively investigated, the contribution of increased serum bilirubin to the impaired resistance to bacterial infection remains to be elucidated . Because bilirubin possesses an antioxidant activity, we hypothesized that bilirubin may scavenge reactive oxygen species (ROS) produced by neutrophils and consequently impair neutrophil bacterial killing . To address this, we evaluated the effects of bilirubin on the bactericidal activity of ROS or of neutrophils in vitro . MATERIALS AND METHODS: The antioxidant activity of bilirubin was determined using an ROS-sensitive fluorophore, dichlorofluorescin diacetate (DCFH-DA) . Bilirubin concentration in the buffer solution was monitored spectorophotometrically after incubation with ROS . The effect of bilirubin on killing of Escherichia coli by ROS or by isolated human neutrophils was determined by counting the viable E . coli after incubation on nutrient agar . RESULTS: The bilirubin concentration in the buffer solution was decreased by the addition of hydrogen peroxide, especially in the presence of peroxidase or ferrous iron . DCFH-DA oxidation by ROS or activated neutrophils was inhibited by bilirubin in a dose-dependent manner . The bactericidal activity of ROS or of isolated neutrophils was significantly attenuated by bilirubin . CONCLUSIONS: Bilirubin impairs bactericidal activity of neutrophils through scavenging ROS . Increased levels of serum bilirubin may well be responsible for the impaired bacterial clearance in patients with hyperbilirubinemia .

Infect Immun, 2001 Mar, 69(3), 1358 - 63
Functional characteristics of a protective monoclonal antibody against serotype A and C lipooligosaccharides from Moraxella catarrhalis; Hu WG et al.; A monoclonal antibody (MAb), designated MAb 8E7 (immunoglobulin G3), specific for Moraxella catarrhalis lipooligosaccharide (LOS) was evaluated for its functional activity in vitro and in a mouse model of colonization . Enzyme-linked immunosorbent assay (ELISA) demonstrated that the MAb 8E7 could be prepared to a high titer against LOS of the homologous strain 035E, and that it had bactericidal activity . MAb 8E7 reacted with M . catarrhalis serotype A and C LOSs but not serotype B LOS, as measured by ELISA and Western blotting . On the basis of published structures of LOSs, this suggests that the epitope recognized by MAb 8E7 is directed to a common sequence of either alpha-GlcNAc-(1-->2)-beta-Glc-(1--> at the branch substituting position 4 of the trisubstituted Glc residue or a terminal tetrasaccharide alpha-Gal-(1-->4)-beta-Gal-(1-->4)-alpha-Glc-(1-->2)-beta-Glc-(1--> at the branch substituting position 6 of the trisubstituted Glc residue . In a whole-cell ELISA, MAb 8E7 reacted with 70% of the 30 wild-type strains and clinical isolates tested . Immuno-electron microscopy demonstrated that MAb 8E7 reacted with a cell surface-exposed epitope of LOS on strain O35E . MAb 8E7 inhibited the adherence of strain O35E to Chang conjunctival epithelial cells by 90% . Passive immunization with MAb 8E7 could significantly enhance the clearance of strain O35E from mouse lungs in an aerosol challenge mouse model . This enhanced bacterial clearance was inhibited when MAb 8E7 was absorbed by M . catarrhalis serotype A LOS, indicating that the M . catarrhalis LOS-directed antibody may play a major role in the enhancement of M . catarrhalis clearance from lungs . These data suggest that MAb 8E7, which recognizes surface-exposed LOS of M . catarrhalis, is a protective antibody against M . catarrhalis.

Biochem Soc Trans, 2000 Dec, 28(6), 857 - 60
A lipid-hydrolysing activity involved in hexenal formation; Matsui K et al.; Short-chain aldehydes such as (3Z)-hexenal and n-hexanal are formed from lipids through sequential actions of lipid-hydrolysing, lipoxygenase and fatty acid hydroperoxide lyase activities . The aldehydes are formed upon wounding of plant tissues, and are reported to have bactericidal and fungicidal activities . Furthermore, it has been reported that the aldehydes can induce expression of a subset of genes involved in disease resistance and that they are involved in a defence response against insect herbivores . Although several genes encoding lipoxygenases and the lyases have been isolated, and characterized to some extent, only little is known about the enzyme accountable for the lipid-hydrolysing step . In this study, we tried to characterize the lipid-hydrolysing activity involved in the short-chain aldehyde formation in Arabidopsis . When Arabidopsis leaves were homogenized, (3Z)-hexenal was formed rapidly within a few minutes . During this time period, the amount of alpha-linolenic acid and C(16:3) rapidly decreased . Such a rapid increase of the aldehyde was repressed almost completely when the leaves were homogenized under a nitrogen stream, and instead free trienoic acids accumulated . A lipase inhibitor, quinacrine, successfully repressed the hydrolysis . It was revealed that trienoic acids in monogalactosyldiacylglycerol were predominantly hydrolysed during the formation of short-chain aldehydes . Collectively, it is suggested that the lipolytic enzyme involved in the short-chain aldehyde formation is a galactolipid-specific lipase.

Liver, 2001 Feb, 21(1), 56 - 63
Major liver resection results in a changed plasma amino acid pattern as reflected by a decreased Fischer ratio which improves by bactericidal/permeability increasing protein; Nijveldt RJ et al.; BACKGROUND/AIMS: Major liver resection results in a high morbidity and mortality, and endotoxin plays a role in post-resection hepatic failure . Severe hepatic failure as seen in hepatitis and cirrhosis may be accompanied by hepatic encephalopathy and is characterized by a typical plasma amino acid pattern reflected by a decreased Fischer ratio . This study was performed to evaluate the plasma amino acid pattern in patients undergoing major liver surgery receiving placebo or the endotoxin-neutralizing agent bactericidal/permeability-increasing protein (rBPI21) . PATIENTS AND METHODS: Forty-eight patients were randomized in this phase II, dose escalation, multicenter trial . Plasma amino acid profiles were determined preoperatively, and on the first (day 1) and third (day 3) postoperative day . RESULTS: In the placebo group the Fischer ratio decreased significantly on both postoperative days . Administration of rBPI21 also resulted in a decreased Fischer ratio on day 1, but not on day 3 . Highly elevated alanine plasma levels were observed on day 1 in placebo-treated patients, whereas rBPI21 prevented this elevation . Plasma alanine levels on day 1 correlated with the duration of post-resection hepatic failure . Conclusions: Major liver resection results in a decreased Fischer ratio and a rise in plasma alanine levels . Plasma levels of alanine on the first postoperative day correlated with the duration of the post-resection hepatic failure . rBPI21 improved the Fischer ratio and prevented the rise of plasma alanine levels.

Vet Microbiol, 2001 Feb 12, 78(3), 241 - 9
Virulence factors in Escherichia coli isolated from the blood of bacteremic neonatal calves; Fecteau G et al.; Twenty-five Escherichia coli isolates from the blood of critically ill bacteremic calves sampled in two separate studies on a calf-rearing farm housing over 15,000 calves, in the San Joaquin Valley, California were studied.Isolates were characterized for O serogroups and for pathotypes as determined by the presence of specific virulence factors including heat-labile enterotoxin (LT), heat-stable enterotoxins a and b (STa, STb), verotoxins 1 and 2 (VT1, VT2), cytotoxic necrotizing factor (CNF), aerobactin, intimin Eae and P, F17 and CS31A fimbrial adhesins, and resistance to bactericidal effects of serum.These isolates constituted a heterogeneous group . However, isolates were mostly aerobactin positive and often resistant to the bactericidal effects of serum . Isolates of pathotypes O78 (n=6), O119:CS31a (n=3), and P positive but O non-typeable (n=3) were associated with a high mortality rate . The remaining isolates belonged to diverse pathotypes, often possessing the adhesins P, F17, CS31A and Eae but belonging to O serogroups other than O78 and O119, and were less frequently associated with mortality.Although no virulence factor common to all isolates was identified, the capacity to use iron by the presence of aerobactin which is important to the capture of iron was a predominant factor . Moreover, certain pathotypes appear to be associated with primary colisepticemia whereas other pathotypes may cause a bacteremia without necessarily leading to septicemia.

