Antimicrobials

Clin Infect Dis, 1998 Jul, 27(1), 23 - 7
The pharmacodynamics of aminoglycosides; Lacy MK et al.; Recently, a more complete understanding of the pharmacodynamics of aminoglycosides has been recognized, indicating that this class of antibiotics exhibits both concentration-dependent bactericidal activity and a postantibiotic effect . This pharmacodynamic information, along with better knowledge of the mechanisms responsible for aminoglycoside toxicity, established the foundation for once-daily aminoglycoside dosing regimens . This new approach to aminoglycoside dosing appears to be safe, efficacious, and cost-effective, resulting in its increasing popularity in clinical practice.

Clin Pharmacokinet, 1998 Jul, 35(1), 65 - 77
Community-based parenteral anti-infective therapy (CoPAT) . Pharmacokinetic and monitoring issues; Williams DN et al.; Community-based parenteral anti-infective therapy (CoPAT) has, over the past 20 years, increased rapidly in many parts of the world including North America, Europe, South America and Australia . CoPAT is a multidisciplinary activity demanding close cooperation between nurses, pharmacists and physicians, as well as with the patient . The selection of an anti-infective drug for use outside the hospital setting must take into account not only the therapeutic effectiveness, cost effectiveness and safety of the drug, but also pharmacological factors such as the dosage schedule and the stability of the drug . Dosage schedules vary with pharmacokinetic factors (e.g . the use of drugs with long half-lives are favoured by CoPAT programmes) and pharmacodynamic features (e.g . once daily gentamicin therapy is attractive and practical because of concentration-dependent bactericidal killing and prolonged post-antibiotic effect) . With selected drugs, the renal and, to a lesser degree, liver function of the patient will influence the dosage schedule . The mode of intravenous (i.v.) drugs administration will vary with volume considerations (limiting the use of syringe-infusion therapy for some drugs), stability issues (prevents drugs that are stable at room temperature for less than 24 hours from being used in multidose computerised delivery systems), as well as patient factors (ability to self administer an i.v . drug) . Monitoring serum anti-microbial concentrations is undertaken to assure effectiveness and avoid toxicity, and is indicated for drugs with a narrow therapeutic window, such as the aminoglycosides . With the advent of the single daily dose administration of aminoglycosides, checking serum concentrations at the mid-point, i.e . 6 to 14 hours following administration of the first dose, is one approach . Because the toxic effects of vancomycin have been overstated, serum concentrations should only be obtained for defined indications primarily to assure therapeutic effectiveness.

Infect Immun, 1998 Aug, 66(8), 3656 - 65
Cloning and expression of the Moraxella catarrhalis lactoferrin receptor genes; Du RP et al.; The lactoferrin receptor genes from two strains of Moraxella catarrhalis have been cloned and sequenced . The lfr genes are arranged as lbpB followed by lbpA, a gene arrangement found in lactoferrin and transferrin receptor operons from several bacterial species . In addition, a third open reading frame, orf3, is located one nucleotide downstream of lbpA . The deduced lactoferrin binding protein A (LbpA) sequences from the two strains were found to be 99% identical, the LbpB sequences were 92% identical, and the ORF3 proteins were 98% identical . The lbpB gene was PCR amplified and sequenced from a third strain of M . catarrhalis, and the encoded protein was found to be 77% identical and 84% similar to the other LbpB proteins . Recombinant LbpA and LbpB proteins were expressed from Escherichia coli, and antisera raised to the purified proteins were used to assess antigenic conservation in a panel of M . catarrhalis strains . The recombinant proteins were tested for the ability to bind human lactoferrin following gel electrophoresis and electroblotting, and rLbpB, but not rLbpA, was found to bind lactoferrin . Bactericidal antibody activity was measured, and while the anti-rLbpA antiserum was not bactericidal, the anti-rLbpB antisera were found to be weakly bactericidal . Thus, LbpB may have potential as a vaccine candidate.

Eur J Biochem, 1998 Apr 15, 253(2), 452 - 61
Bactericidal activity of Lys49 and Asp49 myotoxic phospholipases A2 from Bothrops asper snake venom--synthetic Lys49 myotoxin II-(115-129)-peptide identifies its bactericidal region; Paramo L et al.; Mammalian group-II phospholipases A2 (PLA2) of inflammatory fluids display bactericidal properties, which are dependent on their enzymatic activity . This study shows that myotoxins II (Lys49) and III (Asp49), two group-II PLA2 isoforms from the venom of Bothrops asper, are lethal to a broad spectrum of bacteria . Since the catalytically inactive Lys49 myotoxin II isoform has similar bactericidal effects to its catalytically active Asp49 counterpart, a bactericidal mechanism that is independent of an intrinsic PLA2 activity is demonstrated . Moreover, a synthetic 13-residue peptide of myotoxin II, comprising residues 115-129 (common numbering system) near the C-terminal loop, reproduced the bactericidal effect of the intact protein . Following exposure to the peptide or the protein, accelerated uptake of the hydrophobic probe N-phenyl-N-naphthylamine was observed in susceptible but not in resistant bacteria, indicating that the lethal effect was initiated on the bacterial membrane . The outer membrane, isolated lipopolysaccharide (LPS), and lipid A of susceptible bacteria showed higher binding to the myotoxin II-(115-129)-peptide than the corresponding moieties of resistant strains . Bacterial LPS chimeras indicated that LPS is a relevant target for myotoxin II-(115-129)-peptide . When heterologous LPS of the resistant strain was present in the context of susceptible bacteria, the chimera became resistant, and vice versa . Myotoxin II represents a group-II PLA2 with a direct bactericidal effect that is independent of an intrinsic enzymatic activity, but adscribed to the presence of a short cluster of basic/hydrophobic amino acids near its C-terminal loop.

J Exp Med, 1998 Jul 6, 188(1), 49 - 60
Phase-variable expression of lipopolysaccharide contributes to the virulence of legionella pneumophila; Luneberg E et al.; With the aid of monoclonal antibody (mAb) 2625, raised against the lipopolysaccharide (LPS) of Legionella pneumophila serogroup 1, subgroup OLDA, we isolated mutant 811 from the virulent wild-type strain RC1 . This mutant was not reactive with mAb 2625 and exhibited an unstable phenotype, since we observed an in vitro and in vivo switch of mutant 811 to the mAb 2625-positive phenotype, thus restoring the wild-type LPS . Bactericidal assays revealed that mutant 811 was lysed by serum complement components, whereas the parental strain RC1 was almost serum resistant . Moreover, mutant 811 was not able to replicate intracellularly in macrophage-like cell line HL-60 . In the guinea pig animal model, mutant 811 exhibited significantly reduced ability to replicate . Among recovered bacteria, mAb 2625-positive revertants were increased by fourfold . The relevance of LPS phase switch for pathogenesis of Legionella infection was further corroborated by the observation that 5% of the bacteria recovered from the lungs of guinea pigs infected with the wild-type strain RC1 were negative for mAb 2625 binding . These findings strongly indicate that under in vivo conditions switching between two LPS phenotypes occurs and may promote adaptation and replication of L . pneumophila . This is the first description of phase-variable expression of Legionella LPS.

