Antimicrobial

Microbes Infect, 2000 Jul, 2(8), 907 - 13
Mechanisms of bacterial resistance and response to bile; Gunn JS; Enteric bacteria are resistant to the bactericidal effects of intestinal bile, but these resistance mechanisms are not completely understood . It is becoming increasingly apparent that enteric bacteria have evolved to utilize bile as a signal for the temporal production of virulence factors and other adaptive mechanisms . A greater understanding of the resistance and response of bacteria to bile may assist the development of novel therapeutic, prevention, and diagnostic strategies to treat enteric and extraintestinal infections.

J Bacteriol, 2000 Sep, 182(18), 5225 - 30
Characterization of Escherichia coli DNA lesions generated within J774 macrophages; Schlosser-Silverman E et al.; Macrophages are armed with multiple oxygen-dependent and -independent bactericidal properties . However, the respiratory burst, generating reactive oxygen species, is believed to be a major cause of bacterial killing . We exploited the susceptibility of Escherichia coli in macrophages to characterize the effects of the respiratory burst on intracellular bacteria . We show that E . coli strains recovered from J774 macrophages exhibit high rates of mutations . We report that the DNA damage generated inside macrophages includes DNA strand breaks and the modification 8-oxo-2'-deoxyguanosine, which are typical oxidative lesions . Interestingly, we found that under these conditions, early in the infection the majority of E . coli cells are viable but gene expression is inhibited . Our findings demonstrate that macrophages can cause severe DNA damage to intracellular bacteria . Our results also suggest that protection against the macrophage-induced DNA damage is an important component of the bacterial defense mechanism within macrophages.

J Bacteriol, 2000 Sep, 182(18), 5082 - 90
Sequential inactivation of rdxA (HP0954) and frxA (HP0642) nitroreductase genes causes moderate and high-level metronidazole resistance in Helicobacter pylori; Jeong JY et al.; Helicobacter pylori is a human-pathogenic bacterial species that is subdivided geographically, with different genotypes predominating in different parts of the world . Here we test and extend an earlier conclusion that metronidazole (Mtz) resistance is due to mutation in rdxA (HP0954), which encodes a nitroreductase that converts Mtz from prodrug to bactericidal agent . We found that (i) rdxA genes PCR amplified from 50 representative Mtz(r) strains from previously unstudied populations in Asia, South Africa, Europe, and the Americas could, in each case, transform Mtz(s) H . pylori to Mtz(r); (ii) Mtz(r) mutant derivatives of a cultured Mtz(s) strain resulted from mutation in rdxA; and (iii) transformation of Mtz(s) strains with rdxA-null alleles usually resulted in moderate level Mtz resistance (16 microg/ml) . However, resistance to higher Mtz levels was common among clinical isolates, a result that implicates at least one additional gene . Expression in Escherichia coli of frxA (HP0642; flavin oxidoreductase), an rdxA paralog, made this normally resistant species Mtz(s), and frxA inactivation enhanced Mtz resistance in rdxA-deficient cells but had little effect on the Mtz susceptibility of rdxA(+) cells . Strains carrying frxA-null and rdxA-null alleles could mutate to even higher resistance, a result implicating one or more additional genes in residual Mtz susceptibility and hyperresistance . We conclude that most Mtz resistance in H . pylori depends on rdxA inactivation, that mutations in frxA can enhance resistance, and that genes that confer Mtz resistance without rdxA inactivation are rare or nonexistent in H . pylori populations.

Vet Res, 2000 Jul-Aug, 31(4), 397 - 412
Hyperketonemia and the impairment of udder defense: a review; Suriyasathaporn W et al.; The objective of this study was to review the possible relationships between hyperketonemia and the function of phagocytes with respect to the bovine udder defense mechanism . We hypothesize that an increased incidence of clinical mastitis in high-producing cows is caused by the impairment of the udder defense mechanism during hyperketonemia . First, we review the acute phase of udder defense mechanisms after intramammary infection . The physiological changes of cows in negative energy balance are subsequently discussed . Finally, possible relationships between udder defense and physiological changes during negative energy balance, especially hyperketonemia, are reviewed . The three stages of an acute phase of udder defense are: (1) immediately eliminating invading pathogens by phagocytes, (2) releasing inflammatory substances, especially chemoattractants, and (3) migration of polymorphonuclear leukocytes into the infected udder . Leukocytes from hyperketonemia subjects show a lower capacity of the phagocytic defense mechanism . In addition, the phagocytic and bactericidal capacities of neutrophils are reduced when these cells are acting in the presence of high concentrations of ketone bodies . Lower amounts of cytokine production after bacterial infection are observed in ketotic subjects . The chemotactic capacity of blood leukocytes is impaired in leukocytes obtained from ketotic cows . Lower numbers of blood leukocytes are observed in ketotic cows . In conclusion, the impairment of the udder defense mechanism in negative energy balance cows seems related to hyperketonemia.

Infect Immun, 2000 Sep, 68(9), 5261 - 8
Lipooligosaccharide P(k) (Galalpha1-4Galbeta1-4Glc) epitope of moraxella catarrhalis is a factor in resistance to bactericidal activity mediated by normal human serum; Zaleski A et al.; Moraxella catarrhalis is a respiratory pathogen responsible for acute bacterial otitis media in children and exacerbation of chronic bronchitis in adults . M . catarrhalis strains are frequently resistant to the bactericidal activity of normal human serum . In order to determine if the lipooligosaccharide (LOS) of M . catarrhalis has a role in serum resistance, the UDP-glucose-4-epimerase (galE) gene was identified, cloned, and sequenced and a deletion/insertion mutation was introduced into M . catarrhalis strain 2951 . GalE enzymatic activity, measured in whole-cell lysates, was ablated in M . catarrhalis 2951 galE . Mass spectrometric analysis of LOS isolated with hot phenol-water confirmed that strain 2951 produced a type A LOS . These studies showed that the LOS from 2951 galE had lost two hexose residues due to the galE mutation and that the resultant LOS structure lacked the (Galalpha1-4Galbeta1-4Glc) P(k) epitope found on M . catarrhalis 2951 . Wild-type M . catarrhalis 2951 is resistant to complement-mediated serum bactericidal activity . In contrast, a greater than 2-log(10)-unit reduction in CFU occurred after incubation of 2951 galE in either 50 or 25% pooled human serum (PNHS), and CFU in 10% PNHS decreased by about 1 log(10) unit . These studies suggest that the P(k) epitope of the LOS may be an important factor in the resistance of M . catarrhalis to the complement-mediated bactericidal effect of normal human serum.

Infect Immun, 2000 Sep, 68(9), 4954 - 60
Serum amyloid P component prevents high-density lipoprotein-mediated neutralization of lipopolysaccharide; de Haas CJ et al.; Lipopolysaccharide (LPS) is an amphipathic macromolecule that is highly aggregated in aqueous preparations . LPS-binding protein (LBP) catalyzes the transfer of single LPS molecules, segregated from an LPS aggregate, to high-density lipoproteins (HDL), which results in the neutralization of LPS . When fluorescein isothiocyanate-labeled LPS (FITC-LPS) is used, this transfer of LPS monomers to HDL can be measured as an increase in fluorescence due to dequenching of FITC-LPS . Recently, serum amyloid P component (SAP) was shown to neutralize LPS in vitro, although only in the presence of low concentrations of LBP . In this study, we show that SAP prevented HDL-mediated dequenching of FITC-LPS, even in the presence of high concentrations of LBP . Human bactericidal/permeability-increasing protein (BPI), a very potent LPS-binding and -neutralizing protein, also prevented HDL-mediated dequenching of FITC-LPS . Furthermore, SAP inhibited HDL-mediated neutralization of both rough and smooth LPS in a chemiluminescence assay quantifying the LPS-induced priming of neutrophils in human blood . SAP bound both isolated HDL and HDL in serum . Using HDL-coated magnetic beads prebound with SAP, we demonstrated that HDL-bound SAP prevented the binding of LPS to HDL . We suggest that SAP, by preventing LPS binding to HDL, plays a regulatory role, balancing the amount of LPS that, via HDL, is directed to the adrenal glands.

Free Radic Biol Med, 2000 Jun 1, 28(11), 1611 - 8
Oxidative cellular damage associated with transformation of Helicobacter pylori from a bacillary to a coccoid form; Nakamura A et al.; Exposure to unfavorable conditions results in the transformation of Helicobacter pylori, a gastric pathogen, from a bacillary form to a coccoid form . The mechanism and pathophysiological significance of this transformation remain unclear . The generation of the superoxide radical by H . pylori has previously been shown to inhibit the bactericidal action of nitric oxide, the concentration of which is relatively high in gastric juice . With the use of chemiluminescence probes, both the quality and quantity of reactive oxygen species generated by H . pylori have now been shown to change markedly during the transformation from the bacillary form to the coccoid form . The transformation of H . pylori was associated with oxidative modification of cellular proteins, including urease, an enzyme required for the survival of this bacterium in acidic gastric juice . Although the cellular abundance of urease protein increased during the transformation, the specific activity of the enzyme decreased and it underwent aggregation . Specific activities of both superoxide dismutase and catalase in H . pylori also decreased markedly during the transformation . The transformation of H . pylori was also associated with oxidative modification of DNA, as revealed by the generation of 8-hydroxyguanine, and subsequent DNA fragment . These observations indicate that oxidative stress elicited by endogenously generated reactive oxygen species might play an important role in the transformation of H . pylori from the bacillary form to the coccoid form.

Mol Biotechnol, 1999 Dec 15, 13(3), 247 - 55
Direct and indirect methods of measuring Helicobacter pylori drug susceptibility in vitro; McLaren A; This article outlines a number of methods for the determination of inhibitory and bactericidal activity against H . pylori . Direct methods rely on the ability of bacteria to divide and multiply and ultimately form visible colonies after subjection to antibiotic treatment . Indirect methods rely on the measurement of metabolic activity as a viability marker and are much more rapid, especially taking into account the slow growth and fastidious nature of the organism . Inhibitory concentration measurement does not indicate the bactericidal ability of a drug; inhibition of growth does not necessarily correlate with cell death . Theoretical generation of viable but nonculturable bacteria could bring in to question the validity of direct measurements based on the colony forming ability of an organism posttreatment.

Can J Microbiol, 2000 Jul, 46(7), 623 - 32
Bacterial dynamics in first year sea ice and underlying seawater of Saroma-ko Lagoon (Sea of Okhotsk, Japan) and resolute passage (High Canadian Arctic): inhibitory effects of ice algae on bacterial dynamics; Monfort P et al.; The seasonal development of bacterial abundance in first year bottom ice and underlying seawater were studied at Saroma-ko Lagoon in Hokkaido, Japan, and at Resolute Passage in the High Canadian Arctic during the algal bloom in spring 1992 . The aim of this study was to evaluate whether the high algal concentrations reached during the bloom of ice algae have inhibitory effects on bacterial dynamics . Bacterial abundance (measured as total cell count and colony-forming units CFU) increased with the increase of the algal biomass up to 500 micrograms Chla.L-1 in both locations . Culturable fraction (measured as the percentage of CFU counts versus the total cell counts) was between 7% and 22% at Saroma-ko, and approximately 0.08% at Resolute Passage . When algal biomass exceeded 500 micrograms of Chla.L-1, both bacterial abundance and culturable fraction decreased significantly . There was a maximum threshold of algal biomass (between 500 and 800 micrograms of Chla.L-1) after which bacterial dynamics become negatively coupled to the algal biomass . These results suggest that bactericidal and/or bacteriostatic compounds from these extremely high algal concentrations could explain the decrease in bacterial abundance and culturability in bottom ice observed after the ice algae bloom.

Zentralbl Chir, 2000, 125(5), 450 - 3
{Ciprofloxacin levels in pleural fluid and serum during systemic administration after pneumonectomy}; Padberg WM et al.; Postpneumonectomy empyema represents a frequently lethal complication . It remains unsolved whether prophylactic antibiotics achieve a bactericidal concentration in the pleural cavity after pneumonectomy . 12 patients undergoing pneumonectomy received ciprofloxacin intravenously (2 x 200 micrograms/d) and orally (2 x 500 micromilligrams/d) during the first and second postoperative week, respectively . 1, 6, 9 and 14 days after the operation the ciprofloxacin concentration was measured in the pleural fluid and serum . Already after 24 hours bactericidal levels (0.56 microgram/ml) were found in the pleural fluid, rising to 1.11 micrograms/ml on day 14 under the higher oral dosage . Thus, it could be demonstrated that during the first two weeks after pneumonectomy high concentrations of an antibiotic similar to the levels in the serum can be achieved in the pleural fluid.

J Periodontol, 2000 Jun, 71(6), 1024 - 8
Long-term follow-up of periodontitis in a patient with Chédiak-Higashi syndrome . A case report; Shibutani T et al.; Chediak-Higashi syndrome (CHS) is an extremely rare hereditary disease characterized by leukocyte dysfunction . We report on a 21-year-old woman who presented at the age 9 years with CHS and serious periodontal tissue destruction around erupted teeth . The patient had received systemic, radiographic, immunological, microbial, and clinical periodontal examinations since childhood . The chemotactic activity of neutrophils in the Boyden chamber assay was 22% of the control, and leukocyte bactericidal activity was one-third of the control . Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia were isolated from periodontal pockets . Periodontal treatment including oral hygiene was provided, followed by professional tooth cleaning from the age of 12 to 21 years . However, the mobility of teeth and the inflammation of periodontal tissue progressed . This CHS patient presented with periodontal disease of extremely early onset, which was resistant to periodontal treatment.

J Spinal Cord Med, 2000 Summer, 23(2), 121 - 8
Influence of neurological level on immune function following spinal cord injury: a review; Campagnolo DI et al.; Due to the high incidence of lifelong infections in persons with spinal cord injury (SCI), the authors examined level of injury-related immune characteristics in a cohort of subjects with chronic SCI . Since the sympathetic nervous system and the endocrine system are known to be modulators of immune function, one possible explanation for heightened incidence of infections includes dysregulation of sympathetic outflow tracts in individuals with tetraplegia or high paraplegia . Natural killer cell cytotoxicity (NKCC) and bactericidal function of circulating neutrophils were assayed in a group of 10 individuals with chronic complete cervical SCI, a group of 8 individuals with paraplegia with injuries below the main sympathetic outflow (T-10 and below) and a group of 18 age- and sex-matched controls . In addition, a psychiatric assessment of depression was performed as well as assays of pituitary and adrenal functions . Analyses revealed no significant differences in immune function between all subjects with SCI combined and their matched controls . Further analyses stratifying based on presence or absence of sympathetic dysregulation revealed significantly impaired phagocytic ability and a trend toward reduced NKCC in the group with tetraplegia compared with their controls . Hormonal assays showed that dehydroepiandrosterone (DHEA), and dehydroepiandrosterone sulfate (DS) were higher in individuals with tetraplegia than controls, but no such differences were observed in individuals with paraplegia compared with their controls . The results of this study suggest that individuals sustaining complete cervical SCI experience alterations in immune function, while those with lesions at or below T-10 do not . These findings of level of injury related immune alteration could not be explained by mood differences . This paper is a review of previously published work and the authors' current thinking regarding increased acquisition of infections in this population.

Clin Infect Dis, 2000 Jul, 31(1), 131 - 5 Epub 2000 Jul 25.
Bartonella quintana and urban trench fever; Ohl ME et al.; Contemporary Bartonella quintana infections have emerged in diverse regions of the world, predominantly involving socially disadvantaged persons . Available data suggest that the human body louse Pediculus humanus is the vector for transmission of B . quintana . Descriptions of the clinical manifestations associated with contemporary B . quintana infections have varied considerably and include asymptomatic infection, a relapsing febrile illness, headache, leg pain, "culture-negative" endocarditis, and, in human immunodeficiency virus-infected persons, bacillary angiomatosis . Laboratory diagnosis is most convincing when B . quintana is isolated in blood culture, but growth often takes 20-40 days; problems exist with both sensitivity and specificity of serological assays . On the basis of available information, use of doxycycline, erythromycin, or azithromycin to treat B . quintana infections is recommended . Treatment of uncomplicated B . quintana bacteremia for 4-6 weeks and treatment of B . quintana endocarditis (in a person who does not undergo valve surgery) for 4-6 months are recommended, with the addition of a bactericidal agent (such as a third-generation cephalosporin or an aminoglycoside) during the initial 2-3 weeks of therapy for endocarditis.

Klin Khir, 1993, (1), 14 - 6
{The use of lasers in treating soft-tissue suppurative-inflammatory diseases}; Kalish IuI et al.; The authors have established in vitro, that helium-neon (HN) laser rays at the therapeutic doses had no bactericidal effect . Bactericidal effect of the rays of an ultraviolet (UV) laser is manifested in exposure for 5 min (output power of 3 mW, pulse frequency of 100 Hz, diameter of the irradiated area of 3 mm) . In the clinic, a combined method with the use of HN-laser with radiant power of 20 J/cm2 and UV laser with radiant power of 7 J/cm2 was employed . Duration of treatment of purulent-inflammatory diseases reduced 2-5-fold as compared with that in use of conventional methods of treatment.

J Mol Biol, 2000 Jul 28, 300(5), 1297 - 307
Three-dimensional crystal structure of human eosinophil cationic protein (RNase 3) at 1.75 A resolution; Mallorqui-Fernandez G et al.; Eosinophil cationic protein (ECP; RNase 3) is a human ribonuclease found only in eosinophil leukocytes that belongs to the RNase A superfamily . This enzyme is bactericidal, helminthotoxic and cytotoxic to mammalian cells and tissues . The protein has been cloned, heterologously overexpressed, purified and crystallized . Its crystal structure has been determined and refined using data up to 1 . 75 A resolution . The molecule displays the alpha+beta folding topology typical for members of the ribonuclease A superfamily . The catalytic active site residues are conserved with respect to other ribonucleases of the superfamily but some differences appear at substrate recognition subsites, which may account, in part, for the low catalytic activity . Most strikingly, 19 surface-located arginine residues confer a strong basic character to the protein . The high concentration of positive charges and the particular orientation of the side-chains of these residues may also be related to the low activity of ECP as a ribonuclease and provides an explanation for its unique cytotoxic role through cell membrane disruption .

Infect Immun, 2000 Aug, 68(8), 4759 - 64
Decorin-binding protein A (DbpA) of Borrelia burgdorferi is not protective when immunized mice are challenged via tick infestation and correlates with the lack of DbpA expression by B . burgdorferi in ticks; Hagman KE et al.; Previous studies showed that decorin-binding protein A (DbpA) of Borrelia burgdorferi was a protective immunogen in the murine model of Lyme borreliosis when mice were challenged (needle inoculated) intradermally with in vitro-cultivated spirochetes . In the present study, DbpA-immunized C3H/HeJ mice were not protected from infection when infested with Ixodes scapularis nymphs harboring virulent B . burgdorferi 297 . This lack of protection correlated with the failure to detect DbpA on B . burgdorferi in ticks, suggesting that DbpA is not available as a target for bactericidal antibodies in serum when B . burgdorferi-infected ticks take their blood meal from an immunized host . The failure of DbpA immunization to protect tick-challenged mice contradicts the results of earlier needle inoculation vaccination experiments and suggests that DbpA may not be suitable as a Lyme disease vaccine.

J Biol Chem, 2000 Sep 29, 275(39), 30372 - 7
A mechanism of membrane neutral lipid acquisition by the microsomal triglyceride transfer protein; Read J et al.; The microsomal triglyceride transfer protein (MTP) and apolipoprotein B (apoB) belong to the vitellogenin (VTG) family of lipid transfer proteins . MTP is essential for the intracellular assembly and secretion of apoB-containing lipoproteins, the key intravascular lipid transport proteins in vertebrates . We report the predicted three-dimensional structure of the C-terminal lipid binding cavity of MTP, modeled on the crystal structure of the lamprey VTG gene product, lipovitellin . The cavity in MTP resembles those found in the intracellular lipid-binding proteins and bactericidal/permeability-increasing protein . Two conserved helices, designated A and B, at the entrance to the MTP cavity mediate lipid acquisition and binding . Helix A (amino acids 725-736) interacts with membranes in a manner similar to viral fusion peptides . Mutation of helix A blocks the interaction of MTP with phospholipid vesicles containing triglyceride and impairs triglyceride binding . Mutations of helix B (amino acids 781-786) and of N780Y, which causes abetalipoproteinemia, have no impact on the interaction of MTP with phospholipid vesicles but impair triglyceride binding . We propose that insertion of helix A into lipid membranes is necessary for the acquisition of neutral lipids and that helix B is required for their transfer to the lipid binding cavity of MTP.

Blood, 2000 Jul 1, 96(1), 176 - 81
Bactericidal/permeability-increasing protein (BPI) inhibits angiogenesis via induction of apoptosis in vascular endothelial cells; van der Schaft DW et al.; Bactericidal/permeability-increasing protein (BPI) has been known for some time to function in killing bacteria and in neutralizing the effects of bacterial endotoxin lipopolysaccharide . In the present study, BPI is found to be a novel endogenous inhibitor of angiogenesis . Within the sub-muM range, BPI shows a concentration-dependent inhibition of endothelial cell (EC) proliferation that is mediated by cell detachment and subsequent induction of apoptosis . As measured by flow cytometric analysis of the percentage of subdiploid cells, apoptosis induction was half-maximal at about 250 nmol/L BPI . Apoptosis was confirmed by quantification of cells with nuclear fragmentation . Apoptosis was found to be EC specific . In an in vitro collagen gel-based angiogenesis assay, BPI at 1.8 micromol/L inhibited tube formation by 81% after only 24 hours . Evidence for in vivo inhibition of angiogenesis was obtained, using the chorioallantoic membrane assay in which BPI was seen to be significantly effective at concentrations as low as 180 nmol/L . This newly discovered function of BPI might provide a possible therapeutic modality for the treatment of various pathologic disorders that depend on angiogenesis.

J Clin Microbiol, 2000 Jul, 38(7), 2611 - 21
Antibodies against specific proteins of and immobilizing activity against three strains of Borrelia burgdorferi sensu lato can be found in symptomatic but not in infected asymptomatic dogs; Hovius JW et al.; In an area where Lyme disease is endemic in The Netherlands all dogs had positive titers by whole-cell enzyme-linked immunosorbent assay and appeared to be naturally infected by Borrelia burgdorferi sensu lato . To compare the antibody responses of symptomatic dogs and asymptomatic controls, we performed Western blots and in vitro immobilization assays to study antibody-dependent bactericidal activity . Strains from three different genospecies were employed as the antigen source: B . burgdorferi strain B31, Borrelia garinii strain A87S, and Borrelia afzelii strain pKo . Antibodies against flagellin (p41) and p39 for three strains were found in sera from both symptomatic and asymptomatic dogs and were therefore considered to be markers of exposure . Antibodies against p56 and p30 of strain B31, against p75, p58, p50, OspC, and p<19 of strain A87S, and against p56, p54, p45, OspB, p31, p26, and p<19 of strain pKo were found significantly more frequently in sera from symptomatic dogs younger than 8 years when the first symptoms were observed than in those from age-matched controls (P<0.01) . These antibodies were not found in preclinical sera and appeared during development of disease . Antibodies against OspA of strains B31 and A87S were only seen in acute-phase and convalescent sera from three dogs that recovered from disease . Incubation with 25% normal canine serum did not result in the immobilization of strains B31 and pKo, but partial immobilization of strain A87S (61%+/-24% {standard deviation} at 5 h) occurred . Seven of 15 sera from symptomatic dogs but none of the sera from 11 asymptomatic dogs had antibody-dependent immobilizing activity against one of the strains . Consecutive sera from one of these dogs immobilized two different strains . Antibody-mediated bactericidal serum was not seen before onset of disease, was strongest in the acute phase of disease, and fluctuated during chronic disease . From seven out of eight symptomatic dogs Borrelia DNA was amplified by PCR; in three of them the bactericidal activity was directed against one of the genospecies amplified from that dog; however, four PCR-positive dogs lacked bactericidal activity . In conclusion, dogs with symptomatic canine borreliosis have more-extensive antibody reactivity against Borrelia, as shown by both Western blotting and immobilization assays.

Zh Mikrobiol Epidemiol Immunobiol, 2000 Jan-Feb, (1), 37 - 41
{The properties of Escherichia isolated from the bodies of mice in bacterial translocation after immobilization stress}; Gritsenko VA et al.; As revealed in experiments on mice, 6-hour immobilization stress initiates the process of the translocation of intestinal flora to mesenteric lymph nodes and the blood . This process is accompanied by the infection of parenchymatous organs (the liver, the spleen, the kidneys, the lungs) and the increase of the concentration of E . coli in the proximal sections of the digestive tract (the duodenum and the jejunum) . As the result of the comparative analysis of the phenotypic signs of bacterial isolates obtained from intestinal and "extraintestinal" E . coli populations, the accumulation of clones with highly pronounced seroresistance and such persistence characteristics as anticomplementary and antilysozume signs, as well as resistance to the bactericidal action of leukocytic cation protein with a molecular weight of 11.0-11.5 kD, has been found to occur in the body (the blood, parenchymatous organs and the small intestine).

Ann Agric Environ Med, 2000, 7(1), 33 - 41
Subacute toxicity of orally applied alpha-cypermethrin in Swiss mice; Luty S et al.; The effect of a synthetic pyrethroid - alpha-cypermethrin administered per os for 28 days to Swiss mice was examined on phagocytic and bactericidal activity of neutrophils, and leukocytic image, IL-12 p70 level in blood plasma, as well as histologic and ultrastructural picture of the liver, heart, kidneys, lung and spleen . A synthetic pyrethroid alpha-cypermethrin, {(R,S)-alpha-cyano-3-phenoxybenzyl (R,S)-cis,trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylate}, produced by the Chemical Plant in Jaworzno was used in the study . The preparation for the application per os was used in doses 1/2 LD(50) (25 mg/kg body mass) and 1/5 LD(50) (10 mg/kg body mass) . The results were presented as mean (x) +/- standard error (SEM) and subjected to statistical analysis by the parametric t-Student test . Subacute poisoning of mice with alpha-cypermethrin in doses 1/2 LD(50) and 1/5 LD(50) resulted in decreased bactericidal activity of neutrophils . The dose 10 mg/kg body mass had a stronger stimulatory effect on phagocytic activity than 25 mg/kg body mass . Significantly higher numbers of monocytes and lymphocytes were observed in the blood of male mice poisoned with 1/5 LD(50) alpha-cypermethrin . The administration of alpha-cypermethrin resulted for both doses in the decrease in IL-12 p70 serum secretion . The lowest IL-12 p70 level (pg/ml) was noted among female mice administered 1/2 LD(50) of the preparation . The results of the study may indicate that the pyrethroid in the study had a suppressive effect on Il-12 p70 production . In mice administered 1/5 LD(50) or 1/2 LD(50) of the preparation examined, histopathologic and ultrastructural changes were observed in the liver and kidneys.

Int J Tuberc Lung Dis, 2000 Jun, 4(6), 570 - 6
Efficacy of common antiseptics against mycobacteria; Rikimaru T et al.; SETTING AND OBJECTIVE: Antiseptics are frequently used to prevent mycobacterial infection; however, the reported activities of a number of antiseptics against mycobacteria are not always consistent . The aim of this study was to determine those antiseptics that are useful against mycobacteria . DESIGN: Evaluation of antiseptic activity against mycobacteria in vitro . RESULTS: The effects of different antiseptics on mycobacteria (Mycobacterium avium, M . kansasii and M . tuberculosis) were examined . At concentrations of 0.05%, povidone-iodine (PVP-I) killed 99% or more of all strains tested within 15 seconds, while 0.5% chlorhexidine gluconate and 0.1% benzalkonium chloride showed no bactericidal activity against mycobacteria . M . kansasii and M . tuberculosis were killed after exposure to cresol for 60 seconds at concentrations of 1.0%, but M . avium was unaffected even after 60 seconds . While M . kansasii and M . tuberculosis were killed by treatment with 2.0% glutaraldehyde for 5 minutes, M . avium was highly resistant to this agent . CONCLUSION: PVP-I seems to be a useful antiseptic against mycobacteria . The measured activity of antiseptics should be interpreted carefully, due to the potential for interference by artifacts.

Arch Biochem Biophys, 2000 Jun 15, 378(2), 201 - 9
Structural and functional characterization of BnSP-7, a Lys49 myotoxic phospholipase A(2) homologue from Bothrops neuwiedi pauloensis venom; Soares AM et al.; BnSP-7, a Lys49 myotoxic phospholipase A(2) homologue from Bothrops neuwiedi pauloensis venom, was structurally and functionally characterized . Several biological activities were assayed and compared with those of the chemically modified toxin involving specific amino acid residues . The cDNA produced from the total RNA by RT-PCR contained approximately 400 bp which codified its 121 amino acid residues with a calculated pI and molecular weight of 8.9 and 13,727, respectively . Its amino acid sequence showed strong similarities with several Lys49 phospholipase A(2) homologues from other Bothrops sp . venoms . By affinity chromatography and gel diffusion, it was demonstrated that heparin formed a complex with BnSP-7, held at least in part by electrostatic interactions . BnSP-7 displayed bactericidal activity and promoted the blockage of the neuromuscular contraction of the chick biventer cervicis muscle . In addition to its in vivo myotoxic and edema-inducing activity, it disrupted artificial membranes . Both BnSP-7 and the crude venom released creatine kinase from the mouse gastrocnemius muscle and induced the development of a dose-dependent edema . His, Tyr, and Lys residues of the toxin were chemically modified by 4-bromophenacyl bromide (BPB), 2-nitrobenzenesulfonyl fluoride (NBSF), and acetic anhydride (AA), respectively . Cleavage of its N-terminal octapeptide was achieved with cyanogen bromide (CNBr) . The bactericidal action of BnSP-7 on Escherichia coli was almost completely abolished by acetylation or cleavage of the N-terminal octapeptide . The neuromuscular effect induced by BnSP-7 was completely inhibited by heparin, BPB, acetylation, and CNBr treatment . The creatine kinase releasing and edema-inducing effects were partially inhibited by heparin or modification by BPB and almost completely abolished by acetylation or cleavage of the N-terminal octapeptide . The rupture of liposomes by BnSP-7 and crude venom was dose and temperature dependent . Incubation of BnSP-7 with EDTA did not change this effect, suggesting a Ca(2+)-independent membrane lytic activity . BnSP-7 cross-reacted with antibodies raised against B . moojeni (MjTX-II), B . jararacussu (BthTX-I), and B . asper (Basp-II) myotoxins as well as against the C-terminal peptide (residues 115-129) from Basp-II .

Anasthesiol Intensivmed Notfallmed Schmerzther, 2000 May, 35(5), 316 - 8
{The effect of midazolam and flunitrazepam on the liberation of lysozyme and beta-glucuronidase from neutrophil granulocytes in vitro}; Krumholz W et al.; OBJECTIVE: Polymorphonuclear neutrophile leucocytes (PMNL) play an important role in the defence against bacterial infections . It is known that some anaesthetics are able to disturb PMNL functions . We examined the influence of midazolam and flunitrazepam on the activity of the bactericidal enzymes lysozyme and beta-glucuronidase released from PMNL in vitro . METHODS: As described before {4}, PMNL were isolated from venous blood samples obtained from 10 healthy male volunteers . PMNL stimulation and measurement of lysozyme and beta-glucuronidase activities were conducted according to the description by Metcalf et al . {5} . The BIOMED-system {8} was used for statistical evaluation . RESULTS: Neither midazolam nor flunitrazepam caused any statistically important alteration of lysozyme activity . However, clinically relevant concentrations of both benzodiazepines significantly enhanced beta-glucuronidase activity . The additives of flunitrazepam did not play any role . CONCLUSION: Surprisingly enough, midazolam and flunitrazepam increased the activity of beta-glucuronidase released from PMNL in vitro . At present, this result can neither be explained nor can its importance be estimated . On the other hand, the benzodiazepines did not relevantly influence lysozyme activity.

FEBS Lett, 2000 Jun 16, 475(2), 93 - 6
A new route to peroxynitrite: a role for xanthine oxidoreductase; Godber BL et al.; Peroxynitrite, a potent oxidising, nitrating and hydroxylating agent, results from the reaction of nitric oxide with superoxide . We show that peroxynitrite can be produced by the action of a single enzyme, xanthine oxidoreductase (XOR), in the presence of inorganic nitrite, molecular oxygen and a reducing agent, such as pterin . The effects of oxygen concentration on peroxynitrite production have been examined . The physiologically predominant dehydrogenase form of the enzyme is more effective than the oxidase form under aerobic conditions . It is proposed that XOR-derived peroxynitrite fulfils a bactericidal role in milk and in the digestive tract.

Int J Antimicrob Agents, 2000 Jun, 15(1), 55 - 63
Activity of poloxamer CRL-1072 against drug-sensitive and resistant strains of Mycobacterium tuberculosis in macrophages and in mice; Jagannath C et al.; The present experiments evaluated a new, highly refined poloxamer, CRL-1072, alone and in combination with antibiotics against drug-sensitive and -resistant organisms . In macrophage culture, CRL-1072 reduced the drug concentration inhibiting 99% of control growth of isoniazid (INH) from 10 to 0.15 mg/l (fractional inhibitory concentration=0.07) for a drug-resistant strain . CRL-1072 also increased the susceptibility of drug-resistant strains of Mycobacterium tuberculosis to INH, streptomycin, rifampicin, pyrazinamide, ethambutol, PAS, thiacetazone and ethionamide . Fractional inhibitory concentration values of <0.5 indicated significant synergistic activity . In studies of acute infection in mice, CRL-1072 was only weakly bacteriostatic when used as a single agent but increased the bactericidal activity of INH, streptomycin, rifampicin, pyrazinamide and clindamycin, but not that of ethambutol . CRL-1072 enhanced the bactericidal activity of streptomycin against a streptomycin resistant strain of M . tuberculosis in a murine infection.

Int J Antimicrob Agents, 2000 Jun, 15(1), 49 - 53
A comparison of the bactericidal effects and cytotoxic activity of three types of oxidizing water, prepared by electrolysis, as chemical dental plaque control agents; Shimada K et al.; Acid oxidizing water (AOW), neutral oxidizing water (NtOW) and acid oxidizing water with a low available chlorine concentration (AOW-LC) may be obtained by electrolyzing a solution of tap water containing various quantities of NaCl and HCl . This study compared the bactericidal effects of these waters on cariogenic and periodontopathogenic bacteria and their cytotoxicities against epithelial cells . AOW, NtOW and AOW-LC showed considerable bactericidal effects . The cytotoxicity of AOW-LC was significantly lower than the other solutions tested (P<0.0001) . The results indicated that the three types of oxidizing water had similar activity in inhibiting bacterial plaque formation as conventional chemical plaque-control agents.

J Mol Biol, 2000 Jun 16, 299(4), 1019 - 34
The 1.7 A crystal structure of BPI: a study of how two dissimilar amino acid sequences can adopt the same fold; Kleiger G et al.; We have extended the resolution of the crystal structure of human bactericidal/permeability-increasing protein (BPI) to 1.7 A . BPI has two domains with the same fold, but with little sequence similarity . To understand the similarity in structure of the two domains, we compare the corresponding residue positions in the two domains by the method of 3D-1D profiles . A 3D-1D profile is a string formed by assigning each position in the 3D structure to one of 18 environment classes . The environment classes are defined by the local secondary structure, the area of the residue which is buried from solvent, and the fraction of the area buried by polar atoms . A structural alignment between the two BPI domains was used to compare the 3D-1D environments of structurally equivalent positions . Greater than 31% of the aligned positions have conserved 3D-1D environments, but only 13% have conserved residue identities . Analysis of the 3D-1D environmentally conserved positions helps to identify pairs of residues likely to be important in conserving the fold, regardless of the residue similarity . We find examples of 3D-1D environmentally conserved positions with dissimilar residues which nevertheless play similar structural roles . To generalize our findings, we analyzed four other proteins with similar structures yet dissimilar sequences . Together, these examples show that aligned pairs of dissimilar residues often share similar structural roles, stabilizing dissimilar sequences in the same fold .

J Vet Med A Physiol Pathol Clin Med, 2000 Apr, 47(3), 181 - 92
Age-associated changes in the immune system of German shepherd dogs; Strasser A et al.; In order to look into the ageing of the canine immune system we investigated age-related changes and associated gender-related differences in parameters of innate and acquired immunity in German Shepherd dogs . We obtained the following findings: white blood cell counts, peripheral blood lymphocytes, lymphocyte proliferative activity and interleukin-2 (IL-2) serum concentrations were significantly lower in the group of old animals, whereas the concentrations of gamma-globulins and the functional activity of the complement system were significantly higher in the elderly . Phagocytic and bactericidal activity of polymorphonuclear cells, as well as their 'killing function,' the serum cytokine-like activities of tumour necrosis factor-alpha and the plasma concentrations of immunoglobulin G, as well as of alpha- and beta-globulins, were not significantly affected by age, whereas natural killer-cell activity and the serum cytokine-like activities of IL-1 were significantly higher only in the group of female old animals . With regard to gender-related differences, lymphocyte proliferative activities as well as plasma concentrations of alpha-globulin were significantly higher in the group of female animals, whereas the absolute numbers of segmented neutrophils were significantly lower . Species analogies with regard to ageing as presumed to exist between man and laboratory rodents also seem to be applicable to the dog . The observed age-related changes in the canine immune system are probably among the main causes for the multimorbidity of old age, affecting life expectancy and mortality in the dog and should be recognized and considered by the attending veterinarian.

J Vet Med A Physiol Pathol Clin Med, 2000 Feb, 47(1), 1 - 8
Non-specific immunity and ketone bodies . II: In vitro studies on adherence and superoxide anion production in ovine neutrophils; Sartorelli P et al.; The effects of the ketone bodies beta-OH-butyrate and acetoacetate (2.4 or 4.8 mmol/l), administered singly or simultaneously in vitro, on adherence and superoxide anion (SO) production in ovine neutrophils were investigated by simultaneous assay in 96-well microplates . Because the acetoacetate used was a lithium salt, the effect of 2.4 and 4.8 mmol/l lithium chloride was also tested . Neutrophils from eight non-lactating, non-pregnant ewes were used . SO release from neutrophils was found to be very low in basal conditions and was apparently not stimulated by contact with plastic . Administration of 10(-7) mol/l phorbol myristate acetate (PMA) caused a rapid increase and release of SO production, but smaller than that induced by co-stimulation with plastic and 10(-7) mol/l PMA . LiCl (2.4 and 4.8 mmol/l) significantly increased PMA-stimulated release, but inhibited plastic and PMA co-stimulated SO release . Administration of 2.4 mmol/l ketone bodies inhibited plastic and PMA-costimulated SO release, but the effect of acetoacetate could be due to the lithium component . Administration of 4.8 mmol/l ketone bodies had no effect . Adherence was significantly increased by contact with plastic, and moreover by 10(-7) mol/l PMA . The effect was similar when PMA was acting alone or with plastic . Neither basal nor stimulated adherence were affected by 2.4 or 4.8 mmol/l ketone bodies . LiCl at a concentration of 4.8 mmol/l increased PMA and plastic co-stimulated adherence . The results suggest that, in sheep, only the ketone body beta-OH butyrate at concentrations seen in mild ketosis, could decrease bactericidal activity, while adherence is not affected . In addition to other factors that could impair the efficiency of the immune system in ketotic ruminants, the reduced bactericidal activity may contribute to the higher occurrence of infectious disease in these animals.

Rheumatol Int, 2000, 19(4), 129 - 36
Clinical associations and characterisation of antineutrophil cytoplasmic antibodies directed against bactericidal/permeability-increasing protein and azurocidin; Cooper T et al.; Bactericidal/permeability-increasing protein (BPI) and azurocidin (AZ) are recently described target antigens of antineutrophil cytoplasmic antibodies (ANCA) . In this study, BPI-ANCA were demonstrated most often in patients with ulcerative colitis (36/92, 39%), Crohn's disease (17/66, 26%) and cystic fibrosis (11/14, 79%), but also in patients with rheumatoid arthritis (8/40, 20%), systemic lupus erythematosus (SLE) (111/65, 17%) and mixed connective tissue disease (4/18, 22%) . BPI-ANCA were also common in sera containing antinuclear (ANA) (9/43, 21%) or antidouble-stranded (ds) DNA (7/28, 25%) antibodies . There was no increased frequency of abnormal alpha1-antitrypsin (alphal1AT) phenotypes in patients with BPI-ANCA, and BPI-ANCA were not more common in individuals with an abnormal phenotype . The predominant IgG subclasses were IgG1 and IgG3; IgA but not IgM was present . Both IgG and IgA BPI-ANCA were high affinity antibodies, and the affinity of IgG antibodies did not change with time in the sera tested . Four of the five sera (80%) containing BPI-ANCA did not bind to denatured, reduced BPI, suggesting that most BPI-ANCA recognised conformational epitopes . AZ-ANCA were demonstrated in 2/11 patients (18%) with Wegener's granulomatosis, 3/12 (25%) with cystic fibrosis and 3/14 (21%) with chronic active hepatitis . AZ-ANCA were present in 5/25 sera (25%) with ANA, but the levels were only marginally elevated . AZ-ANCA were uncommon in patients with inflammatory bowel and rheumatological diseases, and in sera containing other autoantibodies . Again, there was no association with abnormal alpha1-AT phenotypes . BPI represents a major ANCA target antigen in patients with rheumatological as well as inflammatory bowel disease and cystic fibrosis, but AZ-ANCA are uncommon.

Int J Biol Macromol, 2000 Jun 13, 27(3), 181 - 6
Chitosan as an enabling excipient for drug delivery systems . I . Molecular modifications; Sabnis S et al.; Chitosan was physicochemically modified for its potential use as a matrix for an implantable antibiotic delivery system that could sustain bactericidal concentrations in the vicinity of an implant or prosthesis . Deacetylation and depolymerization of chitosan were implemented in order to increase the number or accessibility of the reactive amino groups on the polymer backbone for better polymer-drug interaction . The deacetylation process involved reaction of particulate chitosan/depolymerized chitosan with alkali . The rate of deacetylation of chitosan was directly proportional to the reaction temperature up to 80 degrees C; beyond 80 degrees C, rapid degradation of the polymer occurred . The depolymerization of chitosan involved acid digestion of the polymer followed by application of mechanical agitation . This depolymerized product, although water insoluble, possessed a molecular weight that was one to two orders of magnitude lower than that of commercially available chitosans . These products not only exhibited improved reactivity, but also showed increased crystallinity when compared with the parent chitosan . The reactivity was found to be inversely proportional to chitosan's molecular weight . The depolymerization and deacetylation treatments afforded formation of chitosan having a greater number of amino groups available for interactions with the anionic actives.