Vaccine, 2000 Dec 8, 19 Suppl 1, S101 - 7
Vaccines for Moraxella catarrhalis; McMichael JC; Vaccine development for Moraxella catarrhalis is in the antigen identification stage . M . catarrhalis does not appear to synthesize secreted antigens such as exotoxins, nor does it appear to possess a carbohydrate capsule . Modified forms of these antigens are usually good vaccine components . There is some interest in whole bacterial cells and membrane fractions, but the search has largely focused on purified outer surface antigens . All of the present antigens have been selected based on the response seen in animals, although the antibody response seen in people exposed to the bacterium provides some guidance . The antibody response provides information related to the cross-strain preservation of epitopes and whether they are surface exposed . Antigens that elicit antibodies that have complement dependent bactericidal capacity, opsonophagocytic activity or interfere with one of the antigen's known functions such as adhesion or nutrient acquisition are particularly valued . In addition to examining the antibody response, some antigens have been evaluated in a murine pulmonary clearance model . Using these assays and model, several vaccine candidates have been identified . The antigens may be roughly classified by the function they serve the bacterium . One set appears to promote adhesion to host tissues and includes the hemagglutinins, ubiquitous surface protein A1 (UspA1), and possibly the CD protein . A second set is involved in nutrient acquisition . This set includes the lactoferrin binding protein A (LbpA) and lactoferrin binding protein B (LbpB), the transferrin binding protein A (TbpA) and transferrin binding protein B (TbpB), the CD and E porins, and the Catarrhalis outer membrane protein B (CopB) . A third set is comprised of antigens involved in virulence and it includes lipooligosaccharide (LOS) and the ubiquitous surface protein A2 (UspA2) . Antigens of unknown function, such as the 200K protein, may also be vaccine candidates . The antigens that are most suitable will be determined in clinical studies that are only beginning now.

J Immunol, 2001 Feb 1, 166(3), 1951 - 67
A model to predict cell-mediated immune regulatory mechanisms during human infection with Mycobacterium tuberculosis; Wigginton JE et al.; A key issue for the study of tuberculosis infection (TB) is to understand why individuals infected with Mycobacterium tuberculosis experience different clinical outcomes . Elaborating the immune mechanisms that determine whether an infected individual will suffer active TB or latent infection can aid in developing treatment and prevention strategies . To better understand the dynamics of M . tuberculosis infection and immunity, we have developed a virtual human model that qualitatively and quantitatively characterizes the cellular and cytokine control network operational during TB infection . Using this model, we identify key regulatory elements in the host response . In particular, factors affecting cell functions, such as macrophage activation and bactericidal capabilities, and effector T cell functions such as cytotoxicity and cytokine production can each be determinative . The model indicates, however, that even if latency is achieved, it may come at the expense of tissue damage if the response is not properly regulated . A balance in Th1 and Th2 immune responses governed by IFN-gamma, IL-10, and IL-4 facilitate this down-regulation . These results are further explored through virtual deletion and depletion experiments.

J Antimicrob Chemother, 2001 Feb, 47(2), 177 - 82
Sources of variation in studies of the early bactericidal activity of antituberculosis drugs; Sirgel F et al.; The early bactericidal activity (EBA) of antituberculosis drugs can be measured as the daily fall in cfu counts of Mycobacterium tuberculosis in sputum, usually during the first 2 days of treatment . In studies of low potency drugs, it is necessary to compare the treated group of patients with a group who receives no chemotherapy (Nil group) . Over the past 10 years, five Nil groups of between five and 13 patients have been studied in Cape Town and two Nil groups in Hong Kong . There was a suggestion of an increase in variation within the Cape Town groups, as shown by a regression of variance size against study date (P = 0.06), which could not be attributed to technical causes . It might indicate increasing host resistance in the Western Cape epidemic of tuberculosis . Since the weighted mean of all Nil groups at Cape Town was 0.00036, very close to zero, it would seem safe to test means of treated groups against zero thus increasing precision and avoiding ethical problems in delaying treatment . In contrast to Nil groups, the variation found in five groups who received 300 mg isoniazid daily (INH 300) was uniform and homoscedastic, possibly because the additional variation was caused mainly by individual differences in plasma isoniazid concentrations and patient body weights . The mean EBA in the INH 300 series was 0.575 with 95% confidence limits of 0.515 and 0.636.

J Antimicrob Chemother, 2001 Feb, 47(2), 141 - 6
The effect of fosfomycin on neutrophil function; Krause R et al.; Fosfomycin (cis-1,2-epoxypropyl phosphonic acid) is a cell wall synthesis-inhibiting antibiotic . We investigated the effect of fosfomycin on several indices of neutrophil function . Neutrophil phagocytosis was analysed by flow cytometry . Cytosolic calcium kinetics were assessed fluorometrically and neutrophil bactericidal ability was assessed by fluorescence microscopy . Intracellular reactive oxygen intermediate (ROI) production was analysed by flow cytometry and extracellular ROI by cytochrome c reductase assay . After fosfomycin incubation, phagocytosis was unaffected as assessed by the FACS assay . Fosfomycin incubation resulted in enhanced bactericidal ability, in increased intracellular calcium concentrations, elevated extracellular ROI production and decreased chemotaxis but it did not affect intracellular ROI production and chemokinesis.

Aquat Toxicol, 2000 Mar 1, 48(2-3), 211 - 221
Lindane increases in vitro respiratory burst activity and intracellular calcium levels in rainbow trout (Oncorhynchus mykiss) head kidney phagocytes; Betoulle S et al.; Phagocytic cells are the main actors of the fish immune system . They secrete reactive oxygen species (ROS) involved in their bactericidal activity . The effects of lindane on ROS production in rainbow trout phagocytes are contradictory . Here, we study the effects of high concentrations of lindane on ROS production (by chemiluminescence) and on intracellular calcium levels ({Ca(2+)}(i)) (by spectrofluorimetry) in trout phagocytes . In these cells, lindane from 2.5 to 10 microM, increases ROS production and has no effect on {Ca(2+)}(i) . From 25 to 200 microM, lindane leads to a rise in ROS production (maximal value measured: 41152+/-6253 RLU for 100 microM lindane) associated with an increase in {Ca(2+)}(i) (+3149+/-96 nM for 100 microM lindane) and with cytotoxicity which appears 2 min after addition of 100 microM lindane (25.4+/-3.75%; P<0.05) . In the absence of extracellular calcium, ROS production of lindane-treated cells remains significantly higher than in controls (maximal value measured: 1899+/-254 RLU for 25 microM lindane), a significant decrease in {Ca(2+)}(i) is observed in cells treated with 5 or 10 microM lindane (-54+/-35 nM for 10 microM lindane), and an increase in {Ca(2+)}(i) in cells treated with 100 microM lindane (330+/-33 nM) . The rise in {Ca(2+)}(i) induced by lindane is inhibited when cells are preincubated with thapsigargin (Thaps) . We conclude that lindane induces an increase in {Ca(2+)}(i)50 microM) alter Ca(2+) homeostasis in the absence of extracellular Ca(2+), confirming that lindane can affect other intracellular stores of Ca(2+) . At low concentrations (<25 microM), lindane stimulates ROS production by Ca(2+)-independant mechanisms without inducing cytotoxicity . From 25 microM, lindane increases {Ca(2+)}(i) and maximal cytotoxicity appears from 100 microM lindane . Lindane toxicity in fish phagocytes may be associated with high {Ca(2+)}(i) and high ROS production . Thus, ROS are beneficial in protection of the organism but when ROS are produced in excess, they can be toxic for cells and tissues.

J Biomed Mater Res, 2001, 58(1), 97 - 101
Self-sterilizing and self-cleaning of silicone catheters coated with TiO(2) photocatalyst thin films: a preclinical work; Ohko Y et al.; TiO(2) photocatalysts were successfully coated on silicone catheters or medical tubes by pretreatment of the silicone surface with a sulfuric acid solution (5 M) for 3 h . The TiO(2) film adhered to the silicone substrate strongly against tensile and bending stresses . On the TiO(2)-coated silicone-catheters under UV illumination, both the bleaching of methylene blue dye and the photocatalytic bactericidal effect on Escherichia coli (E . coli) cells were confirmed . Thus, this type of catheter can be sterilized and cleaned simply by irradiation with low-intensity UV light and can, therefore, be useful in the protection from catheter-related bacterial infections .