Biochim Biophys Acta, 1998 Jun 24, 1372(1), 78 - 90
Membrane permeabilisation and antimycoplasmic activity of the 18-residue peptaibols, trichorzins PA; Beven L et al.; The membrane permeabilisation properties of six linear natural 18-residue peptaibols, termed trichorzins PA, have been assessed on liposomes and on mollicutes (trivial name, mycoplasmas), a class of parasitic bacteria characterized by a small genome, the lack of a cell wall, a minute cell size, and the incorporation in their plasma membrane of exogenously supplied cholesterol . The trichorzins PA used in this study (PA II, PA IV-VI, PA VIII, and PA IX) differ between them by amino acid or amino alcohol substitutions at positions 4, 7, and 18, and form slightly amphipathic alpha-helices . They proved bactericidal for mollicutes belonging to the genera Acholeplasma, Mycoplasma, and Spiroplasma, with minimal inhibitory concentrations (3.12</=MICs</=50 microM) generally 2 to 4 fold higher than those of alamethicin F50, a related 20-residue peptide (1.56</=MICs</=12.5 microM) . Spiroplasma cells were apparently not protected by the presence of spiralin on their surface . The activities of the six trichorzins PA were not influenced by their sequence variations and no synergistic effect was observed . Consistent with the marginal effect of cholesterol on the incorporation of the trichorzins PA into liposome bilayers, the antibiotic activity was independent of the amount of cholesterol in the membranes of the different mollicutes . The trichorzins PA and alamethicin inhibited the motility of Spiroplasma melliferum, the helical cells being deformed and split into coccoid forms . Membrane potential measurements in Acholeplasma laidlawii and S . melliferum showed that trichorzin PA V and alamethicin F50 very efficiently depolarized the plasma membrane of mollicutes . This was consistent with fluorescence and 23Na NMR measurements on liposomes that revealed the permeabilisation of the lipid bilayer and the nonselective ionophoric activity of the trichorzins PA . These data suggest that the bactericidal activity exhibited by the trichorzins PA on mollicutes is due to the permeabilisation of the plasma membrane .

J Autoimmun, 1998 Apr, 11(2), 185 - 90
Antineutrophil cytoplasmic autoantibodies (ANCA) in children with cystic fibrosis; Sediva A et al.; Anti-neutrophil cytoplasmic antibodies (ANCA) represent a useful diagnostic tool in patients with small vessel vasculitis . Circulating ANCA specific for bactericidal/permeability increasing protein (BPI) have been recently reported in adult patients with cystic fibrosis (CF), an autosomal recessive disorder caused by mutations in the cystic fibrosis transmembrane regulator (CFTR) gene with consequent impaired function of a transmembrane chloride channel . To contribute to the better understanding of the significance of ANCA in this disease, we investigated ANCA presence and antigenic specificity in children with CF . Results were correlated with clinical status, immunological data, age and genotype . The indirect immunofluorescence pattern of a total of 71 children with CF indicated that 31 were c-ANCA positive, while seven were p-ANCA positive . In further ELISA studies of ANCA antigenic specificity, 51 out of 66 investigated samples were positive for BPI, and 14 out of 28 were positive for proteinase 3 (PR3) . We found an association between levels of antibodies against PR3 with age and Pseudomonas infection . We did not, however, find any correlation between CFTR genotypes, Pseudomonas infection or paediatric parameters and the level of anti-BPI antibodies . High positivity of anti-BPI antibodies were seen even among the youngest CF patients, before the development of clinical signs of CF, indicating that formation of ANCA might be a very early event in the disease . Both anti-BPI and anti-PR3 antibodies may play a significant, although variable role, in the pathogenesis of CF.

Bull Acad Natl Med, 1998, 182(2), 285 - 97; discussion 297-8
{Recent aspects of monitoring the dosage of aminoglycosides and vancomycin}; Cheymol G et al.; The narrow range of therapeutic serum concentrations of aminoglycosides and vancomycin and the great variations in their pharmacokinetics from one person to another makes it important to monitor patients at risk that are treated with them . The technique of customizing dosages based on the population pharmacokinetics enables an effective treatment to be rapidly established from a few measurements of serum concentrations . Aminoglycosides may be given as single daily doses because of their concentration-dependent activity . The high peak concentration produces an enhanced, prolonged bactericidal activity, while a low residual concentration reduces the risk of toxicity . The effect of vancomycin is time-dependent . Giving it by continuous i.v . infusion maximizes the time during which the serum antibiotic concentration is effective but non-toxic . Monitoring serum concentrations can help reduce health care costs . But medical training in pharmacokinetics is needed for the optimal use of these therapeutic tools.

Infect Immun, 1998 Jul, 66(7), 3113 - 9
Phenotypic effect of isogenic uspA1 and uspA2 mutations on Moraxella catarrhalis 035E; Aebi C et al.; The UspA surface antigen of Moraxella catarrhalis was recently shown to be comprised of two different proteins (UspA1 and UspA2) which share an internal region containing 140 amino acids with 93% identity (C . Aebi, I . Maciver, J . L . Latimer, L . D . Cope, M . K . Stevens, S . E . Thomas, G . H . McCracken, Jr., and E . J . Hansen, Infect . Immun . 65:4367-4377, 1997) . Isogenic uspA1, uspA2, and uspA1 uspA2 mutants were tested in a number of in vitro systems to determine what effect these mutations, either individually or together, might exert on the phenotype of M . catarrhalis 035E . Monoclonal antibodies specific for UspA1 or UspA2 were used in an indirect antibody accessibility assay to prove that both of these proteins were expressed on the surface of M . catarrhalis . All three mutants grew in vitro at the same rate and did not exhibit autoagglutination or hemagglutination properties that were detectably different from those of the wild-type parent strain . When tested for the ability to adhere to human epithelial cells, the wild-type parent strain and the uspA2 mutant readily attached to Chang conjunctival cells . In contrast, the uspA1 mutant and the uspA1 uspA2 double mutant both attached to these epithelial cells at a level nearly 2 orders of magnitude lower than that obtained with the wild-type parent strain, a result which suggested that expression of UspA1 by M . catarrhalis is essential for attachment to these epithelial cells . Both the wild-type parent strain and the uspA1 mutant were resistant to the bactericidal activity of normal human serum, whereas the uspA2 mutant and the uspA1 uspA2 double mutant were readily killed by this serum . This latter result indicated that the presence of UspA2 is essential for expression of serum resistance by M . catarrhalis.

J Trauma, 1998 Jun, 44(6), 1047 - 54; discussion 1054-5
Recombinant human granulocyte colony-stimulating factor attenuates inflammatory responses in septic patients with neutropenia; Ishikawa K et al.; OBJECTIVE: The objective of this study was to determine the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) administration in septic patients with neutropenia . METHODS: Twenty consecutive septic patients were administered rhG-CSF subcutaneously (2 microg x kg(-1) x d(-1)) for 5 days (group G) . They were compared with 14 septic patients treated earlier without rhG-CSF (group N) . All patients in both groups met the criteria of total leukocyte count (TLC) less than 5,000/mm3 and C-reactive protein (CRP) more than 10 mg/dL . Changes in TLC, absolute neutrophil count (ANC), CRP, respiratory index (RI), Acute Physiology and Chronic Health Evaluation (APACHE) II score, and Goris's Multiple Organ Failure (MOF) index were evaluated . In addition, nucleated cell count (NCC), differentiation in bone marrow aspiration, neutrophil phagocytic and bactericidal activity, serum concentrations of interleukin-6 (IL-6) and IL-8 as inflammatory markers, and plasma concentration of leukocyte elastase (LE) as an indicator of the tissue injury were evaluated in group G . RESULTS: In group G, TLC, ANC, NCC, and neutrophil functions increased significantly, whereas CRP, IL-6, and IL-8 decreased reciprocally . There was no deterioration of LE and RI . Consequently, the APACHE II score and MOF index improved . In group N, however, CRP showed no change concomitant with the APACHE II score and MOF index . CONCLUSION: Administration of rhG-CSF attenuates inflammatory responses without inducing tissue injury in septic patients with neutropenia.