J Gastroenterol Hepatol, 2000 Apr, 15(4), 437 - 42
Anti-neutrophil cytoplasmic antibodies in patients with chronic liver diseases: prevalence, antigen specificity and predictive value for diagnosis of autoimmune liver disease . Swedish Internal Medicine Liver Club (SILK)
Lindgren S, Nilsson S, Nassberger L, Verbaan H, Wieslander J.
BACKGROUND: Anti-neutrophil cytoplasmic antibodies (ANCA) against proteinase 3 are diagnostic of Wegener's granulomatosis, but ANCA occur also in patients with other inflammatory disorders, such as ulcerative colitis, primary sclerosing cholangitis (PSC) and autoimmune hepatitis . As their predictive value for autoimmune liver disease remains unknown, we analysed the prevalence and antigen specificity of ANCA in patients with various chronic liver diseases (CLD) . METHODS: We studied sera from 100 patients with primary biliary cirrhosis (PBC), from 76 with PSC and from 279 with various CLD, consecutively drawn during a 5-year period at the time of liver biopsy . The ANCA were detected by indirect immunofluorescence (IIF) while the antigen specificity was characterized by ELISA by using lactoferrin, neutrophil elastase, cathepsin G and BPI (bactericidal/permeability increasing protein) as antigens . RESULTS: In PBC, ANCA were detected by IIF in 39 patients (39%) . The antigen reactivity by ELISA was lactoferrin in seven, elastase in 15, BPI in 20 and cathepsin G in four patients . Four patients had reactivity against more than one antigen . In PSC, IIF demonstrated ANCA in 49 patients (65%) . The antigen reactivity was lactoferrin in 17, elastase in 14, BPI in 20 and cathepsin G in four patients . Twelve patients showed reactivity against more than one antigen . In CLD, ANCA were observed in sera from 55 patients (20%) . Nineteen of 45 patients (42%) with autoimmune liver disease were ANCA positive versus 36/234 (15%) with non-autoimmune liver disease (P = 0.0002) . Among IIF-positive patients, antibody reactivity against lactoferrin was noted in 14, elastase in 28, BPI in 25 and cathepsin G in five patients . Twenty-one patients had reactivity against more than one antigen . Elastase and BPI antibodies occurred more frequently in patients with autoimmune compared to non-autoimmune liver disease (P < 0.01) . CONCLUSIONS: Anti-neutrophil cytoplasmic antibodies are prevalent in patients with chronic liver diseases, but although they occur more frequently in patients with autoimmune liver disease their specificity and sensitivity for autoimmune liver disease is low . The predominant antigens are lactoferrin, elastase and BPI, but the correlation between IIF findings and ELISA reactivity against these antigens is weak.

Proc Natl Acad Sci U S A, 2000 Jun 6, 97(12), 6885 - 9
Simultaneous activation of NADPH oxidase-related proton and electron currents in human neutrophils; DeCoursey TE et al.; Generation of reactive oxygen species by the NADPH oxidase complex is an important bactericidal weapon of phagocytes . Phorbol myristate acetate (PMA) is a potent agonist for this "respiratory burst" in human neutrophils . Although stoichiometric H(+) efflux occurs during the respiratory burst, efforts to stimulate voltage-gated H(+) channels by PMA in whole-cell patch-clamped phagocytes have been unsuccessful . We have used a modification of the permeabilized-patch configuration that allows control of intracellular pH and preserves second-messenger pathways . Using this method, we show that PMA dramatically enhances and alters voltage-gated proton currents in human neutrophils . PMA produced four alterations in H(+) current properties, each of which increases the H(+) current at any given voltage: (i) a 40-mV negative shift in the H(+) conductance-voltage (g(H)-V) relationship; (ii) faster activation {smaller activation time constant (tau(act))} during depolarizing pulses; (iii) slower deactivation {larger deactivation time constant (tau(tail))} on repolarization; and (iv) a larger maximum H(+) conductance (g(H, max)) . Inward current that directly reflects electron transport by NADPH oxidase was also activated by PMA stimulation . The identity of this electron current was confirmed by its sensitivity to diphenylene iodinium, an inhibitor of NADPH oxidase . Diphenylene iodinium also reversed the slowing of tau(tail) with a time course paralleling the inhibition of electron current . However, the amplitudes of H(+) and electron currents activated by PMA were not correlated . A complex interaction between NADPH oxidase and voltage-gated proton channels is indicated . The data suggest that PMA stimulation modulates preexisting H(+) channels rather than inducing a new H(+) channel.

FEBS Lett, 2000 May 19, 473(3), 269 - 74
alpha-Lactalbumin: structure and function; Permyakov EA et al.; Small milk protein alpha-lactalbumin (alpha-LA), a component of lactose synthase, is a simple model Ca(2+) binding protein, which does not belong to the EF-hand proteins, and a classical example of molten globule state . It has a strong Ca(2+) binding site, which binds Mg(2+), Mn(2+), Na(+), and K(+), and several distinct Zn(2+) binding sites . The binding of cations to the Ca(2+) site increases protein stability against action of heat and various denaturing agents, while the binding of Zn(2+) to the Ca(2+)-loaded protein decreases its stability . Functioning of alpha-LA requires its interactions with membranes, proteins, peptides and low molecular weight substrates and products . It was shown that these interactions are modulated by the binding of metal cations . Recently it was found that some folding variants of alpha-LA demonstrate bactericidal activity and some of them cause apoptosis of tumor cells.

Eur J Contracept Reprod Health Care, 1999 Dec, 4(4), 185 - 6
Improving reproductive sexual health: a primary goal at the beginning of the new millennium; Creatsas G; At the beginning of the millennium, it is important that experts and official organizations direct their strategic plans towards the improvement of reproductive sexual health . Adolescents represent the major target of these plans in an effort to improve their sexual behavior . Sexually transmitted diseases must be reduced since their prevalence is very high in certain eastern European countries . These plans should be considered together with the development of new fertility control methods which provide at the same time bactericidal and antiviral properties . Education, provided especially through the media, should be considered a tool for the improvement of the present situation.

Environ Health Perspect, 2000 May, 108(5), 447 - 52
Quantitative polymerase chain reaction for transforming growth factor-beta applied to a field study of fish health in Chesapeake Bay tributaries; Harms CA et al.; Fish morbidity and mortality events in Chesapeake Bay tributaries have aroused concern over the health of this important aquatic ecosystem . We applied a recently described method for quantifying mRNA of an immunosuppressive cytokine, transforming growth factor-beta (TGF-beta), by reverse transcription quantitative-competitive polymerase chain reaction to a field study of fish health in the Chesapeake Basin, and compared the results to those of a traditional cellular immunoassay macrophage bactericidal activity . We selected the white perch (Morone americana) as the sentinel fish species because of its abundance at all of the collection sites . White perch were sampled from Chesapeake Bay tributaries in June, August, and October 1998 . Splenic mononuclear cell TGF-beta mRNA levels increased and anterior kidney macrophage bactericidal activity decreased, particularly in eastern shore tributaries, from June to August and October . The results of the two assays correlated inversely (Kendall's {Tau} b = -0.600; p = 0.0102) . The results indicated both temporal and spatial modulation of white perch immune systems in the Chesapeake Basin, and demonstrated the utility of quantitative PCR for TGF-beta as a molecular biomarker for field assessment of teleost fish immune status.

Zhonghua Wai Ke Za Zhi, 1997 Jul, 35(7), 389 - 91
{Effect of recombinant bactericidal/permeability-increasing protein on pulmonary cytokine mRNA expression in rats following hemorrhage and resuscitation}; Yao Y et al.; To determine the possible mechanisms underlying beneficial effect of recombinant bactericidal/permeability-increasing protein (rBPI) on acute lung injury response to blood loss, we used reverse transcription polymerase chain reaction to measure pulmonary tumor necrosis factor (TNF), interleukin 6 (IL-6) mRNA expression in a rat model of prolonged hemorrhagic shock (4.00 kPa, 180 min) followed by adequate resuscitation . The results showed that systemic plasma endotoxin concentrations elevated rapidly after a 180-min hemorrhagic insult (P < 0.05), and TNF, IL-6 mRNA expression in the lung were significantly increased at 2, 8 hours after resuscitation respectively . However, treatment with rBPI resulted in almost neutralization of plasma endotoxin values, remarkable reduction of TNF, IL-6 mRNA levels following hemorrhage/resuscitation . Also, it was found that rBPI administration markedly blunted the increase in pulmonary Evans blue dye extravasation, concomitant with a significant decrease in lung myeloperoxidase activity compared with the control group (P < 0.05-0.01) . These data suggest that local proinflammatory cytokine mRNA expression associated with gut origin endotoxemia may be an important mechanism contributing to the development of hemorrhage-induced lung injury . Treatment with rBPI is effective in inhibiting marked TNF, IL-6 mRNA expression and ameliorating acute lung injury secondary to severe hemorrhagic shock.

Ann Clin Lab Sci, 2000 Apr, 30(2), 145 - 58
Review: Free radicals, antioxidants, and the immune system; Knight JA; Oxygen-derived free radicals are important in both natural and acquired immunity . Neutrophil and macrophage phagocytosis stimulates various cellular processes including the "respiratory burst" whereby increased cellular oxygen uptake results in the production of the potent oxidant bactericidal agents, hypochlorous acid and hydroxyl radical . In addition, nitric oxide, a gaseous radical produced by macrophages, reacts with superoxide to form peroxynitrite, also a potent bactericidal agent . Conversely, oxidative stress may be detrimental in acquired immunity by activation of nuclear factor kappa B, which governs gene expression involving various cytokines, chemokines, and cell adhesion molecules, among others . However, antioxidant supplementation essentially reverses several age-associated immune deficiencies, resulting in increased levels of interleukin-2, elevated numbers of total lymphocytes and T-cell subsets, enhanced mitogen responsiveness, increased killer cell activity, augmented antibody response to antigen stimulation, decreased lipid peroxidation, and decreased prostaglandin synthesis.

Rinsho Byori, 2000 Jan, Suppl 111, 117 - 24
{Infections with drug resistant bacteria and their treatment methods--MRSA infections}; Aoki Y; MRSA infection is difficult to treat because of the pathogenecity of causative strains and their resistance to many kinds of antibiotics not all strains of MRSA are pathogenic agents and indications for antibiotic use should be determined strictly . Although several antibiotics show bactericidal activity against MRSA, they are not effective in all patients infected with MRSA . We should select the best drug according to the characteristics of the respective drugs in each patient and search for a better regimen to treat severely infected cases.

J Antimicrob Chemother, 2000 May, 45(5), 681 - 4
Accumulation of KRM-1648 by Mycobacterium aurum and Mycobacterium tuberculosis; Piddock LJ et al.; After exposure to 2 mg/L (14)C-labelled KRM-1648 (a new broad-spectrum benzoxazinorifamycin antibiotic) for 5 min, a steady-state concentration of 31.3 +/- 3 ng/mg cells KRM-1648 and 12 . 6 +/- 0.3 ng/mg cells KRM-1648 was accumulated by wild-type antibiotic-susceptible Mycobacterium aurum (A+) and Mycobacterium tuberculosis (H37Rv), respectively . However, 2 mg/L KRM-1648 was bactericidal for M . tuberculosis . A steady-state concentration of 3 . 7 +/- 0.1 ng/mg cells KRM-1648 was accumulated after exposure to 0.5 mg/L . At pH 4 higher concentrations were accumulated than at pH 7 . A sub-inhibitory concentration of ethambutol increased the concentration of KRM-1648 accumulated, but Tween 80 and reserpine had little or no effect.

World J Surg, 2000 May, 24(5), 499 - 506
Release of endotoxin-binding proteins during major elective surgery: role of soluble CD14 in phagocytic activation; Hiki N et al.; Our previous study demonstrated that soluble CD14 (sCD14) modulates the biologic activity of circulating endotoxin, which appears after surgery . In this study, we examined the behavior of endotoxin-binding proteins, such as sCD14, lipopolysaccharide-binding protein (LBP), and bactericidal/permeability-increasing protein (BPI), in patients' plasma after major abdominal surgery and the phagocytic secretion of sCD14 from peripheral blood mononuclear cells (PBMCs) throughout the observation period . In a prospective study, 15 patients undergoing major abdominal surgery (gastrectomy, n = 3; pancreatectomy, n = 10: colectomy, n = 2) were involved in this study . The endotoxin-binding proteins were perioperatively (preoperatively; postoperative hour 6; days 1, 2, 3, 4, 5, 7, and 10) measured by an enzyme-linked immunosorbent assay (ELISA) . To exclude the hemodilution effect of samples, each parameter was corrected by dividing the respective value by the albumin concentration . The phagocytic activity at each time point was tested as an ex vivo sCD14 secretion from PBMCs in the presence and absence of exogenously added endotoxin, Escherichia coli 055B5 (1 ng/ml) . Significant endotoxemia (0.35 +/- 0.13 EU/ml; p < 0.05) was observed 6 hours after the beginning of surgery . The sCD14/albumin value rapidly increased at 6 hours after surgery, peaked on day 1, and sequentially declined, whereas the BPI/albumin and LBP/albumin ratios increased more gradually and peaked on day 2 . The secretion of sCD14 from 2 x 10(6) PBMCs was significantly enhanced from 6 hours after operation . The increased plasma level of sCD14 may be explained by the parallel-enhanced sCD14 PBMC production . Activated secretion of these endotoxin-binding proteins may play a role in regulating the biologic activity of circulating endotoxin.

Pediatr Surg Int, 2000, 16(3), 165 - 8
Regulation of bacterial translocation by nitric oxide; Nadler EP et al.; Nitric oxide (NO) appears to play a paradoxical role in intestinal physiology . Although NO has potent bactericidal effects, a growing body of evidence suggests that it mediates intestinal injury and breakdown of gut barrier function . Data from our lab and others show an increased incidence of bacterial translocation following endotoxin challenge, and upregulation of inducible NO synthase (iNOS) mRNA and protein in the intestine . These phenomena co-localize with enterocyte apoptosis at the tips of the intestinal villi and immunoreactivity to nitrotyrosine . Electron microscopy reveals swollen mitochondria, implicating these organelles as putative targets for NO or its reactive nitrogen intermediates . We review some of the literature and discuss our current work in trying to define this mechanism.

West Indian Med J, 2000 Mar, 49(1), 20 - 6
An experimental mouse model to study the pathogenicity of Prevotella bivia and investigations of possible virulence; Egwari LO et al.; Induction of subcutaneous abscesses in mice was used to study the pathogenicity of Prevotella bivia both in mono-infection and in mixed cultures with Escherichia coli and Peptostreptococcus spp . Virulence factors such as coaggregation and aggregate formation of cells, haemagglutination activity and tolerance to serum bactericidal activity were investigated for their possible role in P bivia pathogenicity . Monocultures of P bivia, E coli and Peptostreptococcus spp did not induce subcutaneous abscess at concentrations as high as 10(9) colony forming units/millilitre (cfu/ml) . Only E coli persisted at the infection site for up to 7 days post infection but with a marked decline in cell count (8.0 x 10(2) cfu/ml) . The anaerobic organisms did not persist at the infection sites beyond the fifth day . In contrast, mixed cultures of P bivia and E coli or all three organisms potentiated for infective abscess two weeks after infection . Viable cells were recovered from abscesses in greater numbers as the infection progressed . Prevotella bivia was the predominant organism in chronic abscesses while E coli predominated in abscesses in the acute stage of the infection . Prevotella bivia lacked haemagglutination activity against human and sheep erythrocytes and showed marked susceptibility to 50 per cent human serum . These may limit its haematogenous spread . Its ability to form aggregates in molar salt solutions and coaggregate with facultative organisms may account for its persistence in pathological sites.

J Surg Res, 2000 May 1, 90(1), 88 - 93
Effects of intestinal ischemia/reperfusion injury on gastric acid secretion; Castaneda A et al.; BACKGROUND: The mechanism responsible for gastric colonization in critically injured ICU patients remains to be fully elucidated . Moreover, the effects of gut ischemia/reperfusion (I/R) injury on gastric function are unclear . It was our hypothesis that gut I/R injury would cause gastric dysfunction . MATERIALS AND METHODS: Rats were anesthetized and, via laparotomy, the superior mesenteric artery (SMA) was clamped at its aortic origin for 45 min followed by clamp removal . Rats were allowed to awaken and then killed after 6 h of reperfusion . Control rats underwent laparotomy with SMA isolation . Stomachs were removed, gastric fluid was aspirated, and the volume, pH, and protein, bicarbonate, and glucose contents were determined . Serum and antral mucosa were prepared for gastrin radioimmunoassay and the glandular mucosa was assessed for morphologic injury . RESULTS: SMA I/R injury caused significant accumulation of gastric luminal fluid that was alkaline and rich in protein, glucose, and bicarbonate content when compared with sham controls . SMA I/R injury also caused gastric surface epithelial cell injury and significantly increased serum and antral gastrin levels . In additional rats, gut I/R injury inhibited basal acid secretion and blunted the acid secretory response to pentagastrin . CONCLUSIONS: This study demonstrated for the first time that small intestinal I/R injury causes significant gastric dysfunction . The findings suggest that this type of injury, a frequent occurrence in critically injured ICU patients, may predispose patients to gastric colonization due to stasis and loss of the natural bactericidal effects of gastric acid .

J Immunol, 2000 May 1, 164(9), 4804 - 11
Synthetic endotoxin-binding peptides block endotoxin-triggered TNF-alpha production by macrophages in vitro and in vivo and prevent endotoxin-mediated toxic shock; Dankesreiter S et al.; Lipid A, the conserved portion of endotoxin, is the major mediator of septic shock; therefore, endotoxin-neutralizing molecules could have important clinical applications . Here we show that peptides derived from Limulus anti-LPS factor (LALF), bactericidal/permeability increasing protein (BPI) and endotoxin-binding protein, bind to lipid A and block the recombinant LALF/lipid A interaction in vitro . Because their neutralizing capacity in vitro as well as in vivo has been limited, we created hybrid peptides comprising two endotoxin-binding domains . The hybrid molecule LL-10-H-14, containing endotoxin-binding domains from LALF and endotoxin-binding protein, turned out to be the most active peptide within the series of peptides tested here to inhibit the CD14/lipid A interaction and is able in vitro to block the endotoxin-induced TNF-alpha release of murine macrophages up to 90% . Furthermore, LL-10-H-14 not only reduced peak serum levels of TNF-alpha of mice when preinjected but also reduced TNF-alpha levels when given 15 min after the endotoxin challenge . As compared with other peptides, only LL-10-H-14 is able to strongly decrease endotoxin-stimulated TNF-alpha release by human macrophage cell lines as well as by PBMC . Furthermore, the hybrid peptide is protective against endotoxin-provoked lethal shock . As such, LL-10-H-14 could have prophylactic and/or therapeutic properties in humans for the management of septic shock.

J Periodontol, 2000 Mar, 71(3), 368 - 75
Bactericidal activity of a monoclonal antibody against a recombinant 40-kDa outer membrane protein of Porphyromonas gingivalis; Katoh M et al.; BACKGROUND: We have cloned the gene for a 40-kDa outer membrane protein (40-kDa OMP) from Porphyromonas gingivalis 381 . The recombinant (r)40-kDa OMP has become the subject of considerable interest because of its potential role in the development of a vaccine useful for passive immunization . To develop such a vaccine, it is essential to fully understand the functions of anti-r40-kDa OMP antibody in the host defense against P . gingivalis . To that end, we developed a panel of monoclonal antibodies by immunizing mice with purified r40-kDa OMP . The objective of this study was to determine the bactericidal activity on P . gingivalis by the IgG1 monoclonal antibody Pg-ompA2 . METHODS: Bacterial growth measurement, a complement-mediated anti-P . gingivalis assay based on {3H}thymidine uptake, and a 14C-release assay were performed to test the bactericidal activity of Pg-ompA2 to P . gingivalis . RESULTS: In the presence of complement, Pg-ompA2 was lethal to P . gingivalis 381 as well as to the more virulent P . gingivalis strains, including ATCC 53977 and W83 . Using component-deficient complement, we determined that Pg-ompA2 killed P . gingivalis by activating both the classical and alternative complement pathways . CONCLUSIONS: Pg-ompA2 has an in vitro complement-mediated bactericidal activity to P . gingivalis . Pg-ompA2 may contribute to the development of a local immunotherapy that can be applied in the gingival crevice of a patient with P . gingivalis-related periodontitis, or be a vaccine candidate.

Mikrobiologiia, 2000 Mar-Apr, 69(2), 257 - 60
{Effect of chitosan derivatives on the reproduction of Coliphages T2 and T7}; Kochkina ZM et al.; The effect of chitosan derivatives with different degrees of polymerization and deamination, as well as of chitosan 6-O-sulfate and chitosan N-succinate-6-O-sulfate, on the reproduction of coliphages T2 and T7 in Escherichia coli and on the growth of this bacterium was studied . Chitosan derivatives decreased the yield of coliphages and exhibited bactericidal activity . The efficiency of inhibition of viral infection and the bactericidal activity of chitosan were found to be dependent on the degree of its polymerization . At the same time, there was no correlation between the degree of chitosan deamination and the extent of inhibition of viral infection . Anionic chitosan derivatives virtually did not possess antiviral or bactericidal activity . It is assumed that chitosan blocks some stages of phage reproduction . The decrease in the phage-producing ability of E . coli may also be due to the bactericidal effect of chitosan.

Infect Immun, 2000 May, 68(5), 2992 - 4
Bactericidal/permeability-increasing protein prevents mucosal damage in an experimental rat model of chronic otitis media with effusion; Nell MJ et al.; In this study, the efficacy of bactericidal/permeability-increasing protein (BPI) was assessed in a rat model of chronic otitis media with effusion . BPI injection prevented disturbance of the mucociliary clearance system of the middle ear . Hence, it is postulated that BPI can be a new therapy for chronic otitis media with effusion.

Infect Immun, 2000 May, 68(5), 2647 - 54
Protection elicited by native outer membrane protein Oms66 (p66) against host-adapted Borrelia burgdorferi: conformational nature of bactericidal epitopes; Exner MM et al.; Oms66 is a Borrelia burgdorferi outer membrane porin protein whose role in Lyme disease pathogenesis and immunity has not been well established . Oms66 was solubilized from whole-cell lysates of strain B313 (which is derived from B31 but lacks OspA, -B, -C, and -D) and purified to homogeneity by fast-protein liquid chromatography . Purified native Oms66 (nOms66), which retained the ability to form large channels in a planar lipid bilayer model membrane system, and denatured Oms66 (hOms66) were used to immunize New Zealand White rabbits . The resulting Oms66 antisera were tested in a complement-dependent borreliacidal assay in parallel with basal serum and with serum from rabbits immune to reinfection with B . burgdorferi (IRS) . IRS showed high-titer complement-dependent killing of both strains B31 and B313 . Sera from animals immunized with nOms66 showed high-titer complement-dependent killing activity against strain B313 but exhibited no killing of B31 . By comparison, serum generated from immunizations with hOms66 showed no killing activity against either strain . Following adsorption of antiserum to nOms66 with recombinant Oms66 (rOms66), the serum antibodies no longer bound to rOms66 or to nOms66 that had been denatured with 8 M urea . However, the antibodies still bound to nOms66 and killing activity against B313 was retained, thus suggesting that native, conformational epitopes are targets of this bactericidal activity . Six C3H HeJ mice were immunized with nOms66 and were challenged using "host-adapted" B . burgdorferi B31 by skin implantation of infected mouse ear tissue . Four of the six mice were protected against both localized and disseminated infection . These findings indicate that native Oms66 can elicit potent bactericidal activity and significant protective immunity against host-adapted organisms.

Pol Merkuriusz Lek, 2000 Jan, 7(43), 47 - 50
{The role of cytokines and reactive oxygen species in the pathogenesis of sepsis}; Sikora JP; The role of cytokines and reactive oxygen species (ROS) in multiple and not fully explained pathogenesis of sepsis was presented . Close attention was paid to the contribution of inflammatory cytokines (TNF-alpha, IL-1, IL-6, IL-8) to the enhanced phagocyte-derived oxidative metabolism and the activation of respiratory burst . The pleiotropic interaction of these and the other cytokines creating so-called cytokine network was described, among other things in order to express the phagocytic and endothelial receptors . The significant role of polymorphonuclear leucocytes (PMNLs) and macrophages in the pathogenesis of sepsis was outlined, presenting not only their bactericidal activity but immunoregulatory effect connected with cytokine release as well . The significance of T cells cooperating with PMNLs was presented as well . The participation of antiinflammatory cytokines (IL-10, IL-13) and cytokine inhibitors e.g . soluble TNF receptor (sTNFR) and IL-1 receptor antagonist (IL-1 ra) was mentioned; all of them appear in septic patients and are thought to be natural regulators of immunological response in vivo . The key role of ROS generated by the activated phagocytes during sepsis has been outlined; it is proposed that the hypermetabolic response to sepsis results from enhanced ROS generation and so-called oxidant stress is a consequence of the imbalance between their generation and detoxification . The consequences of the action of oxygen free radicals resulting in lipid peroxidation followed by host auto-injury were also described . At the end a possibility of immunotherapy of sepsis connected with the application of pentoxifylline (PTXF) as TNF-alpha inhibitor was recommended to take into consideration.

Gut, 2000 May, 46(5), 725 - 31
Endotoxin, cytokines, and endotoxin binding proteins in obstructive jaundice and after preoperative biliary drainage; Kimmings AN et al.; BACKGROUND: Obstructive jaundice is associated with postoperative complications related to increased endotoxaemia and the inflammatory response . In animals obstructive jaundice is associated with endotoxaemia and cytokine induction, which are reversed by internal biliary drainage . AIMS: To study endotoxaemia and the subsequent inflammatory response in obstructive jaundiced patients and after endoscopic biliary drainage . METHODS: In 15 patients with malignant distal obstructive jaundice, inflammatory and bacteriological parameters were assessed before endoscopic stent placement and after three weeks endoscopic drainage . RESULTS: Drainage reduced bilirubin from 252.5 to 45.1 micromol/l . At baseline low level endotoxaemia was detected (4.3 pg/ml) which was not affected after drainage (4.5 pg/ml) . Serum interleukin 8 (IL-8) and endotoxin binding proteins were increased in jaundice and reduced after drainage (IL-8 113.6 to 20.7 pg/ml; lipopolysaccharide binding protein 24.2 to 16.5 microg/ml; sCD14 17.4 to 7.6 microg/ml; bactericidal/permeability increasing protein 2.9 to 1.8 ng/ml) . Levels of other cytokines, augmented in animals, were only slightly increased and not changed after drainage (tumour necrosis factor (TNF): 21.7 and 18.4 pg/ml; sTNFr p55/75: 2.9/7.0 and 2.7/5.6 ng/ml; IL-6: 4.2 and 6.1 pg/ml; IL-10: 4.5 and 2.7 pg/ml) . Elastase and lactoferrin tended towards reduction after drainage . All bile cultures were positive after stenting . CONCLUSIONS: The effects of obstructive jaundice in humans on endotoxin and cytokines are different from those in animal models . Obstructive jaundice causes alterations in circulating endotoxin binding proteins and IL-8 . Concentrations of other mediators (TNF, previously suggested as being responsible for systemic endotoxaemia effects) are low and not affected by drainage.

J Neurosurg Sci, 1999 Jun, 43(2), 125 - 32; discussion 133
Infections and re-infections in long-term external ventricular drainage . A variation upon a theme; Zingale A et al.; BACKGROUND: The infection of the external ventricular drainage (EDV) remains the main morbidity and mortality associated with this procedure, in the setting of the treatment of hydrocephalus and its complications, leading to excess of hospitalization with annual economic burden . METHODS . In this 3-year retrospective study we selected and reviewed the records of 15 of 143 patients (mean age 34 years with range from 1 months to 70 years; 12 males and 3 females) undergone to prolonged EVD in the setting of management of hydrocephalus (5 patients because of acute ventricular dilation post-intraventricular hemorrhage or post-hemorrhagic HCP, 8 because of V-P shunt infection, 1 because of post-traumatic HCP and 1 because of shunt malfunction by elevated CSF protein) and developing a shunt infection or one or more superinfection . RESULTS: There was a 26% mortality and a 13% morbidity (1 patient had GOS score of 2, 1 score of 3 and 3 score of 5) . The pathogens yielded by CSF culture were normal or transient flora of the patient's skin . The causes of infection were carefully analyzed . CONCLUSIONS: Based on our experience the management of infection in long-term EVD includes: the standardization of the environment of the surgery achieved with a) use of prophylactic antibiotics; b) preparation of the patient and sterile field; c) no touch technique . After implantation of EVD the risk of infection must be minimized by carefully nursing care of EVD, and administration of above prophylactic antibiotics . CSF must be collected for culture and cell count, glucose and protein when clinically indicated . When infection o reinfection is demonstrated by CSF culture then it is advisable to remove the entire hardware and start the antibiotic therapy intravenously and intraventricularly basing on susceptibility tests, CSF penetration of antibiotics, their bactericidal action, toxicity, specificity and cost . Regard to the duration of the therapy, a practical guide is treating for 10-14 days after three consecutive CSF sterile cultures . Thus, convention of EVD to a shunt can be performed within 3 weeks from admission, in the best favourable cases, decreasing the duration of hospital stay and the overall cost of neurosurgical management of the cerebral pathology requiring as therapeutic adjunct and EVD.

Vox Sang, 2000, 78(1), 19 - 27
Leukocyte depletion of red cell components prevents exposure of transfusion recipients to neutrophil elastase; Willy C et al.; BACKGROUND: Polymorphonuclear leukocytes contain a large number of enzymes and bactericidal proteins stored in granules . Neutrophil activation induces degranulation and immediate release of these bioactive substances, including human neutrophil elastase (HNE) also known as elastase-2 (ELA2), which may contaminate whole blood units and blood components . MATERIALS AND METHODS: The HNE concentration was determined in the supernatants of blood components with a commercial enzyme-linked immunosorbent assay (ELISA) . The effect of leukocyte depletion and storage was evaluated by testing whole blood, buffy-coat-reduced, and leukocyte-depleted red cell units . Buffy-coat-derived platelets and plasma were also tested . RESULTS: HNE concentrations at day 1 were about 50 microg/l in all types of red cell components with the exception of leukocyte-depleted red cells (<0.26 microg/l) . In leukocyte-depleted red cells, platelets and plasma, no significant increase was observed during storage . In whole-blood units and buffy-coat-reduced red cells, the HNE concentrations increased steadily and often exceeded 1,000 microg/l when the units expired . CONCLUSION: Leukocyte depletion may limit the inadvertent infusion of bioactive substances derived from polymorphonuclear leukocytes, of which HNE is but one example . The accumulation of HNE in buffy-coat-reduced red cells may be greater than that of whole blood units . HNE accumulates during storage and its quantity may have pathophysiologic significance . Platelets and plasma derived from buffy coats contain some HNE, but leukocyte-depleted red cells virtually none . However, we consider the accumulation of HNE in these components not to be clinically important . The quantities, kinetics, and occurrence in various blood components of HNE contamination differ from those observed with cytokines.

Cardiovasc Res, 2000 Mar, 45(4), 853 - 9
Peri-operative myocardial tissue injury and the release of inflammatory mediators in coronary artery bypass graft patients; Fransen EJ et al.; OBJECTIVE: This study was conducted to evaluate to what extent the ischemia-reperfusion injury resulting from the cardiopulmonary bypass (CPB) and aortic cross-clamping procedures during coronary artery bypass grafting (CABG) contributes to the systemic inflammatory response generally found in these patients . METHODS: Serum levels of enzymes (CK and CK-MB) and non-enzymatic proteins (FABP and myoglobin) as markers of myocardial tissue injury, bactericidal permeability increasing protein (BPI) as an indicator of neutrophil activation, interleukin-6 (IL-6) as inducer of the acute phase response and lipopolysaccharide binding protein (LBP) as parameter of the acute phase response were measured in 15 low-risk CABG patients with cardiopulmonary bypass (CPB), and 17 low-risk CABG patients without CPB . RESULTS: Already 0.5 h after reperfusion significantly increased plasma levels of all markers of myocardial tissue injury were noted in patients having surgery with CPB, but not in non-CPB patients . No significant differences were found between both groups for BPI and IL-6 levels in the early reperfusion period . BPI and IL-6 levels were higher in the non-CPB group on the first post-operative day (P < 0.05) . However, no correlations were found for any marker of peri-operative tissue damage with either early neutrophil activation, or acute phase reactants . CONCLUSIONS: Perioperative myocardial injury resulting from CPB and aortic cross-clamping in low-risk CABG patients does not contribute to the release of inflammatory mediators in these patients.

Electrophoresis, 2000 Feb, 21(3), 665 - 72
Calcium-dependent secretion in human neutrophils: a proteomic approach; Boussac M et al.; Bactericidal, proteolytic and signal proteins released by activated neutrophils play a major role in infection fighting and inflammatory processes . These proteins are mainly stored in organelles called granules until induction of their controlled exocytosis . The present work deals with the characterization of the proteins which are secreted upon activation of human neutrophils by ionomycin and calcium . Proteins were separated by two-dimensional gel electrophoresis and identified by peptide mass fingerprinting . Almost all the previously described soluble components of neutrophil granules could be identified . Moreover, several additional proteins were shown to be secreted by activated neutrophils, namely calgranulins, human cartilage glycoprotein of 39 kDa (HC gp-39), chitotriosidase, and annexin XI . Their subcellular localization and possible functions are discussed.

Inhal Toxicol, 2000 Mar, 12(3), 169 - 86
Inhaled particle-bound sulfate: effects on pulmonary inflammatory responses and alveolar macrophage function; Clarke RW et al.; Acid sulfate-coated solid particles are a significant environmental hazard produced primarily by the combustion of fossil fuels . We have previously described a system for the nascent generation of carbonaceous particles surface coated with approximately 140 microg/m(3) acid sulfate {cpSO(4)(2-); 10 mg/m(3) carbon black (CB) and 10 ppm sulfur dioxide (SO(2)) at 85% relative humidity (RH)} . The effects of inhaled cpSO(4)(2-) on pulmonary host defenses are assessed in the present work . Mice were acutely exposed (4 h) to either 10 mg/m(3) CB, 10 ppm SO(2), or their combination at 10% or 85% RH in a nose-only inhalation chamber . No evidence of an inflammatory response was found following any of the exposures as assessed by total cell counts and differential cell counts from bronchoalveolar lavage fluid . However, alveolar macrophage Fc receptor-mediated phagocytosis decreased only following exposure to 140 microg cpSO(4)(2-), significant suppression occurred after 24 h, maximal suppression occurred at 3 days postexposure, and recovery to preexposure levels required 7-14 days . Intrapulmonary bactericidal activity (IBA) was also suppressed only after exposure to 140 microg cpSO(4)(2-); suppression was maximal at 1 day postexposure and recovered by day 7 . To assess the effects of lower cpSO(4)(2-) concentrations, mice were repeatedly exposed to 1 mg/m(3) CB and 1 ppm SO(2) at 85% RH ( approximately 20 microg/m(3) cpSO(4)(2-) for 4 h/day) for up to 6 days . A significant decrement in IBA was observed following 5 and 6 days of exposure . These studies indicated that acute or repeated exposure to cpSO(4)(2-) could alter pulmonary host defense mechanisms.

J Biol Chem, 2000 Mar 17, 275(11), 7757 - 63
Reduction of nitrite to nitric oxide catalyzed by xanthine oxidoreductase; Godber BL et al.; Xanthine oxidase (XO) was shown to catalyze the reduction of nitrite to nitric oxide (NO), under anaerobic conditions, in the presence of either NADH or xanthine as reducing substrate . NO production was directly demonstrated by ozone chemiluminescence and showed stoichiometry of approximately 2:1 versus NADH depletion . With xanthine as reducing substrate, the kinetics of NO production were complicated by enzyme inactivation, resulting from NO-induced conversion of XO to its relatively inactive desulfo-form . Steady-state kinetic parameters were determined spectrophotometrically for urate production and NADH oxidation catalyzed by XO and xanthine dehydrogenase in the presence of nitrite under anaerobic conditions . pH optima for anaerobic NO production catalyzed by XO in the presence of nitrite were 7.0 for NADH and </=6.0 for xanthine . Involvement of the molybdenum site of XO in nitrite reduction was shown by the fact that alloxanthine inhibits xanthine oxidation competitively with nitrite . Strong preference for Mo=S over Mo=O was shown by the relatively very low NADH-nitrite reductase activity shown by desulfo-enzyme . The FAD site of XO was shown not to influence nitrite reduction in the presence of xanthine, although it was clearly involved when NADH was the reducing substrate . Apparent production of NO decreased with increasing oxygen tensions, consistent with reaction of NO with XO-generated superoxide . It is proposed that XO-derived NO fulfills a bactericidal role in the digestive tract.

Am J Respir Crit Care Med, 2000 Mar, 161(3 Pt 1), 718 - 22
Influenza virus inhibits lysozyme secretion by sputum neutrophils in subjects with chronic bronchial sepsis; Pang G et al.; Neutrophils are central to the control of infection within the bronchial mucosa . To determine whether the link between bacterial and viral infection in the respiratory tract can be partly explained by acute reduction of neutrophil function, we examined the influence of influenza virus on lysozyme secretion by sputum neutrophils obtained from patients with bronchiectasis . Sputum neutrophils infected with influenza A virus had a significantly reduced capacity to secrete lysozyme but not myeloperoxidase . Influenza virus A strains were more effective in inhibiting lysozyme secretion than were influenza B virus strains . Reduction of bactericidal activity was similarly reduced by different strains of influenza A virus, but an influenza virus B strain had no effect . Our results show that downregulation of sputum neutrophil function characterized by lysozyme secretion and bactericidal activity could contribute to reduction in the capacity to control bacterial colonization in the respiratory tract following influenza virus infection.

J Clin Pathol, 1999 Dec, 52(12), 901 - 9
Mononuclear leucocyte function tests in the assessment of the biocompatibility of peritoneal dialysis fluids; Brulez HF et al.; BACKGROUND: Previous studies showed that the currently used dextrose based peritoneal dialysis fluids impair several leucocyte functions . AIMS: To determine which in vitro mononuclear leucocyte (monocyte) function tests most clearly reflect the biocompatibility of peritoneal dialysis fluid . METHODS: Monocytes were tested for phagocytic capacity, bactericidal activity, Fc and C3 receptor expression, and chemiluminescence response, and by analysis of the release of interleukin 8 (IL-8) and tumour necrosis factor alpha (TNF alpha) in the presence of test fluids . Cytokine release was studied in an alternative dynamic in vitro peritoneal dialysis model in which monocytes were exposed to test fluid that was continuously equilibrated with an interstitial fluid-like medium through a microporous membrane . The chemiluminescence response by stressed monocytes was also tested after an 18 h recovery period . All tests were performed during or after exposure to different degrees of glycerol induced osmotic stress and after exposure to a 1% milk-whey derived, polypeptide enriched test fluid . Cells incubated in 0.1% gel Hanks buffer (GH) served as control . RESULTS: Osmotic stress induced impairment of leucocyte function was found by the chemiluminescence assay (mean (SEM): 179 (20)% v 138 (23)% after 30 minutes in 0.5% and 1.5% glycerol, respectively) and by the analysis of IL-8 released by monocytes (44 (9) ng in 0.7% glycerol v 40 (7) ng in 2.0% glycerol) . Only the chemiluminescence assay showed a protective effect of polypeptides on leucocyte function (after > or = 60 minutes) . If monocytes were allowed to recover in culture medium after exposure to test fluids, the changes in chemiluminescence response appeared to be reversible after a 30 minute exposure, but became more pronounced after 60 and 120 minutes . The phagocytosis and bacterial killing assays were less sensitive . The observations carried out with the phagocytosis assay did not correspond with the Fc or C3 receptor density data . CONCLUSIONS: The release of IL-8 by peripheral blood monocytes in a two compartment model and their chemiluminescence response are appropriate assays for the assessment of changes in leucocyte function in response to different peritoneal dialysis fluids.

Mol Cell, 2000 Jan, 5(1), 49 - 57
Signal transduction by a death signal peptide: uncovering the mechanism of bacterial killing by penicillin; Novak R et al.; The binding of bactericidal antibiotics like penicillins, cephalosporins, and glycopeptides to their bacterial targets stops bacterial growth but does not directly cause cell death . A second process arising from the bacteria itself is necessary to trigger endogenous suicidal enzymes that dissolve the cell wall during autolysis . The signal and the trigger pathway for this event are completely unknown . Using S . pneumoniae as a model, we demonstrate that signal transduction via the two-component system VncR/S triggers multiple death pathways . We show that the signal sensed by VncR/S is a secreted peptide, Pep27, that initiates the cell death program . These data depict a novel model for the control of bacterial cell death.

Pediatr Res, 2000 Mar, 47(3), 357 - 61
Activation of human granulocytes by intravenous immunoglobulin preparations is mediated by FcgammaRII and FcgammaRIII receptors; Nemes E et al.; Previous studies from our laboratory have shown that i.v . Ig (IVIG) exposure triggers superoxide anion (O2) generation by and increases bactericidal capacity of human blood granulocytes . However, the molecular interactions between IVIG and granulocytes have not been evaluated before . The objective of this study was to investigate the role of FcgammaRII and FcgammaRIII receptors in the immunomodulatory effects of IVIG concentrates on granulocytes . We found that four different IVIG preparations (concentration range, 1-10 mg/mL) shared the ability to stimulate O2- release in vitro by granulocytes in a dose-dependent manner . Dimers fractionated from IVIG were significantly more potent in inducing activity of the respiratory burst than were monomers . MAb (concentration range, 0.1-10 microg/mL) specific for FcgammaRII and FcgammaRIII receptors inhibited the IVIG-induced O2- release, with a more profound inhibitory effect observed with anti-FcgammaRIII . These findings suggest the involvement of Fcgamma receptors in triggering O2- release by granulocytes exposed to IVIG . We also report that IVIG added to granulocyte suspensions elicited a rapid and vigorous increase in the concentration of cytosolic free calcium, a finding suggesting direct activation and not priming of granulocytes by IVIG through FcgammaRII and FcgammaRIII receptors . The in vitro effects described here might occur in patients treated with IVIG and may, in part, be responsible for inflammatory reactions evoked by infused Ig concentrates as well as the immunomodulatory effect of Ig in patients with autoimmune and inflammatory diseases.