Prog Histochem Cytochem, 2000, 35(4), 259 - 367
Brunner's glands: a structural, histochemical and pathological profile; Krause WJ; Brunner's glands are unique to mammalian species and in eutherians are confined primarily to the submucosa of the proximal duodenum . In the majority of species examined, they begin at the gastrointestinal junction and extend for variable distances distally in the wall of the proximal small intestine . Ducts of individual glands empty either directly into the intestinal lumen or unite with overlying intestinal glands (crypts of Lieberkuhn) dependent on the species . Secretory units of Brunner's glands consist of epithelial tubules that show frequent distal branchings . The secretory units, with the exception of those found in rabbits and horses, consist primarily of a mucin producing cell type . However, other cell types normally associated with the overlying intestinal epithelium may be encountered scattered within the secretory units reflecting the developmental origin of these glands . Secretion from Brunner's glands contributes to a layer of mucus that forms a slippery, viscoelastic gel that lubricates the mucosal lining of the proximal intestinal tract . The unique capacity of this mucus layer to protect delicate underlying epithelial surfaces is due primarily to the gel-forming properties of its glycoprotein molecules . Mucin glycoproteins produced by Brunner's glands consist primarily but not exclusively of O-linked oligosaccharides attached to the central protein core of the glycoprotein molecule . Human Brunner's glands produce class III mucin glycoproteins and are thought to be the product of mucin gene MUC6 which is assigned to chromosome 11 (11p15-11p15.5 chromosome region) . In addition to mucin glycoproteins and a limited amount of bicarbonate, numerous additional factors (epidermal growth factor, trefoil peptides, bactericidal factors, proteinase inhibitors, and surface-active lipids) have been identified within the secretory product of Brunner's glands . These factors, incorporated into the mucus layer, guard against the degradation of this protective barrier and underlying mucosa by gastric acid, pancreatic enzymes, and other surface active agents associated with this region . Yet other factors produced by Brunner's glands function to provide active and passive immunological defense mechanisms, promote cellular proliferation and differentiation, as well as contribute factors that elevate the pH of luminal contents of this region by promoting secretion of the intestinal mucosa, pancreatic secretion and gall bladder contraction . Additional insights concerning the role of Brunner's glands in the mammalian gastrointestinal tract as well as their possible evolution in this class of vertebrates have been gained from a basic understanding of their pathobiology.

Trop Anim Health Prod, 2000 Dec, 32(6), 341 - 51
Studies on cellulitis and other disease syndromes caused by Escherichia coli in broilers in Sri Lanka; Gomis SM et al.; Cellulitis caused by Escherichia coli in broilers results in substantial losses to the broiler industry in North America and Europe due to condemnations at slaughter . The objective of this study was to identify cellulitis in broilers in Sri Lanka and to characterize the E . coli from cellulitis and other colibacillosis lesions . Twenty-four farms from the low- and mid-country were selected and bacterial isolations were obtained from 241 birds . Two hundred and ninety-one gross lesions were observed in these 241 birds and 162 E . coli isolates were obtained . Cellulitis was observed in 21% of the birds . Twenty-one per cent of the birds had multiple lesions due to E . coli . The frequency of detection of other disease syndromes was 162 (67%) birds with pericarditis, 26 (11%) airsacculitis, 24 (10%) hepatitis, 12 (5%) perihepatitis, and 16 (7%) polyserositis (a combination of pericarditis, perihepatitis and airsacculitis) . Serogroups O78, O2, O85 and O88 were distributed among the 32% of typable E . coli and 81% of isolates were assigned to three biotypes . Forty-four per cent of the E . coli isolates produced aerobactin and 88% demonstrated resistance to the bactericidal effect of normal chicken serum . The majority of the E . coli isolates were resistant to the antibiotics commonly used in poultry . All the E . coli isolates were non-haemolytic and 25% of the isolates produced K1 capsule . This study demonstrated the presence of cellulitis in Sri Lanka and this report describes some of the phenotypic characteristics of the E . coli isolates.

Immunol Rev, 2000 Oct, 177, 68 - 78
Leukocyte granule proteins mobilize innate host defenses and adaptive immune responses; Chertov O et al.; "...It is likely that the leukocyte granulations are in fact secretory products, which the cell dissolves and spreads to the environment as needed", Paul Ehrlich, 1900 . Neutrophil granules have long been recognized as mediators of innate host defense . Newly discovered functions for individual granule proteins suggest that granule constituents may also participate in adaptive immune responses . Neutrophil granule-derived cathepsin G, azurocidin/CAP37 and alpha-defensins have been shown to be chemotactic for mononuclear cells and neutrophils . Analysis of the chemotactic activity of alpha-defensins shows that they induce CD45RA+ and CD8 T-lymphocyte cell migration at concentrations 10 to 100-fold below that required for direct bactericidal activity . Additionally, alpha and beta defensins form chemotactic gradients for immature dendritic cells . Recruiting immature dendritic cells to sites of infection is one way for neutrophil granule proteins to initiate adaptive immune responses . Granules found in other leukocytes such as mast cells also contain serine proteases, such as chymase, that are known to chemoattract neutrophils and mononuclear cells . Preliminary evidence suggests that exocytosis of granule-derived products from a variety of leukocytes can mobilize inflammatory cells and immunocytes . Thus, leukocyte granule-derived proteins, more rapidly than chemokines, can mobilize cells that mediate innate host defense and adaptive immunity.

Clin Exp Rheumatol, 2000 Nov-Dec, 18(6), 707 - 12
Contribution of immunofluorescence to the identification and characterization of anti-neutrophil cytoplasmic autoantibodies . The role of different fixatives; Radice A et al.; OBJECTIVE: To study the sera from selected groups of antineutrophil cytoplasmic antibody (ANCA) positive patients by means of the indirect immunofluorescence test (ANCA-IIF) with different fixatives, in order to better discriminate among the various ANCAs (Ag-specificity and disease associations), especially those for which the antigen targets have not yet been identified . METHODS: Eighty pathological serum samples and 15 normal sera were evaluated . Pathological samples included sera from 30 ulcerative colitis (UC) ANCA positive patients, 30 P-ANCA/myeloperoxidase (MPO-ANCA) positive microscopic polyangiitis (MPA) patients, 10 C-ANCA/proteinase 3 (PR3-ANCA) positive Wegener's granulomatosis (WG) patients, and 10 antinuclear antibody (ANA) positive (ANCA negative) systemic lupus erythematosus (SLE) patients . ANCA were detected by IIF on ethanol, methanol and formalin-fixed granulocytes and by ELISAs specific for MPO, PR3, lactoferrin (LF) and bactericidal/permeability-increasing protein (BPI) . Additionally, sera were tested for the presence of antinuclear antibodies on IIF . RESULTS: 96% of serum samples from UC patients, positive by IIF on ethanol-fixed granulocytes, became negative when tested on formalin-fixed neutrophil slides . On the contrary, 95% of sera from vasculitic patients showed a clear diffuse granular cytoplasmic pattern on the same substrate; sera from all 10 SLE patients did not show any reactivity when formalin was used as fixative . On methanol-fixed neutrophils, 100% of UC P-ANCA positive sera were positive with the same pattern versus only 20% of vasculitic P-ANCA positive (MPO positive) . Methanol fixation had no effect on PR3-ANCA and ANA positive sera . CONCLUSION: The comparison of IIF patterns of sera tested on different fixed cells may be useful to distinguish vasculitis-related P-ANCA versus ANA and vasculitis-related P-ANCA versus UC-related P-ANCA.

J Clin Pathol, 2000 Nov, 53(11), 829 - 34
UK NEQAS in antibiotic assays; White LO; How providers of external quality assessment (EQA) programmes relate to and interact with the monitors and watchdog of clinical laboratory performance in the UK is described . With regard to the quality of antibiotic assays, the changes in methodologies and in performance quality between 1971 (when the UK NEQAS for Antibiotic Assays began) and 1999 is reviewed . How improvements in performance and changes of methodology are related is discussed . The findings and conclusions of two experimental pilot EQA distributions (the teicoplanin assay and serum bactericidal test) are also discussed.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 301 - 2
Bacteriostatic and bactericidal activities of moxifloxacin against Coxiella burnetii; Rolain JM et al.; The in vitro activity of moxifloxacin against Coxiella burnetii was compared to those of pefloxacin, ofloxacin, and doxycycline . MICs of moxifloxacin ranged from 0.5 to 1 microg/ml for the Nine Mile, Priscilla, and Q212 strains . Moxifloxacin was not bactericidal against C . burnetii at 4 microg/ml.