Scand J Med Sci Sports, 1998 Apr, 8(2), 91 - 7
Aspects of leukocyte function and the complement system following aerobic exercise in young female gymnasts; Wolach B et al.; Recent studies have reported reduced immunity in trained athletes . Scant information exists on changes in the immune function among trained children . The purpose of this study was to assess the effect of aerobic exercise on the phagocytic process of neutrophils and the complement system in young athletes . Subjects included prepubertal elite female gymnasts (n = 7) and untrained girls (n = 6) aged 10-12 years . Venous blood was withdrawn before, immediately post and 24 h following a 20-min run at a heart rate of 170-180 beats.min-1 . Neutrophil random migration, chemotactic activity, bactericidal function and PMA/FMLP-stimulated superoxide anion release as well as various complement components were assessed . Net chemotaxis was found reduced (P < 0.05) 24 h following exercise (58 +/- 11 vs . 36 +/- 11 cells/field in gymnasts and 47 +/- 7 vs . 42 +/- 8 cells/field in untrained girls pre- and 24 h post-exercise, respectively) . The basal values, as well as post-exercise values of bactericidal activity were lower (P < 0.05) in gymnasts as compared with the control group (0.8 +/- 0.3, 0.8 +/- 0.2 and 0.8 +/- 0.1 log decrease of colonies in gymnasts at pre-, immediately post-, and 24 h post-exercise, respectively and 1.1 +/- 0.1, 1.1 +/- 0.1 and 1.0 +/- 0.2 log decrease of colonies in controls, respectively) . No significant effect on the bactericidal activity was observed in either group following exercise . The addition of homologous sera did not correct the bactericidal activity . PMA-stimulated superoxide anion release decreased (P < 0.05) among gymnasts immediately following exercise (5.7 +/- 0.4 vs . 4.4 +/- 1.0 mmol O2/10(6) PMN.min) and remained low 24 h later . The same trend was observed in FMLP-stimulated neutrophils but the data were not significant . Significantly decreased levels (P < 0.05) of the early complement components (C1Q, C1R) were also found following exercise (1.34 +/- 0.64 vs . 1.27 +/- 0.28 and 1.09 +/- 0.07 vs . 1.02 +/- 0.06 pre- and post-exercise in gymnasts and untrained, respectively) . Furthermore, consistently lower C2 and C3 were observed in gymnasts compared with controls . Neutrophil dysfunction as well as impairment of the complement system seem to occur following exercise.

FEMS Microbiol Lett, 1998 May 15, 162(2), 331 - 7
Variation in Bordetella bronchiseptica lipopolysaccharide during human infection; Gueirard P et al.; We previously reported the case of a human chronic Bordetella bronchiseptica respiratory infection, due to contact with infected rabbits . Lipopolysaccharides of the human isolates, of one rabbit isolate and of isolate from other origins were analyzed with sera from infected mice, rabbit and human . Antigenicity and length of the lipopolysaccharide molecules varied between isolates . We showed a progressive loss of O-chain during infection, associated with an enhanced susceptibility of the isolates to the bactericidal effect of normal serum . This observation suggests the existence of an intracellular niche which selects for strains with distinct lipopolysaccharide types.

Br J Biomed Sci, 1997 Dec, 54(4), 260 - 6
Polymorphonuclear neutrophils in saliva and blood: a comparative study of morphology, function and phenotype; Takubo T et al.; The morphology, phagocytic activity, production of hydrogen peroxide (H2O2) and expression of lymphocyte function-associated antigen-1 (LFA-1; CD11a/CD18), complement receptor type 3 (CR3; CD11b/CD18), p150,95 (CD11c/CD18), and platelet/endothelial cell adhesion molecule-1 (PECAM-1; CD31) by oral polymorphonuclear neutrophils (PMN) are assessed and the results compared with those of blood PMN . There were no differences in the morphology and phagocytic activity between oral and blood PMN . H2O2 production was measured following stimulation with phorbol myristate acetate (PMA) and N-formyl-L-leucyl-L-phenylalanine (FMLP) as indicators of bactericidal activity . There was a significant difference in the H2O2 production by the two groups when stimulated with FMLP; the level of H2O2 production by oral PMN was significantly higher than that by blood PMN . However, there was no significant difference in H2O2 production between oral and blood PMN when stimulated with PMA . The percentage of CD11a- and CD11c-positive concentrated oral PMN was significantly lower than that seen in blood PMN, as was the percentage of CD31-positive cells . Higher H2O2 production by oral PMN following stimulation with FMLP may result in enhanced bactericidal activity . Low expression of CD31 may lead to the accumulation of PMN in the mouth by blocking their return to the bloodstream . These phenomena may be necessary for oral PMN to protect periodontal tissues from bacteria in the mouth.

Clin Infect Dis, 1998 Apr, 26(4), 874 - 7
Activity of amoxicillin/clavulanate in patients with tuberculosis; Chambers HF et al.; Some beta-lactam antibiotics are active in vitro against Mycobacterium tuberculosis . There are anecdotal reports of successful treatment of tuberculosis caused by multiple-drug-resistant strains of M . tuberculosis with regimens that included amoxicillin/clavulanate . Reduction of M . tuberculosis in the sputum of patients with pulmonary tuberculosis during administration of amoxicillin/clavulanate was measured by a quantitative culture method to determine the activity in vivo . Patients were randomized to receive isoniazid, ofloxacin, or amoxicillin/clavulanate for 7 days . Isoniazid was the most effective agent, reducing M . tuberculosis after 2 days at a mean rate (+/- standard deviation) of 0.60 +/- 0.30 log10 cfu/mL per day, compared with 0.32 +/- 0.05 and 0.34 +/- 0.03 for ofloxacin and amoxicillin/clavulanate, respectively . The early bactericidal activity of amoxicillin/clavulanate was comparable to that reported for antituberculous agents other than isoniazid . Further studies of beta-lactam antibiotics with in vitro activity against M . tuberculosis are warranted to define their role in treatment of tuberculosis.

Autoimmunity, 1998, 27(4), 231 - 8
Anti-neutrophil cytoplasm autoantibodies (ANCA) . The need for specific and sensitive assays; Baslund B et al.; Anti-neutrophil cytoplasm antibodies (ANCA) are a group of autoantibodies primarily associated with systemic vasculitis . Hitherto, the method of choice for ANCA detection has been indirect immunofluorescence (IIF) . By this method two major patterns can be seen: a cytoplasmic pattern (cANCA) or a perinuclear pattern (pANCA) . The cANCA pattern is most often caused by antibodies directed against proteinase-3 (PR3) and in rare cases it is caused by anti-myeloperoixdase (MPO) antibodies . The pANCA pattern can de caused by antibodies directed against a large group of proteins i.e . MPO, lactofenin and bactericidal/permeability-increasing protein . Often there is a discrepancy between the results obtained by IIF and those reported from the use of assays with purified antigens . This causes confusion . Until now only anti-PR3 and anti-MPO have been found of any clinical value . Therefore, it would be more proper to use assays with these highly purified antigens instead of an unspecific method like IIF.