J Immunol, 2000 Mar 15, 164(6), 3377 - 84
High susceptibility for cystic fibrosis human airway gland cells to produce IL-8 through the I kappa B kinase alpha pathway in response to extracellular NaCl content; Tabary O et al.; Increasing evidence suggests that in airways from cystic fibrosis (CF) patients, inflammation may precede bacterial infection and be related to an endogenous dysregulation of proinflammatory cytokines in airway epithelial cells . Several investigators have reported that, in CF airway fluids, elevated NaCl concentrations may also contribute to the diseased state by inhibiting the bactericidal properties of airway fluid . Because many proinflammatory cytokines are transcriptionally regulated by the NF-kappa B, we investigated whether an elevated extracellular NaCl content in airway fluids significantly impaired the regulation of the NF-kappa B/I kappa B alpha complex and the chemokine IL-8 production in primary non-CF and CF human bronchial gland epithelial cells . Exposure of non-CF gland cells to hypotonic (85 mM) NaCl solution, compared with isotonic (115 mM) NaCl and hypertonic (170 mM) NaCl solutions, resulted in a significant decrease in IL-8 production that was paralleled by a strong inhibition of activated NF-kappa B associated with an increased cytosolic expression of I kappa B alpha and a decrease in the I kappa B kinase alpha protein level . In CF gland cells, we demonstrated that, compared with the high IL-8 in an hypertonic solution, the release of IL-8 was significantly reduced 2-fold in an isotonic solution and 5-fold in a hypotonic solution . Strikingly, exposure of CF bronchial gland cells to either hypotonic or isotonic milieu did not result in a marked inhibition of the activated NF-kappa B/I kappa B alpha system . This is the first demonstration that primary human CF bronchial gland cells exhibit abnormally high IL-8 production through constitutively activated NF-kappa B and high I kappa B kinase alpha level, whatever the hypo-, iso-, and hypertonic NaCl milieu.

Front Biosci, 2000 Jan 01, 5, D20 - 9
Bacterial resistance to aminoglycosides and beta-lactams: the Tn1331 transposon paradigm; Tolmasky ME; Aminoglycosides (Ags) are a group of antibiotics that exert their bactericidal activity primarily by inhibition of protein synthesis . Aminoglycoside (Ag) molecules bind to the bacterial 30S ribosomal subunit rendering the ribosomes unavailable for translation, which results in cell death . Although these antibiotics are and have been very useful to treat a variety of bacterial infections, in recent years the number of Ag resistant and multiresistant isolates has seriously increased . Mechanisms of resistance to Ag include enzymatic inactivation by acetyltransferases, nucleotidyltransferases (adenylyltransferases), and phosphotransferases, ribosomal alterations, and reduced permeability . Of all Ags, amikacin (Ak) is the most resistant to the action of Ag-modifying enzymes . However, AAC(6')-I type enzymes (a group of 6'-N-acetyltransferases) can utilize Ak as substrate and confer resistance to this antibiotic in addition to other Ags . The gene aac(6')-Ib was found in various bacterial species and various research groups performed mutagenesis studies on this or related enzymes . In one case, aac(6')-Ib was identified in a transposable element, Tn1331, included in pJHCMW1, a plasmid isolated from a clinical K . pneumoniae strain . Tn1331 includes genes encoding two Ag-modifying enzymes (aac(6')-Ib and ant(3")-Ia) and two beta-lactamases (blaTEM and blaOXA-9) . Characterization of other functions of the pJHCMW1 plasmid showed the presence of an RNA-regulated replication origin and a functional oriT . Stability by multimer resolution is achieved by the Tn1331 resolvase.

Br J Sports Med, 2000 Feb, 34(1), 23 - 8
Neutrophil function response to aerobic and anaerobic exercise in female judoka and untrained subjects; Wolach B et al.; OBJECTIVES: Recent studies have indicated reduced immunity in trained athletes . AIM: To assess the effects of aerobic and anaerobic exercise on the phagocytic process in 18-26 year old trained female judoka (n = 8) and untrained controls (n = 7) . METHODS: Each subject participated randomly in two different testing sessions (aerobic, 20 minutes of treadmill running at 70-80% of maximal heart rate; anaerobic, Wingate anaerobic test) . Venous blood samples were drawn before, immediately after, and 24 hours after each session . RESULTS: There were no significant differences in basal values of net chemotaxis (chemotaxis--random migration), bactericidal activity, and superoxide anion release between the judoka and the untrained women . There was a significant decrease in net chemotaxis 24 hours after the aerobic exercise in both the judoka (from 64 (19) to 39 (13) cells/field, p < 0.02) and the untrained controls (from 60 (7) to 47 (12) cells/field, p < 0.05) . Bactericidal activity and superoxide anion release did not change significantly after aerobic exercise in either group . There were no significant changes in net chemotaxis, bactericidal activity, and superoxide anion release after anaerobic exercise in either the judoka or untrained women . CONCLUSIONS: The decrease in net chemotaxis after aerobic, but not after anaerobic, exercise, suggests that net chemotaxis is affected by the combination of exercise intensity and duration, and not by the exercise intensity itself . Similar effects of both exercise sessions in the judoka and the untrained women suggest that training had no effect on neutrophil function response to aerobic and anaerobic exercises.

J Clin Pathol, 1999 Oct, 52(10), 770 - 2
A latex slide agglutination test for rapid detection of antimyeloperoxidase antibody; Ko KH et al.; AIM: To develop and test a new latex slide agglutination test (MPO-LSAT) to detect antimyeloperoxidase (anti-MPO) antibody in serum . METHODS: Latex bead coating was adjusted to give maximum sensitivity by attending to latex size, MPO to latex ratio for coupling, ratio of diluted serum to MPO-latex, reaction time and temperature for coupling, and reaction time for agglutination . Inhibition studies were performed using MPO, proteinase 3, bactericidal/permeability increasing protein, and lactoferrin . RESULTS: There was very good correlation between this test and the conventional anti-MPO enzyme linked immunosorbent assay (ELISA): 81% of sera positive in the ELISA were positive by MPO-LSAT . MPO-LSAT results correlated better with IgM anti-MPO than with IgG anti-MPO . CONCLUSIONS: MPO-LSAT is a simple diagnostic test that is potentially useful in the clinical laboratory as a rapid screening tool for vasculitic diseases.

J Biol Chem, 2000 Feb 18, 275(7), 4687 - 92
Molecular aspects of complement-mediated bacterial killing . Periplasmic conversion of C9 from a protoxin to a toxin; Wang Y et al.; As part of the membrane attack complex complement protein C9 is responsible for direct killing of bacteria . Here we show that in the periplasmic space of an Escherichia coli cell C9 is converted from a protoxin to a toxin by periplasmic conditions missing in spheroplasts . This conversion is independent of the pathway by which C9 enters the periplasm . Both, C9 shocked into the periplasm and plasmid-expressed C9 targeted to the periplasm via a signal sequence are toxic . Toxicity requires disulfide-linked C9 because export into the periplasm of cells defective in disulfide bond synthesis (dsbA and dsbB mutants) is not toxic unless N-acetylcysteine is added externally to promote cystines . A N-terminal fragment, C9{1-144}, is not toxic nor is cytoplasmically expressed C9, even in trxB mutants that are able to form disulfide bonds in the cytoplasm . Importantly, expression of full-length C9 in complement-resistant cells has no effect on their viability . Expression and translocation into the periplasm may provide a novel model to identify molecular mechanisms of other bactericidal disulfide-linked proteins and to investigate the nature of bacterial complement resistance.

Histol Histopathol, 2000 Jan, 15(1), 199 - 205
Current understanding of macrophage type 1 cytokine responses during intracellular infections; Xing Z; Macrophages are important effector cells in cell-mediated immunity against intracellular infection . Among cytokines that macrophages are able to release are IL-12 and TNF alpha . IL-12 is a critical linker between the innate and adaptive cell-mediated immunity, capable of Th1 differentiation and IFN gamma release by T and NK cells . IFN gamma is critically required for the activation of macrophage bactericidal activities . Recently emerging evidence suggests that macrophages are able to release not only IL-12 and TNF alpha but also IFN gamma . However, the mechanisms that control the release of each of these type 1 cytokines in macrophages appear different . While macrophages release TNF alpha in an indiscriminate and IL-12-independent way, the release of IL-12, particularly bioactive IL-12 p70, and IFN gamma is under tight control . We are just beginning to understand what controls the release of IL-12 p70, a question of fundamental importance to understanding the mechanisms underlying the initiation of cell-mediated immunity . Our recent findings have shed more insights into the regulatory mechanisms of macrophage IFN gamma responses . It has become evident that IL-12 is required not only for Th1 differentiation but also for IFN gamma responses by both T cells and macrophages during intracellular infection . In this overview, we have discussed about the current understanding of the regulation of macrophage type 1 cytokine responses during intracellular infection, based upon the recent findings from us and others.

Crit Care Med, 2000 Jan, 28(1), 8 - 15
Prevention of infection in multiple trauma patients by high-dose intravenous immunoglobulins; Douzinas EE et al.; OBJECTIVE: To investigate the activity of intravenous immunoglobulin (IVIG) as a prophylactic agent against infection in trauma victims . DESIGN: Prospective, randomized, double-blind, placebo-controlled study . SETTING: A 20-bed university intensive care unit . PATIENTS: Thirty-nine trauma patients with injury severity scores (ISSs) of 16-50 . INTERVENTIONS: Penicillin was given at the time of admission and continued at least until day 4 . Twenty-one patients received IVIG and 18 patients received human albumin at 1 g/kg in four divided doses (days 1, 2, 3, and 6) . The two groups had similarities in age, gender, Acute Physiology and Chronic Health Evaluation II score, risk of death, and Glasgow Coma Scale score, but differing ISSs (p = .02), at the time of admission . Blood was collected on days 1, 4, and 7 . MEASUREMENTS AND MAIN RESULTS: Clinical variables related to infection were recorded . The complement components C3c, C4 and CH50, IgG, and the fractions of IgG were measured . The serum bactericidal activity (SBA) was assessed at 37 degrees C (98.6 degrees F) and 40 degrees C (104.0 degrees F) at the time of admission and during the course of IVIG administration . Controlling for ISS, IVIG-treated patients had fewer pneumonias (p = .003) and total non-catheter-related infections (p = .04) . Catheter-related infections (p = .76), length of stay in the intensive care unit, antibiotic days, and infection-related mortality did not differ between the two groups . A significantly increased trend in IgG and its subclasses was shown on days 4 and 7 in the IVIG group but not in the control group (p<.000001) . No important differences were noted in complement fractions . The SBA of the groups was similar on day 1, but significantly higher on days 4 and 7 (p<.000001) in the IVIG group, remaining so controlling for complement and ISS . SBA was higher at 40 degrees C (104.0 degrees F) compared with 37 degrees C (98.6 degrees F) (p<.0001) under all three conditions . In both groups, low SBA (on days 1, 4, and 7) was associated with increased risk of pneumonia (p<.01) and non-catheter-related infections (p = .06 for day 1; p<.01 for days 4 and 7) . CONCLUSIONS: Trauma patients receiving high doses of IVIG exhibit a reduction of septic complications and an improvement of SBA . Early SBA measurement may represent an index of susceptibility to infection.

Therapie, 1999 Sep-Oct, 54(5), 607 - 12
In vitro effects of spiramycin and dirithromycin on IL1 beta production by human LPS-stimulated mononuclear cells; Moutard I et al.; Polymorphonuclear neutrophils are the predominant cells in acute inflammatory lesions and their functions and recruitment are regulated by cytokines, including IL1, TNF and IL8 . Antibiotic modulation of inflammatory effects has stimulated investigations of antibiotics for their potential activity as immunomodulators over their primary bactericidal or bacteriostatic activities . This study reports the influence of macrolides, spiramycin and dirithromycin on IL1 beta production . Mononuclear cells, isolated from healthy human volunteers, were preincubated with macrolides (0.1 to 500 micrograms/ml) and stimulated by Escherichia coli lipopolysaccharide . Then, IL1 beta production was detected by western blotting analysis . At therapeutic concentrations, dirithromycin and spiramycin seemed to enhance IL1 beta production by LPS-stimulated cells, with +37 per cent and +28 per cent at 1 microgram/ml respectively . At supratherapeutic concentrations, these drugs seemed to inhibit IL1 beta production through protein kinase C inhibition, with inhibitory concentrations 50 per cent of 378 micrograms/ml for dirithromycin and 234 micrograms/ml for spiramycin . So, macrolides may modulate the host defence system through their influence on cytokine production.

Wien Klin Wochenschr, 1999 Dec 10, 111(22-23), 985 - 9
Functional heterogeneity in the antibodies produced to Borrelia burgdorferi; Benach JL; Antibodies to outer surface molecules of Borrelia burgdorferi (Osp) that have a bactericidal action in the absence of complement have been described . These antibodies are primarily monoclonal to antigenic determinants in OspA and OspB . One of these, CB2, is an IgG1 monoclonal antibody that recognizes an epitope in the carboxy terminus of OspB . The specificity of CB2 is critically dependent on the presence of a lysine (Lys) residue in position 253, not only for binding but also for killing the spirochete . This antibody has been used successfully to select escape variants or mutants that are missing the Lys residue either by a mutation or by deletion as a result of premature stop codons . Other antibodies to OspA, OspB, and p39 have also been characterized with similar properties . Another important feature of CB2 is that its bactericidal action is not dependent on agglutination, since Fab fragments of the whole immunoglobulin molecule can also kill in the absence of complement synergy . The killing action of CB2 is not inhibited by protease inhibitors, and is dependent on the presence of calcium . Upon contact with Borrelia burgdorferi, CB2 causes lysis of the outer membrane and the formation of a spheroplast . The bactericidal mechanism of this antibody is not known . The sequence of the heavy and light chain variable regions of CB2 have striking homology to murine antibodies of the autoimmune repertoire, and some of these antibodies have catalytic properties . In general, catalytic antibodies have enzymatic rates of acceleration that are significantly less than those of proteolytic enzymes . If CB2 were a catalytic antibody, its substrate specificity may be expected to be broader . CB2 does not cleave recombinant OspB, nor does it cleave other protein substrates . Its killing activity is not dependent on proteolysis . Because the bactericidal action of CB2 involves the destruction of the outer membrane, it is possible that a phospholipase could be associated with the mechanism . The mobility of spirochetal lipids is altered after incubation with CB2, and the bactericidal activity is reduced in the presence of phospholipase inhibitors . These studies suggest that the bactericidal mechanism of CB2, and other similar antibodies, is novel.

Biomaterials, 2000 Feb, 21(3), 273 - 81
Bacterial colonization of functionalized polyurethanes; Flemming RG et al.; A protocol was developed for studying the growth of bacteria upon polyurethanes subsequent to the establishment of an adherent bacterial population . An inocula of approximately 10(5) cfu S . aureus were spread on functionalized polyurethanes which included Pellethane, sulfonated Pellethane, phosphonated Pellethane, quaternized amine polyurethanes, and a zwitterionic phosphonated polyurethane . After 24 h incubation, Pellethane, sulfonated Pellethane, and phosphonated Pellethane showed bacterial growth by at least a factor of 10 . In contrast, the zwitterionic phosphonated polyurethane showed a factor of 10 decrease in bacteria after 24 h and the quaternized amine polyurethanes reduced the bacteria to only a few hundred after only 1 h . When treated with bovine serum albumin, Pellethane, sulfonated Pellethane, and phosphonated Pellethane again showed bacterial growth by as much as a factor of 10 over 24 h . The quaternized amine polyurethanes and the zwitterionic phosphonated polyurethane still exhibited bactericidal abilities even when coated with bovine serum albumin, with the zwitterionic material reducing bacteria by more than a factor of 10 over 24 h and the quaternized amine polyurethane reducing the bacteria to only a few hundred after only 1 h . A zone of inhibition study suggested that the bactericidal activity of the zwitterionic phosphonated polyurethane was due to the leaching of cadmium ions . A quaternized amine polyurethane which contained chloride instead of iodide as the counterion to the amine moiety was less bactericidal than the iodide-containing polymer when treated with albumin . Thus, bacteria were able to colonize Pellethane, phosphonated sulfonated Pellethane, and phosphonated Pellethane, but the iodide-containing quaternized amine polyurethane and the zwitterionic polyurethane prevented colonization.

Am J Physiol Gastrointest Liver Physiol, 2000 Jan, 278(1), G105 - 12
Bacterial colonization and healing of gastric ulcers: the effects of epidermal growth factor; Elliott SN et al.; Experimental gastric ulcers are rapidly colonized by various bacteria, resulting in significantly impaired healing . Epidermal growth factor (EGF) is capable of preventing bacterial colonization of the healthy intestinal mucosa . In this study, we examined the possibility that EGF accelerates gastric ulcer healing by reducing bacterial colonization of the ulcer . Gastric ulcers were induced by serosal application of acetic acid . The effect of daily administration of EGF on ulcer healing and bacterial colonization was assessed and compared with the effect of daily treatment with broad-spectrum antibiotics . EGF administration reduced colonization levels and accelerated ulcer healing as effectively as the antibiotic treatment . EGF was without effect on acid secretion or neutrophil infiltration into the ulcer . Bacterial growth was not inhibited in the presence of EGF in vitro . These results demonstrate that EGF reduces bacterial colonization during an established infection of a compromised mucosal surface . This effect may contribute to the ability of EGF to accelerate gastric ulcer healing . This effect is acid independent and not due to an anti-inflammatory effect or to direct bactericidal actions.

J Immunol, 2000 Feb 1, 164(3), 1425 - 31
A bactericidal monoclonal antibody elicits a change in its antigen, OspB of Borrelia burgdorferi, that can be detected by limited proteolysis; Katona LI et al.; mAb CB2, directed against outer surface protein B (OspB), causes bacteriolysis of Borrelia burgdorferi in the absence of complement . How this happens is unknown . We examined the effect of mAb binding on OspB tertiary structure by using limited proteolysis to probe changes in protein conformation . Truncated OspB (tOspB) that lacked N-terminal lipid was cleaved by four enzymes: trypsin, endoproteinase Arg-C, endoproteinase Asp-N, and endoproteinase Glu-C . CB2 affected the cleavage by trypsin and Arg-C, but not by AspN or Glu-C . None of the enzymes cleaved CB2 under these conditions . Both trypsin and Arg-C cleaved tOspB near the N-terminus; CB2 slowed the rate of cleavage, but did not affect the identity of the sites cleaved . Irrelevant mAb had no effect, indicating that the effect was specific . CB2 was active against tOspB of strain B31, but not against tOspB of strain BEP4, to which it does not bind, suggesting that binding was required to elicit the effect on cleavage . With trypsin, CB2 showed a maximal effect at 8 mol of tOspB to 1 mol of mAb . At this ratio, not enough CB2 was present to bind all the tOspB; therefore, either CB2 shows turnover or CB2 acts by binding tOspB and effecting a change in this tOspB such that it, in turn, propagates the effect in other molecules of tOspB . Regardless of the mechanism, these data show that CB2 elicits a change in tOspB that can be measured by its reduced susceptibility to protease cleavage.

Free Radic Biol Med, 1999 Dec, 27(11-12), 1245 - 50
Damage to the cytoplasmic membrane of Escherichia coli by catechin-copper (II) complexes; Hoshino N et al.; In the presence of a nonlethal concentration of Cu(II), washed Escherichia coli ATCC11775 cells were killed by (-)-epigallocatechin (EGC) and (-)-epicatechin (EC) . Cell killing was accompanied by a depletion in both the ATP and potassium pools of the cells, but the DNA double strand was not broken, indicating that the bactericidal activity of catechins in the presence of Cu(II) results from damage to the cytoplasmic membrane . Induction of endogenous catalase in E . coli cells increased their resistance to being killed by the combination of catechins and Cu(II) . In all cases studied, EGC and EC with Cu(II) were found to generate hydrogen peroxide, but its concentration was too low to account for the bactericidal activity . The bactericidal activity of EGC in the presence of Cu(II) was completely suppressed by ethylenediaminetetraacetate, bathocuproine, catalase, superoxide disumutase (SOD), heated catalase, and heated SOD, but not by dimethyl sulfoxide . When catalase, either heated or unheated, was added to the cells incubated with EGC in the presence of Cu(II), it completely inhibited further killing of the cells . These findings suggest that recycling redox reactions between Cu(II) and Cu(I), involving catechins and hydrogen peroxide on the cell surface, must be important in the mechanism of the killing.

Infect Immun, 2000 Feb, 68(2), 449 - 55
Increased Escherichia coli phagocytosis in neutrophils that have transmigrated across a cultured intestinal epithelium; Hofman P et al.; The functionality of polymorphonuclear leukocytes (PMNs) once they migrate into the digestive lumen is still ill defined . More specifically, phagocytic function and bactericidal action of PMNs after transepithelial migration have not received much attention . The aim of the present study is to compare PMN behavior before and after transepithelial migration, in particular (i) phagocytosis and bactericidal activity; (ii) expression of surface molecules, particularly those involved in phagocytosis; and (iii) apoptosis . Cultured human intestinal epithelial T84 cell monolayers were used . The effect of transepithelial migration on phagocytosis was evaluated by immunofluorescence and electron microscopy and by flow cytometric assessment of the engulfment of a strain of Escherichia coli transfected with the green fluorescent protein . Superoxide production by PMNs was investigated by luminol-mediated chemiluminescence . Expression of various surface molecules on PMNs was evaluated by flow cytometry, while PMN apoptosis was assayed by morphologic changes and DNA fragmentation . E . coli phagocytosis by the PMNs was markedly increased after transepithelial migration without modification of superoxide production . CD11b/CD18 and CD47 expression was increased upon PMN transmigration, whereas CD16 expression was decreased and CD29, CD46, CD49e, CD49f, CD55, CD59, CD61, CD95 levels remained unchanged . Apoptosis in transmigrated PMNs was slightly advanced and was observed after 12 h compared to 16 h for nontransmigrated PMNs . In conclusion, the phagocytic capacity of the PMNs is augmented after transepithelial migration, with a dramatic increase in the level of CD11b/CD18 and preservation of the superoxide production . These results suggest a higher bactericidal activity of the PMNs once they have translocated into the digestive lumen.

Bioorg Med Chem, 1999 Nov, 7(11), 2647 - 66
Studies on anti-Helicobacter pylori agents . Part 1: Benzyloxyisoquinoline derivatives; Yoshida Y et al.; The synthesis and optimization of the anti-Helicobacter pylori activity of a novel series of benzyloxyisoquinoline derivatives that was discovered by a random screening process, are described . In the in vitro assay, compound 10c containing a 3-acetamido-2,6-dichlorobenzyl substituent was found to have extremely potent activity against H . pylori and no activity against other common bacteria . The anti-H . pylori activity of 10c was superior to that of amoxicillin (AMPC) (1) and clarithromycin (CAM) (2) . However, 10c did not show in vivo efficacy in a mouse infection model; a feature attributed to the lack of strong bactericidal activity at short contact times.

J Dairy Res, 2000 Nov, 67(4), 503 - 14
Role of endotoxin and TNF-alpha in the pathogenesis of experimentally induced coliform mastitis in periparturient cows; Hoeben D et al.; Twelve cows were experimentally infected in two quarters with 1 x 10(4) cfu Escherichia coli per quarter and six cows were infused with 500 microg endotoxin into two quarters . Six cows infected intramammarily with Esch . coli were treated intravenously with a bactericidal antibiotic 10 h after infection and subcutaneously 20 h later . Blood and milk samples were collected from all cows at regular time intervals . Milk production decreased more rapidly, but was less pronounced, after endotoxin infusion than (during Esch . coli mastitis . The milk production losses in the noninflamed quarters were negligible in endotoxin mastitis, but were substantial during Esch . coli mastitis, probably due to more pronounced systemic effects . Reticulorumen motility was inhibited only during Esch . coli mastitis . Changes in plasma haptoglobin were more pronounced during Esch . coli mastitis, although they occurred sooner during endotoxin mastitis . No changes in plasma activities of enzymes such as lactate dehydrogenase, glutamic-oxaloacetic transaminase and gamma-glutamyl transpeptidase were observed . Concentrations of tumour necrosis factor-alpha increased in both types of mastitis . Absorption of these cytokines into the circulation was highest during Esch . coli mastitis, especially in the untreated control group . We found only minor differences between the treated and untreated Esch . coli groups, but there were larger differences between the Esch . coli groups and the endotoxin group . These differences were probably due to differences in kinetics, composition and amounts of different cytokines released in the mammary gland and subsequently absorption into the circulation . Endotoxin is probably not directly responsible for the systemic changes during coliform mastitis.

J Antimicrob Chemother, 2000 Jan, 45(1), 123 - 7
Serum bactericidal and inhibitory titres in the management of melioidosis; Simpson AJ et al.; A retrospective evaluation of the relationship between serum bactericidal and inhibitory titres and treatment outcome in 195 adult Thai patients with severe melioidosis was conducted . Drug regimens included ceftazidime (52% of patients), co-amoxiclav (24%), imipenem (11%) or the conventional four-drug combination (11%) . Pre- and 1 h post-dose serum samples were collected after 48-72 h of therapy, and serum inhibitory and bactericidal titrations determined . Median post-dose titres were: bactericidal 1:8 (range 0-1:128) and inhibitory 1:16 (range 0-1:128) . Overall mortality was 26% and outcome was not influenced by either inhibitory or bactericidal titres . Pre-dose titres correlated with renal function; renal function was the most important predictor of mortality . Determination of serum inhibitory or bactericidal titres is unhelpful in the management of severe melioidosis.

J Immunol, 2000 Jan 15, 164(2), 959 - 65
Activation of human neutrophils by Mycobacterium tuberculosis H37Ra involves phospholipase C gamma 2, Shc adapter protein, and p38 mitogen-activated protein kinase; Perskvist N et al.; Recent studies have shown that human neutrophils play a significant protective role in mycobacteria infection . When encountered with mycobacteria, neutrophils exhibit the typical early bactericidal responses including phagocytosis and generation of reactive oxygen intermediates (ROI), but the underlying mechanisms are largely unknown . The present study shows that stimulation of neutrophils with an attenuated strain of Mycobacterium tuberculosis H37Ra (Mtb) led to a tyrosine kinase-dependent ROI production in these cells . Stimulation with Mtb induces a rapid and transient tyrosine phosphorylation of several proteins, one of which was identified as phospholipase C gamma 2 (PLC gamma 2) . Several tyrosine-phosphorylated proteins were associated with the PLC gamma 2 precipitates from Mtb-stimulated neutrophils, of which pp46 was characterized as the Shc adapter protein . A role for PLC gamma 2-Shc association in the generation of ROI is supported by the observations that stimulation with Mtb causes the activation of p38 mitogen-activated protein kinase (MAPK), a downstream target of the Shc/Ras signaling cascade, and that the effect of genistein on ROI production coincided with its ability to inhibit both PLC gamma 2-Shc association and p38 MAPK activation . Moreover, pretreatment of neutrophils with a PLC inhibitor markedly suppresses the Mtb-stimulated ROI production as well as p38 MAPK activation in these cells . Taken together, these results indicate that stimulation of neutrophils with Mtb triggers the tyrosine phosphorylation of PLC gamma 2 and its association with Shc, and that such association is critical for the Mtb-stimulated ROI production through activating p38 MAPK.

Artif Organs, 1999 Dec, 23(12), 1055 - 62
The effect of electrolyzed strong acid aqueous solution on hemodialysis equipment; Tanaka N et al.; We reported the high effectiveness of electrolyzed strong acid aqueous solution (ESAAS) in cleaning hemodialysis lines . Although ESAAS has a strong bactericidal action, one concern is its strong acidity . It has a pH of 2.3-2.7, more than 1,000 mV in oxidation-reduction potential (ORP), and 10-50 ppm of available chlorine . The possibility of metal corrosion, degradation of synthetic resins, chorine gas emission, or dissolving calcium carbonate (CaCO3) deposits due to ESAAS's acidity was tested using in vitro experiments . The bactericidal and antiviral effects of various ESAAS's were also tested . Metal corrosion and synthetic resin degradation, although they occurred, were not serious . There were no problems with chlorine gas emission and dissolving of CaCO3 deposits . Each type of ESAAS showed almost the same bactericidal and antiviral effect, but in some cases differences were observed.

J Leukoc Biol, 1999 Dec, 66(6), 915 - 22
Trp-Lys-Tyr-Met-Val-D-Met is a chemoattractant for human phagocytic cells; Bae YS et al.; Trp-Lys-Tyr-Met-Val-D-Met (WKYMVm) is a synthetic peptide that stimulates phosphoinositide (PI) hydrolysis in human leukocytes . The peptide binds to a unique cell surface receptor(s) . Recently we had demonstrated that human neutrophils, monocytes, and B lymphocytes express this peptide-specific receptor and that stimulation of human leukocytes with the peptide leads to activation of the oxidative respiratory system and the bactericidal activity of neutrophils or monocytes . In this study we showed that the peptide induces chemotaxis of phagocytic leukocytes and studied the signaling pathway leading to chemotaxis in human monocytes . The peptide-induced monocyte chemotaxis is pertussis toxin (PTX)-sensitive . This fact correlates with the peptide's stimulation of PI hydrolysis and intracellular Ca2+ ({Ca2+}i) release, which is also PTX-sensitive . We demonstrate that the peptide-specific receptor is different from receptor(s) for monocyte chemoattractant protein-1 (MCP-1) . We also show that intracellular signaling of WKYMVm leading to monocyte chemotaxis is different from that of MCP-1 . The peptide-mediated monocyte chemotaxis is insensitive to protein kinase C (PKC) inhibitor (GF109203X) and butan-1-ol, ruling out PKC and phospholipase D participation in this process . On the other hand, a tyrosine kinase inhibitor (genistein) and RhoA inhibitor (C3 transferase) curtailed the peptide-induced chemotaxis in a concentration-dependent manner, implying the involvement of tyrosine kinase and RhoA, respectively . Treatment of human monocytes with the peptide stimulates tyrosine phosphorylation of several cellular proteins, including p125FAK and Pyk2 and translocation of RhoA from the cytosol to the membrane . We conclude that WKYMVm induces chemotaxis of human phagocytic leukocytes via unique receptors and signaling.

Ann Agric Environ Med, 1999, 6(2), 151 - 9
Studies of toxicity of dermally-absorbed nurelle D 550 EC preparations; Latuszynska J et al.; The aim of the study was the evaluation of the toxic effect of a two-component, preparation Nurelle D 550 EC (500 g of chlorpyrifos and 50 g cypermethrin per 1 l), administrated dermally . Toxicity was evaluated from histological and ultrastructural studies of the internal organs and immunotoxic effects (evaluation of phagocytical and bactericidal activity of neutrophils) . The preparation for dermal application was applied in 2 concentrations (200 mg/kg/day of chlorpyrifos plus 20 mg/kg/day of cypermethrin or 1000 mg/kg/day of chlorpyrifos, plus 100 mg/kg/day of cypermethrin) . The preparation was administrated on the tail skin of female Wistar rats for 4 hours daily for a period of 4 weeks . After 28 days of the experiment, the animals were anaesthetised and blood was taken from the heart to evaluate the granulocyte system . The following organs were taken for histological studies: liver, kidney, lung, heart, spleen, thymus and lymph nodes . Ultrastructural studies were carried out on the lung, liver, kidney and heart . The results of the study showed that dermal application of the pesticide Nurelle D 550 EC resulted in slight morphological and ultrastructural changes in the liver, kidney, lung and heart . The preparation examined slightly elevated the bactericidal activity of neutrophils . The differences, however, were not statistically significant . The phagocytic reaction in animals of both experimental groups did not differ from that observed in control group

Curr Opin Hematol, 2000 Jan, 7(1), 53 - 8
Myeloperoxidase; Winterbourn CC et al.; This review covers recent advances in the biology of myeloperoxidase . Mechanisms of posttranslational processing and how these fail in some of the common deficiency mutants are discussed . We also review the enzymology that points to myeloperoxidase having a number of physiologic substrates in addition to chloride and the evidence that it produces hypochlorous acid in the neutrophil phagosome in sufficient quantities to be bactericidal . Evidence is accumulating that myeloperoxidase-derived oxidants modify biologic macromolecules and cell-regulatory pathways and that they play a role in atherosclerosis . Investigation of disease incidence in relation to a polymorphism in the promoter region of the gene has produced interesting associations . These links with inflammatory diseases can now be pursued further using specific biomarkers of myeloperoxidase activity.

J Agric Food Chem, 1999 Nov, 47(11), 4769 - 76
Effect of hexanal on the shelf life of fresh apple slices; Lanciotti R et al.; In this work the effects of hexanal, as a component of packaging atmosphere, on the shelf life of and evolution of naturally occurring microbial populations in fresh apple slices during storage at 4 and 15 degrees C were evaluated . Although hexanal had no bactericidal effects, in all conditions considered, this volatile molecule significantly extended the shelf life . In fact, the presence of hexanal in the storage atmosphere (at 4 degrees C) totally inhibited mesophilic bacteria and considerably prolonged the lag phase of psychrotrophic bacteria . Also, at 15 degrees C, hexanal strongly inhibited molds, yeasts, and mesophilic and psychrotrophic bacteria . Moreover, hexanal led to a yeast selection favoring species having a reduced spoilage potential due to their prevalent respiratory activity . When added to a modified atmosphere (70% N(2) and 30% CO(2)), this molecule was also very effective in preventing browning reactions for at least 16 days at 15 degrees C . No changes in hue angle values were observed in samples packaged in modified atmosphere with hexanal, even after 16 days of storage at 4 degrees C.

J Appl Microbiol, 1999 Nov, 87(5), 689 - 96
Complement-mediated bactericidal activity of human milk to a serum-susceptible strain of E . coli 0111; Ogundele MO; There has been a lot of controversy concerning the physiological significance of the complement system in human breast-milk . This is mainly due to the observation that human milk contains predominantly non-inflammatory and many anti-inflammatory factors, while simultaneously protecting the infant against a wide range of infectious and other diseases . The present study was carried out to assess the contribution of the complement system to the bactericidal activity of the human colostrum and early lactational milk . Using a serum-sensitive strain of Escherichia coli, different fractions of human breast-milk were assessed for their ability to kill the bacteria, with and without inactivation of their complement components, in comparison to another strain of the bacteria species . Deposition of activated C3 fragments on the killed bacteria, using an established ELISA technique, was demonstrated, further proving that the human milk complement could be activated in vitro . The bactericidal activities of human milk were almost completely abolished by complement heat inactivation at 56 degrees C or by the addition of EDTA.

Protein Sci, 1999 Nov, 8(11), 2392 - 8
Immunochemical evidence that cholesteryl ester transfer protein and bactericidal/permeability-increasing protein share a similar tertiary structure; Guyard-Dangremont V et al.; Cholesteryl ester transfer protein (CETP) plays an important role in plasma lipoprotein metabolism through its ability to transfer cholesteryl ester, triglyceride, and phospholipid between lipoproteins . CETP is a member of a gene family that also includes bactericidal/permeability-increasing protein (BPI) . The crystal structure of BPI shows it to be composed of two domains that share a similar structural fold that includes an apolar ligand-binding pocket . As structurally important residues are conserved between BPI and CETP, it is thought that CETP and BPI may have a similar overall conformation . We have previously proposed a model of CETP structure based on the binding characteristics of anti-CETP monoclonal antibodies (mAbs) . We now present a refined epitope map of CETP that has been adapted to a structural model of CETP that uses the atomic coordinates of BPI . Four epitopes composed of CETP residues 215-219, 219-223, 223-227, and 444-450, respectively, are predicted to be situated on the external surface of the central beta-sheet and a fifth epitope (residues 225-258) on an extended linker that connects the two domains of the molecule . Three other epitopes, residues 317-331, 360-366, and 393-410, would form part of the putative carboxy-terminal beta-barrel . The ability of the corresponding mAbs to compete for binding to CETP is consistent with the proximity of the respective epitopes in the model . These results thus provide experimental evidence that is consistent with CETP and BPI having similar surface topologies.

J Vet Med Sci, 1999 Nov, 61(11), 1249 - 51
Effects of recombinant bovine granulocyte-macrophage colony-stimulating factor on bovine peripheral blood neutrophil functions in vitro and in vivo; Hirai T et al.; Effects of recombinant bovine granulocyte-macrophage colony-stimulating factor (rboGM-CSF) on bactericidal activity of bovine peripheral blood neutrophils in vitro and in vivo were studied . In in vitro experiment, bovine blood neutrophils were cultured for 9 hr in media containing 0.005, 0.05 or 0.5 microg/ml of rboGM-CSF . Neutrophils treated with rboGM-CSF showed significantly higher luminol-dependent chemiluminescence (LDCL) than control cells . In in vivo experiment, neutrophils isolated from cows injected 5.0 microg/kg of rboGM-CSF showed significantly higher Nitrobluetetrazolium (NBT) reduction value than that from control cows 24 hr post injection . Total leukocyte counts of cows injected rboGM-CSF sharply decreased 6 hr post injection and recovered to normal level 2 days post injection . Body temperature of these cows rose 6 hr post injection and back to normal level at 24 hr post injection . It was suggested that rboGM-CSF enhanced bactericidal activity of bovine neutrophils both in vitro and in vivo.

Stomatologiia (Mosk), 1999, 78(6), 9 - 15
{The dynamics of the healing of experimentally reproduced bone defects filled with different polyacrylamide gel-based compounds}; Grigor'ian AS et al.; Time course of healing of standard osseous defects by introduction of polyacrylamide gel (PAG)-based materials was studied in rat experiments . PAG did not prevent the formation of soft-tissue and osseous regenerate in bone defects . Addition of hydroxyapatite, bactericidal agent, and lysozyme to PAG-based composition led in some cases to development of chronic inflammation and giant-cell reaction in osseous wound and to inhibition of the reparative processes.

J Hepatol, 1999 Nov, 31(5), 905 - 12
Organ blood flow after partial hepatectomy in rats: modification by endotoxin-neutralizing bactericidal/permeability-increasing protein (rBPI23); Boermeester MA et al.; BACKGROUND/AIM: Both maintenance of adequate perfusion and regeneration of the remnant liver are important in the recovery of liver function after partial hepatectomy . In previous experiments, we have shown that profound hypotension and liver injury can be attenuated by neutralizing endotoxins . The relative contribution of endotoxemia to changes in liver blood flow and blood flow to other major organs after partial hepatectomy is not known . The aim of this study was to examine the effect of endotoxin neutralization on individual organ blood flows including hepatic artery and splanchnic blood flow after experimental partial hepatectomy and its relation to liver cell proliferation . METHODS: Male Wistar rats underwent either two-thirds partial hepatectomy or sham operation . Treatment consisted of continuous infusion of recombinant N-terminal bactericidal/permeability-increasing protein (rBPI23) or control protein . At 4 h after surgery, organ blood flows were measured using the radiolabeled microsphere technique, and at 24 h, proliferation index in liver tissue was calculated . RESULTS: After partial hepatectomy, blood flows to virtually all organs were significantly lower as compared to values obtained in sham-operated rats . rBPI23 greatly improved hepatic artery flow (p<0.001) but not portal venous flow . These effects of rBPI23 on liver flow preceded an equally enhanced liver cell proliferation (p<0.01) . Endotoxin neutralization led to significantly higher flows to some but not all splanchnic organs . Lung perfusion was significantly improved by rBPI23 . CONCLUSIONS: Neutralization of endogenous endotoxins improves liver blood flow after partial hepatectomy and also periportal and pericentral liver cell proliferation . This proliferation effect may result from an increased hepatic artery flow . Lung, colon, spleen and pancreas flow but not kidney flow was greatly enhanced by rBPI23.

Crit Care Med, 1999 Nov, 27(11), 2459 - 68
Mechanisms for the diminished neutrophil exudation to secondary inflammatory sites in infected patients with a systemic inflammatory response (sepsis); Ahmed NA et al.; OBJECTIVE: To determine the mechanism for the reduced polymorphonuclear neutrophil exudation to secondary inflammatory sites in critically ill patients with infection and systemic inflammatory response (sepsis) . DESIGN: Prospective cohort study . SETTING: Research laboratory and integrated intensive care unit of a tertiary care university-affiliated teaching hospital . PATIENTS: Healthy subjects or critically ill patients with confirmed infection and a systemic inflammatory response (septic patients) . MEASUREMENTS AND MAIN RESULTS: We found that polymorphonuclear neutrophil delivery to a secondary inflammatory site (skin window blisters) is reduced by >70% in humans with sepsis, defined as serious infection and a systemic inflammatory response compared with healthy controls . The expression of the endothelial adhesion molecules intercellular adhesion molecule-1, E-selectin and P-selectin in microvessels from skin biopsies was comparable in the two study groups . Also, CD11a and CD11b levels were equal in circulating polymorphonuclear neutrophils (PMNs) from both study groups . Both adhesion molecules were markedly and equally up-regulated during exudation . Circulating PMNs from septic patients showed marked shedding of L-selectin compared to those of healthy controls, with a corresponding increase in their plasma L-selectin levels . An increased concentration gradient between plasma and exudate fluid was found for tumor necrosis factor-alpha and interleukin-8 in septic patients, but not for C5a . The phagocytic and bactericidal capacity of septic patient circulating PMNs was higher then in healthy control patients, but these differences were lost after exudation . There were no major differences in oxidative burst or intracellular calcium flux of circulating PMNs from the two study groups . Polymorphonuclear neutrophil exudation primed both responses to different extents . CONCLUSIONS: Septic patients deliver fewer PMNs to secondary inflammatory sites . In addition, neutrophil exudation results in loss of the small priming effect for phagocytosis and bactericidal function induced by sepsis . Failure to produce a gradient to C5a and intravascular shedding of L-selectin may be responsible for this sepsis-induced reduction in neutrophil exudation to secondary inflammatory sites.

J Clin Pharmacol, 1999 Nov, 39(11), 1169 - 76
Pharmacokinetics of a recombinant modified amino terminal fragment of bactericidal/permeability-increasing protein (rBPI21) in healthy volunteers; Bauer RJ et al.; Phase I pharmacokinetic and safety studies were conducted in healthy volunteers with rBPI21, a recombinant protein derived from the amino terminal domain of human bactericidal/permeability-increasing protein . rBPI21 was administered as a 30-minute infusion at doses of 0.25 to 4 mg/kg or as a 24- to 48-hour infusion at doses of 2 to 8 mg/kg . For the 30-minute infusions, the clearance of rBPI21 decreased with increasing dose from 8.4 mL/min/kg at 0.25 mg/kg to 3.3 mL/min/kg at 4 mg/kg . For rBPI21 infused over 24 to 48 hours the clearance was 10 to 11 mL/min/kg . The concentration-time profile of rBPI21 was well described by a three-compartmental model with parallel first-order and Michaelis-Menten (saturable) elimination . This model for the clearance of rBPI21 has been useful in estimating starting doses for therapeutic clinical trials.

J Dairy Sci, 1999 Nov, 82(11), 2385 - 92
Effects of the presence of the mammary gland on expression of neutrophil adhesion molecules and myeloperoxidase activity in periparturient dairy cows; Kimura K et al.; Neutrophil function is reduced in periparturient dairy cows . Possible factors that reduce neutrophil function include endocrine changes associated with parturition and metabolic stresses associated with lactogenesis . In this study, mastectomized and intact cows were studied to specifically examine the effects of lactogenesis on neutrophil function in periparturient cows . Expression of adhesion molecules on neutrophils (L-selectin, mediating capture and rolling adhesion, and beta 2-integrin, mediating tight adhesion vital to egress) and neutrophil myeloperoxidase activity (an index of bactericidal activity) were assessed in mastectomized and intact cows . Expression of L-selectin decreased at parturition followed by rapid recovery to prepartum values in both intact and mastectomized cows . Expression of beta 2-integrins increased in intact cows at parturition but not in mastectomized cows . Expression of beta 2-integrins was greater in intact cows than in mastectomized cows throughout the study . Neutrophil myeloperoxidase activity decreased from baseline prepartum values as parturition approached in both intact and mastectomized cows, which suggests the endocrine changes associated with the act of parturition are predominant factors causing loss of neutrophil function . Myeloperoxidase activity recovered to prepartum values within a week of parturition in mastectomized cows; however, myeloperoxidase activity remained depressed in neutrophils obtained from intact cows throughout the first 20 d of lactation . The presence of the mammary gland and its attendant metabolic stresses slowed recovery of neutrophil function, which suggests that the metabolic stress of lactation exacerbated periparturient immunosuppression.