Domest Anim Endocrinol, 2000 Nov, 19(4), 223 - 35
Tumor necrosis factor-alpha and nitrite/nitrate responses during acute mastitis induced by Escherichia coli infection and endotoxin in dairy cows; Blum JW et al.; Concentrations of tumor necrosis factor-alpha (TNF-alpha) and of NO(x) (sum of nitrite and nitrate as indicators of endogenous nitric oxide production) in milk and blood plasma were measured in three mastitis models in dairy cows in early lactation . Escherichia coli P4:O37 bacteria or endotoxin O111:B4 were administered into both left quarters of 12 and 6 cows, respectively . Six of the E . coli-infected cows were treated with a bactericidal antibiotic (Enrofloxacin; Bayer AG, Leverkusen, Germany) i.v . at 10 hr and subcutaneously (sc) at 30 hr after infection . NO(x) concentrations transiently increased maximally 10- to 11-fold in milk of E . coli-infected quarters with or without antibiotic treatment at 24 hr and after endotoxin administration . NO(x) concentrations did not change in milk of unchallenged quarters and in blood plasma . Increases of NO(x) were proceeded by a transient (96- to 149-fold) rise of milk TNF-alpha concentrations, which in endotoxin-administered quarters was maximal at 6 hr and in infected quarters without or with Enrofloxacin treatment at 10 and 14 hr . In blood plasma TNF-alpha concentrations only moderately increased to peaks in endotoxin-administered cows at 6 hr and in E . coli-infected cows at 14 hr postchallenge . In one severely sick, nontreated E . coli-infected cow milk, TNF-alpha response at 14 hr was excessive and followed by a spectacular rise of NO(x) concentration in milk between 48 and 72 hr . In conclusion, a possible clinical relevance of nitric oxide production associated with a rise of intramammary and systemic TNF-alpha during acute mastitis by E . coli infection and endotoxin in lactating dairy cows is indicated, but could not be inhibited by antibiotic treatment.

Biochim Biophys Acta, 2000 Dec 20, 1509(1-2), 65 - 72
Effects of the antibiotic peptide microcin J25 on liposomes: role of acyl chain length and negatively charged phospholipid; Rintoul MR et al.; This paper reports the effects of microcin J25 (MccJ25) on the microviscosity and permeability of phospholipid vesicles of different compositions . The results obtained indicate that MccJ25 interacts with egg L-alpha-phosphatidylcholine (PC) vesicles as demonstrated by peptide intrinsic fluorescence determinations . The interaction depends on the lipid composition of the vesicles . MccJ25 interaction induces a significant fluidity increase of egg PC vesicles . This effect is time and concentration dependent . Both trimethyl ammonium 1,6-diphenyl-1,3,5-hexatriene and 1,6-diphenyl-1, 3,5-hexatriene gave the same results . The microviscosity of L-alpha-phosphatidylcholine dipalmitoyl small unilamellar vesicles (SUVs) was affected while that of L-alpha-phosphatidylcholine dimyristoyl vesicles was not, indicating that the effect was strongly dependent on the chain length of fatty acids . On the other hand, negatively charged L-alpha-phosphatidyl-DL-glycerol (PG) vesicles remarkably inhibited the peptide effect . Nevertheless vesicles composed of L-alpha-phosphatidylethanolamine:PG:cardiolipin (7:2:1), a composition resembling bacterial membrane, were sensitive to the MccJ25 effect . MccJ25 effectively dissipated the valinomycin-induced membrane potential, but induced only a modest leakage (5%) of the trapped Tb(+3)-dipicolinic acid complex . These results indicate that the peptides interact and perturb the bilayer of SUVs . The relationships between this effect and bactericidal action remain to be elucidated.

J Biol Chem, 2001 Apr 13, 276(15), 12292 - 300 Epub 2000 Dec 14.
Complement factor H is a serum-binding protein for adrenomedullin, and the resulting complex modulates the bioactivities of both partners; Pio R et al.; Adrenomedullin (AM) is an important regulatory peptide involved in both physiological and pathological states . We have previously demonstrated the existence of a specific AM-binding protein (AMBP-1) in human plasma . In the present study, we developed a nonradioactive ligand blotting assay, which, together with high pressure liquid chromatography/SDS-polyacrylamide gel electrophoresis purification techniques, allowed us to isolate AMBP-1 to homogeneity . The purified protein was identified as human complement factor H . We show that AM/factor H interaction interferes with the established methodology for quantification of circulating AM . Our data suggest that this routine procedure does not take into account the AM bound to its binding protein . In addition, we show that factor H affects AM in vitro functions . It enhances AM-mediated induction of cAMP in fibroblasts, augments the AM-mediated growth of a cancer cell line, and suppresses the bactericidal capability of AM on Escherichia coli . Reciprocally, AM influences the complement regulatory function of factor H by enhancing the cleavage of C3b via factor I . In summary, we report on a potentially new regulatory mechanism of AM biology, the influence of factor H on radioimmunoassay quantification of AM, and the possible involvement of AM as a regulator of the complement cascade.

Blood Cells Mol Dis, 2000 Dec, 26(6), 567 - 71
A new exon 9 glucose-6-phosphate dehydrogenase mutation (G6PD "Rehovot") in a Jewish Ethiopian family with variable phenotypes; Iancovici-Kidon M et al.; Hereditary nonspherocytic hemolytic anemia (HNSHA) is a rare manifestation of glucose-6-phosphate dehydrogenase (G6PD) gene mutations, caused mainly by mutations located in exon 10 of the G6PD gene and less commonly by mutations in other parts of the gene . A new, exon 9, single-base mutation representing a T --> C transition at cDNA nucleotide 964 was found in three brothers and their carrier mother of Jewish Ethiopian descent . Biochemical characterization of the resultant protein was not performed . Though clinical manifestations included HNSHA in all cases, the severity of hemolysis and the transfusion requirement differed markedly . Severe congenital neutropenia (Kostmann's syndrome)--a disorder never reported before in conjunction with G6PD deficiency--was observed in one case . Levels of white blood cell G6PD activity of the three patients were 0-5% of normal controls . Neutrophil oxidative and bactericidal activities were inherently impaired in the patient with Kostmann's syndrome, but were well preserved in his two siblings .

Biochim Biophys Acta, 2000 Dec 15, 1529(1-3), 257 - 75
Molecular biology and pathophysiological aspects of plasma cholesteryl ester transfer protein; Yamashita S et al.; Plasma cholesteryl ester transfer protein (CETP) facilitates the transfer of cholesteryl ester (CE) from high density lipoprotein (HDL) to apolipoprotein B-containing lipoproteins . Since CETP regulates the plasma levels of HDL cholesterol and the size of HDL particles, CETP is considered to be a key protein in reverse cholesterol transport, a protective system against atherosclerosis . CETP, as well as plasma phospholipid transfer protein, belongs to members of the lipid transfer/lipopolysaccharide-binding protein (LBP) gene family, which also includes the lipopolysaccharide-binding protein (LBP) and bactericidal/permeability-increasing protein . Although these four proteins possess different physiological functions, they share marked biochemical and structural similarities . The importance of plasma CETP in lipoprotein metabolism was demonstrated by the discovery of CETP-deficient subjects with a marked hyperalphalipoproteinemia (HALP) . Two common mutations in the CETP gene, intron 14 splicing defect and exon 15 missense mutation (D442G), have been identified in Japanese HALP patients with CETP deficiency . The deficiency of CETP causes various abnormalities in the concentration, composition, and functions of both HDL and low density lipoprotein . Although the pathophysiological significance of CETP in terms of atherosclerosis has been controversial, the in vitro experiments showed that large CE-rich HDL particles in CETP deficiency are defective in cholesterol efflux . Epidemiological studies in Japanese-Americans and in the Omagari area where HALP subjects with the intron 14 splicing defect of CETP gene are markedly frequent, have shown an increased incidence of coronary atherosclerosis in CETP-deficient patients . The current review will focus on the recent findings on the molecular biology and pathophysiological aspects of plasma CETP, a key protein in reverse cholesterol transport.