Eksp Klin Farmakol, 1998 Mar-Apr, 61(2), 48 - 50
{The effect of the immunostimulating agent trimezon on the cells of the mononuclear phagocytosing system}; Liubin GS et al.; The effect of the immunostimulator trimeson of the 8-aza-gomogonan series on the cells of the mononuclear monocyte system was studied in C3H and ICR mice in vitro . Intraperitoneal injection of trimeson (50 mg/kg 1 x 10(-7)-1 x 10(6) M) inhibits, as a rule, the initial stages of the phagocytizing process but stimulates the bactericidal properties of the peritoneal phagocytes and their mechanisms of processing and destruction of the antigen material . In the system of adaptive transfer of donor splenocytes and peritoneal macrophages to recipients exposed to lethal radiation, trimeson (20 mg/kg intravenously, 50 mg/kg per os) increases the immunogenic function of the mononuclear phagocytes in formation of the primary immune response to sheep erythrocytes.

J Biol Chem, 1998 Jun 5, 273(23), 14071 - 3
Helicobacter pylori generates superoxide radicals and modulates nitric oxide metabolism; Nagata K et al.; During studies of the bactericidal action of nitric oxide (NO), we found that it reversibly inhibited the respiration of Escherichia coli and irreversibly inhibited the respiration of Helicobacter pylori . Peroxynitrite, a reaction product of NO and superoxide, irreversibly inhibited the respiration of both H . pylori and E . coli . H . pylori, but not E . coli, generated substantial amounts of superoxide radicals . These results suggest that NO directly inhibits the respiration of E . coli whereas it rapidly reacts with endogenously generated superoxide radicals in H . pylori . The resulting peroxynitrite inactivates the respiration of H . pylori.

Cell Mol Biol (Noisy-le-grand), 1998 May, 44(3), 521 - 6
Streptomycin bactericidal action is dependent on polyamine endogenous levels in E . coli; Diniello GB et al.; E . coli polyamine-supplemented and depleted cultures showed an important difference in survival to streptomycin; the bactericidal effect of the antibiotic was remarkably higher in cells with normal levels of polyamines . Similar results were observed with kanamycin . Analysis of the polyamine-containing cells pulse-labelled with 35S-methionine during streptomycin action indicated that the amounts of newly-synthesized peptides in various subcellular fractions was different from the amounts formed in the untreated controls; the most dramatic change was found in the residual particulate fraction where the antibiotic treatment caused a 3-fold increase of radioactive proteins . On the contrary, equivalent amounts of labelled peptides were detected in the different fractions prepared from polyamine-depleted bacteria incubated with or without antibiotic . In this case the corresponding residual fraction was only slightly increased . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the different fractions showed some changes elicited by streptomycin in the protein patterns of polyamine-containing bacteria, especially in the residual fractions . The electrophoretic profile corresponding to deprived cells was very similar in all cases . The role of polyamines in the conformation of the outer membrane and in the correct assembly of ribosomes is discussed on account of the enhancing effect of these polycations on the bactericidal action of streptomycin.

J Bone Joint Surg Br, 1998 May, 80(3), 527 - 30
Treatment of osteomyelitis with antibiotic-soaked porous glass ceramic; Kawanabe K et al.; We have developed a new drug delivery system using porous apatite-wollastonite glass ceramic (A-W GC) to treat osteomyelitis . A-W GC (porosity, 70% and 20% to 30%), or porous hydroxyapatite (HA) blocks (porosity 35% to 48%) used as controls, were soaked in mixtures of two antibiotics, isepamicin sulphate (ISP) and cefmetazole (CMZ) under high vacuum . We evaluated the release concentrations of the antibiotics from the blocks . The bactericidal concentration of ISP from A-W GC was maintained for more than 42 days, but that from HA decreased to below the detection limit after 28 days . The concentrations of CMZ from both materials were lower than those of ISP . An in vivo study using rabbit femora showed that an osseous concentration of ISP was maintained at eight weeks after implantation . Osteoconduction of the A-W GC block was good . Four patients with infected hip arthroplasties and one with osteomyelitis of the tibia have been treated with the new delivery system with excellent results.

Dev Comp Immunol, 1998 Jan-Feb, 22(1), 55 - 61
A comparison of the chemiluminescent response of Crassostrea virginica and Morone saxatilis phagocytes to zymosan and viable Listonella anguillarum; Bramble LH et al.; If reactive oxygen species (ROS) produced by hemocytes of the eastern oyster, Crassostrea virginica, impart bactericidal activity, exposure of hemocytes to bacteria should result in increased ROS generation . In an earlier study, this hypothesis was tested using luminol- and lucigenin-augmented chemiluminescence (CL) to measure ROS production . The bacterium Listonella anguillarum did not stimulate a net increase in hemocyte-derived CL, and it was suggested that bacterial antioxidants might suppress hemocyte CL . In the present study a comparison was made, under identical assay conditions, of the zymosan- and bacteria-enhanced luminol CL produced by eastern oyster hemocytes and by striped bass (Morone saxatilis) macrophages, for which L . anguillarum has been shown to be a stimulus in CL reactions . The response to zymosan produced by bass phagocytes was two orders of magnitude greater than that generated by eastern oyster hemocytes . Whereas an increase in net ROS production was not evident when oyster hemocytes were exposed to L . anguillarum, significant stimulation of striped bass macrophage-derived CL occurred . These data suggest that striped bass macrophages have a greater capacity to generate ROS than oyster hemocytes, enabling them to surpass the antioxidant capability of L . anguillarum and produce a luminol CL response.

Antiviral Res, 1998 Apr, 38(1), 25 - 30
Activity of N-chlorotaurine against herpes simplex- and adenoviruses; Nagl M et al.; N-chlorotaurine, an essential weak oxidant produced by stimulated human leukocytes, is known to have bactericidal, fungicidal and vermicidal properties . This study for the first time demonstrates its virucidal activity . By viral suspension tests at incubation times between 5 and 60 min, virus titers of both Herpes simplex virus type 1 and 2 were reduced about 1.3-2.9 log10 and 2.8-4.2 log10 by 0.1 and 1%, (5.5 and 55 mM) N-chlorotaurine, respectively . Virus titer reduction of adenovirus type 5 between 15 and 60 min was 0.5-2.0 and 0.6-4.0 log10, respectively, by the same concentrations of N-chlorotaurine . These findings support a contribution of N-chlorotaurine in destruction of pathogens during inflammatory reactions and also the possibility of its application as an antiviral agent in human medicine.

Australas J Dermatol, 1998 May, 39(2), 96 - 9
Antineutrophil cytoplasmic antibody (ANCA)-positive cutaneous leucocytoclastic vasculitis associated with antithyroid therapy in Graves' disease; Miller RM et al.; Presented is a case of a 27-year-old male with Graves' disease on long-term propylthiouracil treatment who, when changed to carbimazole, rapidly developed a petechial and purpuric eruption on the legs, which subsequently flared on treatment with radioiodine . The clinical diagnosis of leucocytoclastic vasculitis was confirmed on skin biopsy . High-titre antineutrophil cytoplasmic antibodies in a perinuclear pattern (P-ANCA) were identified . No anti-myeloperoxidase activity was noted; therefore, the P-ANCA were classified in the atypical group . The target antigens, as determined by enzyme-linked immunosorbent assay, were lysozyme, lactoferrin and bactericidal/permeability increasing protein . Propylthiouracil and carbimazole are chemically related antithyroid drugs . There are reports of typical and atypical P-ANCA-positive cutaneous vasculitis due to propylthiouracil . Cutaneous vasculitis associated with atypical P-ANCA has not been noted previously to be temporally related to carbimazole use . The consideration of thionamides as possible aetiological agents in cases of P-ANCA-positive drug-induced vasculitis is suggested.