J Agric Food Chem, 1999 Feb, 47(2), 397 - 402
Alkylperoxyl radical-scavenging activity of various flavonoids and other phenolic compounds: implications for the anti-tumor-promoter effect of vegetables; Sawa T et al.; We recently reported that alkylperoxyl radical (ROO(*)) enhanced carcinogenesis in rats treated with carcinogen (Sawa et al . Cancer Epidemiol . Biomarkers Prev . 1998, 7, 1007-1012), and the tumor promoting action of ROO(*) could be reduced by addition of hot-water extracts of vegetables (Maeda et al . Jpn . J . Cancer Res . 1992, 83, 923-928) . Here we described the ROO(*)-scavenging activity of flavonoids and nonflavonoid phenolics and their role in anti-tumor-promoter effects . A model molecular species, ROO(*), was generated from tert-butyl hydroperoxide (t-BuOOH) and heme iron, and the scavenging of t-BuOO(*) was determined by (a) bioassay based on the bactericidal action of ROO(*), (b) luminol-enhanced chemiluminescence, and (c) electron spin resonance . Of 17 authentic plant phenolics tested, 9 compounds (including rutin, chlorogenic acid, vanillin, vanillic acid, neohesperidin, gallic acid, shikimic acid, rhamnetin, and kaempferol) showed remarkably high ROO(*)-scavenging activity . Some of them were detected and quantified in hot-water extracts of mung bean sprouts, used as the model vegetable, and their contents increased after germination, which paralleled very well to the ROO(*)-scavenging capacity of the vegetable extracts . Thus, a diet rich in these radical scavengers would reduce the cancer-promoting action of ROO(*) . Consequently, the carcinogenic potentials of oxygen-related radicals may be suppressed.

Rev Esp Quimioter, 1999 Jun, 12(2), 144 - 8
{Effect of antiinflammatory agents on the accumulation of ofloxacin in human polymorphonuclear leukocytes}; Orero A et al.; The uptake of quinolones is a recently defined pharmacokinetic parameter which is increasing in importance in clinical practice, especially for immunocompromised patients whose polymorphonuclear leukocytes (PMNLs) have their bactericidal systems impaired, or in infections due to bacteria able to survive in the phagocytes . In both situations, antibiotics able to penetrate and be active in the phagocytes are required . The simultaneous administration of an antibiotic and an antiinflammatory drug is frequent, and previous studies have described interactions between the intracellular activity of the quinolones and the presence of phenylbutazone . We studied the effect of the presence of and the pretreatment (37 C for 30 and 60 min) of human PMNLs with the antiinflammatory drugs betamethasone (1 mg/l), hydrocortisone (1 mg/l), phenylbutazone (10 mg/ml), and acetylsalicylic acid (200 mg/l) on the uptake of ofloxacin (10 mg/l) by the PMNLs using a fluorometric method to measure the intra-PMNL concentration of ofloxacin . The presence of betamethasone did not modify the uptake of ofloxacin by PMNLs . Pretreatment of PMNLs with hydrocortisone, phenylbutazone and acetylsalicylic acid produced a significant decrease in the maximum intracellular concentration/extracellular concentration ratios compared with the maximum reached without pretreatment . These results suggest that hydrocortisone, phenylbutazone and acetylsalicylic acid interfere with the uptake of ofloxacin by PMNLs and increase the efflux of ofloxacin from PMNLs.

Biol Neonate, 1999 Dec, 76(6), 331 - 9
Effects of milk and colostrum on superoxide generation of oral polymorphonuclear neutrophils and the changes that occur during storage at low temperature; Kanematsu H et al.; Human oral polymorphonuclear neutrophils (OPMN) generate reactive oxygen species even in the absence of stimulants . Because OPMN from newborn babies are exposed to colostrum and mature milk, the biological properties of these cells including the generation of reactive oxygen species might possibly be affected by the constituents of colostrum and milk . The present work reports the effects of colostrum and mature milk, including the effects of storage at low temperature, on superoxide generation by OPMN . Fresh colostrum and mature milk did not affect either endogenous or formyl-methionyl-leucyl-phenylalanine-induced generation of superoxide by OPMN . However, superoxide generation stimulated by phorbol myristate acetate or arachidonic acid was inhibited by colostrum and mature milk presumably due to binding of the ligands to milk proteins . During the storage of milk at 4 degrees C, free forms of unsaturated long-chain fatty acids increased, and there was concomitant increase in the ability of milk to generate superoxide radicals in OPMN . Kinetic analysis suggested that colostrum and mature milk regulate superoxide generation by OPMN, thereby modulating the bactericidal activity of these cells in the oral cavity .

Shock, 1999 Nov, 12(5), 365 - 72
Ratio of bacteria to polymorphonuclear neutrophils (PMNs) determines PMN fate; Matsuda T et al.; This study was designed to investigate how live Escherichia coli influence the fate of polymorphonuclear neutrophils (PMNs) in vitro . PMNs from 10 healthy volunteers were cocultured with or without live E . coli at different ratios . Heat-killed E . coli (Hk) were also added to PMNs at a ratio of 1:10 . The PMNs were then analyzed by flow cytometry for cell death, reactive oxygen intermediates (ROI) production, and CD16 expression . Morphologic features were also assessed . PMN apoptosis was confirmed by DNA gel electrophoresis . Low doses of E . coli (PMN:E . coli ratios of 1:0.01 and 1:0.1) inhibited PMN apoptosis . In contrast, a high dose of E . coli (PMN:E . coli ratio of 1:10) increased PMN necrosis . ROI production was significantly greater at PMN:E . coli ratios of 1:10 and 1:10 (Hk) than at ratios of 1:0.01 and 1:0.1, or in PMNs cultured alone after a 15 or 30 minute coculture . CD16 expressions were significantly lower in PMNs cocultured with E . coli than in those cultured alone after a 4 or 12-h coculture . Tumor necrosis factor-alpha, interleukin (IL)-1beta and IL-6 levels in cell-free supernatants were also measured . The mean percentages of apoptosis at PMN:E . coli ratios of 1:0.01 and 1:10 (Hk), and in PMNs cultured alone after a 12-h coculture showed significant inverse correlations with these cytokine levels in cell-free supernatants at 12 h . Our results demonstrate that low doses of live E . coli inhibits predominantly PMN apoptosis, whereas a high dose of E . coli increases necrosis . Augmented PMN bactericidal function, via inhibition of PMN cell death, may be beneficial for host defense against bacterial infection and/or sepsis.

Inflammation, 1999 Dec, 23(6), 507 - 21
Increased expression of Fas ligand on Mycobacterium tuberculosis infected macrophages: a potential novel mechanism of immune evasion by Mycobacterium tuberculosis?
Mustafa T, Phyu S, Nilsen R, Bjune G, Jonsson R.
We have studied the location and mechanism of apoptosis within the granulomas in the lungs at various stages of slowly progressive primary murine Mycobacterium tuberculosis infection . Parallel sections were analyzed for detection of mycobacterial antigens, Fas, and Fas ligand (FasL) by immunohistochemistry, and for apoptotic cells by terminal deoxynucleotidyl-transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) method . The frequency of apoptosis was high in the macrophage aggregates as compared to the lymphocyte aggregates and at the interface between them . Five to seven percent of the vacuolated macrophages in the granulomas expressed FasL intensely . These cells contained large amounts of mycobacterial antigens . These findings suggest that M . tuberculosis infection can induce increased expression of FasL in a population of infected macrophages . As a consequence the infected macrophages will be protected from the attack of cytotoxic T cells and activation of bactericidal mechanisms by Th1 type lymphocytes . This constitutes a novel evasion mechanism for M . tuberculosis possibly explaining the chronic course of infection.

Eur J Gastroenterol Hepatol, 1999 Nov, 11(11), 1293 - 8
Alpha1-antitrypsin phenotypes and anti-neutrophil cytoplasmic auto-antibodies in inflammatory bowel disease; Taddei C et al.; OBJECTIVES: Alpha1-antitrypsin (alpha1-AT) is encoded by a highly polymorphic gene with over 75 codominantly expressed alleles at the protease inhibitor (Pi) locus classified as normal, deficient, dysfunctional or null . The aim of this study was to determine in patients with inflammatory bowel disease: (i) the prevalence of anti-neutrophil cytoplasmic auto-antibodies (ANCA) and their antigen specificities; (ii) alpha1-AT Pi phenotypes; and (iii) possible associations between ANCA, disease activity and deficient alpha1-AT alleles . DESIGN: Study of 95 consecutive patients with ulcerative colitis (UC) and 63 patients with Crohn's disease (CD) . METHODS: Diagnosis and disease activity were determined by clinical, endoscopic and histological criteria . ANCA by indirect immunofluorescence (IIF) and Pi phenotyping by isoelectric focusing were performed for all patients . Positive IIF sera were tested in antigen-specific enzyme-linked immunosorbent assay: proteinase 3 (PR3), myeloperoxidase (MPO), lactoferrin (LF), lysozyme, human leucocyte elastase (HLE), cathepsin G and bactericidal/permeability increasing protein (BPI) . RESULTS: Sixty-one patients with UC (64.2%) and only 11 with CD (17.5%) had ANCA (P < 0.001) . Antigen specificities were PR3 (7/61), MPO (3/61), LF (6/61), HLE (1/63) and BPI (10/61) in UC, and PR3 (2/11) and BPI (2/11) in CD . Three PiZ alleles were found, matching the prevalence in the normal French control population . No relationship was found between the presence of ANCA, antibody specificity, disease activity and deficient alpha1-AT alleles . CONCLUSION: ANCA are more frequent in UC than CD and do not correlate with disease activity . ANCA and protease/antiprotease imbalance do not appear to modulate the clinical course of inflammatory bowel disease.

Chest, 1999 Nov, 116(5), 1233 - 9
Impact of blood transfusions on inflammatory mediator release in patients undergoing cardiac surgery; Fransen E et al.; STUDY OBJECTIVES: This study was conducted to investigate whether intraoperative blood transfusions affect the release of proinflammatory mediators in patients undergoing cardiac surgery . Therefore, we measured plasma levels of bactericidal permeability increasing protein (BPI) as a marker of neutrophil activation, interleukin-6 (IL-6), lipopolysaccharide binding protein (LBP), and C-reactive protein (CRP) . In addition, these mediators, except CRP, were also measured in packed red cell units (PCs) administered to these patients . DESIGN: Prospective study . SETTING: Cardiopulmonary surgery department in a university hospital . PATIENTS: One hundred fourteen consecutive patients undergoing cardiac surgery . INTERVENTIONS: Blood samples were taken at induction of anesthesia, at the start of aortic cross-clamping, at aortic unclamping, and at 0.5, 4, 8, and 18 h thereafter . RESULTS: Thirty-six patients received PC intraoperatively . BPI levels in patients who received transfusions were significantly higher at 0.5 and 4 h after aortic unclamping than in patients without transfusions (p < 0.05), and increased with the number of PC administered . IL-6 levels at 0.5, 4, and 18 h after aortic unclamping were also significantly higher in patients who received transfusions (p < 0.01) . BPI was found in all units of packed red cells tested at concentrations up to 15 times preoperative plasma levels in patients . However, PC IL-6 could be detected in none of the samples . Plasma levels of LBP and CRP were similar in both patient groups . LBP was found in very low concentrations in all PC . Patients who received intraoperative transfusions had a worse postoperative performance . CONCLUSIONS: Intraoperative PC transfusions do contribute to the inflammatory response after cardiac surgery both by enhancing part of the response and by directly changing plasma concentrations of inflammatory mediators . Furthermore, these data show that intraoperative PC transfusion is associated with a worse postoperative performance.

Biochim Biophys Acta, 1999 Nov 9, 1461(1), 19 - 26
Tyr-->Trp-substituted peptide 115-129 of a Lys49 phospholipase A(2) expresses enhanced membrane-damaging activities and reproduces its in vivo myotoxic effect; Lomonte B et al.; Myotoxin II is a group II Lys49 phospholipase A(2) (PLA(2)) isolated from the venom of the snake Bothrops asper . Previous studies on a synthetic peptide derived from its heparin-binding, cationic/hydrophobic sequence 115-129 demonstrated a direct functional role of this particular region in the in vitro cytolytic and bactericidal actions of the protein . Nevertheless, no significant myonecrosis has been observed after local intramuscular injection of peptide 115-129 (p115-129) in mice . Since the membrane-damaging action of p115-129 was proposed to depend on its amphiphilic character, the present study examined the effects of substituting its cluster of three tyrosine residues by tryptophan residues, on its toxic/pharmacological activities in vitro and in vivo . This substitution resulted in a drastic enhancement of the membrane-damaging activities of the peptide (p115-W3), together with the clear expression of myotoxic activity in vivo . Both the heparin-binding and antigenic characteristics of p115-129 were essentially conserved in p115-W3, suggesting that the modification did not lead to radical structural alterations . In addition to myotoxicity, cytotoxicity, and bactericidal action, p115-W3 exerted edema-forming activity in the mouse footpad assay . Thus, the synthetic 13-mer p115-W3 reproduced all the known toxic effects of myotoxin II . In spite of its potent membrane-damaging actions, p115-W3 did not acquire direct hemolytic activity upon mouse erythrocytes, an effect which is not present in myotoxin II, but that has been ascribed to the presence of tryptophan in other cationic, membrane-damaging peptides such as mellitin from bee venom . The myotoxic activity of p115-W3 herein described constitutes the first example of a short, PLA(2)-based linear synthetic peptide with the ability to reproduce this effect of a parent protein in vivo . This finding is in clear support of the proposed relevance of the C-terminal region 115-129 in all the membrane-damaging mechanisms exerted by myotoxin II, including the myotoxic mechanism.

Clin Diagn Lab Immunol, 1999 Nov, 6(6), 844 - 50
Changes in endotoxin-binding proteins during major elective surgery: important role for soluble CD14 in regulation of biological activity of systemic endotoxin; Hiki N et al.; Assessment of circulating endotoxin during the perioperative period, which is only demonstrated by the Limulus amebocyte lysate (LAL) test, may be modulated by several endotoxin-binding proteins . Endotoxin-neutralizing capacity (ENC) and the plasma levels of soluble CD14 (sCD14), lipopolysaccharide-binding protein, and bactericidal/permeability-increasing protein (BPI) were determined in 40 patients 6 h prior to skin incision for major abdominal surgery . The bioactivity of plasma endotoxin was tested by the polymyxin B-inhibited stimulatory activity of the plasma samples on healthy monocytes as measured by the release of tumor necrosis factor alpha . Plasma endotoxin levels in almost all patients increased from 0.05 +/- 0.01 to 0.23 +/- 0.03 experimental units (EU) per ml (P < 0.001); more specifically, 17 of 40 samples showed endotoxin levels of greater than 0.2 EU per ml and corresponding reductions in ENC . Soluble CD14 plasma levels were decreased from 5 . 6 +/- 0.3 to 4.6 +/- 0.3 microg per ml (P < 0.05) . ENC was strongly correlated with the sCD14 plasma concentration throughout the period of observation . The addition of sCD14-neutralizing monoclonal anti-sCD14 antibodies reduced ENC both pre- and postoperatively . No correlation could be established between ENC and the plasma levels of BPI, high-density lipoproteins, or low-density lipoproteins determined by measuring the concentrations of apoprotein A and apoprotein B . Biologically active endotoxin was found in only 6 of 17 samples with endotoxin levels greater than 0.2 EU per ml in the LAL test . These samples could be characterized by their perioperative loss of at least 35% of their sCD14 . No change in sCD14 was detected in the remaining 11 samples . The perioperative loss of ENC is partly caused by the loss of sCD14 resulting from its consumption by endotoxin reaching the bloodstream . This study demonstrated the role of sCD14 on the bioactivity of circulating endotoxin in a human model of endotoxemia after major abdominal surgery.

Infect Immun, 1999 Nov, 67(11), 6191 - 3
Syntheses and immunologic properties of Escherichia coli O157 O-specific polysaccharide and Shiga toxin 1 B subunit conjugates in mice; Konadu E et al.; Escherichia coli O157 is the major cause of diarrhea-associated hemolytic uremic syndrome (HUS) . Strains causing HUS contain either Shiga toxin 1 (Stx1) or Stx2, or both . In adult volunteers, conjugate vaccines of detoxified lipopolysaccharide (LPS) elicited bactericidal antibodies to E . coli O157 . Here, the detoxified LPS was conjugated with improved schemes to the nontoxic B subunit of Stx1 . Mice injected with these bivalent conjugates elicited both bactericidal antibodies to E . coli O157 and neutralization antibodies to Stx1.

Infect Immun, 1999 Nov, 67(11), 6181 - 6
The outer membrane of Brucella ovis shows increased permeability to hydrophobic probes and is more susceptible to cationic peptides than are the outer membranes of mutant rough Brucella abortus strains; Freer E et al.; The permeability of the outer membrane (OM) to hydrophobic probes and its susceptibility to bactericidal cationic peptides were investigated for natural rough Brucella ovis and for mutant rough Brucella abortus strains . The OM of B . ovis displayed an abrupt and faster kinetic profile than rough B . abortus during the uptake of the hydrophobic probe N-phenyl-naphthylamine . B . ovis was more sensitive than rough B . abortus to the action of cationic peptides . Bactenecins 5 and 7 induced morphological alterations on the OMs of both rough Brucella strains . B . ovis lipopolysaccharide (LPS) captured considerably more polymyxin B than LPSs from both rough and smooth B . abortus strains . Polymyxin B, poly-L-lysine, and poly-L-ornithine produced a thick coating on the surfaces of both strains, which was more evident in B . ovis than in rough B . abortus . The distinct functional properties of the OMs of these two rough strains correlate with some structural differences of their OMs and with their different biological behaviors in animals and culture cells.

J Immunol, 1999 Nov 1, 163(9), 5013 - 9
Neutrophil dysfunctions, IL-8, and soluble L-selectin plasma levels in rapidly progressive versus adult and localized juvenile periodontitis: variations according to disease severity and microbial flora; Gainet J et al.; We used flow cytometry to analyze the expression of adhesion molecules and the oxidative burst of whole-blood polymorphonuclear neutrophils (PMN) from 26 patients with periodontitis . Three different clinical entities were studied: adult periodontitis (AP), localized juvenile periodontitis (LJP), and rapidly progressive periodontitis (RPP) . Unstimulated PMN from the patients showed reduced Lewis x, sialyl-Lewis x, and L-selectin expression relative to those from healthy control subjects . These alterations were present whatever the severity of periodontal disease . However, PMN from RPP patients showed increased basal H2O2 production and decreased L-selectin shedding . These latter impairments, which correlated with increased IL-8 plasma levels, could contribute to initial vascular damage . In addition, decreased IL-8 priming of H2O2 production by PMN from RPP patients could account for a lower bactericidal capacity of PMN, leading to the large number of bacteria in the subgingival region of RPP patients . Soluble L-selectin plasma levels were also decreased in the RPP group, indicating more severe or diffuse endothelial damage . These abnormalities were not found in the patients with less destructive forms of periodontitis (AP and LJP) . Porphyromonas gingivalis, a bacterial pathogen known to increase IL-8 production by PMN, was found in the periodontal pockets of RPP patients only . These results show links among PMN abnormalities, the clinical form of periodontitis, and the gingival bacterial flora.

Mol Genet Metab, 1999 Oct, 68(2), 283 - 303
Clinical, molecular, and cell biological aspects of Chediak-Higashi syndrome; Introne W et al.; Chediak-Higashi syndrome (CHS) is a rare autosomal recessive disorder characterized by variable degrees of oculocutaneous albinism, easy bruisability, and bleeding as a result of deficient platelet dense bodies, and recurrent infections, with neutropenia, impaired chemotaxis and bactericidal activity, and abnormal NK cell function . Neurologic involvement is variable, but often includes peripheral neuropathy . Most patients also undergo an "accelerated phase," which is a nonmalignant lymphohistiocytic infiltration of multiple organs resembling lymphoma . Death often occurs in the first decade from infection, bleeding, or development of the accelerated phase . The hallmark of CHS is the presence of huge cytoplasmic granules in circulating granulocytes and many other cell types . These granules are peroxidase-positive and contain lysosomal enzymes, suggesting that they are giant lysosomes or, in the case of melanocytes, giant melanosomes . The underlying defect in CHS remains elusive, but the disorder can be considered a model for defects in vesicle formation, fusion, or trafficking . Because the beige mouse demonstrates many characteristics similar to those of human CHS patients, including dilution of coat color, recurrent infections, and the presence of giant granules, it is considered the animal homologue of CHS . The beige gene, Lyst, was mapped and sequenced in 1996, prompting identification of the human LYST gene on chromosome 1q42 . Lyst and LYST show 86.5% sequence homology . LYST encodes a 429 kDa protein with a function that remains unknown, but the source of extensive speculation among students of cell biology .

Hepatogastroenterology, 1999 Jul-Aug, 46(28), 2476 - 82
Immunological function and nutritional status in patients with hepatocellular carcinoma; Iida K et al.; BACKGROUND/AIMS: Infection is a major complication associated with increased morbidity and mortality in patients with hepatocellular carcinoma . We compared the immunological function and nutritional status in 16 patients with hepatocellular carcinoma (13 patients had liver cirrhosis) with those of 21 normal healthy subjects . METHODOLOGY: The immunological function was assessed by chemotaxis and superoxide anion production by neutrophils, phagocytosis and killing activities of neutrophils and monocytes, absolute and relative number of peripheral blood lymphocytes, the percentage of peripheral lymphocyte subsets and serum concentrations of immunoglobulins . RESULTS: Although the phagocytic and bactericidal activities of monocytes and superoxide production of neutrophils were not different between the groups, the phagocytic and bactericidal activities of neutrophils and the percentage of natural killer cells were significantly reduced in patients with hepatocellular carcinoma . In the latter group, the prognostic nutrition index was significantly high compared with normal subjects, indicating a poor nutritional status . The phagocytic and bactericidal activities of neutrophils were low in patients with a poor nutritional status compared to those with a good nutritional status . CONCLUSIONS: Our results suggest that impaired immunological competence and undernourishment may be one of the mechanisms causing increased susceptibility of patients with hepatocellular carcinoma to infection.

Hepatogastroenterology, 1999 Jul-Aug, 46(28), 2273 - 7
Bactericidal/permeability-increasing protein in colonic mucosa in ulcerative colitis; Haapamaki MM et al.; BACKGROUND/AIMS: Increased mucosal concentration of bactericidal/permeability-increasing protein (BPI) has been shown in inflammatory bowel diseases . The purpose of the present study was to investigate the relationship between the mucosal concentration of BPI and the grade of mucosal inflammation in ulcerative colitis . METHODOLOGY: Samples of colonic mucosa from 12 patients with ulcerative colitis and from 8 control patients were studied . The concentration of BPI in tissue extracts was measured by a time-resolved fluoroimmunoassay . The concentration of BPI was compared between samples with histological inflammatory changes of different severity . BPI was localized in tissue sections by immunohistochemistry . RESULTS: The concentration of BPI was higher (p < 0.001) in samples of colonic mucosa from patients with ulcerative colitis (median: 3.2 micrograms/g, range: 0.3-22.6 micrograms/g) than in control samples (0.4 microgram/g, 0.1-0.6 microgram/g,) . Moreover, the concentration of BPI was higher (p = 0.015) in samples with severe inflammation (2.5 mu/g, 0.3-22.6 micrograms/g) than in those with mild inflammation (0.5 mu/g, 0.3-2.5 micrograms/g) . The concentration of BPI in mucosal samples correlated well with the degree of histological inflammation (Spearman R = 0.70, p = 0.01) . BPI was localized in polymorphonuclear leukocytes in the mucosa and stroma of the colonic wall . CONCLUSIONS: The concentration of BPI is increased in the colonic mucosa of patients with ulcerative colitis . The increase in the concentration of BPI in colonic mucosa seems to be closely associated with the inflammatory activity of ulcerative colitis.

Scand J Immunol, 1999 Oct, 50(4), 399 - 404
Inhibitory effect of haptoglobin on granulocyte chemotaxis, phagocytosis and bactericidal activity; Rossbacher J et al.; Human haptoglobin (Hp) is synthesized at hepatic and extrahepatic sites as an acute-phase reactant protein (APP) . We investigated the effects of Hp on granulocyte function . The chemotaxis of freshly isolated human granulocytes and differentiated HL-60 cells in response to the bacterial tripeptide, f-met-leu-phe, was inhibited in the presence of a physiological concentration of Hp, but chemotaxis in the presence of the proinflammatory cytokine interleukin-8 (IL-8) was not inhibited . Phagocytosis of viable Escherichia coli, as well as fluoresceinated nonviable E . coli, was inhibited . Hp also reduced granulocyte intracellular bactericidal activity against E . coli . The observed inhibitory effects of Hp on granulocyte function are similar to those reported for C-reactive protein and suggest that APPs dampen the acute inflammatory response.

Free Radic Res, 1999 Oct, 31(4), 251 - 60
Cross-talk of NO, superoxide and molecular oxygen, a majesty of aerobic life; Inoue M et al.; Because nitric oxide (NO) reacts with various molecules, such as hemeproteins, superoxide and thiols including glutathione (GSH) and cysteine residues in proteins, biological effects and metabolic fate of this gaseous radical are affected by these reactants . Although the lifetime of NO is short particularly under air atmospheric conditions (where the oxygen tension is unphysiologically high), it increases significantly under physiologically low oxygen concentrations . Because oxygen tensions in human body differ from one tissue to another and change depending on their metabolism, biological activity of NO in various tissues might be affected by local oxygen tensions . To elucidate the role of NO and related radicals in the regulation of circulation and energy metabolism, their effects on arterial resistance and energy metabolism in mitochondria, mammalian cells and enteric bacteria were studied under different oxygen tensions . Kinetic analysis revealed that NO-dependent generation of cGMP in resistance arteries and their relaxation were strongly enhanced by lowering oxygen tensions in the medium . NO reversibly suppressed the respiration and ATP synthesis of isolated mitochondria and intact cells particularly under low oxygen tensions . Kinetic analysis revealed that cross-talk between NO and superoxide generated in and around endothelial cells regulates arterial resistance particularly under physiologically low oxygen tensions . NO also inhibited the respiration and ATP synthesis of E . coli particularly under low oxygen tensions . Because concentrations of NO and H+ in gastric juice are high, most ingested bacteria are effectively killed in the stomach . However, the inhibitory effects of NO on the respiration and ATP synthesis of H . pylori are extremely small . Kinetic analysis revealed that H . pylori generates the superoxide radical thereby inhibiting the bactericidal action of NO in gastric juice . Based on such observations, critical roles of the cross-talk of NO, superoxide and molecular oxygen in the regulation of energy metabolism and survival of aerobic and microaerophilic organisms are discussed.

J Infect Dis, 1999 Nov, 180(5), 1597 - 602
Cefotaxime, desacetyl-cefotaxime, and bactericidal activity in spontaneous bacterial peritonitis; Dalmau D et al.; We have prospectively studied 13 episodes of spontaneous bacterial peritonitis (SBP) in 12 patients treated with cefotaxime (CTX) 2 g intravenously every 8 h (mean duration, 5.3 days) . Ascitic fluid was inoculated at the bedside . The cultures were done before, during (day 3 after CTX initiation), and 48-72 h (mean, 56 h) after the end of therapy . All SBP episodes were monomicrobial . During treatment, the concentrations of CTX and desacetyl-cefotaxime (d-CTX) in ascitic fluid were high in all 13 SBP episodes, and d-CTX was still present in 6 patients who had residual ascitic bactericidal titer (ABT) activity after the last dose of CTX . ABTs were >/=1:128 during CTX therapy in 12 episodes and were measurable in 7 patients after the last dose . All patients were cured . The present study provides scientific rationale to the clinical studies that suggest treating SBP episodes with lower doses of antibiotics and shorter treatment duration.

J Am Coll Surg, 1999 Oct, 189(4), 374 - 9
Treatment with recombinant bactericidal/permeability-increasing protein to prevent endotoxin-induced mortality in bile duct-ligated rats; Kimmings AN et al.; BACKGROUND: Operation in patients with obstructive jaundice is associated with substantial morbidity because of increased susceptibility to endotoxin (lipopolysaccharide) and the inflammatory cascade . Different interventions to reduce endotoxemia and cytokine induction, and resulting complications, have been studied . Bactericidal/permeability-increasing protein (BPI) is a naturally occurring endotoxin-binding protein produced in neutrophils . It binds endotoxin, neutralizing the activity and inhibiting cytokine production by mononuclear cells . In experimental endotoxemia in animals and in healthy human volunteers, BPI has shown a protective effect . The aim of this study was to determine whether BPI could protect against increased endotoxin sensitivity in rats with obstructive jaundice and reduce endotoxin-induced mortality . STUDY DESIGN: Male Wistar rats were used . Intraperitoneal Escherichia coli 2mg/kg was given 1 week after sham operation or bile duct ligation (BDL) . Three groups were studied: sham, BDL with placebo, and BDL with 5 mg/kg recombinant BPI21 . RESULTS: BDL rats were jaundiced (mean bilirubin 186 micromol/L; no difference between BDL rats without or with BPI) . Bilirubin remained less than 1 micromol/L in sham-operated rats (p = 0.002) . Endotoxin levels were 3.4pg/mL in sham controls and 3.1 pg/mL in BDL rats before administration of lipopolysaccharide (p = NS) . Two hours after administration, levels were 615.4ng/mL in placebo BDL rats and 10 times less in BPI-treated BDL rats, at 60.2ng/mL (p=0.03) . The same trend was found at 6 hours . At 24 hours, mortality was 1 of 6 in sham-operated rats (15%) versus 8 of 11 in untreated BDL rats (75%) . BPI intervention reduced the death rate to 1 of 12 BDL rats (8%) (p = 0.003) . CONCLUSIONS: Intraperitoneal recombinant BPI21 in rats having BDL reduced endotoxin-induced mortality from 75% to 8%, a death rate comparable to that in nonjaundiced rats . BPI could be an interesting perioperative treatment in clinical obstructive jaundice.

J Pept Sci, 1999 Aug, 5(8), 341 - 51
Structures of trichovirins II, peptaibol antibiotics from the mold Trichoderma viride NRRL 5243; Jaworski A et al.; From the culture broth of the mold Trichoderma viride NRRL 5243 a mixture of polypeptides, named trichovirins (TV), could be isolated and purified by chromatography on XAD-2 adsorber resin and Sephadex LH-20 gel . Chromatography on silica gel using chloroform/methanol 8:2 as eluent provided a mixture of peptides named TV I . Subsequent elution with chloroform/methanol 1:1 yielded a second group of peptides named TV II . That group could be separated into individual components by repetitive HPLC on an octadecylsilyl and a fluorocarbon stationary phase . The sequences of 12 peptides of TV II could be determined by electrospray ionization tandem mass spectrometry of isolated peptides and gas chromatography-mass spectrometry of methanolysates . The N-termini of the 18-mer peptides are acetylated and the C-termini consist of leucinol . Owing to the presence of alpha-aminoisobutyric acid (Aib) residues and the bactericidal and hemolytic activity, the peptides belong to the family of peptaibol antibiotics.

Antimicrob Agents Chemother, 1999 Oct, 43(10), 2356 - 60
Evaluation of rifapentine in long-term treatment regimens for tuberculosis in mice; Lenaerts AM et al.; Besides direct bactericidal activity, long-term effectiveness is one of the most important features to consider when developing new drugs for chemotherapy . In this study, we evaluated the ability of rifapentine (RFP), in monotherapy and combination therapy, to completely eradicate a Mycobacterium tuberculosis infection and to prevent relapse posttreatment in a Swiss mouse model . The combination of RFP, isoniazid (INH), and pyrazinamide (PZA) administered daily resulted in an apparent clearance of M . tuberculosis organisms in the lungs and spleens of infected mice after 10 weeks of treatment . However, 3 months after the cessation of therapy, bacterial regrowth occurred in mice treated for a 12-week period, indicating a relapse of infection . In intermittent treatment regimens of RFP in combination with INH and PZA, sterilization was achieved when mice were treated two to five times per week for 9 weeks . Bacterial growth was still observed in the once-weekly treatment group . Our results show that mouse models can predict important parameters for new drugs . We stress the necessity for long-term posttreatment observation in animal models for the routine evaluation of new drugs for antituberculosis chemotherapy.

Vaccine, 1999 Aug 20, 18(1-2), 109 - 18
Evaluation of a 74-kDa transferrin-binding protein from Moraxella (Branhamella) catarrhalis as a vaccine candidate; Chen D et al.; An outer membrane protein from Moraxella catarrhalis with a mass of 74-kDa was isolated and evaluated as a vaccine candidate . The 74-kDa protein binds transferrin, and appears to be related to the other proteins from the organism that are reported to bind transferrin . The 74-kDa protein possessed conserved epitopes exposed on the bacterial surface . This is based on the reactivity with whole bacterial cells as well as complement dependent bactericidal activity of sera from mice immunized with the isolated proteins from the O35E and TTA24 isolates . However, there was divergence in the degree of antibody cross-reactivity with the protein from one strain to another . This serotypic divergence was reflected in both the complement-dependent bactericidal activities of the antibodies elicited in mice and the capacity of immune mice to clear the bacteria in a murine pulmonary model . Antibodies affinity purified from human plasma lacked bactericidal activity even though they were reactive with all the tested isolates . The 74-kDa protein appears to be a good vaccine candidate, but more studies are needed to understand its antigenic variability and whether antibodies toward it are protective.

Wiad Lek, 1999, 52(3-4), 168 - 73
{Acne vulgaris as a therapeutic problem}; Brzezinska-Wcislo L; Acne vulgaris, the most frequent skin disease of adolescence, because of its location, particularly long lasting course and in, case of some patients, leaving deformations in a form of scars can lead to disturbances in the psychosocial sphere . Medications used in the topical and systemic therapy of acne should reveal such proportions as: anti-seborrhoeic, bactericidal, bacteriostatic, antiphogistic, comedoleitis, anti-comedoformative . Both, in the topical and systemic therapy of acne vulgaris antibiotics such as tetracyclines, erythromycin and clindamycin are used . Other medicaments prescribed for the topical therapy of acne are azelaic acid and benzoil peroxide, retinoic acid--tretinoin and 13-cis-Retinoic acid--isotretinoin . Isotretinoin--13-cis-Retinoic acid used in the systemic therapy is the most potent drug--treating acne which has not reacted to any conventional methods of therapy.

Przegl Lek, 1999, 56(4), 267 - 9
{The effect of Cordafen (nifedipine) on endocytosis and bactericidal activity of polymorphonuclear granulocytes in peripheral blood}; Misiewicz A et al.; The study was performed in patients taking Cordafen Polfa 30 mg a day . Just after three days of its administration, impaired basal metabolism of polymorphonuclear granulocytes in NBT test was found . After seven days of Cordafen use was demonstrated . Cordafen has no effect on oxygen metabolic activity in granulocytes stimulated by latex particles.

JPEN J Parenter Enteral Nutr, 1999 Sep-Oct, 23(5 Suppl), S59 - 61
Glutamine as an immunoenhancing nutrient; Saito H et al.; New strategies for immunonutritional support include administration of special nutrients such as glutamine . Glutamine is important in several key metabolic processes of immune cells and enterocytes . Exogenous glutamine augments the functions of lymphocytes and macrophages . Neutrophils also reportedly utilize glutamine at a significant rate . Our recent studies demonstrated that glutamine enhances neutrophil function . This article focuses on the effects of glutamine on neutrophil function in surgical stress . Enteral glutamine administration enhanced peritoneal and hepatic bacterial clearance in our rat peritonitis model . Furthermore, IV glutamine supplementation improved the outcome of animals with severe surgical stress . Our in vitro study revealed that supplemental glutamine augmented the bacterial killing function of neutrophils from postoperative patients . Glutamine increased phagocytosis of the neutrophils . In addition, glutamine dose-dependently increased production of reactive oxygen intermediates (ROI) by neutrophils . Thus, our studies suggest that glutamine supplementation may improve bactericidal function of neutrophils by increasing both phagocytosis and ROI production . In conclusion, glutamine plays an important role in neutrophil function . Glutamine may be useful for the prevention, and treatment, of severe infection in critical illness and trauma.

Appl Environ Microbiol, 1999 Sep, 65(9), 4094 - 8
Bactericidal activity of photocatalytic TiO(2) reaction: toward an understanding of its killing mechanism; Maness PC et al.; When titanium dioxide (TiO(2)) is irradiated with near-UV light, this semiconductor exhibits strong bactericidal activity . In this paper, we present the first evidence that the lipid peroxidation reaction is the underlying mechanism of death of Escherichia coli K-12 cells that are irradiated in the presence of the TiO(2) photocatalyst . Using production of malondialdehyde (MDA) as an index to assess cell membrane damage by lipid peroxidation, we observed that there was an exponential increase in the production of MDA, whose concentration reached 1.1 to 2.4 nmol . mg (dry weight) of cells(-1) after 30 min of illumination, and that the kinetics of this process paralleled cell death . Under these conditions, concomitant losses of 77 to 93% of the cell respiratory activity were also detected, as measured by both oxygen uptake and reduction of 2,3,5-triphenyltetrazolium chloride from succinate as the electron donor . The occurrence of lipid peroxidation and the simultaneous losses of both membrane-dependent respiratory activity and cell viability depended strictly on the presence of both light and TiO(2) . We concluded that TiO(2) photocatalysis promoted peroxidation of the polyunsaturated phospholipid component of the lipid membrane initially and induced major disorder in the E . coli cell membrane . Subsequently, essential functions that rely on intact cell membrane architecture, such as respiratory activity, were lost, and cell death was inevitable.

Antimicrob Agents Chemother, 1999 Sep, 43(9), 2176 - 82
The structural gene for microcin H47 encodes a peptide precursor with antibiotic activity; Rodriguez E et al.; Microcin H47 is a bactericidal antibiotic produced by a naturally occurring Escherichia coli strain isolated in Uruguay . The microcin genetic system is located in the chromosome and extends over a 10-kb DNA segment containing the genes required for microcin synthesis, secretion, and immunity . The smallest microcin synthesis gene, mchB, was sequenced and shown to encode a highly hydrophobic peptide . An mchB-phoA gene fusion, which directed the synthesis of a hybrid bifunctional protein with both PhoA and microcin H47-like activities, was isolated . The results presented herein lead us to propose that microcin H47 is indeed a ribosomally synthesized peptide antibiotic and that its peptide precursor already has antibiotic activity of the same specificity as that of mature microcin.

Biochem Biophys Res Commun, 1999 Sep 7, 262(3), 647 - 50
Singlet oxygen ((1)Delta(g)O(2)) as the principal oxidant in myeloperoxidase-mediated bacterial killing in neutrophil phagosome; Tatsuzawa H et al.; Intraphagosomal viability of wild type E . coli and lycopene (a powerful (1)O(2) quencher)-producing transformant E . coli was investigated using human polymorphonuclear leukocytes as the cells for phagocytosis of opsonized viable bacteria . While the viability of both wild type and the transformant E . coli decreased very rapidly in the phagosome, but the viability of the lycopene-transformant in phagosomes was about 1.7 times higher than that of wild type E . coli after 5 min of incubation . The results were very similar to the results obtained when E . coli strains were exposed to (1)O(2) generated in myeloperoxidase-H(2)O(2)-Br(-) system (a pure (1)O(2) generating system) at pH 4.5 . The reason for HOCl, which may be generated in the myeloperoxidase-H(2)O(2)-Cl(-) system under physiological conditions but does not become involved in bactericidal action, could be explained by the near neutral pH in phagosomes at which bacterial killing by chlorination is extensively attenuated . This is the first report which proved (1)O(2)-mediated bacterial killing in neutrophil-bacterial phagosomal system .

J Infect, 1999 Jul, 39(1), 81 - 7
Anti-neutrophil cytoplasmic autoantibodies (ANCA) to bactericidal/permeability-increasing (BPI) protein recognize the carboxyl terminal domain; Dunn AC et al.; OBJECTIVES: to identify the region of bactericidal/permeability-increasing protein (BPI) recognized by anti-BPI ANCA . METHODS: sera from 140 patients with a variety of clinical diagnoses (20 systemic vasculitis, 12 cystic fibrosis, 22 bronchiectasis/chronic obstructive airways disease, three diabetes mellitus, 13 chronic renal failure, 12 primary sclerosing cholangitis, eight ulcerative colitis, three Crohn's disease, seven cancer, and 40 other or unknown diagnoses) known to be reactive against native (nBPI), were screened by solid phase enzyme linked immunosorbent assay (ELISA) against a panel of recombinant fusion proteins; holo BPI (rBPI), recombinant lipopolysaccharide binding protein (rLBP), an N-terminal fragment of rBPI (rBPI21 ) and 'fusion' proteins containing the C- or N-terminal ends of BPI spliced with N-or C-ends of LBP, respectively . RESULTS: a strong correlation was seen between the degree of reactivity to rBPI and the BPI C-terminal fusion protein, r=0.69, P < 0.001, as well as between nBPI and rBPI protein, r=0.55, P < 0.001, but not between nBPI and the N-terminal region of BPI (rBPI21), or proteins containing only the N-terminal fragment . Binding to proteins containing the BPI C-terminus was confirmed to be specific by fluid phase inhibition ELISA and Western blot analyses . CONCLUSIONS: together these data suggest that circulating autoantibodies to BPI from patients with different diseases recognize the C-terminal region of BPI.

J Med Entomol, 1999 Jul, 36(4), 426 - 34
Morphological and cytochemical characterization of female Anopheles albimanus (Diptera: Culicidae) hemocytes; Hernandez S et al.; Hemocytes of 2- to 3-d-old female Anopheles albimanus Wiedemann are described by morphology, cytochemistry, and functional criteria . Supplemented Grace's insect medium in a modified Foley's perfusion method was used to obtain hemolymph from An . albimanus . Morphological analysis indicated 3 types of hemocytes were present, prohemocytes, plasmatocytes, and granular cells . Prohemocytes were small round cells with a high nuclear/cytoplasmic ratio . Plasmatocytes were the most abundant cell types in the hemolymph, and appeared as small to large and spindle-shaped cells with round or elongate nucleus, variable number of vacuoles, small granules, and pseudopodia . Granular cells were small to large and round with a large number of cytoplasmic granules, vacuoles, and numerous filopodia . Ultrastructurally, prohemocytes were undifferentiated with abundant free ribosomes and with few small electron-dense granules . Plasmatocytes were rich in mitochondria, rough endoplasmic reticulum, free ribosomes, small electron-dense granules, numerous peripheral vacuoles and with an important organelle polarization . Granular cells contained numerous large electron-dense granular inclusions and vacuoles . Cytochemical studies showed that plasmatocytes and granular cells have cationic bactericidal proteins . Only granular cells showed phenoloxidase and probably lysosomal activities . In vitro functional studies demonstrated that both plasmatocytes and granular cells were able to attach to glass slides, and only plasmatocyte had phagocytic activity and motility . These results characterize the hemocytes of An . albimanus and suggest that plasmatocytes and granular cells may have a role in defense responses to foreign organisms.