Clin Pharmacokinet, 2000 Nov, 39(5), 335 - 43
Pharmacokinetics and pharmacodynamics of cephalosporins in cerebrospinal fluid; Lutsar I et al.; Largely because of their low lipophilicity, cephalosporins poorly penetrate through the blood-brain barrier, achieving relatively low cerebrospinal fluid (CSF) concentrations . However, the minimum bactericidal concentrations (MBCs) of the extended spectrum cephalosporins for common meningeal pathogens are generally low; thus, therapeutic CSF drug concentrations several-fold greater than the MBC can be achieved with currently recommended dosage regimens . However, the effectiveness of cephalosporin therapy is unreliable in patients with meningitis caused by highly penicillin-resistant pneumococci . As in other body sites, the bactericidal activity of cephalosporins in CSF predominantly depends on the time their concentrations exceed the MBC of infecting organisms (t>MBC) . Experimental studies show that, for maximal efficacy, t>MBC values greater than 90% of the dosage interval are required in meningitis . Such values are usually achieved in humans with currently recommended dosage regimens because the half-lives of cephalosporins are 2- to 3-fold longer in CSF than in serum . Several advanced generation cephalosporins have shown equal efficacy in clinical trials, but only cefotaxime, ceftriaxone and ceftazidime are currently approved for the treatment of patients with bacterial meningitis.

Acta Medica (Hradec Kralove), 2000, 43(3), 91 - 4
Long-term alteration of immune functions following low level exposure to sarin in rats; Kassa J et al.; 1 . Long term alteration of immune functions caused by low doses of nerve agent sarin were studied in rats exposed to sarin by inhalation . The alteration of immune functions by sarin was monitored by using two methods (the evaluation of in vitro spontaneous as well as stimulated proliferation of spleen cells and in vitro bactericidal activity of peritoneal macrophages) at 3, 6 and 12 months following sarin exposure . 2 . The results indicate that not only symptomatic but also asymptomatic dose of sarin is able to alter some immune functions at six and twelve months following exposure to sarin . 3 . Thus, not only organophosphorus insecticides but also nerve agents such as sarin can be potentially immunotoxic even at very low doses that do not cause clinically manifested intoxication following the inhalation exposure . The ability of sarin at low doses to alter immune functions seems to be really long term (up to 12 months following the exposure).

Biol Pharm Bull, 2000 Nov, 23(11), 1267 - 74
Liposomes as immunomodulator--inhibitory effect of liposomes on NO production from macrophages; Aramaki Y; Liposomes have been artificially made into membranous vesicles composed essentially of naturally occurring phospholipids and have been found to serve as a carrier of drugs and an immunological adjuvant . After being intravenously injected, they are quickly removed from the blood circulation and trapped by Kupffer cells of the liver and macrophages of spleen . However, the changes liposomes exert in these cells with which liposomes interact remain unresolved . To clarify this point is very important to assure the safe use of liposomes as drug carriers . Macrophages have many unique functions, and nitric oxide (NO) produced by NO synthase (NOS) which is induced in response to some cytokines and bacterial products such as lipopolysaccharide (LPS) is responsible for the bactericidal, tumoricidal and immune regulatory activities . On the other hand, overexpressed NO is implicated in the development of atherosclerosis, DNA injury, and hypotension associated with septic shock . This article focuses on the effects of liposomes on NO production from LPS-stimulated mice peritoneal macrophages in vitro; we found that liposomes composed of phosphatidylserine inhibit NO production . We also discuss the mechanism of the inhibitory activity of liposomes.

Infect Immun, 2000 Dec, 68(12), 7175 - 9
Characterization of bactericidal immune responses following vaccination with acellular pertussis vaccines in adults; Weingart CL et al.; Sera from six adults, collected before and after acellular pertussis vaccination, and from a placebo control were examined for the ability to elicit two bactericidal immune defenses, (i) antibody-dependent complement-mediated bacterial lysis and (ii) opsonization and phagocytosis by human neutrophils . The samples were chosen based on low preimmunization titers and strong postimmunization responses to various combinations of vaccine antigens . All but two prevaccination samples demonstrated activity indicative of complement-mediated lysis . Preimmunization activity could have been due to prior infection or childhood immunization . Immunization did not result in improved bactericidal activity for any of the individuals, and in two cases immunization caused a statistically significant decrease in complement-mediated lysis . Similarly, opsonization with the postimmunization sera failed to enhance attachment or phagocytosis of bacteria by neutrophils, and one postimmunization sample with a strong response to filamentous hemagglutinin caused an inhibition of phagocytosis that was statistically significant compared to that observed for the no-serum control . In summary, booster immunization of adults with acellular pertussis vaccines was not found to increase bactericidal activity over preimmunization levels . Identifying ways to promote bactericidal immune responses might improve the efficacy of acellular pertussis vaccines.

Antimicrob Agents Chemother, 2000 Dec, 44(12), 3288 - 97
Persistence of Chlamydia trachomatis is induced by ciprofloxacin and ofloxacin in vitro; Dreses-Werringloer U et al.; An in vitro cell culture model was used to investigate the long-term effect of ciprofloxacin and ofloxacin on infection with Chlamydia trachomatis . Standard in vitro susceptibility testing clearly indicated successful suppression of chlamydial growth . To mimic better in vivo infection conditions, extended treatment with the drugs was started after infection in vitro had been well established . Incubation of such established chlamydial cultures with ciprofloxacin and ofloxacin not only failed to eradicate the organism from host cells, but rather induced a state of chlamydial persistence . This state was characterized by the presence of nonculturable, but fully viable, bacteria and the development of aberrant inclusions . In addition chlamydia exhibited altered steady-state levels of key chlamydial antigens, with significantly reduced major outer membrane protein and near constant hsp60 levels . Resumption of overt chlamydial growth occurred after withdrawal of ciprofloxacin, confirming the viability of persisting chlamydia . In vitro ciprofloxacin results are consistent with clinical data, thereby providing an explanation for treatment failures of ciprofloxacin . Parallel in vitro studies with ofloxacin suggest a better correlation between clinical and laboratory-defined efficacy, although the clinical studies on which this assessment is based did not include monitoring of chlamydial persistence . The data presented here clearly demonstrate that under at least some circumstances, standard determination of MICs and minimal bactericidal concentrations for C . trachomatis allows no more than a simple definition of whether an antibiotic has some anti chlamydial activity; however, such testing is not always sufficient to verify that the antibiotic will eliminate the organism in vivo.

J Biol Chem, 2001 Feb 23, 276(8), 5908 - 15 Epub 2000 Nov 16.
A hydrophobic cluster at the surface of the human plasma phospholipid transfer protein is critical for activity on high density lipoproteins; Desrumaux C et al.; The plasma phospholipid transfer protein (PLTP) belongs to the lipid transfer/lipopolysaccharide binding protein (LT/LBP) family, together with the cholesteryl ester transfer protein, the lipopolysaccharide binding protein (LBP) and the bactericidal permeability increasing protein (BPI) . In the present study, we used the crystallographic data available for BPI to build a three-dimensional model for PLTP . Multiple sequence alignment suggested that, in PLTP, a cluster of hydrophobic residues substitutes for a cluster of positively charged residues found on the surface of LBP and BPI, which is critical for interaction with lipopolysaccharides . According to the PLTP model, these hydrophobic residues are situated on an exposed hydrophobic patch at the N-terminal tip of the molecule . To assess the role of this hydrophobic cluster for the functional activity of PLTP, single point alanine mutants were engineered . Phospholipid transfer from liposomes to high density lipoprotein (HDL) by the W91A, F92A, and F93A PLTP mutants was drastically reduced, whereas their transfer activity toward very low density lipoprotein and low density lipoprotein did not change . The HDL size conversion activity of the mutants was reduced to the same extent as the PLTP transfer activity toward HDL . Based on these results, we propose that a functional solvent-exposed hydrophobic cluster in the PLTP molecule specifically contributes to the PLTP transfer activity on HDL substrates.

