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