Ann Pharmacother, 1999 Jul-Aug, 33(7-8), 868 - 70
Interaction between ciprofloxacin and rifampin; Temple ME et al.; OBJECTIVE: To discuss the necessity of dose adjustment for ciprofloxacin or rifampin during their concurrent use . DATA SOURCES: A MEDLINE search (1966-December 1998) was completed using key terms rifampin and fluoroquinolone . English-language journals were considered . DATA SYNTHESIS: Studies in elderly patients after 14 days of therapy with oral ciprofloxacin and rifampin did not demonstrate significant differences in the pharmacokinetics of ciprofloxacin as compared with those in patients receiving ciprofloxacin alone . Similar results were found in intravenous drug abusers . In comparison to the pharmacokinetics of both ciprofloxacin and rifampin when given alone, the serum bactericidal activity of rifampin, when given with ciprofloxacin in healthy elderly volunteers, was reduced but still evident . Ciprofloxacin pharmacodynamics were not significantly altered . Serum bacterial titers of ciprofloxacin and pefloxacin increased twofold when given with rifampin, although their clinical significance is unknown . RECOMMENDATION: No strong evidence of a significant interaction exists to support dose adjustment for ciprofloxacin or rifampin during their concurrent use.

Int Arch Occup Environ Health, 1999 Aug, 72(5), 323 - 9
Priming of cytokine release and increased levels of bactericidal permeability-increasing protein in the blood of animal facility workers; Borm PJ et al.; OBJECTIVE: To investigate cellular immune responses in laboratory animal workers who are exposed to high levels of animal allergens but also to other biologically active substances containing lipopolysaccharides (LPS), i.e., endotoxins . METHODS: A survey among 20 animal facility workers and 20 matched (gender, smoking) controls was conducted using exposure measurements (endotoxin, colony-forming units of bacteria and fungi) and a questionnaire on respiratory symptoms . Blood samples were taken to determine the ex vivo whole-blood release of tumor necrosis factor-alpha (TNF) and interleukin-8 (IL-8) as well as plasma levels of LPS-binding protein (LBP), bactericidal permeability-increasing protein (BPI), the 75-kDa soluble TNF receptor (sTNFR75), and total/specific IgE . RESULTS: Although exposure to endotoxin was low (range 0.05-2.8 ng/m(3)), a significant (P < 0.05) increase in plasma BPI (4-fold) and srTNF75 (1.2-fold) was found, suggesting a response to inhalation of LPS . Also, the capacity of blood leukocytes to release TNF and IL-8 in response to ex vivo exposure to workplace dust was increased . Data were not confounded by specific allergies, levels of IgE, smoking, or respiratory symptoms . CONCLUSIONS: A profound effect on the cellular immune response was seen in animal workers with low endotoxin exposure and a high antigenic load . It remains to be determined which other biologically active substance(s) are involved in this effect.

J Trace Elem Med Biol, 1999 Jul, 13(1-2), 68 - 75
Short exposure of polymorphonuclear leucocytes to sodium fluoride suppresses the response to fMLP; Knoll-Kohler E et al.; Fluoridated dental care products are used to prevent dental decay . Up to now, there are no data available on whether the fluoride (F-) component of these products affects the bactericidal activity of salivary polymorpho-nuclear leucocytes, which are involved in the protection of the oral mucosa against infection . Therefore, after determining the concentration/time profile of F- in mixed saliva of healthy subjects after topical application of 0.5 g of a 1.25% F- containing gel, unstimulated and fMLP-stimulated polymorphonuclear leucocytes (PMNs) were shortly exposed to these F- concentrations and the generation of superoxide and hypochloric acid were measured, as well as the liberation of lysomal enzymes, and correlated with the cellular Ca2+ and cAMP-levels . The results show that F-, at concentrations as retained in saliva, did not activate the oxidative burst in unstimulated PMNs . In fMLP-activated PMNs, F-suppressed the receptor-mediated increase in the oxidative burst and the liberation of fl-glucuronidase by reduction of the availability of extracellular Ca2+ and, thus, the influx of Ca2+ necessary to couple completely the fMLP signal to effector pathways . These F- concentrations neither altered the liberation of Ca2+ from internal stores nor induced a rise in cAMP . The possible clinical consequences of these results for xerostomic patients with respect to the generation of HOSCN/OSCN/SCN in saliva an important non-immune factor for oral health, are dicussed.

Drug Dev Ind Pharm, 1999 Aug, 25(8), 937 - 44
Design and biopharmaceutical evaluation of nitrofurantoin-loaded Eudragit RS100 micropellets; Bedi S et al.; Nitrofurantoin, a synthetic bactericidal drug, was encapsulated with Eudragit RS 100 polymer by a coacervation phase separation technique using variable proportions of polyisobutylene (0% to 3%) as a protective colloid . The micropellets were evaluated by scanning electron microscopy (SEM), particle size distribution, wall thickness, and loss of wall polymer were determined . The in vitro release experiments were carried out over the entire pH range of the gastrointestinal tract, the data obtained from the dissolution profiles were compared in the light of different kinetic models, and the regression coefficients were compared . The in vivo studies were performed on female human volunteers . A linear correlation was obtained from in vitro-in vivo studies.

J Clin Invest, 1999 Aug, 104(3), 327 - 35
Role of the tyrosine kinase pyk2 in the integrin-dependent activation of human neutrophils by TNF; Fuortes M et al.; Secretion of inflammatory products from neutrophils can be induced by a combination of signals from ligated integrins and receptors for soluble, physiological agonists such as TNF . Here we identify pyk2 in primary human neutrophils; localize it to focal adhesions and podosomes; and demonstrate its tyrosine phosphorylation, activation, and association with paxillin during stimulation of adherent cells by TNF . Tyrphostin A9 emerged as the most potent and selective of 51 tyrosine kinase inhibitors tested against the TNF-induced respiratory burst . Tyrphostin A9 inhibited TNF-induced tyrosine phosphorylation of pyk2 without blocking the cells' bactericidal activity . Wortmannin, an inhibitor of phosphatidylinositol-3-kinase, potently blocked the TNF-induced respiratory burst and selectively inhibited tyrosine phosphorylation of pyk2 . Thus, pyk2 appears to play an essential role in the ability of neutrophils to integrate signals from beta(2) integrins and TNF receptors.

J Trauma, 1999 Jul, 47(1), 136 - 41
In vitro elution of antibiotic from antibiotic-impregnated biodegradable calcium alginate wound dressing; Lin SS et al.; OBJECTIVE: The authors investigated the calcium alginate dressing as a drug-delivery system for the treatment of various surgical infections . METHODS: Cytotoxicity of the calcium alginate dressing to fibroblasts and HeLa cells was evaluated by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MITT) colorimetric assay . The calcium alginate dressing was mixed with vancomycin, and lyophilized or not lyophilized to form two types of antibiotic dressings . The antibiotic dressings were placed in 2 mL of phosphate buffered saline (PBS) or in PBS containing 0.01% calcium ions, and incubated at 37 degrees C . The PBS was changed daily, and the removed solutions were stored at -70 degrees C until the antibiotic concentration in each sample was determined by high performance liquid chromatography assay . RESULTS: The results suggested that the antibiotic dressings present no obvious toxic risk to their use as a drug-delivery system . The concentration of vancomycin in each sample was well above the breakpoint sensitivity concentration (the antibiotic concentration at the transition point between bacterial kill . ing and resistance to the antibiotic) for more than 14 days . The release was most marked during the first 48 hours . The concentration of calcium ions in PBS and the lyophilization of the manufacture process of antibiotic dressings prolonged the antibiotic diffusion duration . The diameter of the sample inhibition zone ranged from 10 to 11 mm, and the relative activity of vancomycin ranged from 62.88% to 92.18% . CONCLUSION: All antibiotic dressings released bactericidal concentrations of the antibiotics in vitro for the period of time needed to treat surgical infections . This study offers a convenient method to meet the specific antibiotic requirement for different patients.

Microbios, 1999, 98(389), 7 - 14
Efficacy of disinfectants and heat against Escherichia coli O157:H7; Oie S et al.; The bactericidal activity of disinfectants and hot water against ten Escherichia coli O157:H7 strains, which were isolated from faeces of patients with enterohaemorrhagic E . coli infection, were evaluated and showed different DNA patterns . After exposure to 0.1% benzalkonium chloride, 0.1% chlorhexidine gluconate containing a nonionic surfactant, and 80% (v/v) ethanol, 99.99% of viable bacterial cells were killed at 20 degrees C within 15 s irrespective of the presence or absence of 0.1% albumin . On the other hand, after exposure to hot water, 99.99% of the bacterial cells were killed within 15 s at 70 degrees C . These results suggest that benzalkonium chloride, chlorhexidine gluconate containing a nonionic surfactant, ethanol, and hot water at 70 degrees C or more are effective for disinfection of E . coli O157:H7 in hospitals.

Clin Exp Immunol, 1999 Jul, 117(1), 183 - 9
Heat treatment of normal human sera reveals antibodies to bactericidal permeability-inducing protein (BPI); Brownlee AA et al.; Heat treatment of normal sera to 56 degrees C for 30 min, a common procedure for the inactivation of viruses, e.g . HIV, reveals the presence of antibodies to neutrophil cytoplasm antigens (ANCA), as detected by indirect immunofluorescence on ethanol-fixed human neutrophils and by antigen-specific ELISA for BPI . Reactivity was not seen to the other common vasculitis-associated antigens proteinase 3 (PR3) or myeloperoxidase (MPO) . The effect of temperature was maximal at 56 degrees C, with substantial antibody demonstrable after only 5 min at this temperature . In experiments using polyethylene glycol (PEG)6000 to remove immune complexes, the effect of heating could be abrogated by preincubation with 8% PEG, which suggested that these anti BPI antibodies might be complexed in sera . After passage of normal plasma over a protein G column, the acid-eluted fraction contained elevated levels of antibodies to BPI but not to other vasculitis-associated antigens such as PR3 or MPO, nor to glomerular basement membrane (GBM), the Goodpasture antigen which is recognized by the pathogenically important human antibodies shown to mediate nephritis in transfer experiments . Moreover the levels of anti-BPI in the IgG fraction could be augmented by preincubation with glycine pH 2.5 for 30 min . This anti-BPI activity could be inhibited by addition of the unbound material from the protein G column and this inhibitory material was not heat-labile at 56 degrees C . The molecular specificity of this autoreactivity was confirmed using recombinant BPI in coincubation experiments and the epitope localized to the C or N terminal moieties by the use of recombinant fusion proteins.

J Med Microbiol, 1999 Jul, 48(7), 637 - 48
Bactericidal activity of rat lung lavage fluid against Bordetella pertussis; Al-Fellah GN et al.; Cell-free lung lavage fluid (LLF) from healthy normal rats killed phase I (wild-type, virulent) Bordetella pertussis at 37 degrees C in vitro . B . parapertussis was also killed by the LLF, but phase IV (avirulent mutant) B . pertussis and some other common bacterial species, including B . bronchiseptica, were not . Transmission electron microscopy of thin sections of the phase I B . pertussis showed extensive structural damage and cell lysis . None of the other mammalian species tested had LLF with bactericidal activity against B . pertussis as high as that of the rat . Rats killed with halothane yielded LLF with higher bactericidal activity than when CO2 was used . Ultracentrifugation of LLF at 55,000 g gave a surfactant (pellet) fraction that had c . 95% of the bactericidal activity and which was biochemically distinct from the 5% of activity in the supernate fraction . Phospholipids and fatty acids appeared to be involved in LLF bactericidal activity, but not complement or lysozyme . Arachidonic acid was the most active of the fatty acids tested . Artificial surfactant, as used in premature infants, had no bactericidal effect on B . pertussis.

Arch Insect Biochem Physiol, 1999, 41(3), 144 - 7
Lung local defense in experimental diabetes mellitus and the effect of 11,20-dihydroxyecdysone in combination with maninil; Najmutdinova DK et al.; Disorders of lung local defensive mechanisms were noted in rats with alloxane-induced diabetes mellitus, which showed itself as neutrophilic infiltration and decrease of macrophagal bactericidal action in the lungs . All this contributed to the development and chronization of bronchopulmonary inflammatory diseases . Use of maninil caused further inhibition of cellular and humoral lung immunity indices . Addition of 11,20-dihydroxyecdysone (turkesterone) to the therapy resulted in positive changes in the lung defense mechanisms . Arch .

Crit Care Med, 1999 Jun, 27(6), 1153 - 8
Efficacy of liposomal antibiotic therapy in a rat infusion model of Escherichia coli peritonitis; Martineau L et al.; OBJECTIVE: To compare the potential therapeutic effect of liposomal vs . free cefoxitin . DESIGN: Randomized, controlled study, using a rat model of peritonitis . SETTING: Government research facility . SUBJECTS: Male Sprague-Dawley rats . INTERVENTIONS: Rats were infused intraperitoneally with 6.5 x 10(8) colony forming units of Escherichia coli over 12 hrs . Animals were then randomized to receive intravenous saline, free cefoxitin, liposomal cefoxitin, or plain liposomes twice daily until they were killed . MEASUREMENTS AND MAIN RESULTS: Free cefoxitin significantly reduced the number of E . coli after 24 hrs compared with saline treatment in both liver and spleen . However, liposomal cefoxitin further decreased the bacterial content by five-fold to ten-fold in these organs . Minimal bactericidal effect was observed in animals injected with plain liposomes . Although administration of liposomal cefoxitin for 7 days further reduced bacterial counts in liver and spleen, there was no apparent beneficial bactericidal effect of free cefoxitin over saline at 7 days . There was approximately a ten-fold reduction in bacterial content in the lungs after 24 hrs in all three treatments, but no further reduction was observed after 7 days . There was no difference in 7-day survival rate in animals treated with plain liposomes or saline (45% vs . 39%) . Although survival tended to increase with free cefoxitin treatment (64%), this outcome was significantly improved with the use of liposomal cefoxitin (82%) . CONCLUSIONS: Liposomal cefoxitin enhanced bacterial killing in liver and spleen in this model of E . coli peritonitis . It also improved survival outcome relative to no treatment but not compared with free cefoxitin.

Arch Biochem Biophys, 1999 Jul 15, 367(2), 311 - 6
Reactive oxygen species are partially involved in the bacteriocidal action of hypochlorous acid; Dukan S et al.; Hypochlorous acid (HOCl) is probably the most widely used disinfectant worldwide and has an important role in inflammatory reaction and in human resistance to infection . However, the nature and mechanisms of its bactericidal activity are still poorly understood . Bacteria challenged aerobically with HOCl concentrations ranging from 9.5 to 76 microM exhibit higher ability to form colonies anaerobically than aerobically . Conversely, aerobic plating greatly increased lethality after an anaerobic HOCl challenge, although anaerobic survival did not depend on whether HOCl exposure was aerobic or anaerobic . Even a short transient exposure to air after anaerobic HOCl challenge reduced anaerobic survival, indicative of immediate deleterious effects of oxygen . Exposure to HOCl can cause lethal DNA damage as judged by the fact that recA sensitivity to HOCl was oxygen dependent . Antioxidant defenses such as reduced glutathione and glucose-6-phosphate dehydrogenase were depleted or inactivated at 10 microM HOCl, while other activities, such as superoxide dismutase, dropped only above 57 microM HOCl . Cumulative deficiencies in superoxide dismutase and glucose-6-phosphate dehydrogenase rendered strains hypersensitive to HOCl . This indicates that part of HOCl toxicity on Escherichia coli is mediated by reactive oxygen species during recovery .

Nat Med, 1999 Jul, 5(7), 788 - 92
Protective effects of C5a blockade in sepsis; Czermak BJ et al.; Sepsis in humans is a difficult condition to treat and is often associated with a high mortality rate . In this study, we induced sepsis in rats using cecal ligation and puncture (CLP) . In rats depleted of the complement factor C3, CLP led to very short survival times (about 4 days) . Of the rats that underwent CLP ('CLP rats') that were C3-intact and treated with preimmune IgG, most (92%) were dead by 7 days . Blood neutrophils from these rats contained on their surfaces the powerful complement activation product C5a . This group had high levels of bacteremia, and their blood neutrophils when stimulated in vitro had greatly reduced production of H2O2, which is known to be essential for the bactericidal function of neutrophils . In contrast, when companion CLP rats were treated with IgG antibody against C5a, survival rates were significantly improved, levels of bacteremia were considerably reduced, and the H2O2 response of blood neutrophils was preserved . Bacterial colony-forming units in spleen and liver were very high in CLP rats treated with preimmune IgG and very low in CLP rats treated with IgG antibody against C5a, similar to values obtained in rats that underwent 'sham' operations (without CLP) . These data indicate that sepsis causes an excessive production of C5a, which compromises the bactericidal function of neutrophils . Thus, C5a may be a useful target for the treatment of sepsis.

Chemotherapy, 1999 Jul-Aug, 45(4), 249 - 52
In vitro bactericidal activities of a new oral cephalosporin, E1100, and morphologic changes on Escherichia coli; Mikamo H et al.; Escherichia coli is one of the most common aerobic bacteria in pelvic inflammatory diseases . Oral cephalosporins have been widely used against those infections . We investigated in vitro morphologic changes induced on E . coli by a new oral cephalosporin, E1100, and its bactericidal activity on this organism . Morphologic changes were observed by electron microscopy . E1100 induced morphologic changes (filamentation) and exerted a bactericidal activity on E . coli . The filamentation induced by E1100 was time and concentration dependent.

Antimicrob Agents Chemother, 1999 Jul, 43(7), 1805 - 7
Efficacy of gatifloxacin in experimental Escherichia coli meningitis; Lutsar I et al.; The effectiveness of gatifloxacin therapy (15 mg/kg every 5 h {q5h}) was compared with that of meropenem (75 mg/kg q5h) and cefotaxime (75 mg/kg q5h) therapy in experimental meningitis caused by a beta-lactamase-producing strain of Escherichia coli . Gatifloxacin therapy was more rapidly bactericidal than cefotaxime but similar to meropenem therapy (bacterial killing rates at 5 h, 0.83 +/- 0.26, 0 . 46 +/- 0.3, and 0.73 +/- 0.17 CFU/ml/h, respectively; P = 0.03 for gatifloxacin versus cefotaxime) . At 10 h, seven of eight animals treated with gatifloxacin had <10 CFU/ml in their cerebrospinal fluid, compared with one of seven treated with cefotaxime therapy (P = 0.01) . Gatifloxacin was at least as effective as currently available antibiotics in this model of E . coli meningitis.

Antimicrob Agents Chemother, 1999 Jul, 43(7), 1638 - 43
Effective treatment of acute and chronic murine tuberculosis with liposome-encapsulated clofazimine; Adams LB et al.; The therapeutic efficacy of liposomal clofazimine (L-CLF) was studied in mice infected with Mycobacterium tuberculosis Erdman . Groups of mice were treated with either free clofazimine (F-CLF), L-CLF, or empty liposomes twice a week for five treatments beginning on day 1 (acute), day 21 (established), or day 90 (chronic) postinfection . One day after the last treatment, the numbers of CFU of M . tuberculosis in the spleen, liver, and lungs were determined . F-CLF at the maximum tolerated dose of 5 mg/kg of body weight was ineffective; however, 10-fold-higher doses of L-CLF demonstrated a dose response with significant CFU reduction in all tissues without any toxic effects . In acutely infected mice, 50 mg of L-CLF/kg reduced CFU 2 to 3 log units in all three organs . In established or chronic infection, treated mice showed no detectable CFU in the spleen or liver and 1- to 2-log-unit reduction in the lungs . A second series of L-CLF treatments cleared M . tuberculosis in all three tissues . L-CLF appears to be bactericidal in the liver and spleen, which remained negative for M . tuberculosis growth for 2 months . Thus, L-CLF could be useful in the treatment of tuberculosis.

FEMS Microbiol Lett, 1999 Jun 15, 175(2), 211 - 6
Inhibitory effect of lipopolysaccharide on apoptotic cell death in macrophages infected with Actinobacillus actinomycetemcomitans; Muro M et al.; We have reported that the macrophage-like cell line J774.1, when infected with the periodontopathic bacterium Actinobacillus actinomycetemcomitans, undergoes apoptosis . In this study, we examined whether stimulation of J774.1 cells with lipopolysaccharide (LPS) before the infection affects the subsequent apoptosis . Cytotoxicity on the LPS-stimulated cells was about half of the unstimulated control cells . DNA fragmentation in the LPS-stimulated cells was also significantly lower than in the control cells, whereas it was increased to a level similar to that of the control cells by addition of a nitric oxide (NO) inhibitor . In addition, significantly smaller numbers of live A . actinomycetemcomitans were recovered from the LPS-stimulated macrophages at 8 h after the infection as compared with the control cells . These findings suggest that the inhibitory effect of LPS on apoptosis results from an enhanced NO-mediated bactericidal activity.

J Surg Res, 1999 Jul, 85(1), 136 - 41
Antiendotoxin agents share molecular homology within their lipopolysaccharide binding domains; Kellogg TA et al.; BACKGROUND: The purpose of this study was to determine whether antiendotoxin agents exhibit molecular homology within their lipopolysaccharide (LPS) binding domains, suggesting a common mechanism of action . We hypothesized that the presence of positively charged basic amino acids or a paucity of negatively charged acidic amino acids, or both, would be a critical characteristic of that portion of the molecule that binds to the highly negatively charged deep core/lipid A (DCLA) region of LPS . MATERIALS AND METHODS: We analyzed the amino acid sequences of the variable light (VL) and heavy (VH) chain complementarity-determining regions (CDRs) of anti-DCLA monoclonal antibodies (mAbs) 1B6, 5A5, and 7C5 and compared them with (1) the CDRs of three irrelevant control mAbs and (2) the LPS binding region of bactericidal permeability-increasing protein (BPI) . We purified and amplified the specific nucleotide sequences of the variable regions using reverse transcriptase polymerase chain reaction . DNA was sequenced by dideoxy termination, and protein sequences were deduced and analyzed . The percentages of acidic, basic, polar, and hydrophobic amino acids within VH and VL chain CDRs were determined . RESULTS: We identified a paucity of negatively charged acidic amino acids exclusively within VL chain CDRs of anti-DCLA mAbs (P < 0.005) . Although increased, the number of positively charged basic residues was not statistically significantly different; neither was the number of polar or hydrophobic amino acids . Conclusions . Our data suggest that the near absence of negatively charged acidic residues is critical for LPS binding . This characteristic appears to reside exclusively in the VL chain CDRs of anti-DCLA mAbs .

Am J Gastroenterol, 1999 Jun, 94(6), 1546 - 50
High-level disinfection of gastrointestinal endoscopes: are current guidelines adequate?
Kovacs BJ, Chen YK, Kettering JD, Aprecio RM, Roy I.
OBJECTIVE: For a germicide to obtain a high level disinfection (HLD) claim, FDA requires demonstration of a 6-log reduction of mycobacterial inoculum under worst case conditions . The purpose of this study was to assess the adequacy of current guidelines for high level disinfection of GI endoscopes using alkaline glutaraldehyde in simulated-use testing . METHODS: Various gastrointestinal endoscopes were contaminated with Mycobacterium chelonae in 46 experiments . Quantitative cultures were obtained from each endoscope channel separately after each step: inoculation, standardized manual cleaning, immersion in 2% glutaraldehyde (Cidex) for 10, 20, or 45 min at room temperature, 70% isopropanol rinse, and drying . RESULTS: Manual cleaning alone achieved a 4-log reduction . After 10 min of glutaraldehyde exposure, but before alcohol rinse, two of 10 experiments failed to achieve a 6-log reduction . However, after alcohol rinse, all 10 experiments achieved HLD . After 20 min of glutaraldehyde exposure, but before alcohol rinse, one of 18 experiments failed to achieve a 6-log reduction . After alcohol rinse, all 18 experiments achieved HLD . After 45 min of glutaraldehyde exposure, but before alcohol rinse, one of 18 experiments failed to achieve a 6-log reduction . After alcohol rinse, all 18 experiments achieved HLD . Thus, if the entire reprocessing protocol including manual cleaning, glutaraldehyde exposure, alcohol rinse, and drying was taken into account, the required 6-log reduction of mycobacteria was achieved with a minimum of 10 min of glutaraldehyde exposure at room temperature . CONCLUSIONS: Current guidelines for high level disinfection using glutaraldehyde are appropriate . Alcohol rinse is a valuable adjunctive step for drying and for its bactericidal effects.

J Exp Med, 1999 Jun 7, 189(11), 1847 - 52
Neutrophil-specific granule deficiency results from a novel mutation with loss of function of the transcription factor CCAAT/enhancer binding protein epsilon; Lekstrom-Himes JA et al.; Neutrophil-specific granule deficiency (SGD) is a rare disorder characterized by recurrent pyogenic infections, defective neutrophil chemotaxis and bactericidal activity, and lack of neutrophil secondary granule proteins . CCAAT/enhancer binding protein (C/EBP)epsilon, a member of the leucine zipper family of transcription factors, is expressed primarily in myeloid cells, and its knockout mouse model possesses distinctive defects, including a lack of neutrophil secondary granule proteins . Sequence analysis of the genomic DNA of a patient with SGD revealed a five-basepair deletion in the second exon of the C/EBPepsilon locus . The predicted frame shift results in a truncation of the 32-kD major C/EBPepsilon isoform, with loss of the dimerization domain, DNA binding region, and transcriptional activity . The multiple functional defects observed in these early neutrophil progenitor cells, a consequence of C/EBPepsilon deficiency, define SGD as a defect in myelopoiesis and establish the requirement for C/EBPepsilon for the promyelocyte-myelocyte transition in myeloid differentiation.

J Lipid Res, 1999 Jun, 40(6), 1123 - 30
Structure and phospholipid transfer activity of human PLTP: analysis by molecular modeling and site-directed mutagenesis; Huuskonen J et al.; The plasma phospholipid transfer protein (PLTP) is an important regulator of high density lipoprotein (HDL) metabolism . We have here, based on sequence alignments of the plasma LPS-binding/lipid transfer protein family and the X-ray structure of the bactericidal/permeability increasing protein (BPI), modeled the structure of PLTP . The model predicts a two-domain architecture with conserved lipid-binding pockets consisting of apolar residues in each domain . By site-directed mutagenesis of selected amino acid residues and transient expression of the protein variants in HeLa cells, the pockets are shown to be essential for PLTP-mediated phospholipid transfer . A solid phase ligand binding assay was used to determine the HDL-binding ability of the mutants . The results suggest that the observed decreases in phospholipid transfer activity of the N-terminal pocket mutants cannot be attributed to altered HDL-binding, but the C-terminal lipid-binding pocket may be involved in the association of PLTP with HDL . Further, the essential structural role of a disulfide bridge between cysteine residues 146 and 185 is demonstrated . The structural model and the mutants characterized here provide powerful tools for the detailed analysis of the mechanisms of PLTP function.

Antisense Nucleic Acid Drug Dev, 1999 Apr, 9(2), 117 - 23
Cytotoxic hammerhead ribozymes; Levitz R et al.; Small catalytic RNA molecules of the hammerhead ribozyme type were found to have cytotoxic effects unrelated to their intended activity . An expression library of ribozyme sequence variants was constructed in a recA-deficient strain of Escherichia coli such that individual library members differed in regions designed to form base pairs with human immunodeficiency virus-1 (HIV-1) tat mRNA . The parental ribozyme and many variants exhibited a bacteriostatic effect . One variant studied in detail was also bactericidal . When its expression was induced, ribozyme-dependent inhibition of bacterial growth was not observed in recA+ or recA+ lexA3 (Ind-) cells, suggesting that the recombination function of the RecA protein, not the absence of the SOS response, is sufficient to alleviate the cytotoxic effect . These data document the need for careful testing for toxic effects during intracellular studies of ribozyme action.

Diagn Microbiol Infect Dis, 1999 Jun, 34(2), 147 - 52
Immunological defense mechanisms in tuberculosis and MAC-infection; Hartmann P et al.; Protective immunity to mycobacterial infections develops in immunocompetent hosts after activation of alpha beta- and gamma delta-T cells in association with the generation of a protection-specific cytokine profile that stimulates the bactericidal potential of the macrophages . The maintenance of a delicate balance between Th1 and Th2 response is decisive for infection control and prevention of exacerbation of disease . Mycobacterial infection in the immunocompromised host is mainly due to the diminished cellular immune function . In addition, nontuberculous mycobacteria isolated from AIDS patients have special virulence factors that promote development of disease by further compromising the function of an already damaged cytokine network.

Clin Chem Lab Med, 1999 Mar, 37(3), 341 - 9
Supplemental immune globulins in sepsis; Werdan K; Intravenous immune globulins are widely used as supplemental treatment of sepsis, septic shock and systemic inflammation in the critically ill, although this indication has at best been validated in part . Likely beneficial mechanisms of action may include the improvement of serum bactericidal activity due to neutralizing and opsonizing immunoglobulin (Ig)G- and IgM-antibodies, as well as stimulation of phagocytosis and neutralization of bacterial endo- and exotoxins; another attractive mode of action may represent immune globulin-mediated modification and specific suppression of proinflammatory cytokine release from endotoxin- and superantigen-activated blood cells . For the "entire group of patients with sepsis and septic shock" a reduction in mortality by intravenous immune globulin could not be documented; however, in the score-based immunoglobulin in sepsis (SBITS)-study with 653 patients included, a moderate improvement in sepsis morbidity and multiple organ dysfunction syndrome was demonstrated . In defined sepsis subgroups, a reduction in mortality by intravenous immune globulin has been seen in individual small, not yet confirmed trials . Finally, the incidence of some severe infections in well characterized "patients at risk" and "operations at risk" is reduced by intravenous immune globulin prophylaxis . Thus, intravenous immune globulin is not a "magic bullet"of sepsis treatment, but it may reduce morbidity and thereby represent a useful piece of stone in the therapeutic mosaic of sepsis treatment.

J Immunol, 1999 Jun 1, 162(11), 6580 - 8
Active sites in complement components C5 and C3 identified by proximity to indels in the C3/4/5 protein family; Low PJ et al.; We recently suggested that sites of length polymorphisms in protein families (indels) might serve as useful guides for locating protein:protein interaction sites . This report describes additional site-specific mutagenesis and synthetic peptide inhibition studies aimed at testing this idea for the paralogous complement C3, C4, and C5 proteins . A series of C5 mutants was constructed by altering the C5 sequence at each of the 27 indels in this protein family . Mutants were expressed in COS cells and were assayed for hemolytic activity and protease sensitivity . Mutants at five indels showed relatively normal expression but substantially reduced sp . act., indicating that the mutations damaged sites important for C5 function . Twenty-three synthetic peptides with C5 sequences and 10 with C3 sequences were also tested for the ability to inhibit C hemolytic activity . Three of the C5 peptides and one of the C3 peptides showed 50% inhibition of both C hemolytic and bactericidal activities at a concentration of 100 microM . In several cases both the mutational and peptide methods implicated the same indel site . Overall, the results suggest that regions important for function of both C3 and C5 lie proximal to residues 150-200 and 1600-1620 in the precursor sequences . Additional sites potentially important for C5 function are near residue 500 in the beta-chain and at two or three sites between the N-terminus of the alpha'-chain and the C5d fragment . One of the latter sites, near residue 865, appears to be important for proteolytic activation of C5.

Sao Paulo Med J, 1998 Nov-Dec, 116(6), 1873 - 8
Chédiak-Higashi syndrome: presentation of seven cases; Carnide EM et al.; CONTEXT: Chediak-Higashi Syndrome (CHS) is a rare autosomal recessive disease characterized by recurrent infections, giant cytoplasmic granules, and oculocutaneous albinism . OBJECTIVE: To describe clinical and laboratory findings from CHS patients . DESIGN: Case report . SETTING: The patients were admitted into the Allergy and Immunology Unit of the Instituto da Crianca, a tertiary public care institution . CASES REPORT: Seven patients had oculocutaneous albinism, recurrent infections and giant cytoplasmic granules in the leukocytes . One patient had low IgG levels and three showed impaired bactericidal activity of neutrophils . Six patients died of infectious complications during the accelerated phase . Therapy included ascorbic acid and antibiotics . Chemotherapy was used for the accelerated phase in two patients . Bone marrow transplantation (BMT) was proposed for one patient . DISCUSSION: The authors emphasize the need for early diagnosis and therapy of CHS . BMT should be indicated before the accelerated phase of the disease has developed.

Curr Opin Pediatr, 1999 Jun, 11(3), 237 - 40
Nutritional support of the pediatric surgical patient; Amii LA et al.; This review discusses the important developments in pediatric surgical nutrition over the past year . Sepsis and total parenteral nutrition-associated cholestasis remain complex problems for patients on total parenteral nutrition . Investigations suggest that total parenteral nutrition may compromise bactericidal activity, increasing the risk of sepsis . Sepsis possibly sensitizes the liver to cholestatic injury . Small volume enteral feeds may restore immune system function . Current research does not support an association between phytosterols in parenteral lipid solutions and total parenteral nutrition-associated cholestasis . Methionine has been identified as a potential hepatotoxin . Ursodeoxycholic acid and S-adenosyl-L-methionine are the most promising treatments of total parenteral nutrition-associated cholestasis . Small bowel transplant is now a reasonable option for patients with irreversible intestinal failure . Patient and graft survival rates have improved with FK-506 (Tacrolimus) immunosuppression . Isolated intestinal grafts have the best survival rate (92% at 1 year) . Most surviving graft recipients are weaned off of total parenteral nutrition . The Cox Proportional Hazard model may help to identify candidates for small bowel transplant . This equation predicts the duration of dependence on total parenteral nutrition . Patients with irreversible intestinal failure can then be referred for early small bowel transplantation.

J Protein Chem, 1999 Feb, 18(2), 193 - 8
Role of cysteine residues in human plasma phospholipid transfer protein; Qu SJ et al.; Phospholipid transfer protein (PLTP) belongs to a family of human plasma lipid transfer proteins that bind to small amphophilic molecules . PLTP contains cysteines at residues 5, 129, 168, and 318 . Bactericidal/permeability-increasing protein, which is a member of the same gene family, contains an essential disulfide bond between Cys135 and Cys175; these residues, which correspond to Cys129 and Cys168 in PLTP, are conserved among all known members of the gene family . To identify the importance of these and the remaining cysteine residues to PLTP secretion and activity, each was replaced by a glycine by site-directed mutagenesis . The mutant as well as wild-type PLTP cDNAs were cloned into the mammalian expression vector pSV.SPORT1, and the PLTP cDNAs were transfected to COS-6 cells for expression . PLTP Cys129 --> Gly and PLTP Cys168 --> Gly were secretion incompetent . Neither PLTP mass nor activity was detectable in cell lysates and culture medium . Relative to wild-type PLTP, PLTP Cys5 --> Gly and PLTP Cys318 --> Gly exhibited similar specific activities but partially impaired PLTP synthesis and secretion . Intracellular PLTP appeared as two bands of 75 and 51 kDa corresponding to reported molecular masses for the glycosylated and nonglycosylated forms . The specific activities of PLTP Cys5 --> Gly and PLTP Cys318 --> Gly were similar in the cell lysates and medium, suggesting that glycosylation does not affect transfer activity.

J Colloid Interface Sci, 1999 Jun 1, 214(1), 106 - 108
Removal of Formaldehyde by Activated Carbons Containing Amino Groups; Tanada S et al.; Formaldehyde has been used for disinfection and antisepsis in hospitals due to its bactericidal action, but it is toxic to humans . Hence, we developed adsorbates for the removal of formaldehyde . The adsorbate was prepared by the amination of an activated carbon surface . The removal efficiency and the adsorption mechanism of formaldehyde onto the aminated activated carbon were studied . The concentrated sulfuric acid and nitric acid treatment introduced nitro groups onto the surface of the activated carbon . The nitro groups were reduced by the reaction of powdered iron and hydrochloric acid to the amino groups . The amount of formaldehyde adsorbed onto the activated carbon increased with the amination of the activated carbon because of the increasing interaction between the surface of the activated carbon and the formaldehyde .

Proc Natl Acad Sci U S A, 1999 May 11, 96(10), 5464 - 9
Plant ribosome recycling factor homologue is a chloroplastic protein and is bactericidal in escherichia coli carrying temperature-sensitive ribosome recycling factor; Rolland N et al.; We have isolated a protein, mature RRFHCP, from chloroplasts of spinach (Spinacia oleracea L.) that shows 46% sequence identity and 66% sequence homology with ribosome recycling factor (RRF) of Escherichia coli . RRF recycles ribosomes through disassembly of the posttermination complex . From the cDNA analysis and from the amino-terminal sequencing of the isolated protein, the mature RRFHCP was deduced to have a Mr of 21,838 with 193 aa . It lacks the 78-aa chloroplast targeting sequence encoded by the RRFHCP cDNA sequence . The RRFHCP synthesized in vitro was imported into isolated chloroplasts with simultaneous conversion to the mature RRFHCP . Transcription of the gene coding for RRFHCP was not dependent on light, yet it was limited mostly to photosynthetic tissues in which only one transcript size was detected . Mature RRFHCP exerted a bactericidal effect on E . coli carrying temperature-sensitive RRF at the permissive temperature whereas wild-type E . coli was not affected.

Eur J Surg, 1999 Mar, 165(3), 253 - 8
Effects of laparotomy, and carbon dioxide and air pneumoperitoneum, on cellular immunity and peritoneal host defences in rats; Daphan CE et al.; OBJECTIVE: To assess the effects of laparotomy, and insufflation of carbon dioxide and air, on the immune system in rats . DESIGN: Randomised laboratory study . SETTING: Teaching hospital, Turkey . ANIMALS: 77 Wistar rats randomly allocated to 2 groups one of which was sensitised with dinitrofluorobenzene (DNFB, n = 43) and one of which was not (n = 34) . INTERVENTIONS: The DNFB group was sensitised and subdivided into control (n = 8), laparotomy alone (n = 7), and insufflation with carbon dioxide (CO2) for 30 and 60 mins (n = 7 in each) or room air for 30 and 60 mins (n = 7 in each) . A week later DNFB was reapplied to the ears . In the group not sensitised with DNFB the animals were subdivided similarly, the corresponding numbers in each group being, 6, 6, 6, 6, 5, and 5 . MAIN OUTCOME MEASURES: Delayed type hypersensitivity (DTH) measured by ear swelling in the DNFB group, and peritoneal bactericidal activity, total free peritoneal cell counts (TPC), and cell types in the non-sensitised group . RESULTS: There were significant differences in the degree of ear swelling in the DNFB group between control and laparotomy groups (p = 0.0001) and between control and both insufflations of air (p = 0.002 and p = 0.0003, respectively) . In the non-sensitised group peritoneal bactericidal activity was significantly increased after 7 hours in the 60 mins air insufflation group (p = 0.04) . At 24 hours there were no differences among the groups . TPC were not affected . The number of peritoneal polymorphonuclear leucocytes (PMN) was significantly higher in the laparotomy alone group than in the control or any of the insufflation groups (p < 0.05) . CONCLUSIONS: Laparotomy and air insufflation depressed cell-mediated immunity . Peritoneal bactericidal activity was affected only after 60 minutes of air insufflation.

Am Surg, 1999 May, 65(5), 411 - 6
Can oral metronidazole substitute parenteral drug therapy in acute appendicitis? A new policy in the management of simple or complicated appendicitis with localized peritonitis: a randomized controlled clinical trial; Banani SA et al.; To demonstrate the efficacy of oral metronidazole (OM) in simple or complicated appendicitis with localized peritonitis, a randomized prospective study was carried out in 1083 patients, ranging in age from 4 to 50 years (mean age, 21.38) . The patients were randomly divided into two groups . The study group (SG) (524 patients) received OM (500 mg for adults, 7-10 mg/kg if less than 15 years) 2-3 hours before operation . The drug was continued 4 to 5 hours after operation, every 8 hours, for three doses if the appendix was mild to severely inflamed . In the case of complicated appendicitis (114 patients), the same dose was given for 3 to 6 days, depending on the absence or presence of pus . Ceftizoxime was administered to the control group (CG) (559 patients) 2 to 3 hours before operation and then postoperatively every 6 hours for three doses if the appendix was mild to severely inflamed . The complicated cases in the CG (120 patients) received a combination of penicillin, chloramphenicol, and gentamicin for 3 to 6 days, depending on the absence or presence of pus . The serum concentration of metronidazole measured in 43 patients was at bactericidal level in 40 (mean +/- SD standard deviation, 10.65 +/- 4.89 microg/mL) . The rate of wound infection was not significantly different in the SG and the CG with the same degree of pathology (3.17% vs 2.96% if uncomplicated; 15.78% vs 14.16% if complicated, respectively) . Pelvic collection occurred in four adults and one child in the CG with perforated appendicitis (4.16%) . The same complication developed in two adults and two children in the SG with perforated appendicitis (3.5%) . All six adults and one of the children in the SG had to be re-explored, whereas the remaining two children responded to conservative management (OM and gentamicin) . In uncomplicated cases, hospital stay and hospital charge were both almost the same in both groups . However, length of hospitalization was nearly 1 day shorter and hospital cost per day was about 30 per cent less in complicated cases in the SG as compared with the CG . Conclusively, OM may not only substitute parenteral antibiotics in acute appendicitis as a prophylactic agent, but it may also be used as a cost-effective drug and is more convenient to the patient.

Tohoku J Exp Med, 1999 Feb, 187(2), 157 - 71
Reduced immune function and malnutrition in the elderly; Kawakami K et al.; An important observation in elderly subjects is their susceptibility to infection associated with a decline in host immune function . Nutrition is also an important factor that influences host defense against infection . We, therefore, evaluated the relationship between nutritional status in 155 healthy subjects ranging in age from 20 to 99 years and various immunological parameters, including the phagocytic and bactericidal activities of neutrophils and monocytes, superoxide production and chemotaxis of neutrophils, lymphocyte subsets, blastoid transformation and serum immunoglobulins . Aging was associated with increased phagocytic activity of neutrophils but not bactericidal activity, superoxide production or chemotaxis of neutrophils . Aging was also associated with a significant decrease in the number of lymphocytes as well as a decline in mature T cells and helper/inducer T cells but with increased numbers of activated T cells, suppressor T cells and natural killer cells . In addition, blastoid transformation in response to phytohemagglutinin (PHA) and concanavalin A (Con A) was significantly reduced in aged subjects . A poor nutritional status was noted in individuals 60 years of age or older . The nutritional status did not influence neutrophil function but correlated significantly with the number of lymphocytes and degree of blastoid formation with PHA and Con A stimulation . Our results suggest that the cell-mediated immunity in elderly subjects is reduced as a result of malnutrition, and that improvement of the nutritional status may enhance the immune function, likely contributing to their successful aging.