Am J Respir Crit Care Med, 1998 May, 157(5 Pt 1), 1436 - 40
Once-weekly rifapentine-containing regimens for treatment of tuberculosis in mice; Grosset J et al.; The bactericidal activities of several once-weekly rifapentine (P)-containing combination regimens against Mycobacterium tuberculosis, and their ability to prevent the selection of rifampin (R)-resistant mutants, were compared with those of the standard six-times-weekly regimen consisting of R, isoniazid (H), and pyrazinamide (Z) in a mouse experiment . Mice were infected intravenously with 1.3 x 10(7) cfu of M . tuberculosis strain H37Rv, and 8 wk of treatment began on Day 14 after infection, when mice were randomly allocated to an untreated control group and nine treatment groups of 30 mice each . At the end of 8 wk of treatment, all the tested regimens showed promising bactericidal activities . Once-weekly P alone was less bactericidal than six-times-weekly R alone; likewise, the once-weekly P-containing combined regimens were less bactericidal than the six-times-weekly standard regimen . However, the difference in killing was about 1 log10, which represented only a fraction of the overall 4 log10 to 5 log10 magnitude of killing effects . The addition of streptomycin (S) improved the bactericidal effect of once-weekly PHZ, and the effect of once-weekly PHZS was further enhanced when it was preceded by 2 wk of daily HZS . The latter regimen achieved the same level of activity as the standard six-times-weekly regimen . All of the once-weekly P-containing combined regimens were able to prevent the selection of R-resistant mutants, whereas monotherapy with R or P selected resistant mutants in approximately 50% of animals.

Chest, 1998 May, 113(5), 1290 - 5
Systemic inflammation present in patients undergoing CABG without extracorporeal circulation; Fransen E et al.; STUDY OBJECTIVES: This study was conducted to evaluate to what extent the cardiopulmonary bypass (CPB) procedure in patients undergoing coronary artery bypass grafting (CABG) contributes to the systemic inflammatory response . Therefore, we measured bactericidal permeability increasing protein (BPI) as an indicator of neutrophil activation, interleukin 6 as inducer of the acute phase response, and lipopolysaccharide binding protein and C-reactive protein as parameters of the acute phase response in patients undergoing CABG either with or without the use of CPB . DESIGN: Prospective study . SETTING: Cardiopulmonary surgery department in a university hospital . PATIENTS: Sixteen patients undergoing elective CABG were included . Eight patients underwent surgery with CPB, and eight patients underwent surgery without CPB (non-CPB) . INTERVENTIONS: In the CPB group, blood samples were taken upon induction of anesthesia, at the start of aortic cross-clamping, at aortic unclamping, and 0.5, 4, 8, and 18 h thereafter . In the non-CPB group, blood samples were taken upon induction of anesthesia, and 0.5, 4, 8, and 18 h after completion of the bypass graft anastomoses . MEASUREMENTS AND RESULTS: BPI release from neutrophil granules markedly increased during surgery in CPB patients but not in non-CPB patients . The increase in acute phase reactants, however, was the same in both patient groups . CONCLUSIONS: These data indicate that the acute phase response in CABG patients, which has historically been ascribed to the CPB procedure, is predominantly caused by the surgical procedure per se . Early neutrophil activation, however, is seen only when extracorporeal circulation is used.

FEMS Microbiol Lett, 1998 May 1, 162(1), 117 - 22
Physiological consequences of the over-production of E . coli truncated molecular chaperone DnaJ; al-Herran S et al.; In Escherichia coli one of the main molecular chaperones is DnaJ (hsp40) which mediates in a variety of highly conserved cellular process including protein folding reactions and assembly/disassembly of protein complexes . In this study we have investigated the toxicity of the over-production of DnaJ and two truncated polypeptides by examining growth rates, cell morphology and cell viability . Full-length DnaJ (1-375 amino acids) and truncated polypeptides, corresponding to the last 176 (containing the substrate binding domain) and 266 (containing the zinc finger-like domain) amino acids of the C-terminus of DnaJ, DnaJ delta 1-199 and delta 1-108 respectively, were over-produced via IPTG induction . High levels of synthesis were determined by SDS-PAGE and Western blotting using anti-DnaJ antibodies . The over-production of full-length DnaJ resulted in a low degree of filamentation and a decrease in cell viability . However, over-production of DnaJ truncated polypeptides, especially DnaJ delta 1-108, was bactericidal and resulted in a loss of viability and defective septation.

Antimicrob Agents Chemother, 1998 May, 42(5), 1115 - 20
Bactericidal activity of a single-dose combination of ofloxacin plus minocycline, with or without rifampin, against Mycobacterium leprae in mice and in lepromatous patients; Ji B et al.; To develop a fully supervisable, monthly administered regimen for treatment of leprosy, the bactericidal effect of a single-dose combination of ofloxacin (OFLO) and minocycline (MINO), with or without rifampin (RMP), against Mycobacterium leprae was studied in the mouse footpad system and in previously untreated lepromatous leprosy patients . Bactericidal activity was measured by the proportional bactericidal method . In mouse experiments, the activity of a single dose of the combination OFLO-MINO was dosage related; the higher dosage of the combination displayed bactericidal activity which was significantly inferior to that of a single dose of RMP, whereas the lower dosage did not exhibit a bactericidal effect . In the clinical trial, 20 patients with previously untreated lepromatous leprosy were treated with a single dose consisting of either 600 mg of RMP plus 400 mg of OFLO and 100 mg of MINO or 400 mg of OFLO plus 100 mg of MINO . The OFLO-MINO combination exhibited definite bactericidal activity in 7 of 10 patients but was less bactericidal than the RMP-OFLO-MINO combination . Both combinations were well tolerated . Because of these promising results, a test of the efficacy of multiple doses of ROM in a larger clinical trial appears justified.

Antimicrob Agents Chemother, 1998 May, 42(5), 1022 - 7
Modulation of gyrase-mediated DNA cleavage and cell killing by ATP; Li TK et al.; An uncoupler of oxidative phosphorylation, 2,4-dinitrophenol, and an aconitase inhibitor, fluoroacetic acid, both of which are known to lower the cellular ATP pool, protected Escherichia coli cells from the bactericidal actions of gyrase poisons including quinolone antibiotics, nalidixic acid and ciprofloxacin, and the epipodophyllotoxins VP-16 and VM-26 . Using purified E . coli DNA gyrase, we examined the effect of ATP on gyrase-mediated DNA cleavage in the presence of these gyrase poisons . ATP was shown to stimulate gyrase-mediated DNA cleavage from 10- to more than 100-fold in the presence of these gyrase poisons . ADP antagonized the stimulatory effect of ATP . Consequently, gyrase-mediated DNA cleavage induced by gyrase poisons is modulated by the ATP concentration/ADP concentration ({ATP}/{ADP}) ratio . Coumermycin A1, an inhibitor of the ATPase subunit of DNA gyrase, like ADP, also effectively antagonized the stimulatory effect of ATP on gyrase-mediated DNA cleavage induced by gyrase poisons . Furthermore, coumermycin A1, like DNP and fluoroacetic acid, also protected cells from the bactericidal action of gyrase poisons . In the aggregate, our results are consistent with the notion that the {ATP}/{ADP} ratio, through its modulatory effect on the gyrase-mediated DNA cleavage, is an important determinant of cellular susceptibility to gyrase poisons.

