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-