Ther Apher, 1998 Feb, 2(1), 3 - 12
Extracorporeal endotoxin removal by polymyxin B immobilized fiber cartridge: designing and antiendotoxin efficacy in the clinical application; Shoji H et al.; We have developed an extracorporeal hemoadsorption cartridge, the PMX cartridge, to eliminate endotoxin from peripheral blood circulation . As an adsorbent, a polymyxin B covalently immobilized fiber (PMX-F) was developed . After the optimization of the condition of immobilization, fixed polymyxin B maintained its ability to adsorb endotoxin and its bactericidal activity . PMX-F could detoxify many kinds of endotoxin in vitro . Fixed polymyxin B was estimated to interact with the lipid A portion of endotoxin . Utilization of fibrous adsorbents enabled us to design the PMX cartridge with a large surface area and low blood pressure drop in the blood flow compartment and to apply it safely to the direct hemoperfusion procedure . In Japan, the PMX cartridge is now being clinically applied as one of the therapeutical interventions for sepsis, septic shock, and septic multiple organ failure . In multicenter clinical studies, the blood endotoxin level has been significantly decreased . Accompanied with elimination of endotoxin, hemodynamic abnormalities such as low blood pressure and low systemic vascular resistance were significantly improved . In more recent multicenter studies, the average number of failed organs; severity of illness score, such as Goris score; and vasopressor dosage were significantly decreased . The PMX cartridge is expected to be effective in the intervention for the treatment of septic shock . Endotoxin may be one of the therapeutical targets for the treatment of sepsis.

Vestn Otorinolaringol, 1999, (2), 48 - 9
{Reasons for the application of medical ozone in the treatment of chronic purulent mesotympanitis}; Shakov VIu et al.; Ozone-oxygen mixture (ozone concentration 600 microg/l) was used in irrigations of the tympanic cavity in 52 patients with chronic purulent mesotympanitis (CPM) . The course of the treatment consisted of 5-7 procedures . 18 CPM controls received standard treatment without the irrigations . The course of the ozone-oxygen irrigations produced good results: ear discharge and tympanic mucosa inflammation stopped in 81% of the irrigated patients, levels of myeloperoxidase which marks inflammation reduced significantly, bactericidal effects of the mixture were observed for all the detected pathogens, antibiotic sensitivity of the bacteria rose . Fast antiinflammatory result of ozone therapy was due not only to its bactericidal effect but also via its antihypoxic and immunomodulating mechanisms.

Clin Diagn Lab Immunol, 1999 May, 6(3), 420 - 4
Gamma interferon treatment of patients with chronic granulomatous disease is associated with augmented production of nitric oxide by polymorphonuclear neutrophils; Ahlin A et al.; Treatment with gamma-interferon (IFN-gamma) is associated with reduced frequency and severity of infections in chronic granulomatous disease (CGD), but the mechanism is unknown . Since the inducible nitric oxide (NO) synthase can be amplified by IFN-gamma in murine macrophages, for example, we hypothesized that IFN-gamma might modulate NO release from polymorphonuclear neutrophils (PMNs) in patients with CGD . Eight patients with CGD and eight healthy controls were studied . Each patient was given either 50 or 100 microg of IFN-gamma per m2 on two consecutive days . The production of NO from N-formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated PMNs was assessed as the NG-monomethyl-L-arginine-inhibitable oxidation of oxyhemoglobin to methemoglobin in the presence of catalase and superoxide dismutase . Prior to IFN-gamma treatment, the PMNs from CGD patients produced 372 +/- 27 (mean +/- standard error of the mean) pmol of NO/10(6) PMNs at 45 min, while the control PMNs produced 343 +/- 44 pmol . On day 1 after IFN-gamma treatment, NO production increased to 132% +/- 25% of that for controls, and on day 3 it reached 360% +/- 37% (P < 0.001) of that for controls . On day 8, the values still remained higher, 280% +/- 78% more than the control values . Likewise, the bactericidal capacity of PMNs increased on day 3 . The present data show that IFN-gamma treatment of CGD patients is associated with an increased production of NO from PMNs when activated by fMLP . Since these PMNs lack the capacity to produce superoxide anions, it is conceivable that this increase in NO release could be instrumental in augmenting host defense.

Kansenshogaku Zasshi, 1999 Mar, 73(3), 239 - 43
{Effect of antibiotics and antibody on phagocytic bactericidal activity of polymorphonuclear neutrophils of Bordetella pertussis}; Muraoka H et al.; The phagocytic bactericidal activity of the polymononucler neutrophils (PMNs) that were collected from healthy volunteer with and without antibody against Bordetella pertussis was investigated . Furthermore, these activity against B . pertussis under observing penicillins or macrolides antibiotics was investigated . Although no efficacy to B . pertussis strain by the PMNs in serum without antibody, but the viable cells of B . pertussis decreased to 1/1,000 1 hr after incubation and was not detected after 4 hrs . In particular, the viable cells of B . pertussis by the PMNs in serum with antibody was markedly reduced when azithromycin was present . These results suggests that the synergistic action of macrolide antibiotics and antibody-mediated phagocytic bactericidal activity on B . pertussis may have clinical relevance.

Hum Reprod, 1999 Apr, 14(4), 1101 - 6
Generation of peroxynitrite and apoptosis in placenta of patients with chorioamnionitis: possible implications in placental abruption; Nakatsuka M et al.; The reaction of nitric oxide (NO) and superoxide results in the formation of peroxynitrite, a potent and relatively long-lived oxidant . In infectious diseases, these molecules are not only bactericidal but also toxic to host cells . Chorioamnionitis is often complicated by premature rupture of membranes and can be associated with placental abruption . These diseases are significant causes of premature low-birth-weight deliveries and consequently the morbidity and mortality of neonates . Lipopolysaccharide, bacterial endotoxin, is known to be elevated in the amniotic fluid of patients with chorioamnionitis . Lipopolysaccharide is known to induce the formation of NO and superoxide . We report here that nitrite/nitrate, stable metabolites of NO, were increased in serum from patients with chorioamnionitis . Immunohistochemical studies demonstrated enhanced expression of inducible NO synthase and formation of nitrotyrosine, a footprint of peroxynitrite, in the placentae from patients with chorioamnionitis and also in patients with placental abruption . Furthermore, apoptotic cell death was also increased in the placentae from patients with both diseases . These results suggest that chorioamnionitis and a portion of placental abruption may share a common cascade of placental injury . Nitric oxide and its metabolities may play an important role in this cascade.

Kansenshogaku Zasshi, 1999 Feb, 73(2), 156 - 62
{A study of human beta-defensin-1 and human beta-defensin-2 in airway mucosal defense}; Hiratsuka T et al.; Human beta-defensin-1 (hBD-1) and human beta-defensin-2 (hBD-2) are new members of the defensin family . In this study, we investigated their gene expressions in the respiratory epithelial surface of the human lung and their bactericidal activities against Escherichia coli . Both hBD-1 and hBD-2 gene transcripts were detected in human lung tissue and cellular component of bronchoalveolar lavage fluid by reverse transcription-polymerase chain reaction . Synthetic hBD-1 and hBD-2 had dose-dependent bactericidal activities against E . coli . The concentration for the 50% colony reduction of hBD-2 was 0.46 nmol/ml, that for HNP-1 2.15 nmol/ml, and hBD-1 99.3 nmol/ml under conditions nearly the same as in human bronchial airway surface liquid . We prepared an antiserum against hBD-2 and established a highly sensitive radioimmunoassay and quantified plasma hBD-2 concentrations in normal subjects and patients with pneumonia . The plasma concentration of hBD-2 in normal individuals was 8.3 +/- 0.9 fmol/ml (mean +/- SE) . The mean hBD-2 plasma concentration for the 12 patients with bacterial pneumonia in the acute stage was 34.2 +/- 3.4 fmol/ml, significantly higher than normal individuals, and returned to the normal range after recovery . The presence of hBD-1 and hBD-2 in the airway tract and their bactericidal activity suggest that they function in innate airway mucosal defense.

J Endod, 1999 Feb, 25(2), 105 - 8
An evaluation of the CO2 laser for endodontic disinfection; Le Goff A et al.; The goal of this study was to evaluate the bactericidal action of the CO2 laser on animal teeth infected with an endodontic bacterial species . After instrumentation, 24 freshly extracted incisors were inoculated with a known concentration of Actinomyces odontolyticus and incubated anaerobically for 18 h . The incisors were separated into three groups: group 1--untreated control teeth; group 2--teeth treated with 3% NaOCl; and group 3--teeth lased with a CO2 laser at 5 W using three successive 9.9 s irradiation periods with 10 s between treatments . For each of the three groups, 60 microliters samples were removed using gel loading capillary pipette tips, and the diluted samples were plated in triplicate on Columbia agar plates . After a 5- to 6-day incubation, the colony-forming units were counted, and the quantitative results were subjected to an analysis of variance . The results of this analysis indicated an average 85% decrease in the colony-forming units in the laser-treated group, compared with the control group . According to Fisher and Scheffe tests, the differences in the averages between the control and laser groups were statistically significant (p < 0.05) . The NaOCl treatment was statistically superior to the CO2 laser treatment.

J Clin Microbiol, 1999 May, 37(5), 1370 - 5
Characterization of nonenterotoxigenic Escherichia coli strains producing F17 fimbriae isolated from diarrheic lambs and goat kids; Cid D et al.; Forty-five ovine and caprine nonenterotoxigenic Escherichia coli strains producing F17-related fimbriae were characterized with respect to the fimbrial structural subunit and adhesin subtypes produced . In addition, several characteristics related to the virulence of strains producing F17 fimbriae were studied . Most of the strains (73%) possessed the f17cA structural subunit gene, whereas the f17aA and f17dA genes were detected only on three (6%) and two (4%) strains, respectively . The f17bA gene was not detected . All but one of these strains possessed the f17G genes of the adhesin subfamily II . The only strain having the f17G gene of subfamily I possessed the structural subunit gene f17dA . Sequencing of the f17A and f17G genes of four selected strains confirmed the association of f17cA and f17dA structural subunit genes with the f17G genes of the adhesin subfamily II . These results indicated that adhesins of the subfamily II are prominent among ovine and caprine isolates and that they are indistinctly associated with the F17 structural subunit subtypes on these field strains . CS31A- and CNF2-related genes were not detected . Most of the strains adhered in vitro to ovine intestinal brush borders (36 of 45) and agglutinated the erythrocytes of different species in the presence of D-mannose (39 of 45) . F17-positive strains produced colicin V (57%) and were resistant to the bactericidal effect of serum (91%) in significantly higher percentages than F17-negative strains (34% produced colicin V, and 66% were serum resistant) . Thus, most of the studied ovine and caprine strains showed phenotypic characteristics of septicemic strains.

J Hepatol, 1999 Mar, 30(3), 413 - 8
Reversible Kupffer cell suppression in biliary obstruction is caused by hydrophobic bile acids; Sung JJ et al.; BACKGROUND/AIM: Biliary obstruction is associated with suppressed Kupffer cell clearance of bacteria and intracellular bactericidal activity of the phagocytes . We studied the superoxide generation of Kupffer cell in biliary obstruction and after incubation with bile acids to elucidate the mechanism of impaired intracellular killing of these phagocytes . METHODS: Kupffer cells were extracted from rats with common bile duct ligation or sham-operation . The extracted cells were tested for superoxide production immediately after extraction, 2 h and 24 h post-extraction . Superoxide generation from Kupffer cells after incubation with five bile acids (cholic acid, taurocholic acid, deoxycholic acid, chenodeoxycholic acid and ursodeoxycholic acid) at two different concentrations (0.1 mM and 1.0 mM) were also studied . Cell viability was monitored by trypan blue exclusion . RESULTS: Kupffer cells extracted from common duct-ligated animals had significantly lower superoxide production (-1.37+/-0.24 nmol O2(-)/10(6) cells) compared to that from sham-operated rats (2.54+/-0.58 nmol O2(-)/10(6) cells) and non-operated rats (2.15+/-0.76 nmol O2(-)/10(6) cells) (p<0.05) . After 2 h of resting in culture medium, these cells recovered significantly in superoxide production to 2.72+/-0.63 nmol O2(-)/10(6) cells (p<0.01) . A dose-related reduction in superoxide production was demonstrated when Kupffer cells were incubated with bile acids . Hydrophobic bile acids (deoxycholic acid and chenodeoxycholic acid) caused more significant suppression than with hydrophilic bile acids (cholic acid, taurocholic acid, ursodeoxycholic acid) . The drop in superoxide production after bile acid treatment was not due to cell death . Washing in Hank's balanced salt solution resulted in partial recovery of Kupffer cell superoxide production . CONCLUSION: High blood levels of hydrophobic bile acids are likely to be the cause of impaired intracellular bactericidal activity of Kupffer cells in biliary obstruction.

Gen Pharmacol, 1999 Feb, 32(2), 185 - 8
Anticancer drugs inhibit induction of NO synthase in rat in vivo; Inagaki R et al.; Nitric oxide (NO), synthesized by inducible NO synthase (iNOS) in immunoreactive cells, plays important roles in their activities such as bactericidal and tumoricidal functions . We examined the distribution of iNOS and evaluated the effects of anticancer drugs, 4'-epi-doxorubicin (EPI-DXR) and mitomycin C (MMC), on iNOS induction by lipopolysaccharide in rats . Ascites cells and bone marrow showed the highest induction of iNOS in the tested organs . Administration of EPI-DXR to rats strongly inhibited iNOS induction in lung, ascites, and bone marrow, but only slightly in liver and spleen . MMC administration inhibited the induction in the most immune reactive organs.

Antimicrob Agents Chemother, 1999 Apr, 43(4), 758 - 62
Recombinant bactericidal/permeability-increasing protein (rBPI21) in combination with sulfadiazine is active against Toxoplasma gondii; Khan AA et al.; The activity of recombinant bactericidal/permeability-increasing protein (rBPI21), alone or in combination with sulfadiazine, on the intracellular replication of Toxoplasma gondii was assessed in vitro and in mice with acute toxoplasmosis . rBPI21 markedly inhibited the intracellular growth of T . gondii in human foreskin fibroblasts (HFFs) . Following 72 h of exposure, the 50% inhibitory concentration of rBPI21 for T . gondii was 2.6 micrograms/ml, whereas only slight cytotoxicity for HFF cells was observed at the concentrations tested . Subsequent mathematical analyses revealed that the combination of rBPI21 with sulfadiazine yielded slight to moderate synergistic effects against T . gondii in vitro . Infection of mice orally with C56 cysts or intraperitoneally (i.p.) with RH tachyzoites resulted in 100% mortality, whereas prolongation of the time to death or significant survival (P = 0.002) was noted for those animals treated with 5 to 20 mg of rBPI21 per kg of body weight per day . Treatment with rBPI21 in combination with sulfadiazine resulted in significant (P = 0.0001) survival of mice infected i.p . with tachyzoites but not of mice infected orally with T . gondii cysts . These results indicate that rBPI21 is active in vitro and in vivo against T . gondii and that its activity is significantly enhanced when it is used in combination with sulfadiazine . To our knowledge, this is the first report of the activity of rBPI21 against a protozoan parasite.

Vet Microbiol, 1999 Feb 12, 64(4), 323 - 32
Relationship between resistance to complement, virulence and outer membrane protein patterns in pathogenic Escherichia coli O2 isolates; Chaffer M et al.; To establish a possible relationship between resistance to complement, virulence and outer membrane protein banding patterns, ten E . coli O2 strains isolated from chickens with colibacillosis were studied for: (1) resistance to the bactericidal effect of complement by a quantitative microtiter method, (2) virulence, as determined by chicken lethality test, and (3) outer membrane protein banding patterns yielded by SDS-polyacrylamide gel electrophoresis . The ten isolates were classified into three groups: (1) Group 1, consisting of four isolates showed: (a) high resistance to complement, (b) high virulence, and (c) different pattern between 35 and 40 kDa with a weak peptide band at 35 kDa . (2) Group 2, consisting of one isolate showed: (a) high resistance to complement, (b) low virulence, and (c) a weak peptide band at 35 kDa . (3) Group 3, consisting of five isolates showed: (a) low resistance to complement, (b) low virulence, and (c) identical OMP pattern between 35 and 40 kDa exhibiting a strong peptide band at 35 kDa . The results suggest that high resistance to complement may be necessary but no sufficient for virulence and that OMP banding patterns may be a marker for virulence.

J Antimicrob Chemother, 1998 Dec, 42(6), 825 - 9
Killing kinetics of intracellular Afipia felis treated with amikacin; Le Pocher H et al.; Afipia felis is a facultative intracellular bacterium which multiplies in macrophages following inhibition of phagosome-lysosome (P-L) fusion . When A . felis-infected cells are incubated for 72 h with various antibiotics, only aminoglycosides are found to be bactericidal . We therefore studied the killing of intracellular A . felis by amikacin, and its relationship with the restoration of P-L fusion . Amikacin reduced the number of A . felis from 8.5 x 10(5) to 3.5 x 102 cfu/mL within 94 h . P-L fusion was restored after 30-40 h of incubation with amikacin . Both mechanisms may participate in the intracellular killing of bacteria.

Antimicrob Agents Chemother, 1999 Mar, 43(3), 661 - 6
Fluoroquinolone action against clinical isolates of Mycobacterium tuberculosis: effects of a C-8 methoxyl group on survival in liquid media and in human macrophages; Zhao BY et al.; When the lethal action of a C-8 methoxyl fluoroquinolone against clinical isolates of Mycobacterium tuberculosis in liquid medium was measured, the compound was found to be three to four times more effective (as determined by measuring the 90% lethal dose) than a C-8-H control fluoroquinolone or ciprofloxacin against cells having a wild-type gyrA (gyrase) gene . Against ciprofloxacin-resistant strains, the C-8 methoxyl group enhanced lethality when alanine was replaced by valine at position 90 of the GyrA protein or when aspartic acid 94 was replaced by glycine, histidine, or tyrosine . During infection of a human macrophage model by wild-type Mycobacterium bovis BCG, the C-8 methoxyl group lowered survival 20- to 100-fold compared with the same concentration of a C-8-H fluoroquinolone . The C-8 methoxyl fluoroquinolone was also more effective than ciprofloxacin against a gyrA Asn94 mutant of M . bovis BCG . In an M . tuberculosis-macrophage system the C-8 methoxyl group improved fluoroquinolone action against both quinolone-susceptible and quinolone-resistant clinical isolates . Thus, a C-8 methoxyl group enhances the bactericidal activity of quinolones with N1-cyclopropyl substitutions; these data encourage further refinement of fluoroquinolones as antituberculosis agents.

Antimicrob Agents Chemother, 1999 Mar, 43(3), 537 - 42
Reduced pyrazinamidase activity and the natural resistance of Mycobacterium kansasii to the antituberculosis drug pyrazinamide; Sun Z et al.; Pyrazinamide (PZA), an analog of nicotinamide, is a prodrug that requires conversion to the bactericidal compound pyrazinoic acid (POA) by the bacterial pyrazinamidase (PZase) activity of nicotinamidase to show activity against Mycobacterium tuberculosis . Mutations leading to a loss of PZase activity cause PZA resistance in M . tuberculosis . M . kansasii is naturally resistant to PZA and has reduced PZase activity along with an apparently detectable nicotinamidase activity . The role of the reduction in PZase activity in the natural PZA resistance of M . kansasii is unknown . The MICs of PZA and POA for M . kansasii were determined to be 500 and 125 micrograms/ml, respectively . Using {14C}PZA and {14C}nicotinamide, we found that M . kansasii had about 5-fold-less PZase activity and about 25-fold-less nicotinamidase activity than M . tuberculosis . The M . kansasii pncA gene was cloned on a 1.8-kb BamHI DNA fragment, using M . avium pncA probe . Sequence analysis showed that the M . kansasii pncA gene encoded a protein with homology to its counterparts from M . tuberculosis (69.9%), M . avium (65.6%), and Escherichia coli (28.5%) . Transformation of naturally PZA-resistant M . bovis BCG with M . kansasii pncA conferred partial PZA susceptibility . Transformation of M . kansasii with M . avium pncA caused functional expression of PZase and high-level susceptibility to PZA, indicating that the natural PZA resistance in M . kansasii results from a reduced PZase activity . Like M . tuberculosis, M . kansasii accumulated POA in the cells at an acidic pH; however, due to its highly active POA efflux pump, the naturally PZA-resistant species M . smegmatis did not . These findings suggest the existence of a weak POA efflux mechanism in M . kansasii.

Wien Klin Wochenschr, 1998 Dec 23, 110(24), 886 - 93
Growth inhibitory and bactericidal efficacy of sera from Lyme borreliosis patients on B . burgdorferi strains; Kraiczy P et al.; Two B . afzelii strains EB1 and FEM1, classified in normal human sera (NHS) as serum-resistant, and an intermediate serum-sensitive B . burgdorferi s.s . strain 297, were tested in regard of their serum sensitivity in immune sera (IS) of patients at all stages of Lyme borreliosis by a growth inhibition assay (GIA) . Fifty-four per cent (13/24) of the tested IS were GIA positive, while the sera of patients in stage III disease inhibited the growth more frequently than did the patients with sera of stage II or stage I disease . Growth inhibition was predominantly directed against strain FEM1 (12/24), less against strain EB1 (4/24) and strain 297 (2/24) . A growth inhibiting effect on two strains was only detectable for two IS and merely one stage III serum inhibited all three strains . Positive results in the GIA required fresh serum and resulted in the killing of the borreliae . The detection of the deposited complement components C3 and C9 on the surfaces of the inhibited strains by means of immunofluorescence assays confirmed the role of complement . In Westernblot analyses of strain FEM1, it was striking that GIA-positive IS reacted 3- to 5-fold more often with proteins of molecular masses of 48.9-, 38.6-, 27.5-, 25-, 23.1- (OspC), 21.7-, and 16-kDa, than did GIA-negative IS . Furthermore, two proteins of approximately 20- and 31.2-kDa reacted exclusively with GIA-positive IS . Antibodies reacting with these proteins could play a role in the growth inhibition of NHS-resistant borrelial strains, OspC.

Shock, 1999 Feb, 11(2), 77 - 81
Postinjury suppression of human neutrophil cytokine production results from the stabilization of inhibitory kappaB; Zallen G et al.; Postinjury neutrophil (PMN) dysfunction is a well recognized event that may be responsible for increased infections . PMN cytokine production is an important component of their bactericidal capacity . When PMNs are stimulated, inhibitory factor kappaB (IkappaB) is degraded, allowing nuclear factor kappaB (NFkappaB) to translocate to the nucleus and promotes genes for the transcription of the interleukin-8 (IL-8) and tumor necrosis factor (TNF) genes . We hypothesize that similar to their late postinjury depressed superoxide production, postinjury PMNs manifest suppressed cytokine production, which is mediated by stabilization of IkappaB levels . METHODS: Twelve severely injured patients with an injury severity score (ISS) of 24 (+/-4.6) were studied as well as 10 elective surgical patients as a control . PMNs were isolated and incubated for 24 h in RPMI . PMNs were stimulated with lipopolysaccharide (LPS; 100 ng) or PAF (200 nm) and fMLP (1 microM) and release of IL-8, TNF, and interleukin-1 receptor antagonist (IL-1ra) were measured . Postinjury PMNs were also stimulated with LPS (100 ng), and IkappaB breakdown was measured at 0, 30, and 60 min using gel electrophoresis . RESULTS: Postinjury PMNs displayed a significant suppression of both IL-8 and TNF on postinjury Days 1-3, while the release of IL-1ra was preserved throughout the entire study period . In contrast, elective surgical patients demonstrated no decrease in IL-8 or TNF . Furthermore, IkappaB levels were preserved in the postinjury PMNs as compared with normal control PMNs . CONCLUSION: Postinjury PMNs have a suppressed release of both IL-8 and TNF following injury that did not occur in elective surgical patients . Furthermore, the NFkappaB/IkappaB-independent IL-1ra did not show suppression of release . In addition, stabilization of IkappaB following severe injury leads to decreased PMN IL-8 and TNF production . This genetic reprogramming may help explain PMN dysfunction and subsequent infections seen in severely injured patients.

Infect Immun, 1999 Mar, 67(3), 1424 - 31
Characterization of human bactericidal antibodies to Bordetella pertussis; Weiss AA et al.; The Bordetella pertussis BrkA protein protects against the bactericidal activity of complement and antibody; however, some individuals mount an immune response that overcomes this bacterial defense . To further characterize this process, the bactericidal activities of sera from 13 adults with different modes of exposure to B . pertussis (infected as adults, occupational exposure, immunized with an acellular vaccine, or no identified exposure) against a wild-type strain and a BrkA complement-sensitive mutant were evaluated . All of the sera killed the BrkA mutant, suggesting past exposure to B . pertussis or cross-reactive organisms . Several samples had no or minimal activity against the wild type . All of the sera collected from the infected and occupationally exposed individuals but not all of the sera from vaccinated individuals had bactericidal activity against the wild-type strain, suggesting that some types of exposure can induce an immune response that can overcome the BrkA resistance mechanism . Adsorbing serum with the wild-type strain removed the bactericidal antibodies; however, adsorbing the serum with a lipopolysaccharide (LPS) mutant or an avirulent (bvg mutant) strain did not always result in loss of bactericidal activity, suggesting that antibodies to either LPS or bvg-regulated proteins could be bactericidal . All the samples, including those that lacked bactericidal activity, contained antibodies that recognized the LPS of B . pertussis . Bactericidal activity correlated best with the presence of the immunoglobulin G3 (IgG3) antibodies to LPS, the IgG subtype that is most effective at fixing complement.

Infect Immun, 1999 Mar, 67(3), 1310 - 6
The levels and bactericidal capacity of antibodies directed against the UspA1 and UspA2 outer membrane proteins of Moraxella (Branhamella) catarrhalis in adults and children; Chen D et al.; The UspA1 and UspA2 proteins from Moraxella catarrhalis share antigenic epitopes and are promising vaccine candidates . In this study, the levels and bactericidal activities of antibodies in sera from healthy adults and children toward UspA1 and UspA2 from the O35E strain were measured . Human sera contained antibodies to both proteins, and the levels of immunoglobulin G (IgG) antibodies were age dependent . Adult sera had significantly higher titers of IgG than child sera (P < 0.01) . The IgG3 titers to the UspA proteins were higher than the IgG1 titers in the adults' sera, while the IgG1 titers were higher than the IgG3 titers in the children's sera (P < 0.05) . The IgG antibodies in the sera from 2-month-old children appeared to be maternally derived, since the mean titer was significantly higher than that in sera from 6- to 7-month-old children (P < 0.05) . Serum IgA antibodies to both UspA1 and UspA2 were low during the first 7 months of age but thereafter gradually increased along with the IgG titers . Analysis of sera absorbed with UspA1 or UspA2 showed that the antibodies to UspA1 and UspA2 were cross-reactive with each other and associated with serum bactericidal activity . Examination of affinity-purified human antibodies confirmed that naturally acquired antibodies to UspA1 and UspA2 were bactericidal and cross-reactive . These results support using UspA1 and UspA2 in a vaccine to prevent M . catarrhalis infections.

Rocz Akad Med Bialymst, 1998, 43, 292 - 7
Nitroblue tetrazolium test in patients with renal tumor in the course of embolization; Mantur M et al.; The main role of neutrophilic granulocytes is phagocytosis and destruction of phagocytized bacteria . The nitroblue tetrazolium test (NBT), used to evaluate the bactericidal function of granulocytes, was assessed in 45 patients with renal tumour before and 14 days after renal artery embolization . No statistically significant spontaneous test NBT differences were found, compared with control group both before and 14 days after embolization . Following stimulation granulocytes in vitro by LPS of E . coli the ability of granulocytes to reduce NBT was lowered . The differences, as compared with control group, were statistically significant (p < 0.05) . Our results indicates considerable impairment of lower metabolic reserve in patients with renal tumour necessary for maximum stimulation.

Oftalmologia, 1998, 42(2), 9 - 12
{The synthesis and effects of nitric oxide at the retinal level}; Cruce M et al.; Nitric oxide (NO) is synthetized from L-arginine by the aid of an enzyme--NO synthase (NOS) . This enzyme has there isoforms, which are either constitutive; neural NOS (nNOS) and endothelial NOS (eNOS) or inducible (iNOS) . These three isoforms are expressed also in the retina . NO produced in small amounts by nNOS and eNOS is involved in neurotransmission in the retina and in the regulation of retinal arteriolar tonicity . NO produced in large quantities by iNOS is a bactericidal agent but can also generate inflammation of the retina and even retinal degeneration.

Scand J Gastroenterol, 1998 Dec, 33(12), 1284 - 8
Recognition of bactericidal/permeability-increasing protein by perinuclear anti-neutrophil cytoplasmic antibody-positive sera from ulcerative colitis patients: prevalence and clinical significance; Vecchi M et al.; BACKGROUND: The aim of this study was to evaluate a) the role of bactericidal/permeability-increasing protein (BPI) as a possible antigen determining perinuclear anti-neutrophil cytoplasmic antibody (p-ANCA) reactivity in ulcerative colitis and b) the prevalence and clinical correlates of anti-BPI antibodies in patients with ulcerative colitis on the basis of their p-ANCA status . METHODS: p-ANCA and anti-BPI antibodies were evaluated by means of indirect immunofluorescence and enzyme-linked immunosorbent assay methods in a group of 112 ulcerative colitis patients (including 42 patients subjected to proctocolectomy) well defined as far as their clinical features and p-ANCA status . RESULTS: Anti-BPI antibodies were detected in 24% of non-operated patients and were significantly more frequent in p-ANCA-positive patients (32% versus 5% in p-ANCA-negative patients; P < 0.015) . The prevalence of anti-BPI antibodies was similar in non-operated and operated patients and was high in men, in patients with an extensive and aggressive disease, and in patients developing pouchitis after surgery . CONCLUSIONS: These data indicate that BPI is a neutrophil antigen frequently recognized by p-ANCA-positive ulcerative colitis sera . The presence of anti-BPI antibodies appears to identify further immunologic and clinical heterogeneity in ulcerative colitis.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1999 Jan, 87(1), 83 - 7
Bactericidal effect of electrolyzed neutral water on bacteria isolated from infected root canals; Horiba N et al.; OBJECTIVE: The purposes of this study were to examine the time-related changes in pH, oxidation-reduction potential, and concentration of chlorine of electrolyzed neutral water and to evaluate the bactericidal effect of electrolyzed neutral water against bacteria from infected root canals . STUDY DESIGN: Various properties of electrolyzed neutral water--pH value, oxidation-reduction potential, and concentration of chlorine--were measured at different times after storage of the water in the open state, the closed state, or the closed-and-dark state . The bactericidal effect of the various electrolyzed neutral water samples was then tested against 17 strains of bacteria, including 15 strains isolated from infected canals, as well as against 1 strain of fungus . Each bacterial or fungal suspension was mixed with electrolyzed neutral water, and the 2 substances were reacted together for 1 minute . After incubation for 1 to 7 days, the bactericidal effect of the electrolyzed neutral water was determined . RESULTS: The pH value and oxidation-reduction potential of electrolyzed neutral water remained almost unchanged when the water was stored in a dark, closed container . However, the concentration of chlorine decreased from 18.4 ppm to 10.6 ppm . Electrolyzed neutral water showed a bactericidal or growth-inhibitory effect against the bacteria . CONCLUSIONS: The results indicate that electrolyzed neutral water maintains a constant pH and oxidation-reduction potential when kept in a closed container without light and that it exhibits a bacteriostatic/bactericidal action against isolates obtained from infected root canals.

Rev Mal Respir, 1998 Dec, 15(6), 699 - 711
{Ozone and the immune system}; Lacroix G et al.; This review focuses on the effect on health of changes in the immune system secondary to ozone exposure and on various mechanistic hypotheses put forward . Beyond the problems related to the variability of study criteria (e.g . age, sex, concentration and duration of different types of exposure, the slightly volatile nature of ozone and the complexity of the immune system), ozone may induce immunostimulation as shown by intensified allergic phenomena or immunosuppression expressed by increased sensitivity to bacterial infections . Different functions of the immune response (for example macrophage and polynuclear phagocytic and bactericidal activity, NK activity, cytokine and antibody production ...) are affected . In terms of risk, the consequences of these changes depend on their intensity, their perennial nature and their association with particular genetic characteristics or other forms of external aggression, for example infection . The effect of exposure to a mixture of pollutants with unknown interactions should also be taken into consideration . Finally, the problem of normal but possibly exaggerated immune response to a compound whose allergenicity may have been modified by ozone must also be taken into account.

Int J Antimicrob Agents, 1998 Nov, 10(4), 285 - 9
Bactericidal activity of lansoprazole and three macrolides against Helicobacter pylori strains tested by the time-kill kinetic method; Fera MT et al.; The bactericidal activities of macrolides (clarithromycin, roxithromycin and azithromicyn) and lansoprazole, alone and in combination, against Helicobacter pylori strains were evaluated . It was found that the association of lansoprazole and clarithromycin resulted in a marked synergism, while the combination of roxithromycin or azithromycin with lansoprazole had synergistic and additive effects.

Pancreas, 1999 Jan, 18(1), 21 - 7
Bactericidal/permeability-increasing protein and group I and II phospholipase A2 during the induction phase of human acute pancreatitis; Kemppainen E et al.; Activated endogenous mediators of inflammation have important roles in the pathogenesis and complications of acute pancreatitis (AP) . These mediators include bactericidal/ permeability-increasing protein (BPI) and phospholipase A2 (PLA2) . The time course of their activation during human AP is not known . The aim of this study was to evaluate the kinetics of BPI, group I (pancreatic) and group II (synovial type) PLA2 during human AP with temporally defined onset, as being induced by endoscopic retrograde cholangiopancreatography (ERCP) . Serum samples of 273 consecutive patients undergoing ERCP were collected before and at 3, 6, and 24 h after ERCP . Twenty-four (8.7%) patients developed ERCP-induced pancreatitis . Seven of them were graded to have a severe disease . Forty randomly selected patients undergoing ERCP without evidence of pancreatitis served as controls . The serum concentrations of BPI and groups I and II PLA2 were measured by specific immunoassays . The mean concentration of BPI increased from 14 to 26 microg/L at 24 h after ERCP in patients with AP . In the control group, BPI values remained unchanged, and the difference was statistically significant (p<0.001) . The increase of BPI was seen in 22 of 28 patients with AP at 3 h after the onset of the disease . BPI values were higher in severe post-ERCP pancreatitis than in mild disease (p = 0.07; NS) . The serum concentrations of group II PLA2 before ERCP were consistently higher in the control patients than in the patients with pancreatitis, 65.8 and 14.2 microg/L, respectively . High baseline values in the control group were associated with preexisting infectious diseases . Thereafter, the mean concentration decreased in the control group to 44 microg/L and increased in the pancreatitis group up to 27.5 microg/L . The difference was statistically significant (p = 0.007) . Increased group II PLA2 values were seen in 10 of 17 patients with mild AP and in five of seven patients with severe disease . There were no significant differences in group I or II PLA2 values in patients with mild or severe AP . The serum concentration of group I PLA2 increased in the patients with post-ERCP pancreatitis from 5.4 to 37.5 microg/L at 24 h . The difference was statistically significant, (p< 0.001) as compared with controls . In conclusion, in acute pancreatitis, the increase of BPI in serum starts at 3 h after the onset of the disease, and the concentration seems to correlate with the severity of the disease . Increased group II PLA2 concentrations also were seen in patients with mild AP . The kinetics of group I PLA2 resembles that of other pancreatic enzymes.

Blood Cells Mol Dis, 1998 Dec, 24(4), 544 - 51
Leukocyte function in chronic myeloproliferative disorders; Wolach B et al.; The myeloproliferative disorders (MPD) are clonal diseases that originate from a transformed stem cell and involve all myeloid lineage . The affected cells have both proliferative and functional impairment . Therefore, we evaluated and compared neutrophil function in 31 patients with polycythemia vera (PV), idiopathic myelofibrosis (MF), chronic myeloid leukemia (CML), and essential thrombocytosis (ET) . Neutrophil chemotaxis, random migration, bactericidal activity and superoxide anion release in these patients were simultaneously compared to those of 31 healthy controls . In this study, chemotactic activity was significantly impaired in patients with PV and CML as compared to controls (M+/-SE: 42 +/- 6 vs . 69+/- 5 cells/field; p<0.005 and 47+/-7 vs . 68+/- 5; p<0.05, respectively) . The assessment of the bactericidal activity of neutrophils showed no impairment in most of the patients . In the CML group, the serum had a very strong "lytic" effect on bacteria, possibly due to the high levels of serum lysozyme (22 +/- 2 microgram/ml) . The superoxide anion release was found to be normal in most of the patients . Nevertheless, in 25% of PV patients the superoxide production was impaired (less than 60% of the simultaneous controls) . In ET most patients had normal neutrophil function . Regarding the effect of treatment, neutrophil chemotactic activity was found to be significantly reduced in the hydrea-treated patients, as compared to the non- treated patients (p<0.001) or healthy controls (<0.0001) . We conclude that disturbances in neutrophil function are present in patients with various MPDs, except ET . This probably reflects abnormal maturation of ancessors of the damaged stem cells . Nevertheless, we should keep in mind that therapy itself could affect neutrophil functions . This matter should be studied more extensively . Although infections are not common in MPD disorders, they occasionally occur . It is possible that impairment in the phagocytic function contribute to the development of infections in patients with myeloproliferative disorders.

Diagn Microbiol Infect Dis, 1998 Nov, 32(3), 229 - 35
Comparison of various in vitro susceptibility methods for testing Stenotrophomonas maltophilia; Carroll KC et al.; A total of 57 clinical isolates were screened by disk diffusion for a related pharmacodynamic study . Testing was performed using National Committee for Clinical Laboratory Standards guidelines, except that results were interpreted at 16 to 18 h and 48 h . Of the 57 isolates, 19 were randomly chosen for additional comparative susceptibility testing of five methods (disk diffusion, Etest, Alamar colorimetric broth microdilution, Vitek, and MicroScan) and an in-house broth microdilution method . The two diffusion methods (disk and Etest) had the closest correlation . The commercial broth microdilution methods and the in-house microdilution method generated inconsistent results for all agents except trimethoprim-sulfamethoxazole . Vitek compared poorly with both diffusion and microbroth dilution methods . The most significant discrepancies were evident with all methods when the incubation period was extended to 48 h . When results were interpreted at 48 h, the incidence of resistance for all bactericidal agents was approximately double the resistance observed at 16 to 18 h . The bacteriostatic agents, trimethoprim-sulfamethoxazole and doxycycline, demonstrated the greatest in vitro activity and were least influenced by extended incubation with diffusion methods . Because correlative in vivo and in vitro studies have not revealed an effective therapeutic regimen for serious S . maltophilia infections, susceptibility results with all testing methods should be interpreted with caution when choosing therapy for patients with life-threatening infections . Susceptibility testing for this heterogeneous group remains controversial and routine testing, with the possible exception of doxycycline (or minocycline) and trimethoprim-sulfamethoxazole, should be avoided . Our data support that if testing is done with bactericidal agents, consideration should be given to interpretation after 48-h incubation.

J Comput Aided Mol Des, 1998 Nov, 12(6), 543 - 56
Patenting computer-designed peptides; Patel S et al.; The problem of designing new peptides that possess specific properties, such as bactericidal activity, is of wide interest . Recently, attention has focused on the use of Computer-Aided Molecular Design techniques in parallel with more traditional 'synthesise and test' methods . These techniques may typically use Genetic Algorithms to optimise molecules based on Neural Network models that predict activity . In this paper we describe a successful application of this Molecular Design methodology that has resulted in novel bactericidal peptides of real value . A key issue for commercial utilisation of such results is the ability to protect the intellectual property rights associated with the discovery of new molecules . Typically peptide patents use structural templates of amino acid hydrophobicity-hydrophilicity that define highly regular peptide patent spaces . In an extension of established patenting practice we describe a patent application that uses a Neural Net predictive model to define the regions of peptide space that we claim within the patent . This formalism makes no a priori assumptions about the regularity of the patent space . A preliminary comparative investigation of the shape and size of this and other bactericidal peptide patent spaces is conducted.

J Pharm Pharmacol, 1998 Nov, 50(11), 1317 - 20
Rapid killing of Mycobacterium terrae by N-chlorotaurine in the presence of ammonium is caused by the reaction product monochloramine; Nagl M et al.; We have studied the activity of the weak endogenous oxidant N-chlorotaurine against Mycobacterium terrae . The study revealed slow killing of more than 2h duration by 1% (55 mM) N-chlorotaurine . In the presence of ammonium chloride, however, killing times decreased to a few minutes, even by 0.1% N-chlorotaurine . This phenomenon is explained by formation of the lipophilic and therefore more bactericidal monochloramine as a result of transhalogenation of ammonia by N-chlorotaurine.

Eur J Pediatr, 1998 Dec, 157(12), 987 - 91
Antineutrophil cytoplasmic antibodies in children; Sediva A et al.; To define the diagnostic meaning of antineutrophil cytoplasmic antibodies (ANCA) positivity in children, we analysed 1485 consecutive sera sent for routine immunological investigation to our department from January to August 1996 . Using this large screening, we identified the most typical clinical disorders associated with ANCA in childhood . Out of 1485 indirect immunofluorescence (IIF) tests for ANCA, 143 were ANCA positive, 70 had a cytoplasmic IIF pattern (c-ANCA), and 73 a perinuclear IIF pattern (p-ANCA) . The ANCA associated diseases in childhood were cystic fibrosis (CF) (31 c-ANCA, 7 p-ANCA positive out of 71 CF children investigated), juvenile chronic arthritis (JCA) (21 p-ANCA positive out of 78), auto-immune hepatitis (AIH) (4 c-ANCA and 12 p-ANCA positive out of 19), and ulcerative colitis (UC) (2 c-ANCA, 5 p-ANCA positive out of 15) . In cases of c-ANCA positivity we determined the antigenic specificity of ANCA for proteinase 3 and/or bactericidal/permeability increasing protein . Borderline anti-proteinase 3 levels were found in CF, and in high levels in one boy with Wegener granulomatosis . Bactericidal/permeability increasing protein was characteristic target antigen in children with CF . p-ANCA positive children were further tested for the specificity for myeloperoxidase, which was detected mostly in children with JCA . CONCLUSION: The spectrum of diseases associated with ANCA in children includes, besides the diagnostic associations typical for adults, several typical pediatric entities, mainly juvenile chronic arthritis and cystic fibrosis.