Am J Hematol, 1998 May, 58(1), 8 - 15
Neonatal neutrophil inflammatory responses: parallel studies of light scattering, cell polarization, chemotaxis, superoxide release, and bactericidal activity; Wolach B et al.; Neutrophil dysfunction among newborn infants, especially those born prematurely, is well recognized, but the mechanism responsible for this phenomenon is yet to be clarified . In this study, we evaluated the stimulus response coupling in neutrophils from 90 healthy newborns and 96 healthy adults in an effort to establish whether defective neonatal neutrophil function is a result of impaired signal perception or immature responsiveness . Measurement of rapid- and slow-light scattering responses (LSR) to 1 microM FMLP stimulation revealed that neonatal neutrophils have about one-half the corresponding responsiveness of adult cells (rapid-LSR: 6.1 +/- 3.1 arbitrary light intensity units vs . 12.0 +/- 2.8, P < .001; and slow-LSR: 5.0 +/- 2.5 vs . 9.1 +/- 2.0; P < .001) . The same markedly reduced activity was observed in newborn neutrophil chemotaxis and bactericidal activity in comparison with adult cells . Nevertheless, low FMLP concentrations (less than 1 nM) induced no difference in cell polarization between newborn and adult neutrophils, yet at higher FMLP concentrations, the newborn revealed significantly reduced cell polarization . Our data suggest that newborn infants bear a fully functional FMLP signal perception but lack the full capacity of inflammatory responsiveness.

Am J Med Sci, 1998 May, 315(5), 307 - 13
Complement component C9 enhances the capacity of beta-lactam antibiotics to kill Escherichia coli in vitro and in vivo; Jung E et al.; Complement component C9 is required for rapid complement-mediated killing of Escherichia coli . In this report, the influence of supplemental C9 on the bactericidal and protective effects of beta-lactam antibiotics in neonates was assessed . By rocket immunoelectrophoresis, the intrinsic C9 concentrations of pooled serum from both human and rat neonates was less than 20% of adult levels . Supplemental C9 purified from human plasma enhanced the capacity of ampicillin-treated serum from human neonates to impair the survival of E coli O7:K1:NM (P < 0.02) . Similarly, supplemental C9 enhanced the capacity of cefotaxime-treated neonatal rat serum to impair the survival of E coli O1:K1:NM (P < 0.05) . Moreover, the intraperitoneal administration of C9 enhanced the survival of cefotaxime-treated neonatal rats that were septic with E coli (P < 0.05) . These observations may contribute to the development of new strategies, such as augmentation of complement component serum concentrations, to reduce the morbidity and mortality of neonatal E coli sepsis.

Kansenshogaku Zasshi, 1998 Mar, 72(3), 211 - 7
{Bactericidal and anti-toxin activities of catechin on enterohemorrhagic Escherichia coli}; Okubo S et al.; We examined the bactericidal activity of catechin, an astringent ingredient of tea, on enterohemorrhagic Escherichia coli (EHEC) O157:H7 and the anti-toxin activity of catechin on vero toxin (VT), the main pathogenic factor of EHEC O157:H7 . To examine bactericidal activity, we added 1 X 10(4) CFU/ml bacteria to 1.25 to 20 W/V% of green tea extract or the PBS solution containing 25 to 400 micrograms/ml of (-) epigallocatechin gallate (EGCg), which is the main catechin ingredient of green tea leaf, and counted the number of live bacteria at various intervals . After 3 to 5 hours, no live bacteria were seen in 1.25 to 2.5 (regular drinking concentration) % green tea extract . In the high concentrations of 100 to 400 micrograms/ml EGCg the number of live bacteria decreased with time and after 24 hours no survivors were seen . In the low concentrations of 25 to 50 micrograms/ml EGCg, however, no change was observed in the number of live bacteria during 5 hours . After 24 hours the bacteria in 50 micrograms/ml were killed and the number of bacteria in 25 micrograms/ml decreased to one tenth of that at the start . To examine the anti-toxin activity, we mixed equal volumes of 2 ng/0.1 ml VT2 and 0.5 to 2 mg/0.1 ml catechin in vitro and incubated them at 37 degrees C for various times . Then we inoculated 0.2 ml of the mixture intraperitonealy to BALB/c mice . One mg of catechin inhibited by 100% the lethal toxicity of 2 ng of VT2 (LD 100) to mice . The inhibition of lethal toxicity of VT2 by catechin depended on the incubation time . The rate of inhibition was 0, 40 and 100% for 9, 12 and 18-24 hours incubation, respectively . These results suggest that catechin has not only bactericidal activity on EHEC O157:H7 but also anti-toxin activity on vero toxin.

Proc Natl Acad Sci U S A, 1998 Apr 14, 95(8), 4321 - 6
Translocation of inserted foreign epitopes by a channel-forming protein; Jakes KS et al.; Certain bacterial protein toxins are able to insert themselves into, and at least partially across, lipid bilayer membranes in the absence of any auxiliary proteins, by using unknown mechanisms to overcome the high energy barrier presented by the hydrophobic bilayer core . We have previously shown that one such toxin, colicin Ia, translocates a large, hydrophilic part of itself completely across a lipid bilayer in conjunction with the formation of an ion-conducting channel . To address the question of whether the colicin can translocate any arbitrary amino acid sequence, we have altered the translocated segment by inserting, singly, two different foreign epitopes . Colicins containing either epitope retain significant bactericidal activity and form channels of normal conductance in planar bilayers . Furthermore, antibodies added on the side of the bilayer opposite that to which the colicin was added interact specifically with the corresponding epitopes, producing an inhibition of channel closing . Thus, the inserted epitopes are translocated along with the rest of the segment, suggesting that a surprisingly small part of colicin Ia, located elsewhere in the molecule, acts as a nonspecific protein translocator.

Rocz Akad Med Bialymst, 1997, 42(1), 141 - 7
The bactericidal activity of blood platelets in patients infested with Entamoeba histolytica/Entamoeba dispar; Matowicka-Karna J et al.; Blood platelets participate actively in organism defensive reactions and particularity in antiparasitic immunity . Platelets stimulation can be the result of contact with a parasite, the presence of immunoglobulins, complement and lymphokines . The aim of the study was to evaluate the blood platelets bactericidal activity in the course of amoebiasis . The examination was carried out in the group of 22 patients with E . histolytica/E . dispar, diagnosed on the basis of cyst forms presence in faeces . The results show that blood platelets bactericidal activity is significantly impaired in the course of amoebiasis.

J Antimicrob Chemother, 1998 Mar, 41 Suppl B, 23 - 7
The comparative in-vitro activity of roxithromycin and other antibiotics against Bordetella pertussis; Brett M et al.; In spite of vaccination programmes, whooping cough epidemics continue to occur . The disease affects all age groups, although its severity is greatest in the young, with infants being particularly vulnerable . Erythromycin is generally accepted as the drug of choice both for treatment and for prophylaxis during epidemics . Roxithromycin is a macrolide with pharmacokinetic advantages over erythromycin; it is well absorbed, produces high serum concentrations, has a long half-life and penetrates respiratory secretions well . There are no accepted standards for testing the sensitivity of Bordetella pertussis to antibiotics, and reports of the activity of roxithromycin and erythromycin are variable . Using Isosensitest agar supplemented with 5% horse blood and an inoculum of 10(4) cfu, 88 strains of B . pertussis were tested for their sensitivity to roxithromycin, erythromycin, rifampicin and trimethoprim/sulphamethoxazole . The range of MICs was 0.12-0.5 mg/L for both roxithromycin and erythromycin . Roxithromycin was bactericidal, with an MBC of 1 mg/L (as compared with 0.5 mg/L for erythromycin) . Since roxithromycin is well tolerated by children when used for respiratory tract infections, the good in-vitro activity against B . pertussis, combined with its favourable pharmacokinetics, suggest it may be a good candidate for use in the treatment and prophylaxis of whooping cough.