J Exp Med, 2000 Nov 6, 192(9), 1237 - 48
Natural resistance to intracellular infections: natural resistance-associated macrophage protein 1 (Nramp1) functions as a pH-dependent manganese transporter at the phagosomal membrane; Jabado N et al.; Mutations at the natural resistance-associated macrophage protein 1 (Nramp1) locus cause susceptibility to infection with antigenically unrelated intracellular pathogens . Nramp1 codes for an integral membrane protein expressed in the lysosomal compartment of macrophages, and is recruited to the membrane of phagosomes soon after the completion of phagocytosis . To define whether Nramp1 functions as a transporter at the phagosomal membrane, a divalent cation-sensitive fluorescent probe was designed and covalently attached to a porous particle . The resulting conjugate, zymosan-FF6, was ingested by macrophages and its fluorescence emission was recorded in situ after phagocytosis, using digital imaging . Quenching of the probe by Mn(2+) was used to monitor the flux of divalent cations across the phagosomal membrane in peritoneal macrophages obtained from Nramp1-expressing (+/+) and Nramp1-deficient (-/-) macrophages . Phagosomes from Nramp1(+/+) mice extrude Mn(2+) faster than their Nramp(-/-) counterparts . The difference in the rate of transport is eliminated when acidification of the phagosomal lumen is dissipated, suggesting that divalent metal transport through Nramp1 is H(+) dependent . These studies suggest that Nramp1 contributes to defense against infection by extrusion of divalent cations from the phagosomal space . Such cations are likely essential for microbial function and their removal from the phagosomal microenvironment impairs pathogenesis, resulting in enhanced bacteriostasis or bactericidal activity.

J Antimicrob Chemother, 2000 Nov, 46(5), 811 - 4
Bactericidal effect of antibiotics on Bartonella and Brucella spp.: clinical implications; Rolain JM et al.; The species Bartonella and Brucella are phylogenetically closely related bacteria, both of which can produce chronic infections in humans that are difficult to cure with antibiotics . MICs of antibiotics for both species correlate poorly with the in vivo efficacy of the antibiotics . In this study we have determined MBCs of several antibiotics for this group of pathogens . Only the aminoglycosides were bactericidal, and this correlates well with the usefulness of these antibiotics for the therapy of human brucellosis and chronic Bartonella spp . infections such as endocarditis . Our data indicate that current clinical experience in treating brucellosis may help to define better the optimum antibiotic therapy for Bartonella-related diseases.

Ann Rheum Dis, 2000 Nov, 59(11), 898 - 903
Characterisation of autoantibodies to neutrophil granule constituents among patients with reactive arthritis, rheumatoid arthritis, and ulcerative colitis; Locht H et al.; OBJECTIVE: To study the frequency and distribution of antineutrophil cytoplasmic autoantibodies (ANCA) among patients with reactive arthritis (ReA), rheumatoid arthritis (RA), and ulcerative colitis (UC) using different immunological methods . METHODS: Fifty serum samples from patients with reactive arthritis (26 with acute disease and 24 with chronic disease-that is disease of more than one year) were analysed for ANCA with indirect immunofluorescence, enzyme linked immunosorbent assay (ELISA) with six different neutrophil granule proteins as antigens, and immunoblotting on whole neutrophil extract and extracts of azurophil and specific granules . Thirty serum samples from patients with RA and UC served as controls in ELISA and indirect immunofluorescence . RESULTS: Sixteen per cent of patients with ReA were positive in immunofluorescence compared with 30% of patients with RA, and 70% of patients with UC . Thirty two per cent of patients with ReA were positive in ELISA . Antibodies directed against lactoferrin occurred in 20%, antibodies against bactericidal permeability increasing protein (BPI), elastase, cathepsin G, myeloperoxidase, and proteinase 3 were found in 8%, 2%, 2%, 8%, and 6%, respectively . Overall, 50% of RA sera and 53% of UC sera were positive in one or more ELISA assays, the corresponding figures for antibodies against individual antigens were for RA 7%, 3%, 0%, 13%, 47%, 17% and for UC 13%, 20%, 0%, 23%, 10%, and 17% . In immunoblotting, bands corresponding to lactoferrin and BPI were recognised in 44% and 22% of ReA sera . CONCLUSION: Antibodies against neutrophil granule antigens are often found in patients with ReA, primarily among those with chronic disease . The different methods detect various subsets of antibodies, with immunoblotting being the most and immunofluorescence the least sensitive.

Biol Trace Elem Res, 2000 Mar, 73(3), 269 - 78
Cytochrome c oxidase, Cu,Zn-superoxide dismutase, and ceruloplasmin activities in copper-deficient bovines; Cerone SI et al.; The activity of several cuproenzymes in relation to the immune system was examined in serum and blood cells from bovines with molybdenum-induced copper deficiency . Five female cattle were given molybdenum (30 ppm) and sulfate (225 ppm) to induce experimental secondary copper deficiency . Ceruloplasmin activity was determined in serum . The Cu,Zn-superoxide dismutase and cytochrome c oxidase activities were measured in peripheral blood lymphocytes, neutrophils, and monocyte-derived macrophages . Copper deficiency was confirmed from decreased serum copper levels and the animals with values less than 5.6 micromol/L were considered deficient . The content of intracellular copper decreased between 40% and 70% in deficient cells compared with the controls . In copper-deficient animals, the serum ceruloplasmin activity decreased to half of the control value . Both of them, the Cu,Zn-superoxide dismutase and the cytochrome c oxidase activities, undergo a significant reduction in leukocytes, showing differences among diverse cell populations . We concluded that the copper deficiency alters the activity of several enzymes, which mediate antioxidant defenses and ATP formation . These effects may impair the cell immune functionality, affecting the bactericidal capacity and making the animals more susceptible to infection.

Antimicrob Agents Chemother, 2000 Nov, 44(11), 2969 - 78
Stages of polymyxin B interaction with the Escherichia coli cell envelope; Daugelavicius R et al.; The effects of polymyxin B (PMB) on the Escherichia coli outer (OM) and cytoplasmic membrane (CM) permeabilities were studied by monitoring the fluxes of tetraphenylphosphonium, phenyldicarbaundecaborane, and K(+) and H(+) ions . At concentrations between 2 and 20 microgram/ml, PMB increased the OM permeability to lipophilic compounds and induced a leakage of K(+) from the cytosol and an accumulation of lipophilic anions in the cellular membranes but did not cause the depolarization of the CM . At higher concentrations, PMB depolarized the CM, forming ion-permeable pores in the cell envelope . The permeability characteristics of PMB-induced pores mimic those of bacteriophage- and/or bacteriocin-induced channels . However, the bactericidal effect of PMB took place at concentrations below 20 microgram/ml, indicating that this effect is not caused by pore formation . Under conditions of increased ionic strength, PMB made the OM permeable to lipophilic compounds and decreased the K(+) gradient but was not able to depolarize the cells . The OM-permeabilizing effect of PMB can be diminished by increasing the concentration of Mg(2+) . The major new findings of this work are as follows: (i) the OM-permeabilizing action of PMB was dissected from its depolarizing effect on the CM, (ii) the PMB-induced ion-permeable pores in bacterial envelope were registered, and (iii) the pore formation and depolarization of the CM are not obligatory for the bactericidal action of PMB and dissipation of the K(+) gradient on the CM.

Infect Immun, 2000 Nov, 68(11), 6250 - 6
Antigenic structure of outer membrane protein E of Moraxella catarrhalis and construction and characterization of mutants; Murphy TF et al.; Outer membrane protein E (OMP E) is a 50-kDa protein of Moraxella catarrhalis which possesses several characteristics indicating that the protein will be an effective vaccine antigen . To study the antigenic structure of OMP E, eight monoclonal antibodies were developed and characterized . Three of the antibodies recognized epitopes which are present on the bacterial surface . Fusion peptides corresponding to overlapping regions of OMP E were constructed, and immunoblot assays were performed to localize the areas of the molecule bound by the monoclonal antibodies . These studies identified a surface-exposed epitope in the region of amino acids 80 through 180 . To further study the protein, two mutants which lack OMP E were constructed . In bactericidal assays, the mutants were more readily killed by normal human serum compared to the isogenic parent strains . These results indicate that OMP E is involved in the expression of serum resistance of M . catarrhalis.