Arch Pharm Res, 1998 Jun, 21(3), 326 - 9
Studies on the synthesis of naphthoquinoids; Park OS et al.; Four derivatives of 6-oxo-3,4,4a,5-tetrahydro-3-hydroxy-2,2-dimethylnaphtho-1,2-pyran (1), known as bactericidal, bacteriostatic, fungicidal, fungistatic agents, were synthesized to investigate the effect of substituents on the aromatic ring.

J Osaka Dent Univ, 1998 Apr, 32(1), 9 - 15
Effect of antibiotics on rat leukocyte function; Miyata T et al.; Many antibiotics are used to treat infection in clinical practice . Actions of these drugs involve specific immune enhancement and improved overall phagocytic and bactericidal capacities of polymorphonuclear leukocytes and macrophages at the site of infection . In this study, we examined the effects of antibiotics on chemotaxis and phagocytosis by reacting macrophages with 7 different antibiotics, specifically ampicillin (ABPC), cephalexin (CEX), cefotiam (CTM), amikacin (AMK), clindamycin (CLDM), tetracycline (TC) and bleomycin (BLM) . At 1 microgram/ml, there were significant differences in chemotaxis between control and experimental groups treated with agents other than ABPC and CEX (CLDM: p < 0.05, AMK: p < 0.01, CTM, TC and BLM: p < 0.001) . At 10 micrograms/ml, there were significant differences in chemotaxis between the control group and all treated groups (ABPC and CLDM: p < 0.01, CEX, CTM, AMK, TC and BLM: p < 0.001) . At 100 micrograms/ml, all antibiotics significantly inhibited chemotaxis (p < 0.001) . Phagocytosis was evaluated by determining both the ratio and index of phagocytosis . There was a significant difference in the phagocytosis ratio between the control group and the group treated with 10 micrograms/ml of BLM (p < 0.001) . At 100 micrograms/ml, all agents but ABPC significantly reduced phagocytosis in a dose-dependent manner . Agents other than ABPC similarly reduced the phagocytosis index . Significant differences were noted between the control group and groups treated with BLM or AMK (BLM: 10 micrograms/ml: p < 0.01, 100 micrograms/ml: p < 0.001, AMK: 100 micrograms/ml: p < 0.01).

Lett Appl Microbiol, 1998 Dec, 27(6), 328 - 30
Bactericidal activity of catechin-copper (II) complexes on Escherichia coli ATCC11775 in the absence of hydrogen peroxide; Kimura T et al.; Washed Escherichia coli ATCC11775 cells were killed by (-)-epigallocatechin (EGC) in the presence of a non-lethal concentration of Cu2+ (1 mumol l-1) without additional H2O2, but not by (-)-epicatechin (EC) . EGC alone (< 0.1 mmol l-1) did not reduce the viability of the cells . The survival curve obtained in the presence of EGC and Cu2+ was similar to that obtained in the presence of (-)-adrenaline (EN) and Cu2+.

Am J Med, 1998 Dec, 105(6), 484 - 7
Clinical and immunological study of 7 patients with minocycline-induced autoimmune phenomena; Elkayam O et al.; PURPOSE: Prolonged treatment with minocycline for acne vulgaris has been associated with the development of arthralgia, arthritis, and other autoimmune phenomena . We characterized the clinical, laboratory, and immunological profiles of seven patients with this syndrome . SUBJECTS AND METHODS: Clinically the patients were studied with special emphasis on prior minocycline treatment, presenting symptoms, physical findings, course, and outcome . Laboratory tests included fluorescent antinuclear and antineutrophil cytoplasmic (ANCA) antibodies, as well as antibodies to myeloperoxidase, bactericidal permeability increasing protein, elastase, cathepsin G, lactoferrin, cardiolipin, and histone . RESULTS: All 7 patients presented with polyarthritis or arthralgia, morning stiffness, and fever after 6 to 36 months of minocycline treatment . The skin was involved in five patients (three with livedo reticularis and two with subcutaneous nodules) . Two patients had chronic active hepatitis . Increased titers of perinuclear ANCA (p-ANCA) were detected in all seven patients; five patients had fluorescent antinuclear antibodies, two had antihistone autoantibodies and one had anticardiolipin antibodies . Antigenic characterization of p-ANCA disclosed antibodies to bactericidal permeability increasing protein in one patient, to elastase in three patients, and to cathepsin G in five patients . Symptoms resolved in five patients upon discontinuation of minocycline; the other two patients were treated with corticosteroids and also achieved remissions . CONCLUSION: Minocycline-induced autoimmune syndrome is characterized by reversible polyarthralgia or arthritis, morning stiffness, fever, frequent skin involvement, occasional chronic active hepatitis, and increased titers of p-ANCA with various minor p-ANCA-related antigens.

Blood, 1999 Jan 1, 93(1), 357 - 62
The use of allele-specific recombinant Fc gamma receptor IIIb antigens for the detection of granulocyte antibodies; Bux J et al.; The Fcgamma receptor IIIb (FcgammaRIIIb) for the Fc domain of IgG is expressed exclusively on neutrophils . The FcgammaRIIIb bears allotypic polymorphisms referred to as NA1, NA2, and SH, which are known for their frequent involvement in alloimmune and autoimmune neutropenias as well as in transfusion reactions . The bactericidal capacity of isolated neutrophils is easily activatable, and activation results in self-desintegration, thus preventing storage of neutrophils . As a result, only freshly isolated granulocytes can be used for antibody screening, often making it impossible to use typed panel cells . To provide a readily available source of typed panel cells, we therefore established stable mammalian cells expressing recombinant NA1, NA2, and SH antigens . We isolated mRNA from typed neutrophils and then transcribed it in cDNA . The cDNA that codes for the different forms of the FcgammaRIIIb was amplified by polymerase chain reaction and was subsequently subcloned into the mammalian expression vector pcDNA3 . Chinese hamster ovary (CHO) cells were transfected with allele-specific constructs, and stable cell lines expressing FcgammaRIIIb were selected by flow cytometry . Because human sera show high background fluorescence with transfectants in flow cytometry, the monoclonal antibody-specific isolation of granulocyte antigens (MAIGA) assay was performed . By MAIGA assay, we tested 14 well-characterized human alloantibodies directed against the antigens NA1, NA2, and SH; 5 FcgammaRIIIb-specific isoantibodies; and 12 FcgammaRIIIb-reactive autoantibodies . Except one NA1- and one SH-specific alloantibody, all other antibodies could be identified by the use of CHO transfectants . In contrast to neutrophils, fixed CHO cells can be stored at 4 degrees C for at least 4 weeks or stored frozen for a longer period . This longer shelf life of the transfected CHO cells compared with isolated neutrophils will simplify the detection of the clinically most important FcgammaRIIIb-reactive alloantibodies and autoantibodies.

Proc Natl Acad Sci U S A, 1998 Dec 22, 95(26), 15688 - 93
A mAb recognizing a surface antigen of Mycobacterium tuberculosis enhances host survival; Teitelbaum R et al.; Murine mAbs reactive with the surface of Mycobacterium tuberculosis were assayed for their ability to affect the course of infection in mice challenged with virulent organisms . An IgG3 mAb (9d8) specific for arabinomannan and reactive with purified antigen from a clinical isolate of M . tuberculosis conferred partial protection on mice after respiratory challenge (30-60% survival >75 days; P </= 0.05) . Control mice pretreated with an irrelevant mAb of the same isotype succumbed to tuberculosis within 30 days . Mice with gene disruptions in interferon gamma and major histocompatibility complex Class II also were partially protected from challenge . The protective mAb was neither bactericidal nor inhibitory of infection or bacterial replication . Nevertheless, it profoundly altered the nature of the granulomas in the infected lungs . Mice treated with mAb 9d8 and challenged with M . tuberculosis localized the pathogen within granuloma centers, suggesting that the mAb conferred protection by enhancing a cellular immune response.

Ann Agric Environ Med, 1998, 5(2), 109 - 16
Toxicity of dermally applied alpha-cypermethrin in rats; Luty S et al.; The aim of the study was to assess the immunotoxic effect of dermally applied alpha-cypermethrin in rats based on phagocytic and bactericidal activity of neutrophils of peripheral blood, and the general toxic effect based on histological and ultrastructural examination of internal organs . The preparation was dermally applied in doses of 50 mg/kg and 250 mg/kg . It was administered to the tail skin of female Wistar rats, 4 hours daily for 28 days . After the experiment, the animals were anaesthetized and heart blood was taken in order to evaluate the activity of granulocyte system . The following organs were taken for histological examinations: brain, lung, heart, liver, spleen, kidneys, thymus and lymphatic nodes . Lung, liver, kidney and heart were used for ultrastructural studies . The results of the study showed that bactericidal and phagocytic activity of neutrophils was stimulated after administration of 50 mg/kg alpha-cypermethrin . Dermal application of the preparation resulted in slight histological changes in liver, kidney, lung and brain . Pathological changes in heart were observed only on the level of ultrastructure.

J Clin Immunol, 1998 Nov, 18(6), 414 - 20
Minimally invasive surgery induces endotoxin-tolerance in the absence of detectable endotoxemia; Lemaire LC et al.; Lipopolysaccharide (LPS) tolerance is characterized by an impaired proinflammatory cytokine production upon restimulation of mononuclear cells with LPS . LPS is considered the primary activator for this phenomenon . In response to major injury and extensive abdominal surgery, an immune reaction comparable to LPS tolerance has been described . Therefore, it was investigated whether primary stimuli other than LPS could induce cytokine downregulation . In eight patients who underwent a laparoscopic cholecystectomy, blood was obtained before and after induction of anaesthesia and 2, 6, and 24 hr postoperatively . Ex vivo stimulation of whole blood resulted in a transient reduction (nadir 2 hr postoperatively) of tumor necrosis factor-alpha, interleukin (IL)-1 beta, and interferon-gamma release, while IL-1 receptor antagonist production increased . Stress hormones, LPS-binding protein, and bactericidal/permeability-increasing protein do not seem to be involved . This study shows that minimally invasive surgery, in the absence of endotoxemia, can induce LPS desensitization . These data suggest that prior endotoxemia is not essential for the development of LPS tolerance.

Ann Agric Environ Med, 1998, 5(1), 57 - 64
Toxicity of dermally absorbed dichlorvos in rats; Luty S et al.; Toxicity of dermally absorbed dichlorvos was studied in rats, based on its effects on internal organs, and on phagocytic and bactericidal activity of the neutrophile system . The studies were conducted on 30 female rats of Wistar strain . The animals were divided into three groups, two of which were experimentally exposed to dermal absorption of dichlorvos (37.5 mg/kg - 1/2 LD50; or 7.5 mg/kg - 1/10 LD50;), and one control group which was exposed to dermal absorption of the solvent . The animals were exposed to dermal absorption for 4 hours daily for a period of 4 weeks . After 28 days, the rats were anaesthetized, blood was drawn from the heart to evaluate the activity of the neutrophilic system, and the internal organs excised for histological and ultrastructural studies . Dermally absorbed dichlorvos caused histopathological changes in lungs, lymphatic glands and thymus, as well as histopathological and ultrastructural changes in liver, kidneys and heart muscle . Dichlorvos stimulated the bactericidal and phagocytic activity of neutrophils.

Cancer Lett, 1998 Sep 25, 131(2), 209 - 14
Antitumour activity of diallyl sulfide on polycyclic aromatic hydrocarbon-induced mouse skin carcinogenesis; Singh A et al.; Diallyl sulfide (DAS), a major flavour component of garlic, is known to modulate xenobiotic metabolism and possess antitoxic, bactericidal, antineoplastic, hypolipidemic and hypocholesteromic effects . In the present study, the anticarcinogenic activity of DAS on a 7,12-dimethylbenzanthracene (DMBA)- or benzo{a}pyrene (B(a)P)-induced mouse skin model of carcinogenesis was evaluated . DAS was applied topically either 1 h prior to or 1 h after the administration of DMBA or B(a)P . A significant protection from neoplasia was observed in DAS- and DMBA/B(a)P-exposed animals when DAS was applied topically compared to the animals exposed only to DMBA/B(a)P . In the animals where DAS was applied 1 h prior to the application of DMBA, a lower magnitude of neoplasia was recorded in terms of the cumulative number of tumours and average number of tumours per mouse during the entire period of study (28 weeks) compared to the animals exposed to DAS 1 h later, while in B(a)P-exposed animals, the antitumorigenic potential of DAS was more evident in the mice treated with DAS 1 h after the B(a)P exposure compared to the animals treated with DAS 1 h prior to B(a)P . The antitumour activity of DAS was of a much higher magnitude in B(a)P-induced carcinogenesis in comparison to animals exposed to DMBA in terms of tumour incidence, cumulative number of tumours and average number of tumours per mouse . The results suggest that DAS has a protective effect in PAH-induced mouse skin carcinogenesis.

J Leukoc Biol, 1998 Dec, 64(6), 817 - 27
Platelet-activating factor induces a concentration-dependent spectrum of functional responses in bovine neutrophils; Swain SD et al.; We characterized the dose response of bovine neutrophils to platelet-activating factor (PAF) with respect to the following functions: calcium flux and membrane potential changes, actin polymerization, degranulation, and the production and/or priming of the oxidative burst . PAF at very low concentrations (10(-10) and 10(-9) M) caused changes in intracellular calcium and membrane potential in bovine neutrophils, whereas moderate PAF concentrations (> or = 10(-7) M) resulted in increased actin polymerization . Degranulation responses to PAF were more complex: low concentrations (10(-9) M) caused secretory granule degranulation, moderate doses (> or = 10(-7) M) caused specific granule degranulation, whereas azurophil degranulation only occurred at high (10(-5) M) PAF concentrations . Treatment of bovine neutrophils with PAF at concentrations > or = 10(-7) M also caused up-regulation of the adhesion molecules Mac-l and L-selectin . PAF stimulation resulted in a very weak {compared to phorbol myristate acetate (PMA)} oxidative burst in bovine neutrophils, and only at high (10(-6) M) concentrations . Unlike human neutrophils, bovine neutrophils were poorly primed by PAF treatment . Only high concentrations of PAF (10(-5) M) caused an increased rate of PMA-stimulated superoxide production, although lower doses of PAF did reduce the lag time preceding the PMA-induced oxidative burst . The overall pattern that can be inferred is that lower concentrations of PAF promote neutrophil sensitivity and interaction by selective degranulation, up-regulation of adhesion molecules, and increased actin polymerization . In contrast, higher PAF concentrations can promote, albeit weakly, more direct bactericidal responses, such as the release of reactive oxygen species and granule enzymes . The ability of PAF to modulate a graded response in bovine neutrophils would allow the cell to respond proportionally to the severity of a stimulus.

Int J Immunopharmacol, 1998 Nov, 20(11), 615 - 24
A comparative investigation of the restorative effects of roxithromycin on neutrophil activities; Noma T et al.; The effects of roxithromycin, a macrolide antibiotic, on neutrophil activities were investigated in six seriously handicapped patients with severe mental retardation . Neutrophil activities were evaluated by flow cytometry using a heparinized blood analysis method . All six patients showed decreased levels of neutrophil phagocytosis, intracellular killing, and CD11b expression . Treatment with roxithromycin in vitro selectively restored the decreased phagocytic and bactericidal activities of neutrophils in these patients . There was no significant restorative effect with cefaclor, ofloxacin, or aztreonam . These results suggest the need to consider therapeutic effects of antibiotics on neutrophil functions in patients at increased risk for bacterial infections due to decreased neutrophil activities.

FEBS Lett, 1998 Nov 20, 439(3), 329 - 33
Inactivation of bacterial respiratory chain enzymes by singlet oxygen; Tatsuzawa H et al.; To distinguish the bactericidal action of singlet oxygen (1O2) from hypohalous acids, wild-type and lycopene transformant E . coli strains were exposed to each of the oxidants and then bacterial viability was investigated . 1O2 was generated by chemical and enzymatic systems at pH 4.5 . ExpoSure of wild-type E . coli to 1O2 caused a significant loss of E . coli viability due to inactivation of membrane respiratory chain enzymes by 1O2 . This action of 1O2 could be attenuated by lycopene in the bacterial cell membrane . In the lycopene transformant strain of E . coli, inactivation of NADH oxidase and succinate oxidase by hypohalous acids were significantly suppressed, but E . coli viability was unaffected . Based on these findings, we suggest that phagocytic leukocytes produce 1O2 as a major bactericidal oxidant in the phagosome.

Clin Exp Immunol, 1998 Dec, 114(3), 462 - 7
A mixed population of immature and mature leucocytes in umbilical cord blood results in a reduced expression and function of CR3 (CD11b/CD18); Reddy RK et al.; Neonatal neutrophils express less membrane and cytoplasmic CR3 (iC3b-receptor, Mac-1, alphaM beta2-integrin) than do adult neutrophils, and it has been suggested that this renders neonatal neutrophils deficient in diapedesis and bactericidal activity . The reason(s) for this deficiency are unknown . In this study, CR3 expression and the CR3-dependent respiratory burst activity of individual neonatal neutrophils are quantified in comparison with adult leucocytes using flow cytometry . Monocytes and neutrophils are defined as CD14highCD15low and CD14lowCD15high, respectively . Although neonatal neutrophils bore less CR3 on average than did adult neutrophils, neonatal neutrophils were more heterogeneous and many neonatal neutrophils expressed adult levels of CR3 . Because of higher neutrophil concentrations in cord versus adult blood, the calculated number of neutrophils in cord blood expressing high amounts of CR3 was equivalent to that of adult blood . Similar findings were made with monocytes . The size of the CR3-dependent respiratory burst stimulated by particulate beta-glucan correlated directly with the expression of CR3 by individual neutrophils . With neonatal and adult neutrophils having comparable CR3 densities, the respiratory burst activities were equivalent . Wright-Giemsa differential staining of the subset of neonatal neutrophils with low CR3 levels isolated by fluorescence-activated cell sorting showed a higher proportion of immature cells than the sorted population expressing high CR3 levels . Therefore, higher proportions of immature cells in cord blood probably explain previous reports of deficient CR3 expression and function . The typical neutrophilia of cord blood may compensate for this apparent deficiency by providing adult concentrations of mature neutrophils.

J Dairy Sci, 1998 Nov, 81(11), 2841 - 9
Properties of a heparin-binding peptide derived from bovine lactoferrin; Shimazaki K et al.; A heparin-binding peptide was isolated from a proteolytic hydrolysate of bovine lactoferrin by affinity chromatography using an immobilized heparin column . Analysis of amino acid sequences at the N-terminus showed that this heparin-binding peptide is derived from the region beginning at the 17th amino acid residue of the bovine lactoferrin sequence . The molecular mass of this peptide was 3195.5 as measured by matrix-assisted laser desorption-time of flight mass spectrometry . This peptide is the same as the bactericidal peptide lactoferricin B . In an aqueous environment, this peptide displays mainly a beta-sheet structure and an unordered structure as assessed by measurements of circular dichroism spectra . When this peptide was mixed with heparin, a distinct spectral change was induced because of conformational alteration of the peptide . This spectral change was reversible . Analysis of data from peptide synthesis indicated that binding by the sequence Arg28-Met29-Lys30-Lys31 of bovine lactoferrin is significant and that there is a synergistic contribution from Lys18-Cys19-Arg20-Arg21, and Arg38-Arg39.

Am J Infect Control, 1998 Dec, 26(6), 572 - 7
Wound management in an era of increasing bacterial antibiotic resistance: a role for topical silver treatment; Wright JB et al.; BACKGROUND: Antibiotic-resistant bacteria represent an increasing concern in wound infections . Wound colonization with these organisms normally results in aggressive management of the wound complicated by a greatly limited choice of therapeutic antibiotics . Silver and other noble metals are recognized as potential allies in combating these organisms in wounds . METHODS: Three types of topical silver applications were tested to determine their bactericidal efficacies against clinical isolates of antibiotic-resistant organisms . The silver-based applications represent 3 methods of applying silver to wounds: as a liquid (silver nitrate), incorporated in a cream (silver sulfadiazine) and as a dressing coating (silver-coated dressings) . The reduction in the viable bacterial population recovered from test articles after exposure to silver provided a comparative measure of the bactericidal efficacies of these silver applications . RESULTS: All of the products demonstrated an ability to reduce the number of viable bacteria . However, the methods varied in their efficacy against antibiotic-resistant bacteria, with the silver-coated dressing being the most efficacious and silver nitrate the least efficacious . CONCLUSIONS: Silver was demonstrated to be effective at killing the antibiotic-resistant strains tested . The silver-coated dressing was particularly rapid at killing the tested bacteria and was effective against a broader range of bacteria . Silver may be a useful prophylactic or therapeutic agent for the prevention of wound colonization by organisms that impede healing, including antibiotic-resistant bacteria.

Appl Environ Microbiol, 1998 Dec, 64(12), 5057 - 60
The bactericidal activity of pediocin PA-1 is specifically inhibited by a 15-mer fragment that spans the bacteriocin from the center toward the C terminus; Fimland G et al.; A 15-mer peptide fragment derived from pediocin PA-1 (from residue 20 to residue 34) specifically inhibited the bactericidal activity of pediocin PA-1 . The fragment did not inhibit the pediocin-like bacteriocins sakacin P, leucocin A, and curvacin A to nearly the same extent as it inhibited pediocin PA-1 . Enterocin A, however, was also significantly inhibited by this fragment, although not as greatly as pediocin PA-1 . This is consistent with the fact that enterocin A contains the longest continuous sequence identical to that of pediocin PA-1 in the region spanned by the fragment . The fragment inhibited pediocin PA-1 to a much greater extent than did the other 29 possible 15-mer fragments that span pediocin PA-1 . The results suggest that the fragment-by interacting with the target cells and/or pediocin PA-1-interferes specifically with pediocin-target cell interaction.

Am J Med, 1998 Nov, 105(5), 393 - 9
Antineutrophil cytoplasmic antibodies in primary sclerosing cholangitis: defined specificities may be associated with distinct clinical features; Roozendaal C et al.; PURPOSE: The clinical significance of antineutrophil cytoplasmic autoantibodies (ANCA) in primary sclerosing cholangitis has not been established . We investigated whether analysis of the antigenic specificities of ANCA is useful for delineating clinical subsets of the disease . METHODS: Sixty-nine patients with primary sclerosing cholangitis were studied . The presence of ANCA was analyzed by indirect immunofluorescence . Antibodies directed against specific antigens--proteinase 3, myeloperoxidase, elastase, bactericidal/permeability-increasing protein, cathepsin G, and lactoferrin--were identified by enzyme-linked immunosorbent assay . RESULTS: ANCA were detected by indirect immunofluorescence in 46 (67%) patients . In antigen-specific enzyme-linked immunosorbent assays, 37 (55%) of the 69 patients had antibodies to one or more antigens: 32 (46%) had antibodies to bactericidal/permeability-increasing protein, 16 (23%) had antibodies to cathepsin G, and 15 (22%) had antibodies to lactoferrin . Only 3 patients had antibodies to proteinase 3 . Antibodies to myeloperoxidase or elastase were not detected . Twenty (29%) patients had antibodies to different antigens simultaneously . ANCA as detected by indirect immunofluorescence were not significantly associated with the presence of cirrhosis nor with the coexistence of inflammatory bowel disease . However, antibodies to bactericidal/permeability-increasing protein and cathepsin G were both associated with the presence of cirrhosis, and antibodies to lactoferrin were more frequently detected in patients with primary sclerosing cholangitis in conjunction with ulcerative colitis than in those without inflammatory bowel disease . CONCLUSION: Defined specificities of ANCA in primary sclerosing cholangitis may be related to particular clinical features of the disease.

Nephrol Dial Transplant, 1998 Nov, 13(11), 2821 - 4
Antineutrophil cytoplasmic autoantibodies (ANCA) and their target antigens in Chinese patients with lupus nephritis; Zhao MH et al.; BACKGROUND: ANCA have been found in patients with systemic lupus erythematosus (SLE); however, the prevalence of ANCA and their target antigens is still not certain . This study is to investigate the prevalence of ANCA and their target antigens in Chinese patients with lupus nephritis . METHODS: Ninety-five serum samples were collected from 95 renal-biopsy-proven lupus nephritis patients . Indirect immunofluorescence using ethanol-fixed leukocytes as substrate and ELISA using six highly purified known ANCA antigens as solid-phase ligands were performed . The specific ANCA antigens included proteinase 3, myeloperoxidase, bactericidal/permeability-increasing protein, human leukocyte elastase, cathepsin G, and lactoferrin . The prevalence of ANCA in patients with (n=65) and without (n=30) active renal pathological lesions was also compared to reveal whether ANCA correlates with disease activity . RESULTS: (i) None of the sera recognized proteinase 3, myeloperoxidase, and human leukocyte elastase, and only one serum recognized bactericidal/permeability-increasing protein . The striking finding was that 59/95 (62.1%) sera recognized cathepsin G and the titres of some sera reached 1/3200 . Eight of 95 sera (8.4%) recognized lactoferrin . (ii) The percentage of anti-cathepsin G antibody positive samples in patients with active renal lesions was significantly higher than in patients without active lesions (73.4 vs 36.7%, P<0.0001), whereas, anti-lactoferrin antibodies had no correlation with active renal lesions . (iii) By indirect immunofluorescence, only 22% of the 95 sera were ANCA positive . CONCLUSIONS: Our results suggest that the majority of lupus nephritis patients have ANCA and that the major target antigens is cathepsin G . Anti-cathepsin G antibodies seem to be correlated with renal disease activity.

J Clin Apheresis, 1998, 13(3), 103 - 7
Bacterial decontamination of blood stem cell apheresis products; Gagnon JA et al.; High-dose chemotherapy using autologous bone marrow or mobilized blood as the source of stem cells for haematologic rescue, is being widely used for a variety of haematological malignancies and solid tumours . To collect sufficient numbers of haematopoietic stem cells for successful engraftment, standard apheresis procedures are performed . Newer techniques and refinements of the procedure allow using only 1 to 2 apheresis products (AP) for autografting . Bacterial contamination of the AP, although very rare, sometimes occurs and may lead to generalized infection in the recipient . The apheresis must be repeated, sometimes even including time-consuming and costly mobilization . At our institution, the patients' blood stem cells are usually mobilized with chemotherapy followed by daily s.c . haematopoietic growth factor injections or with growth factor alone . An apheresis machine is used for collection through a central venous line and the AP is routinely checked for bacterial contamination . Results are only available after the product has been processed and cryopreserved . In the last 5 years, we observed bacterial contamination in four of our AP . Therefore, we investigated the possibility of in vitro antibiotic decontamination . Using standard antibiograms, we determined the sensitivities of the contaminating bacteria . By incubating the products with the specific antibiotics at bactericidal concentrations, we were able to sterilize the probes from the contaminating bacteria . In the concurrently performed controls without the active substance, bacteria were still detectable . We conclude that in selected cases, in vitro decontamination using pretested antibiotics, may be a feasible, cost-effective, and easy alternative to performing additional apheresis procedures.

Biochem Biophys Res Commun, 1998 Nov 18, 252(2), 492 - 6
Affinities of different proteins and peptides for lipopolysaccharide as determined by biosensor technology; de Haas CJ et al.; Biosensor technology was employed to study the specific interactions of different lipopolysaccharide (LPS)-binding proteins and peptides with LPS, using an LPS-coated surface . Two methods to immobilize biotinylated LPS to streptavidin-coated sensor chips (SA-chips) were evaluated . Biotinylated LPS in PBS or biotinylated LPS, pretreated with EDTA and sodium-desoxycholate, were injected across an SA-chip, resulting in a 'high-' and 'low- mass' LPS chip, respectively . While the 'high mass' LPS chip appeared to be unstable, the 'low mass' LPS chip resulted in reproducible binding curves for bactericidal/permeability-increasing protein (rBPI21) with a binding affinity corresponding to the literature (Kd: 3.75 nM) . New Kd values were obtained for serum amyloid P component (SAP, Kd: 3.9 nM), a recently discovered new LPS-binding protein, and cationic protein 18 (CAP18, Kd: 0.58 nM) . Moreover, binding affinities of bioactive BPI- and SAP-derived peptides could be determined . This study shows for the first time the applicability of biosensor technology to study interactions of proteins and peptides with LPS, using an LPS-coated sensor chip .

Biol Pharm Bull, 1998 Oct, 21(10), 1057 - 61
Bactericidal action of 4,4'-(alpha,omega-polymethylenedithio)bis-(1-alkylpyridinium iodide)s; Maeda T et al.; Bactericidal action of novel bis-quaternary ammonium compounds (bis-QACs), 4,4'-(alpha,omega-polymethylenedithio)bis(1-alkylpyridinium iodide)s (4DTBP-m,n) was studied . The bactericidal activity of 4DTBP-m,n in water was not affected by the molecular hydrophobicity unlike general mono-QAC, N-dodecylpyridinium iodide (P-12), while the bacteriostatic activity in the medium was reduced with their hydrophobicity . This result suggested that the hydrophobic materials in the medium interact with 4DTBP-m,n and cover their active moiety . Since the bactericidal activity using the measurement system supplemented with peptone was influenced by the molecular hydrophobicity, this speculation was supported . The plots of the bacteriostatic activities of 4DTBP-m,n against the surface hydrophobicities of various bacteria accord to the straight line as in the case of P-12 . The slope of the line of 4DTBP-6,12 was comparatively smaller than that of 4DTBP-6,8, indicating that the compounds having longer alkyl group tend to reduce their activities against the bacteria with hydrophobic cell surface because of their interaction with the hydrophobic materials . The novel bis-QACs have an ability to liberate rapidly and abundantly the turbid materials from cells, that is, a bacterioclastic activity . The bacterioclastic activity of P-n was influenced by the length of alkyl group, while 4DTBP-6,n had almost the same activity regardless of its length . Observation by scanning electron microscope (SEM) revealed that 4DTBP-6,8 fatally damaged Escherichia coli cells, and that the morphological alteration of the cells caused by the bis-QAC was greatly different from that of the usual QAC . Therefore, the effective bacterioclastic action, and excellent bactericidal action is due to the unique dimeric structure of 4DTBP-m,n.

Antimicrob Agents Chemother, 1998 Nov, 42(11), 2978 - 84
Fluoroquinolone action against mycobacteria: effects of C-8 substituents on growth, survival, and resistance; Dong Y et al.; Fluoroquinolones trap gyrase on DNA as bacteriostatic complexes from which lethal DNA breaks are released . Substituents at the C-8 position increase activities of N-1-cyclopropyl fluoroquinolones against several bacterial species . In the present study, a C-8-methoxyl group improved bacteriostatic action against gyrA (gyrase-resistant) strains of Mycobacterium tuberculosis and M . bovis BCG . It also enhanced lethal action against gyrase mutants of M . bovis BCG . When cultures of M . smegmatis, M . bovis BCG, and M . tuberculosis were challenged with a C-8-methoxyl fluoroquinolone, no resistant mutant was recovered under conditions in which more than 1, 000 mutants were obtained with a C-8-H control . A C-8-bromo substituent also increased bacteriostatic and lethal activities against a gyrA mutant of M . bovis BCG . When lethal activity was normalized to bacteriostatic activity, the C-8-methoxyl compound was more bactericidal than its C-8-H control, while the C-8-bromo fluoroquinolone was not . The C-8-methoxyl compound was also found to be more effective than the C-8-bromo fluoroquinolone at reducing selection of resistant mutants when each was compared to a C-8-H control over a broad concentration range . These data indicate that a C-8-methoxyl substituent, which facilitates attack of first-step gyrase mutants, may help make fluoroquinolones effective antituberculosis agents.

J Immunol, 1998 Nov 1, 161(9), 4983 - 91
Fusion of azurophil granules with phagosomes and activation of the tyrosine kinase Hck are specifically inhibited during phagocytosis of mycobacteria by human neutrophils; N'Diaye EN et al.; Pathogenic mycobacteria parasitize macrophages and reside within phagosomes, which do not fuse with lysosomal granules . Mycobacteria are also internalized by neutrophils, which possess at least two types of granules, specific and azurophil granules, the latter being specialized lysosomes . Here, we investigated the ability of mycobacteria to inhibit the fusion of these granules with their phagosomes in human neutrophils . It was found that when pathogenic (Mycobacterium kansasii and Mycobacterium avium) or nonpathogenic (Mycobacterium smegmatis and Mycobacterium phlei) mycobacteria were internalized by neutrophils, they induced the inhibition of azurophil granule fusion with phagosomes even when they were serum opsonized . In contrast, secretion of specific granule content and production of O2-, both of which contribute to the neutrophil bactericidal response, were triggered . Hck is a Src family tyrosine kinase associated with azurophil granules . During internalization of zymosan, azurophil granules fused with phagosomes and Hck was activated and translocated to the phagosomal membrane, whereas in neutrophils engulfing mycobacteria, Hck did not translocate and remained unactivated . The activation of the tyrosine kinase Fgr was not affected . These results indicate that 1) pathogenic and nonpathogenic mycobacteria trigger similar bactericidal responses in neutrophils, 2) phagocytosis and fusion of azurophil granules can be uncoupled by mycobacteria, and 3) Hck could be one of the key elements of the azurophil secretory pathway that are altered during phagocytosis of mycobacteria.

Diagn Microbiol Infect Dis, 1998 Sep, 32(1), 39 - 44
The bactericidal activity of clarithromycin versus ampicillin alone and in combination with omeprazole and/or bismuth against clarithromycin-susceptible and clarithromycin-resistant strains of Helicobacter pylori; Coudron PE et al.; We evaluated the in vitro bactericidal effect of clarithromycin versus ampicillin alone and in combination against clarithromycin-sensitive and clarithromycin-resistant strains of Helicobacter pylori . No combination containing clarithromycin achieved complete bactericidal effect against clarithromycin-resistant strains . Complete bactericidal effect was achieved for all strains by triple-agent combinations that contained bismuth, omeprazole, and relatively high concentrations of ampicillin . These in vitro results demonstrate the additive bactericidal activity provided by a combination of agents for the eradication of H . pylori . Bismuth may play a particularly important role in the bactericidal effect.

Nippon Geka Gakkai Zasshi, 1998 Aug, 99(8), 523 - 7
{Sepsis and organ failure--its pathogenesis and treatment}; Hanasawa K et al.; Sepsis, septic shock, and multiple organ dysfunction are heterogeneous and sophisticated clinical syndromes which result from the interplay of mediators of cellular function and inflammation . Secondary mediators such as lipids (prostaglandin, thromboxane, platelet-activating factor), peptides (bradykinin, vasoactive intestinal peptide), amines (histamine, serotonin) and complements are key mediators which lead to the state of shock in human sepsis . Endotoxin may also cause multiple organ dysfunction syndrome (MODS) . New antiendotoxin treatment and the strategy for sepsis including endotoxemia are reviewed . Monoclonal antibodies directed at core epitopes and lipid A (E5, HA1-A) could not reproduce the beneficial effects . Bactericidal/permeability increasing protein (BPI)) . Endotoxin neutralizing protein (ENP)) and E5531 may have potential in the treatment of sepsis . Another treatment using extracorporeal endotoxin removal is reported . Polymyxin B immobilized fiber (PMX), commercialized as Toraymyxin, is now widely used in Japan for severe sepsis and septic MODS . PMX treatment improves the symptoms related to the septic state, a hemodynamic disorders, and cytokine levels including tumor mecrosis factor, interleukin (IL)-6, and IL-10, with a decrease in endotoxin levels . Phase II, III, and IV clinical trials with extracorporeal endotoxin removal by PMX revealed that endotoxin removal is helpful in the treatment of septic patients.

Shock, 1998 Oct, 10(4), 278 - 84
Growth hormone and insulin-like growth factor I augment bactericidal capacity of human polymorphonuclear neutrophils; Inoue T et al.; Effects of growth hormone (GH) and insulin-like growth factor (IGF)-I on bactericidal capacity of human polymorphonuclear neutrophils (PMNs) were investigated . Venous blood was collected from healthy volunteers . In Experiment 1, PMNs were isolated, incubated with GH or IGF-I, and cocultured with Escherichia coli . E . coli-killing capacity, viability, and CD11b and CD16 expressions of PMNs were then assessed . Both GH and IGF-I enhanced E . coli killing by PMNs . GH preserved PMN viability during E . coli killing, whereas IGF-I enhanced PMN CD11b expression before coculture with E . coli . In Experiment 2, whole blood was washed and incubated with GH or IGF-I . PMNs in washed whole blood were then analyzed for phorbol myristate acetate (PMA)-stimulated CD11b, CD35, and CD16 expressions and production of reactive oxygen intermediates (ROI), as well as phagocytosis with/without anti-CD11b antibody . IGF-I enhanced PMN expressions of CD11b and CD35, but not CD16, stimulated with PMA . Both hormones enhanced phagocytosis, which was abrogated by anti-CD11b antibody, and intracellular ROI production by PMNs . These results indicate that both GH and IGF-I augment human PMN bactericidal capacity, via increased phagocytosis and intracellular ROI production . Preservation of PMN viability by GH and enhanced complement receptor expression by IGF-I may also be associated with augmented PMN bactericidal capacity . Although PMN activation has potentially harmful aspects, these results encourage additional studies to confirm the clinical relevance of exogenous GH or IGF-I for the prevention or management of septic complications in perioperative or critically ill patients especially with low circulating GH and/or IGF-I levels.

Mikrobiol Z, 1998 May-Jun, 60(3), 70 - 9
Treatment and prophylaxis of some infectious diseases of swine and cattle using of natural alpha-interferons; Kishko IaH et al.; On the basis of experimental studies which proved stimulating action on absorbing and bactericidal function of blood phagocytes and antibody genesis the methods of therapy and prophylaxis of wide spread infections in modern live-stock (transmissive gastroenteritis of pigs, parainfluenza of calves, colibacteriosis of swine and cattle) were created . It was established, that optimal one-time therapeutical dose of homologous alpha-IFN for intramuscular administration to new-born pigs was 2000-4000 IU per head, prophylactic dose was 1000-2000 IU . Therapeutical doses of alpha-IFN for calves did not exceed 4000-6000 IU, prophylactic--4000 IU . As a rule, it is necessary to introduce the preparation three times with 48 h intervals . With the account of these data the large-scale commissional trials of these preparations were carried out.

J Trauma, 1998 Oct, 45(4), 649 - 55
Impact of stomach and colon injuries on intra-abdominal abscess and the synergistic effect of hemorrhage and associated injury; Croce MA et al.; BACKGROUND: Colon wounds are recognized to be highly associated with intra-abdominal abscess (IAA) after penetrating trauma, whereas gastric wounds are thought to contribute minimally to abscess because of the bactericidal effect of low pH . This study evaluated the impact of stomach or colon wounds, the contribution of other risk factors, and associated abdominal injuries on IAA . METHODS: Patients with penetrating colon or stomach wounds during a 10-year period were reviewed and stratified by age, Injury Severity Score, transfusions, and associated abdominal injuries . Early deaths (<48 hours) from hemorrhage were excluded . Outcomes analyzed were IAA and death . RESULTS: A total of 812 patients were identified . There were 32 late deaths (4%), of which 28% were attributable to IAA and multiple organ failure . IAA rates for isolated stomach or colon wounds were 0 and 4.2%, respectively . The presence of associated injuries increased IAA rates to 7.5 and 8.8%, respectively . Independent predictors of IAA determined by multivariate analysis included age, transfusions, gunshot wounds, and associated injuries to the liver, pancreas, and kidney . CONCLUSION: Gastric injuries are equivalent to colon wounds in their contribution to IAA . Contamination from either organ without associated injury is minimally associated with IAA, but injury to both appears synergistic . The immunosuppressive effects of age and hemorrhage, in addition to significant associated injury, enhance the development of IAA.

Can J Microbiol, 1998 Jul, 44(7), 692 - 7
Genetic analysis of microcin H47 immunity; Rodriguez E et al.; Microcin H47 is a bactericidal antibiotic produced by a natural Escherichia coli isolate . The genetic system encoding microcin production and immunity consists of at least seven clustered genes . Four of these are devoted to microcin biosynthesis and two genes are required for its secretion into the extracellular medium . The product of the seventh gene, mchI, determines the cell's self-immunity . This gene was shown to encode a highly hydrophobic 69-residue peptide . Analysis of the MchI amino acid sequence, as well as the characterization of MchI-PhoA hybrid proteins, indicated that the microcin immunity product is probably exported out of the cytoplasm and remains an integral membrane peptide . This localization of the immunity peptide points to the cellular membrane as the site of action of microcin H47.

Adv Exp Med Biol, 1998, 443, 23 - 32
Direct detection and quantitative determination of bovine lactoferricin and lactoferrin fragments in human gastric contents by affinity mass spectrometry; Kuwata H et al.; Lactoferricin (Lfcin) is a bioactive fragment of lactoferrin derived from the bactericidal and putative lymphocyte receptor binding domain(s) located within the N-lobe of lactoferrin . Although known to be liberated from at least three species of lactoferrin, conditions leading to Lfcin generation in vivo and factors affecting its distribution are still not known . Recently, we have developed a method of surface-enhanced laser desorption/ionization (SELDI) affinity mass spectrometry using n-butyl terminal groups for surface-enhanced affinity capture (SEAC) to quantify not only Lfcin generated in vivo but also other lactoferrin fragments . Unlike previous efforts to detect lactoferrin and Lfcin with specific antibodies, the SELDI affinity assay distinguished lactoferrin, lactoferrin fragments, Lfcin and unrelated peptides without their interference with each other . To evaluate Lfcin generation in vivo, the experimental design involved feeding 200 mL of 10 mg/mL (1.22 x 10(-4) mol/L) bovine lactoferrin to an adult . Gastric contents were recovered 10 min after ingestion . Lfcin produced in vivo was directly captured by the SEAC device . The amount of Lfcin in the gastric contents was 16.91 +/- 2.65 micrograms/mL (5.350 +/- 0.838 x 10(-6) mol/L) . However, a large proportion of the ingested lactoferrin was not completely digested . Lactoferrin fragments containing the Lfcin region were analyzed by in situ hydrolysis with pepsin after being captured by the SEAC device . As much as 5.740 +/- 0.702 x 10(-5) mol/L of the partially degraded lactoferrin fragments were found to contain the Lfcin region, including peptide domains 17-43, 17-44, 12-44, 9-58, and 16-76 of bovine lactoferrin . These results show that bovine Lfcin can be produced in the human stomach after ingestion of an infant formula supplemented with bovine lactoferrin . It is now important to determine whether Lfcin is generated in the intestinal tract of formula-fed and breast-fed infants, and geriatric patients consuming foods enriched with lactoferrin.