LDA J, 1997 Fall, 56(3), 16 - 8
Efficacy of topical disinfectants; Oeschger MP et al.; A comparison of bactericidal activity of two surface disinfectants showed that both were highly effective . The glutaraldehyde-based disinfectant showed higher efficacy on roughed surfaces than the alcohol-based disinfectant . The study used a novel, and generally applicable, method to evaluate the surface killing potential of disinfectants.

Int J Biochem Cell Biol, 1997 Dec, 29(12), 1401 - 18
IL-6-regulated transcription factors; Akira S; Through the cloning of two transcription factors named NF-IL6 and STAT3/APRF, two types of IL-6 signal transduction pathways from the cell surface to the nucleus have been revealed . NF-IL6 is phosphorylated and activated by a Ras-dependent MAP kinase cascade, while STAT3/APRF is directly tyrosine-phosphorylated by JAK kinases that associate with the cytoplasmic portion of the receptor, and translocates to the nucleus and activates transcription (JAK-STAT pathway) . STAT3 is also tyrosine phosphorylated in response to epidermal growth factor (EGF), granulocyte colony-stimulating factor (G-CSF), leptin and other IL-6-type cytokines including ciliary neurotrophic factor (CNTF), oncostatin M and leukemia inhibitory factor (LIF) . Mice deficient in the genes for NF-IL6 and STAT3 were generated . NF-IL6 mice were highly susceptible to facultative intracellular bacteria owing to ineffective killing of the pathogens by the macrophages . Futhermore, the tumor cytotoxicity of macrophages from NF-IL6 KO mice was severely impaired . These results demonstrate a crucial role of NF-IL6 in macrophage bactericidal and tumoricidal activities . The target disruption of STAT3 resulted in embryonic lethality prior to gastrulation, demonstrating that STAT3 is essential for the early development of mouse embryos.

Lett Appl Microbiol, 1998 Mar, 26(3), 194 - 8
Effects of tea tree oil on Escherichia coli; Gustafson JE et al.; Tea tree oil (TTO) stimulates autolysis in exponential and stationary phase cells of Escherichia coli . Electron micrographs of cells grown in the presence of TTO showed the loss of electron dense material, coagulation of cell cytoplasm and formation of extracellular blebs . Stationary phase cells demonstrated less TTO-stimulated autolysis and also had greater tolerance to TTO-induced cell death, compared to exponentially grown cells . It was also revealed that subpopulation of stationary phase cells demonstrated increased tolerance to TTO-bactericidal effects.

Clin Exp Immunol, 1998 Apr, 112(1), 10 - 6
Catalase and alpha-enolase: two novel granulocyte autoantigens in inflammatory bowel disease (IBD); Roozendaal C et al.; In IBD, the target antigens of anti-neutrophil cytoplasmic autoantibodies (ANCA) have not been fully identified, which limits the analysis of the diagnostic significance as well as of the possible pathophysiological role of these antibodies . In this study, we identify the target antigens of ANCA in large groups of patients with ulcerative colitis (UC) and Crohn's disease (CD) . Apart from antibodies against lactoferrin and bactericidal/permeability-increasing protein (BPI), which have been reported before, antibodies against two novel granulocyte antigens were identified: antibodies against a 57/56-kD doublet were found in 38% of samples from UC patients and in 26% of samples from CD patients, whereas antibodies against a 47-kD protein were found in 10% of samples from UC patients and in 18% of samples from CD patients . Partial purification and amino acid sequence analysis identified the 57-kD protein as catalase and the 47-kD protein as alpha-enolase . This study is the first to report catalase and alpha-enolase as granulocyte antigens for autoantibodies in IBD.

Diagn Microbiol Infect Dis, 1998 Feb, 30(2), 135 - 7
Timed kill kinetic studies of levofloxacin, ofloxacin, and ciprofloxacin against Moraxella catarrhalis; Barrett MS et al.; Levofloxacin bactericidal activity was compared to ciprofloxacin and ofloxacin against 10 strains of Moraxella catarrhalis . The cidal action (by kill-curve analysis) was slightly more rapid for levofloxacin, but all tested fluoroquinolones were considered bactericidal for all strains tested, including those producing BRO-1 and 2 beta-lactamases.

Int J Antimicrob Agents, 1997 Jan, 9(3), 169 - 73
The activity of rifabutin against Mycobacterium leprae in armadillos; Dhople AM et al.; The activity of rifabutin (LM 427) against Mycobacterium leprae was evaluated in armadillos inoculated earlier with human-derived M . leprae . Rifabutin was administered daily at a dose of 6 mg/kg body weight/day . The effect of rifabutin on M . leprae harvested from armadillos was determined by measuring the intracellular levels of ATP (an indicator of metabolic activity) of M . leprae and also their ability to multiply in the mouse footpads and in vitro in DH medium . Within 2 weeks of initiating the treatment, ATP levels declined to 21% of the original (pre-treatment level) and these M . leprae failed to multiply in the footpads of mice as well as in the in vitro culture system . This suggests that rifabutin was able to kill all M . leprae within 2 weeks . After 8 weeks the treatment was terminated and results showed that M . leprae from the treated armadillos remained non-viable in the mouse footpad system as well as in the in vitro system, indicating bactericidal action of rifabutin . The results suggest that rifabutin can be a substitute for rifampin in the leprosy multi-drug therapy regimen.

PDA J Pharm Sci Technol, 1998 Jan-Feb, 52(1), 37 - 42
Bacterial cell size and surface charge characteristics relevant to filter validation studies; Mittelman MW et al.; There are two recognized mechanisms whereby organisms are retained by liquid filters; namely, sieve-retention and adsorption . The efficiency of each may be influenced by the organism, suspending milieu, and by the filtration conditions . Validations of sterilizing filtrations require the use of organism suspensions in product-specific media . However, where the product is bactericidal to the challenge organism(s), surrogate solutions may be required . The ideal surrogate solution would minimize adsorptive retention, ensuring that the sterilizing action of the filter under consideration is the consequence of sieve-retention . This review explores the impact that various physicochemical factors may have on bacterial cell size and cell surface characteristics . An understanding of interactions among challenge bacteria, suspending fluid, and filter medium is essential for the development of surrogate solutions that provide a "worst case" mileu for filter validation studies or a "placebo," non-inhibitory challenge solution.

Helicobacter, 1998 Mar, 3(1), 21 - 7
Growth inhibitory and bactericidal activities of lansoprazole compared with those of omeprazole and pantoprazole against Helicobacter pylori; Nakao M et al.; BACKGROUND: Helicobacter pylori plays a role in the pathogenesis of both duodenal and gastric ulcers . The aim of this study was to evaluate the effect of the proton pump inhibitor (PPI), lansoprazole, commonly used in eradication regimens, on growth, bactericidal activity and morphology of H . pylori in vitro in comparison with other PPIs . MATERIALS AND METHODS: Growth inhibitory activity of each of the PPIs was evaluated by determining minimum inhibitory concentrations using an agar dilution method . Bactericidal activity was determined by analysis of the viable cells in culture at various time points after incubation with different concentrations of the PPIs . Bacterial morphology was examined using scanning electron microscopy of fixed cells after exposure to the test substances . Urease activity in cell extracts of H . pylori that had been incubated with increasing concentrations of the PPIs was determined by colorimetry . RESULTS: The growth inhibitory activity of lansoprazole was significantly more potent than that of omeprazole or pantoprazole (MIC90 6.25 vs . 25 and 100 micrograms/ml, respectively) . Exposure of H . pylori to lansoprazole produced loss of viability and an aberrant bacterial morphology, which was more extensive than seen with omeprazole or pantoprazole . Lansoprazole dose dependently inhibited urease activity; its effectiveness was comparable with omeprazole but more potent than pantoprazole . CONCLUSIONS: The mechanism of action that leads to loss of viability of H . pylori cells appears to differ between the three PPIs investigated; lansoprazole was the most potent of the three agents in terms of growth inhibition and disruption of bacterial morphology.