Eur J Surg, 2000 Sep, 166(9), 700 - 5
Systemic inflammatory response in acute cholangitis and after subsequent treatment; Kimmings AN et al.; OBJECTIVE: To measure the concentrations of endotoxin and inflammatory mediators during an attack of acute cholangitis and see what effect endoscopic treatment had on these mediators . DESIGN: Prospective study . SETTING: University teaching hospital The Netherlands . SUBJECTS: Ten patients with acute cholangitis . INTERVENTIONS: Measurements were made during the attack and 1 week after endoscopic treatment . MAIN OUTCOME MEASURE: Changes in clinical variables, and severity of biliary obstruction . Concentrations of endotoxin, cytokines, and endotoxin binding proteins, in plasma . RESULTS: The causes of cholangitis were obstructed endoprosthesis (n = 4) and stones (n = 6) . The median bilirubin concentration during the attack was 70.0 micromol/L (range 14-156) and 14.5 micromol/L (range 9-80) after treatment (p < 0.05) . Median (range) plasma endotoxin concentrations were 3.6 pg/ml (3.2-107) and 3.6 (2.4-5), respectively . Concentrations of cytokines were high during the acute attack and significantly lower after treatment: median tumour necrosis factor (TNF) fell from 44.6 pg/ml (range 1.2-403) to 7.3 (0-53); soluble TNF receptor p55 from 4.9 ng/ml (2.7-13.8) to 3.6 (1.4-8.2) and TNF receptor p75 from 11.6 ng/ml (7.1-40.6) to 8.1 (2.9-31.3); interleukin 6 (IL-6) fell from 690 pg/ml (34.1-4594) to 8.2 (0-39.3), IL-8 from 226.2 pg/ml (31.6-712.7) to 21.4 (4.2-63.5) and IL-10 from 33.4 pg/ml (2.7-5605) to 4.7 (0-16.7) (p < 0.03) . Values for lipopolysaccharide binding protein and soluble CD14 also fell significantly (p < 0.01) from 86.5 (43.4-200) to 21.5 (11.3-37.5) and from 200 (59-200) to 47.8 (0.47-200), respectively . The concentration of bactericidal permeability increasing protein did not change significantly, being 7.1 (2-18.9) during the acute attack and 4.6 (0.8-17.7) a week later . CONCLUSION: There is a considerable systemic inflammatory response during cholangitis, which is dramatically reduced one week after endoscopic treatment.

J Hosp Infect, 2000 Sep, 46(1), 73 - 6
Evaluation of the bactericidal efficiency of a 2% alkaline glutaraldehyde solution on Mycobacterium xenopi; Dauendorffer JN et al.; Mycobacterium xenopi (M . xenopi) has been implicated in hospital-acquired infections associated with colonization of hospital water systems . M . xenopi is considered to be as resistant as other atypical mycobacteria, which are known to be resistant to many disinfecting treatments . However, the efficacy of disinfectants on this organism has not yet been studied . Therefore we decided to evaluate its susceptibility to 2% alkaline glutaraldehyde solution, which is commonly used in hospitals . Tests were conducted using five strains of M . xenopi: three isolated from human samples, an environmental strain and a collection strain . We used a membrane filtration assay and counted surviving bacteria before and after several exposure times (5, 15, 30 and 60 min) with the disinfecting solution . The log10 reduction factor of organisms achieved within 60 min contact ranged from 2.5 to 7.5 . This showed M . xenopi to be more resistant to disinfectants than M . tuberculosis or M . smegmatis and suggested that environmental strains may be more resistant to alkaline glutaraldehyde than those isolated from human samples.

J Appl Microbiol, 2000 Sep, 89(3), 517 - 25
Bactericidal action of high-power Nd:YAG laser light on Escherichia coli in saline suspension; Ward GD et al.; Infra-red light (1064 nm) from a high-power Nd:YAG laser caused more than 90% loss of viability of Escherichia coli during exposures that raised the temperature of PBS suspensions of the bacteria to 50 C in a thermocouple-equipped cuvette . In contrast, there was minimal loss of viability after heating the same suspensions to 50 degrees C in a water-bath, or in a PCR thermal cycler . The mechanism of laser killing at 50 degrees C was explored by differential scanning calorimetry, by laser treatment of transparent and turbid bacterial suspensions, and by optical absorbency studies of E . coli suspensions at 1064 nm . Taken together, the data suggested that the bactericidal action of Nd:YAG laser light at 50 degrees C was due partly to thermal heating and partly to an additional, as yet undefined, mechanism . Scanning electron microscopy revealed localized areas of surface damage on laser-exposed E . coli cells.

J Gen Physiol, 2000 Oct, 116(4), 587 - 98
Protein translocation across planar bilayers by the colicin Ia channel-forming domain: where will it end?
Kienker PK, Jakes KS, Finkelstein A.
Colicin Ia, a 626-residue bactericidal protein, consists of three domains, with the carboxy-terminal domain (C domain) responsible for channel formation . Whole colicin Ia or C domain added to a planar lipid bilayer membrane forms voltage-gated channels . We have shown previously that the channel formed by whole colicin Ia has four membrane-spanning segments and an approximately 68-residue segment translocated across the membrane . Various experimental interventions could cause a longer or shorter segment within the C domain to be translocated, making us wonder why translocation normally stops where it does, near the amino-terminal end of the C domain (approximately residue 450) . We hypothesized that regions upstream from the C domain prevent its amino-terminal end from moving into and across the membrane . To test this idea, we prepared C domain with a ligand attached near its amino terminus, added it to one side of a planar bilayer to form channels, and then probed from the opposite side with a water-soluble protein that can specifically bind the ligand . The binding of the probe had a dramatic effect on channel gating, demonstrating that the ligand (and hence the amino-terminal end of the C domain) had moved across the membrane . Experiments with larger colicin Ia fragments showed that a region of more than 165 residues, upstream from the C domain, can also move across the membrane . All of the colicin Ia carboxy-terminal fragments that we examined form channels that pass from a state of relatively normal conductance to a low-conductance state; we interpret this passage as a transition from a channel with four membrane-spanning segments to one with only three.

Am J Otol, 2000 Sep, 21(5), 625 - 30
Inhibition of endotoxin effects on cultured human middle ear epithelium by bactericidal permeability-increasing protein; Nell MJ et al.; HYPOTHESIS/BACKGROUND: Endotoxin can induce morphologic changes to middle ear epithelium, which can disturb the mucociliary clearance system (MCS) and lead to otitis media with effusion (OME) . The bactericidal/permeability-increasing (BPI) protein is a major component of neutrophil granules and binds with high affinity to endotoxin . In this study, the capacity to inhibit the effects of endotoxin by rBPI21, a recombinant amino-terminal analog derived from BPI, was investigated on cultured human middle ear epithelium using light microscopy and scanning- and transmission electron microscopy . METHODS: Human middle ear epithelium was air-exposed cultured on a collagenous underlayer with different additions of endotoxin and rBPI21 to the culture medium . The tissue specimens were inspected after 4 weeks for the number of ciliated and secretory cells, thickness of the mucosal layer, and cell size . RESULTS: The morphologic changes induced by endotoxin were increased thickness of the mucosal layer and increased number of secretory cells . These changes were significantly diminished or even absent when endotoxin was added with rBPI21 to the culture medium . CONCLUSION: rBPI21 can inhibit morphologic changes in the middle ear epithelium due to endotoxin . Hence, the authors believe that rBPI21 can be a new therapeutic agent in the treatment of OME.

Infect Immun, 2000 Oct, 68(10), 5668 - 72
Identification of constituents of human neutrophil azurophil granules that mediate fungistasis against Histoplasma capsulatum; Newman SL et al.; Previously we demonstrated that human neutrophils mediate potent and long-lasting fungistasis against Histoplasma capsulatum yeasts and that all of the fungistatic activity resides in the azurophil granules . In the present study, specific azurophil granule constituents with fungistatic activity were identified by incubation with H . capsulatum yeasts for 24 h and by quantifying the subsequent growth of yeasts via the incorporation of {(3)H}leucine . Human neutrophil defensins HNP-1, HNP-2, and HNP-3 inhibited the growth of H . capsulatum yeasts in a concentration-dependent manner with maximum inhibition at 8 microg/ml . At a concentration of 4 microg/ml, all possible paired combinations of defensins exhibited additive fungistatic activity against H . capsulatum yeasts . Cathepsin G and bactericidal-permeability-increasing protein (BPI) also mediated fungistasis against H . capsulatum in a concentration-dependent manner . The fungistatic activities of combinations of cathepsin G and BPI were additive, as were those of combinations of cathepsin G or BPI with HNP-1, HNP-2, and HNP-3 . Lysozyme and elastase exhibited modest antifungal activity, and azurocidin and proteinase 3 exhibited no significant fungistasis against H . capsulatum yeasts . Thus, defensins, cathepsin G, and BPI are the major anti-H . capsulatum effector molecules in the azurophil granules of human neutrophils.