Presse Med, 1998 May, 27 Suppl 1, 28 - 32
{Efficacy of pneumococcal vaccination}; Hessel L; POLYVALENT VACCINE: Pneumococcal vaccination is based on the antigenic properties of polysides carried on the bacterial capsule which induce production of bactericidal serotype-specific antibodies . The 23 serotypes contained in the current vaccine cover 85 to 90% of all strains observed in pneumococcal infections in Europe . PROVEN EFFICACY: Prospective and retrospective studies have demonstrated that pneumococcal vaccination can prevent 60 to 80% of invasive pneumococcal infections in immunocompetent elderly subjects and/or persons in high-risk groups with underlying disorders . TARGET POPULATION: Immunocompetent persons in the 60-65 year and older age group comprise the target population for pneumococcal vaccination . Incidence and mortality of pneumococcal infections is high in this group . Simple vaccination strategies should be implemented.

Aust Dent J, 1998 Aug, 43(4), 244 - 9
Candida-associated denture stomatitis . Aetiology and management: a review . Part 3 . Treatment of oral candidosis; Webb BC et al.; Treatment of oral candidosis with topical antifungal agents such as nystatin and amphotericin B is effective initially . However, medication can produce side effects in some patients and when therapy is stopped the condition can recur . Alternative treatment involving the use of antiseptics and disinfecting agents has been shown to play an important role in the control of dental plaque . The use of sodium hypochlorite as an overnight denture soak has been shown to eliminate denture plaque and recent investigations have demonstrated that microwave irradiation of dentures at a specified setting and exposure time is bactericidal and candidacidal.

Z Gastroenterol, 1998 Aug, 36(8), 625 - 33
p-ANCA target antigens in ulcerative colitis; Folwaczny C et al.; INTRODUCTION: Data about the nature and pathophysiological relevance of the target antigens reacting with perinuclear antineutrophil cytoplasmatic autoantibodies (p-ANCA) in ulcerative colitis are inconsistent and partly conflicting . In the majority of the previous studies only one singular potential target antigen was investigated . The present study aimed on the simultaneous assessment of five different p-ANCA subtypes in patients with ulcerative colitis and attempted to detect reactivity against one of the previously described antigens and to correlate specificity for different target antigens with clinical features of the disease . METHODS: Sera from 61 patients with ulcerative colitis and from 56 healthy controls were tested using indirect immunofluorescence and enzyme-linked immunosorbent assays specific for elastase, lactoferrin, cathepsin G, myeloperoxidase and bactericidal permeability increasing protein (BPI) . p-ANCA subtypes were correlated with clinical features of ulcerative colitis like disease extent or presence of extraintestinal manifestations . Moreover, a possible correlation to current immunosuppressive therapy was evaluated . RESULTS: In 46% (28/61) of patients with ulcerative colitis and in 4% (2/56) of the controls p-ANCA were detected . p-ANCA subtypes were distributed as follows: 26% (16/61) anti-BPI . 16% (10/61) anticathepsin G, 15% (9/61) antielastase, 7% (4/61) antilactoferrin, 5% (3/61) antimyeloperoxidase . Presence of anticathepsin G antibodies was negatively correlated with immunosuppressive therapy . No further correlations between p-ANCA subtypes and clinical characteristics were observed . DISCUSSION: p-ANCA subtypes in inflammatory bowel disease react with a variety of different target antigens and are not correlated with clinical features of the disease.

Am J Respir Crit Care Med, 1998 Oct, 158(4), 1026 - 31
Prediction of clinical severity and outcome of ventilator-associated pneumonia . Comparison of simplified acute physiology score with systemic inflammatory mediators; Froon AH et al.; Systemic kinetics of three inflammatory mediators (bactericidal/permeability-increasing protein {BPI}, soluble intercellular adhesion molecule {sICAM}, and soluble E-selectin {sE-selectin}) were studied during the development of ventilator-associated pneumonia (VAP) (n = 42), diagnosed on quantitative cultures of bronchoscopic samples . From a pool of collected samples, nested samples were used to measure mediators on Days -4, -2, 0, and +2, relative to diagnosis . Correlations between systemic levels of mediators and clinical severity of infection (VAP with or without severe sepsis or septic shock) and patient outcome (mortality at Day 10 after diagnosis) were studied . Predictive values of inflammatory mediators were compared with daily Simplified Acute Physiology Score II (SAPS II) values and the logarithmic number of bacteria in bronchoscopic samples . During the development of VAP, increasing SAPS II scores and rising systemic mediator levels were only found in patients in whom VAP was accompanied with severe sepsis or septic shock . Values of SAPS II and plasma levels of BPI and sE-selectin, but not sICAM, increased from the day of diagnosis on in patients who died within 10 d of diagnosis . Systemic levels of inflammatory mediators did not better predict clinical severity or patient outcome than daily SAPS II scores . The logarithmic number of bacteria in bronchoscopic samples poorly correlated with circulating levels of inflammatory mediators, severity of infection, and patient outcome . Our findings show that a clinical scoring system (SAPS II score) is at least as good as a predictor for the clinical severity of infection and patient outcome, and provide new information on the kinetics of inflammatory mediators during the development of VAP.

Biochim Biophys Acta, 1998 Oct 15, 1375(1-2), 52 - 60
Ionic channels formed by a primary amphipathic peptide containing a signal peptide and a nuclear localization sequence; Chaloin L et al.; The peptide SP-NLS (Ac-Met-Gly-Leu-Gly-Leu-His-Leu-Leu-Leu-Ala10-Ala-Ala-Leu-Gln-Gly- Ala -Lys-Lys-Lys-Arg20-Lys-Val-NH-CH2-CH2-SH) is composed of a hydrophobic signal sequence (SP, Met-1 to Ala-16) followed by a polycationic nuclear localization sequence (NLS, Lys-17 to Val-22) terminated by a cysteamide group . Designed to act as drug carrier this primary amphipathic peptide proved cytotoxic and bactericidal when used at high concentrations, probably by inducing the formation of ion channels . In this work, we show that indeed SP-NLS exhibits a pore-forming activity when incorporated into planar lipid bilayers and Xenopus laevis oocyte plasma membranes, with conductance values of 25 pS in 0.1 M NaCl . In both membranes, the insertion of the peptide was voltage-triggered whereas the induced conductances proved almost voltage-independent . Moreover, SP-NLS ion channels were selective for monovalent cations (K+>Na+>Li+>tetraethylammonium+>choline+) . The ion channel activity of this type of peptides thus provides some insight on their toxicity but also on the mechanism involved for their membrane crossing process.

Acta Crystallogr D Biol Crystallogr, 1998 Jul 1, 54 ( Pt 4), 510 - 21
Structure of a basic phospholipase A2 from Agkistrodon halys Pallas at 2.13 A resolution; Zhao K et al.; The basic phospholipase A2 isolated from the venom of Agkistrodon halys Pallas (Agkistrodon blomhoffii Brevicaudus) is a hemolytic toxin and one of the few PLA2's capable of hydrolyzing the phospholipids of E . coli membranes in the presence of a bactericidal/permeability-increasing protein (BPI) of neutrophils . The crystal structure has been determined and refined at 2.13 A to an R factor of 16.5% (F > 3sigma) with excellent stereochemistry . A superposition of the two molecules in the asymmetric unit gives an r . m.s . deviation of 0.326 A for all Calpha atoms . The refined structure allowed a detailed comparison with other PLA2 species of known structures . The overall architecture is similar to those of other PLA2's with a few significant differences . One of which is in the region connecting the N-terminal helix and the Ca2+-binding loop . Unexpectedly, the conformation of the peptide plane Cys29-Gly30 in the Ca2+-binding loop is very different to that of other PLA2's . The amide NH of Gly30 does not point toward the proposed site for stabilization of the tetrahedral intermediate oxyanion of the substrate analogue . The structure includes four residues which occur less frequently in other PLA2's . His1, Arg6 and Trp70 located at the interfacial recognition site may play an important role in the interaction with aggregated substrates, while Trp77 contributes to the hydrophobic interactions between the beta-wing and the main body of the molecule . This structure analysis reveals that two clusters of basic residues are located at or near the interfacial recognition site, forming an asymmetric positively charge distribution . In contrast to the acidic isoform, the present enzyme is a dimer in the crystalline state . The special phospholipid hydrolysis behaviors are discussed in the light of the structure determined.

Int J Food Microbiol, 1998 Aug 18, 43(1-2), 135 - 8
Mathematical modeling to predict the bactericidal effect of processed vinegar on Escherichia coli O157:H7; Tsujihata S et al.; The combined effects of acetic acid, temperature and sodium chloride on Escherichia coli O157:H7 inactivation were examined in processed vinegar . To express their effects, quadratic polynomial models were applied . The logarithm transformation of sodium chloride concentration provided a better fit of the data than the use of a non-transformation value . On the basis of this finding, the polynomial models should be distinguished into two types, i.e . the non-sodium chloride model and the sodium chloride model, both of which had high R2 values (0.988 and 0.978, respectively).

Antimicrob Agents Chemother, 1998 Oct, 42(10), 2650 - 5
Pharmacodynamics of gatifloxacin in cerebrospinal fluid in experimental cephalosporin-resistant pneumococcal meningitis; Lutsar I et al.; The purpose of this study was to evaluate the cerebrospinal fluid (CSF) pharmacodynamics of a new fluoroquinolone, gatifloxacin (AM-1155), in experimental pneumococcal meningitis . The penetration of gatifloxacin into CSF, calculated as the percentage of the area under the concentration-time curve (AUC) in CSF over the AUC in blood, was 46 to 56% . Gatifloxacin showed linear pharmacokinetics in CSF, and 1 h after intravenous dosages of 7.5, 15, or 30 mg/kg of body weight, peak CSF concentrations were 0.46 +/- 0.08 (mean +/- standard deviation), 0.94 +/- 0.16, and 1.84 +/- 0.5 microg/ml, respectively . The elimination half-life of gatifloxacin in CSF was 3 . 8 to 5.6 h (compared with 2.7 to 3.2 h in blood) . There was a significant interrelationship among the highest measured values of gatifloxacin in blood and CSF/minimal bactericidal concentration (Cpeak/MBC), the time antibiotic concentrations exceeded the MBC (T > MBC), and AUC/MBC (r = 0.94); in single-dose experiments, each correlated significantly with the bacterial killing rate . Divided-dose regimens, resulting in greater T > MBC values but lower Cpeak/MBC ratios, were more effective in terms of bacterial clearance compared with corresponding single-dose regimens . Gatifloxacin therapy was as effective as currently recommended regimens (e.g., a combination of ceftriaxone and vancomycin) against this highly cephalosporin-resistant pneumococcal strain . The bactericidal activity of gatifloxacin in CSF was closely related to the AUC/MBC ratio, but maximal activity was achieved only when drug concentrations exceeded the MBC for the entire dosing interval.

Otolaryngol Pol, 1997, 51 Suppl 25, 349 - 52
{The evaluation of granulocyte function before and after surgical treatment in patients with chronic inflammation of the middle ear and palatine tonsils}; Zalewski P et al.; The aim examinations was the evaluation of granulocytes (PMNL) pre- and postsurgical treatment in patients with chronic inflammation of middle ear and palatine tonsils . The examined 40 persons and divided into three groups: I--18 patients with chronic inflammation of middle ear, II--10 ones with chronic tonsillitis and III--12 healthy persons . There were evaluated: "in vivo" and "in vitro" migration, bactericidal index and absorption of S . aureus labeled 14C isotope (phagocytic index) in the own modification . Preoperative treatment in patients with chronic inflammation of middle ear and palatine tonsils in comparison to the healthy were noticed: characteristic increase of the migration area, the MIF liberation and the phagocytic index, decrease in the bactericidal index (but more in group II) . After a year postsurgical treatment the function of granulocytes in patients of group II was more similar to the healthy than in group I.

Arch Histol Cytol, 1998 Aug, 61(3), 199 - 214
Immunoelectron microscopic identification of lysozyme-expressing cells in human labial salivary glands; Miyazaki T et al.; Although human labial gland secretions contain serous components such as the bactericidal enzyme, lysozyme, the presence of serous cells in this gland has yet to be clearly visualized under the electron microscope . The present study identifies lysozyme-expressing cells of the labial glands using microwave-fixed, Epon-Araldite-embedded specimens, which showed excellent preservation of both ultrastructural detail and antigenicity for post-embedding immunogold labeling of lysozyme . Ultrastructurally, all of the secretory cells of the glands appeared to be a mucous-type and have a serial maturation relationship, consistent with a previous report by TANDLER et al . (1969a): their secretory granules were electron-lucent and exhibited reactivity for mucus staining by the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method . We classified them into two immature types (I, II) and two mature types (I, II) . Their distinctive features were the following: 1) relatively small (0.5-1 microm) secretory granules and well-developed basal rough endoplasmic reticulum for the immature types; 2) larger (1-2 microm) secretory granules and well-developed Golgi apparatus, which showed intense PA-TCH-SP reactivity in the 2-3 trans-cisterns, for the mature types; 3) few secretory granules in the immature type I; and 4) the darkest appearance for the mature type II . Immunogold labeling with anti-lysozyme showed specific labeling of the two immature-type cells, in which gold particles were found mainly over the secretory granules and Golgi apparatus, and moderately over the rough endoplasmic reticulum . In the secretory granules, the labeling was distributed throughout the contents and was present even if they showed strong PA-TCH-SP reactivity; in the Golgi area, it was seen over the stacked cisternae, trans-Golgi networks, and condensing vacuoles . No specific labeling was seen in the mature-type cells or in the duct cells . These immature- and mature-type cells were almost equivalent to the "serous demilune or acinus" and "mucous tubule" cells, respectively, at the light-microscopic level . These results indicate that the traditional "immature mucous-type cells" of the human labial glands produce lysozyme and should be classified as seromucous cells.

Khirurgiia (Mosk), 1998, (8), 20 - 2
{The treatment of wounds of the anal canal and perineum}; Protsenko VM et al.; Analysis of processes of wound healing in 215 patients after operations for hemorrhoids, anal fissures and rectal fistulas was carried out . Clinico-cytologic control of healing of postoperation wound has shown that ultra-violet irradiation of wound surface is most effective at early stages of wound healing due to its pronounced bactericidal and antiinflammatory action . Low frequency laser irradiation intensifies tissue reparative and hastens healing of wounds . The proposed method for treatment of wounds by successive use of ultra-violet and laser irradiation of wound surface combined with conventional ointment medications, improves the results of treatment and decreases the rate of purulent complications 2 times, promotes rapid healing of wounds.

Ann Thorac Cardiovasc Surg, 1998 Aug, 4(4), 209 - 13
The role of aggressive medical therapy along with early surgical intervention in the cure of Brucella endocarditis; Uddin MJ et al.; Timing of surgical intervention in Brucella endocarditis remains controversial . In this report, we review our experience in an attempt to collect some information on the best approach for this entity . From June 1992 to December 1996, 5 male patients between the ages of 20 and 35 years with Brucella endocarditis were operated on in our centre . Three of them had native valve endocarditis (NVE) and 2 with prosthetic valve endocarditis (PVE) . All patients belonged to New York Heart Association (NYHA) class III-IV . All had developed anti Brucella antibodies with serum agglutination titres of > 320 and the sera tested from 3 patients were Enzyme Linked Immunosorbent Assay (ELISA) positive for anti-Brucella IgM and/or IgG antibodies . In 3 cases 2D-echocardiography showed large vegetation on the affected valve . Blood cultures were positive in 4 patients, 2 of them (one each of NVE and PVE) had the valve material culture positive for Brucella . All cases were treated with a combination of doxycycline, refampicine and gentamicin before surgery . Major indication for surgical intervention was severe haemodynamic instability which developed during the course of antibiotic therapy either early (3 cases) or late (2 cases) . All patients became asymptomatic at the end of 7 days postoperatively . On the follow-up for a period of 8-51 months, all patients were in NYHA class I-II without evidence of recurrence of infection . These data suggest that in either NVE or PVE Brucella, medical therapy alone may not be sufficient due to the eventual haemodynamic deterioration secondary to valve tissue destruction or dysfunction of the prosthetic valve caused by the infective process . Therefore, a combination of aggressive medical therapy with multiple bactericidal antibiotics and early surgical intervention may result in a successful outcome, but further studies are needed to reach a reliable conclusion.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2235 - 9
Pharmacokinetic profiles of high-dose intravenous ciprofloxacin in severe sepsis . The Baragwanath Ciprofloxacin Study Group; Lipman J et al.; The pharmacokinetics of 400 mg of ciprofloxacin given intravenously (i.v.) every 8 h (q8h) in severely septic adults was documented in a multidisciplinary, tertiary referral intensive care unit (ICU) . Sixteen evaluable patients (three pharmacokinetic profiles) without renal dysfunction and with severe sepsis were studied . Ciprofloxacin at a dosage of 400 mg given i.v . q8h was administered over 1 h . Plasma samples for assay (high-pressure liquid chromatography) were taken at timed intervals (preinfusion, at the end of infusion, and at 1, 2, 3, 5, and 7 h postinfusion) for first-dose kinetics (day 0 {D0}), D2, and between D6 and D8 . All pharmacokinetic variables were calculated by noncompartmental methods . Standard intensive care was provided . Peak ciprofloxacin concentrations were as follows: D0, 6 . 01 +/- 1.93 mg/liter; D2, 6.68 +/- 2.01 mg/liter; and D6 to D8 6.45 +/- 1.54 mg/liter . Trough levels were as follows: D0, 0.6 +/- 0.5 mg/liter; D2, 0.7 +/- 0.4 mg/liter; and D6 to D8 0.6 +/- 0.4 mg/liter . The areas under the concentration curves (8 h) were as follows: D0, 13.3 +/- 3.8 mg . h/liter; D2, 16.8 +/- 5.4 mg . h/liter; and D6 to D8, 15.5 +/- 4.7 mg . h/liter . No drug-related serious adverse events occurred . For 17 of 18 patients enrolled in the study, the causative organisms were susceptible to ciprofloxacin . One patient developed renal failure (non-drug related) after the administration of three doses of ciprofloxacin . One patient was infected with ciprofloxacin-resistant organisms on enrollment . Nine of 16 evaluable patients had clinical cures, and 8 had bacteriological cures . One patient developed a ciprofloxacin-resistant superinfection . In two patients the clinical course was indeterminate . Two bacteriological failures occurred . We conclude that in critically ill adults ciprofloxacin at a dosage of 400 mg given i.v . q8h is safe . Its pharmacokinetic profile provides bactericidal activity against most organisms encountered in an ICU . Except for some initial accumulation on D2, no further accumulation occurred in patients without renal failure . Ciprofloxacin should be administered i.v . at a dosage of 400 mg q8h for severe sepsis.

J Biomed Mater Res, 1998 Fall, 43(3), 338 - 48
Concise review of mechanisms of bacterial adhesion to biomaterial surfaces; An YH et al.; This article reviews the mechanisms of bacterial adhesion to biomaterial surfaces and the factors affecting the adhesion . The process of bacterial adhesion includes an initial physicochemical interaction phase (phase one) and a late molecular and cellular interaction phase (phase two), which is a complicated process affected by many factors, including the characteristics of the bacteria themselves, the target material surface, and the environmental factors, such as the presence of serum proteins or bactericidal substances.

Curr Opin Struct Biol, 1998 Aug, 8(4), 426 - 34
The implications of the structure of the bactericidal/permeability-increasing protein on the lipid-transfer function of the cholesteryl ester transfer protein; Bruce C et al.; The cholesteryl ester transfer protein (CETP) is evolutionarily related to the bactericidal/permeability-increasing protein (BPI) . The recently solved structure of BPI shows an elongated, boomerang-shaped molecule, with two hydrophobic pockets opening to its concave side . These pockets each contain a phospholipid molecule . A model of CETP, based on the recently solved crystal structure of BPI, provides the basis for interpreting functional studies on CETP . In this model, C-terminal residues 461-476, which were shown to be required for neutral lipid transfer between plasma lipoproteins, from an amphipathic helix covering the opening of the N-terminal pocket . A possible lipid-transfer mechanism for CETP, with the initial step involving the disordering of lipids in the lipoprotein surface, followed by the flipping and entry of a lipid molecule into the hydrophobic lipid-binding pocket, is hypothesized in light of structural evidence and recent studies.

Zentralbl Bakteriol, 1998 Jul, 288(1), 103 - 10
Killing of Pseudomonas pseudomallei by polymorphonuclear leukocytes and peritoneal macrophages from chicken, sheep, swine and rabbits; Markova N et al.; Differences in the kinetics of Pseudomonas pseudomallei killing by peritoneal macrophages (PM) and polymorphonuclear leucocytes (PMNL) from chickens, sheep, swine and rabbits were found . P . pseudomallei was rapidly killed by porcine PM and PMNL . However the bacterial killing by ovine and lapine PM and PMNL proceeded at a slower rate . In contrast, chicken PM and PMNL ingested and killed the lowest number of P . pseudomallei bacteria . The differences in the bactericidal activity of PM and PMNL from different animal species correlated with the level of their acid phosphatase and glycolytic activity.

J Immunol, 1998 Sep 1, 161(5), 2552 - 60
Differential regulation of lipopolysaccharide (LPS) activation pathways in mouse macrophages by LPS-binding proteins; Amura CR et al.; LPS binding to its receptor(s) on macrophages induces the synthesis of inflammatory mediators involved in septic shock . While the signaling mechanism(s) remains to be fully defined, the human LPS-binding protein (LBP) is known to regulate responses to LPS by facilitating its binding to CD14 on human monocytes . The structurally related bactericidal permeability increasing protein (BPI) differs from LBP by inhibiting LPS-induced human monocyte activation . We have demonstrated that, unlike the human monocyte response to LPS, both LBP and BPI inhibited LPS-stimulated TNF-alpha production in mouse peritoneal macrophages . In contrast, LPS-dependent nitric oxide release was not affected by LBP . LPS induces the phosphorylation of a number of proteins in a dose and time-dependent manner, however, the pattern of LPS-induced phosphorylation was not reduced by either LBP or BPI under conditions that result in selective TNF-alpha inhibition . Further, activation of the transcription factor NF-kappaB in response to LPS was also not modified by either LBP or BPI . Finally, no differences were detected in TNF-alpha or inducible nitric oxide synthase mRNA accumulations induced by LPS in the presence or absence of either protein, whereas a slight decreased mRNA stability was observed in the group with LPS treatment . These results would suggest that many of the early signaling events contribute to LPS-induced macrophage signaling at a point preceding the divergence of pathways that differentially regulate TNF-alpha and NO production.

Br J Clin Pharmacol, 1998 Aug, 46(2), 163 - 7
Constant rate infusion of vancomycin in premature neonates: a new dosage schedule; Pawlotsky F et al.; AIMS: Since vancomycin's bactericidal action has been shown to be time-dependent, a constant rate infusion over 24 h might result in a better bactericidal efficacy . The purpose of this study was to define a new dosage schedule in prematures . METHODS: Two vancomycin 24 h constant rate infusion schedules were tested in two groups of neonates . Postconceptional age (PCA) was 27 to 41 weeks in group 1 (n=24) and 28 to 51.5 weeks in group 2 (n=29) . Group 1 neonates received continuous infusion of 10 to 30 mgkg(-1) day(-1), adjusted for PCA and weight . Group 2 was designed to take into account the significant relationship observed in group 1 between vancomycin clearance standardized on weight and PCA and consisted of a constant loading dose of 7 mg kg(-1) followed by continuous infusion of 10 to 40 mg kg(-1) day(-1) adjusted for PCA and weight . RESULTS: Mean vancomycin serum concentration at steady state was 11+/-3.1 mg1(-1) in group 1 and 15.4+/-6.2 mg1(-1) in group 2 . Fifty-six percent of group 1 values vs 88% of group 2 values were between 10 and 30 mg at steady state (P<0.01) . Both regimens were well tolerated . CONCLUSIONS: A loading dose of vancomycin followed by constant rate infusion of the appropriate dose adjusted for PCA and weight might improve vancomycin concentrations in neonates.

Annu Rev Nutr, 1998, 18, 297 - 330
Plasma lipid transfer proteins, high-density lipoproteins, and reverse cholesterol transport; Bruce C et al.; Cholesteryl ester transfer protein (CETP) and phospholipid transfer protein (PLTP) are members 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 CETP and 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 human CETP genetic deficiency states and CETP transgenic mice indicates that CETP plays a major role in the catabolism of high-density lipoprotein (HDL) cholesteryl esters and thereby influences the concentration, apolipoprotein content, and size of HDL particles in plasma . PLTP also appears to have an important role in determining HDL levels and speciation . Recent data indicate that genetic CETP deficiency is associates with an excess of coronary heart disease in humans, despite increased HDL levels . Also, CETP expression is anti-atherogenic in many mouse models, even while lowering HDL . These data tend to support the reverse cholesterol transport hypothesis, i.e., that anti-atherogenic properties of HDL are related to its role in reverse cholesterol transport . Recently, another key molecule involved in this pathway was identified, scavenger receptor BI; this mediates the selective uptake of HDL cholesteryl esters in the liver and thus constitutes a pathway of reverse cholesterol transport parallel to that mediated by CETP . Reflecting its role in reverse cholesterol transport, the CETP gene is up-regulated in peripheral tissues and liver in responses to dietary or endogenous hypercholesterolemia . An analysis of the CETP proximal promoter indicates that it contains sterol regulatory elements highly homologous to those present in 3-hydroxy-3-methylglutaryl-coenzyme A reductase; the CETP gene is transactivated by the binding of SREBP-1 to these elements . A challenge for the future will be the manipulation of components of the reverse cholesterol transport pathway, such as CETP, PLTP, or scavenger receptor BI for therapeutic benefit.

Vaccine, 1998 Oct, 16(17), 1688 - 92
Clinical and immunological assessment of a candidate Lyme disease vaccine in healthy adults: antibody persistence and effect of a booster dose at month 12; Van Hoecke C et al.; A candidate Lyme vaccine was administered to 20 adult volunteers following a 0, 1, 2 months vaccination schedule, with a booster at 12 months . An immune response, assessed as 'LA-2 equivalent' antibody titres using an inhibition ELISA, was induced in all vaccinees which persisted until the booster . Titres were increased 25-fold following the booster and persisted through month 24 . There was a good correlation between 'LA-2 equivalent' antibody titres and a bactericidal assay (r2 = 0.86) . Local symptoms were mild, resolving spontaneously within 72 h, with no reports of rash, arthralgia or other systemic symptoms . This Lyme vaccine was safe, well-tolerated and elicited an antibody response in all volunteers which persisted at least 12 months after the booster.

Infect Immun, 1998 Sep, 66(9), 4374 - 81
Isolation and characterization of two proteins from Moraxella catarrhalis that bear a common epitope; McMichael JC et al.; The UspA1 and UspA2 proteins of Moraxella catarrhalis are potential vaccine candidates for preventing disease caused by this organism . We have characterized both proteins and evaluated their vaccine potential using both in vitro and in vivo assays . Both proteins were purified from the O35E isolate by Triton X-100 extraction, followed by ion-exchange and hydroxyapatite chromatography . Analysis of the sequences of internal peptides, prepared by enzymatic and chemical cleavage of the proteins, revealed that UspA1 and UspA2 exhibited distinct structural differences but shared a common sequence including an epitope recognized by the monoclonal antibody 17C7 . By sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), purified UspA1 exhibited a molecular weight of approximately 350, 000 when unheated and a molecular weight of 100,000 after being heated for 10 min at 100 degreesC . In contrast, purified UspA2 exhibited an apparent molecular weight of 240,000 by SDS-PAGE that did not change with the length of time of heating . Their sizes as determined by gel filtration were 1,150,000 and 830,000 for UspA1 and UspA2, respectively . Preliminary results indicate the proteins have separate functions in bacterial pathogenesis . Purified UspA1 was found to bind HEp-2 cells, and sera against UspA1, but not against UspA2, blocked binding of the O35E isolate to the HEp-2 cells . UspA1 also bound fibronectin and appears to have a role in bacterial attachment . Purified UspA2, however, did not bind fibronectin but had an affinity for vitronectin . Both proteins elicited bactericidal antibodies in mice to homologous and heterologous disease isolates . Finally, mice immunized with each of the proteins, followed by pulmonary challenge with either the homologous or a heterologous isolate, cleared the bacteria more rapidly than mock-immunized mice . These results suggest that UspA1 and UspA2 serve different virulence functions and that both are promising vaccine candidates.

Infect Immun, 1998 Sep, 66(9), 4183 - 92
The transferrin binding protein B of Moraxella catarrhalis elicits bactericidal antibodies and is a potential vaccine antigen; Myers LE et al.; The transferrin binding protein genes (tbpA and tbpB) from two strains of Moraxella catarrhalis have been cloned and sequenced . The genomic organization of the M . catarrhalis transferrin binding protein genes is unique among known bacteria in that tbpA precedes tbpB and there is a third gene located between them . The deduced sequences of the M . catarrhalis TbpA proteins from two strains were 98% identical, while those of the TbpB proteins from the same strains were 63% identical and 70% similar . The third gene, tentatively called orf3, encodes a protein of approximately 58 kDa that is 98% identical between the two strains . The tbpB genes from four additional strains of M . catarrhalis were cloned and sequenced, and two potential families of TbpB proteins were identified based on sequence similarities . Recombinant TbpA (rTbpA), rTbpB, and rORF3 proteins were expressed in Escherichia coli and purified . rTbpB was shown to retain its ability to bind human transferrin after transfer to a membrane, but neither rTbpA nor rORF3 did . Monospecific anti-rTbpA and anti-rTbpB antibodies were generated and used for immunoblot analysis, which demonstrated that epitopes of M . catarrhalis TbpA and TbpB were antigenically conserved and that there was constitutive expression of the tbp genes . In the absence of an appropriate animal model, anti-rTbpA and anti-rTbpB antibodies were tested for their bactericidal activities . The anti-rTbpA antiserum was not bactericidal, but anti-rTbpB antisera were found to kill heterologous strains within the same family . Thus, if bactericidal ability is clinically relevant, a vaccine comprising multiple rTbpB antigens may protect against M . catarrhalis disease.

Biol Res, 1997, 30(4), 149 - 60
Construction of a gene encoding the insect bactericidal protein attacin . Studies on its expression in Escherichia coli; Santibanez E et al.; Attacin, a bactericidal small protein is produced by the giant silk moth Hyalophora cecropia . This paper deals with our efforts to clone the attacin cDNA in a bacterial vector to express it in Escherichia coli and produce the protein in sufficient amount, for further studies . We chose two inducible expression vector/bacterial cell systems: pPL-lambda/N99cI+ cells which is able to be induced by nalidixic acid, and pET3d/BL21(DE3) cells carrying a T7 RNA polymerase gene which is IPTG-inducible . After cloning in the pPL-lambda system and under no addition of the inducer, isolated transformants carried this plasmid with at least 2 concurrent deletions that drastically affected attacin expression, even though attacin gene seems to be intact as deduced by its PCR amplification . It was concluded that basal attacin expression occurred in this system and bacterial growth was limited . Plasmid deletions may have emerged by selection pressure as a way to avoid bactericidal expression and allow bacteria survival . The second cloning attempt was done in pET3d vector/BL21 cells, that should not express the cloned sequence (they lack T7 RNA polymerase gene) . Transformed BL21 cells gave 3 recombinant plasmids, 2 of them presented a C deletion that generated an early stop signal in the attacin coding region . The third clone, pET-ATT18, carrying an intact gene, was transferred to BL21(DE3)-IPTG inducible cells in order to be expressed . Attacin was undetectable in stained gels or by Western blot analysis . However, expression was visualized in grown cells after 30 min of IPTG induction and 5 min of {35S}-methionine labeling, as a 22.5 kDa protein band by using gel electrophoresis and fluorography . This low level of expression drastically affected bacterial growth . Considering that attacin has no lytic activity, these results suggest that this molecule should block bacterial growth directly at the cytoplasm by an unknown mechanism, since no signal peptide coding sequence was incorporated in this gene construction, precluding periplasmic or external destination of this protein.

Drug Chem Toxicol, 1998 Aug, 21(3), 355 - 70
Inhibitory actions of glycyrrhizic acid on arylamine N-acetyltransferase activity in strains of Helicobacter pylori from peptic ulcer patients; Chung JG; Arylamine N-acetyltransferase (NAT) activities with 2-aminofluorene (2-AF) and p-aminobenzoic acid (PABA) as substrates were determined in Helicobacter pylori, collected from patients with peptic ulcers . The NAT activity was determined using an acetyl CoA recycling assay and high pressure liquid chromatography . Inhibition of growth studies from H . pylori demonstrated that glycyrrhizic acid elicited dose-dependent bactericidal effect in H . pylori cultures, i.e.; the greater the concentration of glycyrrhizic acid, the greater the inhibition of growth of H . pylori . Cytosols or suspensions of H . pylori with and without selected concentrations of glycyrrhizic acid co-treatment showed different percentages of 2-AF and PABA acetylation . The data indicated that there was decreased NAT activity associated with increased glycyrrhizic acid in H . pylori cytosols and intact cells . For the cytosol and intact bacteria examinations, the apparent values of Km and Vmax were decreased after co-treated with 80 M glycyrrhizic acid . This report is the first demonstration of glycyrrhizic acid inhibition of arylamine NAT activity and glycyrrhizic acid inhibition of growth in the bacterium H . pylori.

Arch Immunol Ther Exp (Warsz), 1998, 46(3), 183 - 92
Effect of general and regional anesthesia on some neutrophil functions; Ciepichal J et al.; Transient immunosuppression in patients during anesthesia and surgery may cause postoperative infections . In the immunity against pathogens the processes of non-specific defence are of special importance and engage neutrophils as the major cell type . The aim of this study was to monitor activity of neutrophils during and after surgery, applying different methods of anesthesia . The status of neutrophils was determined by evaluation of chemotactic, phagocytic and chemiluminescence (bactericidal) activities . The study included 78 patients, who underwent surgery due to pathological injury-effected changes or due to varicose veins of extremities . Blood samples for determination of the neutrophil activity were taken just before anesthesia, when the symptoms of anesthesia occurred, 15 min after starting of surgery, 24 h and 5-7 day after surgery . In majority of patients we observed neutrophilic leukocytosis . There was a statistically significant correlation between leukocytosis and percentage of neutrophils in the subsequent days after surgery, depending on a kind and extension of surgical intervention . In addition, we have found a decrease in chemotactic, phagocytic and bactericidal activity after all kinds of anesthesia . The decrease in the phagocytic activity lasted longer, usually at least 24 h after surgery, whereas chemotaxis and chemiluminescence returned quickly to control levels . In the spinal anesthesia the chemiluminescence was not altered; this phenomenon could result from blockade of the sympathetic nerve system and activation of neutrophils metabolism . The brachial plexus blockade had the statistically smallest effect on the function of neutrophils . In the long-term anesthesia (longer than 2 h), particularly using halothane, we have observed depressed values of chemotaxis, phagocytosis and chemiluminescence which lasted longer than in other studied group of patients . We conclude that monitoring of the neutrophil functions is a useful form of studying immune reactions of patients to anesthesia and surgery.

Zh Mikrobiol Epidemiol Immunobiol, 1998 May-Jun, (3), 3 - 6
{Characterization of the recombinant strain Francisella tularensis R1A}; Kislichkin NN et al.; For the first time F.tularensis recombinant strain R1A, obtained by the transfer of genes responsible for virulence in F.tularensis strain B 399 A-Cole into recipient cells of the R-form, was studied . Strain R1A was characterized by morphological, tinctorial and biochemical properties, similar to those of F.tularensis virulent strains, and became resistant to the bactericidal action of animal sera, as well as heat resistant (tr42) . The results of animal experiments revealed that strain R1A proved to be highly virulent for noninbred white mice, faintly virulent for guinea pigs and avirulent for rabbits . In contrast to the R-form, recombinant strain R1A exhibited high antigenic activity, as well as partially protected guinea pigs from 100 DCL of F.tularensis highly virulent strain B A-Cole . The totality of its properties places recombinant strain R1A in an intermediate position between F.tularensis virulent and vaccine strains.

Microbios, 1998, 93(375), 115 - 27
Inhibitory actions of ellagic acid on growth and arylamine N-acetyltransferase activity in strains of Helicobacter pylori from peptic ulcer patients; Chung JG; Arylamine N-acetyltransferase (NAT) activity in Helicobacter pylori was inhibited by ellagic acid, a possible chemopreventive drug . The NAT activity was determined using an acetyl CoA recycling assay and high pressure liquid chromatography . Inhibition of growth studies using H . pylori demonstrated that ellagic acid elicited a dose-dependent bactericidal effect in H . pylori cultures, i.e . the greater the concentration of ellagic acid, the greater the inhibition of growth of H . pylori . The IC50 value was 1 mM for inhibition of growth of H . pylori . Cytosols or suspensions of H . pylori with and without selected concentrations of ellagic acid co-treatment showed different percentages of 2-aminofluorene and p-aminobenzoic acid acetylation . The data indicated that there was decreased NAT activity associated with increased ellagic acid in H . pylori cytosols and intact cells . For the cytosol and intact bacteria examinations, the apparent values of K(m) and Vmax decreased after co-treatment with 1 mM ellagic acid . This report is the first demonstration of ellagic acid inhibition of arylamine NAT activity and ellagic acid inhibition of growth in the bacterium H . pylori.

J Pediatr Surg, 1998 Jul, 33(7), 1104 - 7
Pharmacokinetics of once-daily gentamicin dosing in pediatric patients; Bass KD et al.; BACKGROUND/PURPOSE: To achieve cost-effective health care in adults, once-daily aminoglycosides administration has been used and judged to be safe and efficacious . A similar strategy in children requires the characterization of pharmacokinetic parameters and the development of a therapeutic monitoring protocol for this antibiotic regimen . METHODS: A prospective, controlled, randomized (2:1) study was undertaken in 50 pediatric patients between June 1995 and September 1997 . Children between 6 months and 18 years who required gentamicin therapy based on independent clinical assessment were eligible if they had normal renal function, no aminoglycoside allergies, were not neutropenic, or did not have cystic fibrosis . Measurements included a peak, 4-hour, 8-hour, and trough gentamicin levels to determine volume of distribution (Vd) and elimination constant (Ke) . Ototoxicity and nephrotoxicity were monitored by pre- and postaudiology examinations and serial calculated creatinine clearance determinations, respectively . RESULTS: Thirty-three patients received 7.5 mg/kg every 24 hours, and 17 patients received 2.5 mg/kg every 8 hours . Most frequent indications for treatment were ruptured appendicitis (n = 19) followed by wound infections caused by trauma (n = 4), but the spectrum of treatment was broad including enteric, genitourinary, central nervous system, biliary, ophthalmologic, and orthopedic infections . Pharmacokinetic data indicated that 24-hour dosing resulted in higher peak levels compared with 8-hour dosing (20.4 +/- 45.4 v 7.2 +/- 6.2 mg/L, P < .0001) and lower trough levels (0.29 +/- .02 v 0.69 +/- 0.13, P < .0001), whereas rate of elimination constant and volume of distribution were not significantly different . No nephrotoxicity or ototoxicity has been noted in either group . CONCLUSIONS: These data confirm that once-daily dosing of gentamicin is a safe method of treatment that provides equivalent pharmacokinetics compared with traditional dosing and enhances bactericidal effect based on higher peak levels, avoids toxicity, and allows cost savings.

J Antimicrob Chemother, 1998 Jun, 41(6), 643 - 8
Bactericidal activity and postantibiotic effect of clarithromycin and 14-hydroxyclarithromycin, alone and in combination, against Legionella pneumophila; Martin SJ et al.; Time-kill curves using simulated peak and trough serum concentrations, and postantibiotic effect (PAE) were determined for clarithromycin and 14-hydroxyclarithromycin alone and in combination against four strains of Legionella pneumophila . Both compounds were bactericidal at both peak and trough concentrations . The combination at trough concentrations demonstrated lower killing activity than the parent drug in three of the four strains . PAE ranges were 7.28-17.3 h for clarithromycin, 6.78-14.77 h for the metabolite, and 5.15-13.23 h for the combination.

Protein Sci, 1998 Jul, 7(7), 1643 - 6
Detecting distant relatives of mammalian LPS-binding and lipid transport proteins; Beamer LJ et al.; In mammals, a family of four lipid binding proteins has been previously defined that includes two lipopolysaccharide binding proteins and two lipid transfer proteins . The first member of this family to have its three-dimensional structure determined is bactericidal/permeability-increasing protein (BPI) . Using both the sequence and structure of BPI, along with recently developed sequence-sequence and sequence-structure similarity search methods, we have identified 13 distant members of the family in a diverse set of eukaryotes, including rat, chicken, Caenorhabditis elegans, and Biomphalaria galbrata . Although the sequence similarity between these 13 new members and any of the 4 original members of the BPI family is well below the "twilight zone," their high sequence-structure compatibility with BPI indicates they are likely to share its fold . These findings broaden the BPI family to include a member found in retina and brain, and suggest that a primitive member may have contained only one of the two similar domains of BPI.

Cell Immunol, 1998 Jul 10, 187(1), 34 - 7
Bovine lactoferrin stimulates the phagocytic activity of human neutrophils: identification of its active domain; Miyauchi H et al.; Bovine LF (bLF) at concentrations in the range of 50-250 micrograms/ml enhanced the phagocytic activity of human neutrophils as determined by measuring the incorporation of FITC-labeled latex beads by flow cytometry . The stimulatory effect of bLF was not abrogated by hydrolysis with pepsin . Bovine lactoferricin (bLFcin), which is a bactericidal fragment purified from a pepsin hydrolysate of bLF (bLFH), also enhanced the phagocytic activity, whereas, in contrast, the fraction of bLFH depleted of bLFcin showed no stimulatory effect . The phagocytosis-enhancing activity of bLF still remained after washing the neutrophils, following exposure to bLF . Also, bLF pretreatment of the latex beads stimulated their uptake . These results demonstrate that bLF is effective in promoting the phagocytic activity of human neutrophils . This activity appears to be due to its bLFcin domain and may involve dual mechanisms of direct binding to neutrophils and opsonin-like activity.

Mol Microbiol, 1998 Jun, 28(6), 1335 - 43
Discovery of critical Tol A-binding residues in the bactericidal toxin colicin N: a biophysical approach; Raggett EM et al.; Colicins translocate across the Escherichia coli outer membrane and periplasm by interacting with several receptors . After first binding to outer membrane surface receptors via their central region, they interact with TolA or TonB proteins via their N-terminal regions . Finally, the toxic C-terminal region is inserted into or across the cytoplasmic membrane . We have measured the binding of colicin N to TolA by isothermal titration microcalorimetry (ITC) and tryptophan fluorescence . The isolated N-terminal domain exhibits a higher affinity for TolA (Kd = 1 microM) than does the whole colicin (18 microM), and similar behaviour has been observed when the N-terminal domain of the g3p protein of the bacteriophage fd, which also binds TolA, is examined in isolation and in situ . This may indicate a similar mechanism in which a cryptic TolA binding site is revealed after primary receptor binding . The isolated colicin N N-terminal domain appears to be unstructured in circular dichroism and fluorescence studies . We have used mutagenesis and ITC to characterize the TolA binding site and have shown it to be of a different sequence and much further from the N-terminus than previously thought.

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