Infect Immun, 2000 Oct, 68(10), 5619 - 27
Gallium disrupts iron metabolism of mycobacteria residing within human macrophages; Olakanmi O et al.; Mycobacterium tuberculosis and M . avium complex (MAC) enter and multiply within monocytes and macrophages in phagosomes . In vitro growth studies using standard culture media indicate that siderophore-mediated iron (Fe) acquisition plays a critical role in the growth and metabolism of both M . tuberculosis and MAC . However, the applicability of such studies to conditions within the macrophage phagosome is unclear, due in part to the absence of experimental means to inhibit such a process . Based on the ability of gallium (Ga(3+)) to concentrate within mononuclear phagocytes and on evidence that Ga disrupts cellular Fe-dependent metabolic pathways by substituting for Fe(3+) and failing to undergo redox cycling, we hypothesized that Ga could disrupt Fe acquisition and Fe-dependent metabolic pathways of mycobacteria . We find that Ga(NO(3))(3) and Ga-transferrin produce an Fe-reversible concentration-dependent growth inhibition of M . tuberculosis strains and MAC grown extracellularly and within human macrophages . Ga is bactericidal for M . tuberculosis growing extracellularly and within macrophages . Finally, we provide evidence that exogenously added Fe is acquired by intraphagosomal M . tuberculosis and that Ga inhibits this Fe acquisition . Thus, Ga(NO(3))(3) disruption of mycobacterial Fe metabolism may serve as an experimental means to study the mechanism of Fe acquisition by intracellular mycobacteria and the role of Fe in intracellular survival . Furthermore, given the inability of biological systems to discriminate between Ga and Fe, this approach could have broad applicability to the study of Fe metabolism of other intracellular pathogens.

Antimicrob Agents Chemother, 2000 Oct, 44(10), 2919 - 21
Bactericidal activities of HMR 3647, moxifloxacin, and rifapentine against Mycobacterium leprae in mice; Consigny S et al.; Bactericidal activities of HMR 3647 (HMR), moxifloxacin (MXFX), and rifapentine (RPT) against Mycobacterium leprae, measured by the proportional bactericidal technique in the mouse footpad system, were compared with those of the established antileprosy drugs clarithromycin (CLARI), ofloxacin (OFLO), and rifampin (RMP) . Administered in five daily doses of 100 mg/kg of body weight, HMR appeared slightly more bactericidal than CLARI . In a single dose, MXFX at 150 mg/kg was more active than the same dose of OFLO and displayed exactly the same level of activity as RMP at 10 mg/kg; the combination MXFX-minocycline (MINO) (MM) was more bactericidal than the combination OFLO-MINO (OM); RPT at 10 mg/kg was more bactericidal than the same dose of RMP and even more active than the combination RMP-OFLO-MINO (ROM); the combination RPT-MXFX-MINO (PMM) killed 99.9% of viable M . leprae and was slightly more bactericidal than RPT alone, indicating that the combination PMM showed an additive effect against M . leprae.

Int J Tuberc Lung Dis, 2000 Sep, 4(9), 796 - 806
Role of individual drugs in the chemotherapy of tuberculosis; Mitchison DA; During the course of chemotherapy, certain drugs are predominant in their bactericidal activities . Isoniazid is responsible for an initial kill of about 95% of organisms during the first 2 days of treatment . Its bactericidal role is then replaced by rifampicin and pyrazinamide during the intensive phase . In the continuation phase with an isoniazid/rifampicin regimen, rifampicin is the only effective drug against persisters, as shown by the similarity of response by patients with initially isoniazid-resistant or sensitive strains . If the continuation phase regimen does not contain rifampicin but does contain isoniazid, the dominant bactericidal drug is isoniazid . In this case, the response of patients with initial isoniazid resistance is appreciably less good than in those with sensitive organisms . The review suggests exploration in randomised control trials of a continuation phase of rifampicin (or rifapentine) alone . It also suggests the importance of the dose size of rifampicin and the need for exploring a higher dose . Finally, it emphasises the importance of finding drugs that act on persisting organisms that are phenotypically but not genetically resistant to rifampicin.

J Antimicrob Chemother, 2000 Sep, 46(3), 363 - 8
Inactivation of a hepadnavirus by electrolysed acid water; Tagawa M et al.; Glutaraldehyde is used as a disinfectant for endoscopes, but is an irritant and so should be replaced by an alternative . Electrolysed acid water (EAW) has a bactericidal effect, and an endoscopic washing device using EAW has been developed in Japan . To investigate the effect of EAW on the infectivity of viruses, we treated duck hepatitis B virus (DHBV), which has similar properties to hepatitis B virus, with EAW, and determined the number of remaining infectious virus particles in a bioassay system . One-day-old Pekin ducks were inoculated with duck serum containing 10(5.5) ID(50) DHBV; the serum had previously been incubated with 100 volumes of EAW or ion-exchanged water at room temperature for 7 min . DHBV infection was indicated by detection of viral DNA in duck serum samples 1-8 weeks after inoculation . Treatment of serum with EAW diminished DHBV infectivity whereas treatment with ion-exchanged water did not . The virus load was estimated to have been reduced to 10(1)-10(3) ID(50) during the first 1 min and to <10(0.5) ID(50) in the next 6 min of incubation when compared with the control . Thus, EAW directly inactivates DHBV and its clinical application is recommended.

Bull Exp Biol Med, 2000 May, 129(5), 484 - 6
Effect of peroxidase on the development of experimental leprosy in mice after intraplantar infection; Maslov AK et al.; Lyophilized horseradish peroxidase (activity 100 U/mg) administered per os in a dose of 100-200 mg/kg fodder enhanced bactericidal activity of phagocytes in mice experimentally infected with Mycobacterium leprae, which manifested in suppression of M . leprae growth in comparison with untreated controls.

Acta Gastroenterol Latinoam, 2000, 30(3), 177 - 86
{Analytical review of multicenter studies with polycresulene for hemorrhoidal pathologies}; Espinosa DJ; Seven centres investigated the therapeutic efficacy and tolerability of policresulene associated to cinchocaine administered locally as ointment, suppositories or both formulations in 2287 patients with hemorrhoid pathology . The studies were conducted with a standardised protocol and case report forms and with the same score criteria for rating efficacy and tolerability according to the physicians and the patients . Highly satisfactory results were achieved in 1904 patients (83.2%) according to the investigators criteria . Patients rated the outcome most satisfactory for 1881 cases (82.2%) . The following were found to be the principal indications: external and internal hemorrhoids associated with bleeding, acute anal fissures, rhagades and perforated or incised perianal thrombosis, anal eczema and anal pruritus, proctitis and wound treatment after proctologic surgery . None of the investigators found any serious adverse event . Mild to moderate adverse reactions in 10% of the patients were local discomfort, pruritus, burning or irritation . Such symptoms occurred at the beginning of treatment . The favourable effects of policresulene are attributed to its unique mechanism of action . The highly acid characteristics of the substance causes a selective coagulation of the necrotic tissues leaving healthy tissues unaffected . The desquamation and remotion of the necrotic tissues induces rapid wound cleansing, and a reactive hyperemia of the treated area enhancing epithelization . Its highly acid pH produces a marked bactericidal action on the most common pathogens and C . albicans as well . Policresulene has hemostyptic properties producing vasoconstriction of the myofibrils of the blood vessels arresting profuse bleeding from large areas . The local anesthetic cinchocaine contributes to the initial pain relief . None of the formulations contains corticosteroids which makes this preparations also suitable for long term treatment periodsPublication Types:

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