J Antimicrob Chemother, 1998 Feb, 41(2), 171 - 7
Flavodoxin-dependent pyruvate oxidation, acetate production and metronidazole reduction by Helicobacter pylori; Kaihovaara P et al.; Helicobacter pylori flavodoxin was purified to homogeneity from cell extracts of strain NCTC 11637 . The molecular weight of the protein was estimated by gel electrophoresis to be 18 kDa . Oxidized flavodoxin showed an absorption spectrum with maxima at 378 nm and 453 nm, and it was reduced to a neutral form of flavin semiquinone by the electrons generated in the oxidation of pyruvate . This coenzyme A dependent pyruvate:flavodoxin oxidoreductase activity of H . pylori was also detected as a reduction of methyl viologen or cytochrome c by bacterial extracts . The apparent Km of pyruvate was 310 microM . Anaerobically incubated bacteria (10{9}) of strain NCTC 11637 produced acetate (96 +/- 16 nmol/h) from pyruvate concomitantly reducing metronidazole (17 +/- 5 nmol/h) . In anaerobic conditions both sensitive and resistant H . pylori strains reduced metronidazole, and there was a significant positive correlation between acetate production and metronidazole activation (r = 0.77, P < 0.01, n = 11) . In the presence of atmospheric oxygen, H . pylori excreted twice as much acetate but metronidazole was not activated . These results suggest that the pyruvate:flavodoxin oxidoreductase complex catalyses pyruvate oxidation in H . pylori . Electrons generated in this reaction are transferred to flavodoxin and under anaerobic conditions further to metronidazole (imidazoles) thus reducing the drug to its bactericidal form.

Cent Eur J Public Health, 1998 Feb, 6(1), 51 - 6
Long-term monitoring of the immune reactivity of stainless steel welders; Hanovcova I et al.; The immune reactivity of stainless steel welders (n = 22-53) was evaluated in a three year's study . The results (phagocytic activity, cellular and humoral immunity) were statistically compared with those in control group of non-exposed persons from the same plant (n = 14-23) and with long-term laboratory reference values (LRV) (n = 14-311) . In welders several changes were found when compared to the LRV: in humoral response there were higher prealbumin, lysozyme, circulating immune complexes and lower IgG . In phagocytic tests there were lower ingestion, bactericidal activity and higher metabolic activity of peripheral mononuclear leucocytes . In cellular immunity the marked lymphocytosis, higher counts of T-lymphocytes, CD4+ and CD8+ lymphocytes were noticed . After lowering the concentrations of metals in the working area there were trends to normal values in some parameters {relative numbers of T-lymphocytes, relative number of CD4+ lymphocytes, phagocytic activity, metabolic activity of leucocytes (INT index), IgA, complement C3, transferrin} . The extent and the length of the exposure to welding fumes, smoking and changed conditions at working place were followed as well.

J Cell Biochem, 1998 Mar 1, 68(3), 355 - 65
Analysis of differential gene expression in rat tibia after an osteogenic stimulus in vivo: mechanical loading regulates osteopontin and myeloperoxidase; Miles RR et al.; The skeleton has the ability to alter its mass, geometry, and strength in response to mechanical stress . In order to elucidate the molecular mechanisms underlying this phenomenon, differential display reverse transcriptase-polymerase chain reaction (DDRT-PCR) was used to analyze gene expression in endocortical bone of mature female rats . Female Sprague-Dawley rats, approximately 8 months old, received either a sham or bending load using a four-point loading apparatus on the right tibia . RNA was collected at 1 h and 24 h after load was applied, reverse-transcribed into cDNA, and used in DDRT-PCR . Parallel display of samples from sham and loaded bones on a sequencing gel showed several regulated bands . Further analysis of seven of these bands allowed us to isolate two genes that are regulated in response to a loading stimulus . Nucleotide analysis showed that one of the differentially expressed bands shares 99% sequence identity with rat osteopontin (OPN), a noncollagenous bone matrix protein . Northern blot analysis confirms that OPN mRNA expression is increased by nearly 4-fold, at 6 h and 24 h after loading . The second band shares 90% homology with mouse myeloperoxidase (MPO), a bactericidal enzyme found primarily in neutrophils and monocytes . Semiquantitative PCR confirms that MPO expression is decreased 4- to 10-fold, at 1 h and 24 h after loading . Tissue distribution analysis confirmed MPO expression in bone but not in other tissues examined . In vitro analysis showed that MPO expression was not detectable in total RNA from UMR 106 osteoblastic cells or in confluent primary cultures of osteoblasts derived from either rat primary spongiosa or diaphyseal marrow . Database analysis suggests that MPO is expressed by osteocytes . These findings reinforce the association of OPN expression to bone turnover and describes for the first time, decreased expression of MPO during load-induced bone formation . These results suggest a role for both OPN and MPO expression in bone cell function.

Microb Pathog, 1998 Jan, 24(1), 57 - 63
Antibodies to lipopolysaccharide block adherence of Shiga toxin-producing Escherichia coli to human intestinal epithelial (Henle 407) cells; Paton AW et al.; Shiga toxin-producing Escherichia coli (STEC) are a diverse group of organisms known to cause diarrhoea, haemorrhagic colitis and haemolytic uraemic syndrome (HUS) in humans . During the early stage of infection, numbers of STEC in the gut may be very high (of the order of 10(9)/g faeces), but as disease progresses, the numbers may drop rapidly such that STEC are undetectable within a week . Convalescent sera from patients recovering from HUS frequently contain high levels of antibody to E . coli lipopolysaccharide (LPS) of the infecting serotype, and it is possible that a local immune response to LPS contributes to elimination of the organism from the gut . We have recently demonstrated that STEC strains isolated from HUS cases have enhanced adherence to a human intestinal epithelial cell line (Henle 407) compared with STEC strains from non-human sources . In this study, we examined the capacity of STEC strains belonging to O-antigen types O111 and O157 to adhere to human intestinal epithelial (Henle 407) cells in the presence or absence of anti-LPS . Adherence was inhibited by up to 95% by anti-LPS of the homologous, but not heterologous serotype . This effect was not an artefact of serum bactericidal or agglutinating activity . Preincubation with purified homologous or heterologous LPS did not prevent adherence, suggesting that LPS was not acting as an adhesin per se . Nevertheless, these findings raise the possibility that oral administration of preparations containing anti-LPS may interfere with colonization of the human gut by STEC, and therefore could be of potential therapeutic value if administered early in the course of infection .

EMBO J, 1998 Feb 16, 17(4), 1141 - 51
Evidence for in vivo ribosome recycling, the fourth step in protein biosynthesis; Janosi L et al.; Ribosome recycling factor (RRF) catalyzes the fourth step of protein synthesis in vitro: disassembly of the post-termination complex of ribosomes, mRNA and tRNA . We now report the first in vivo evidence of RRF function using 12 temperature-sensitive Escherichia coli mutants which we isolated in this study . At non-permissive temperatures, most of the ribosomes remain on mRNA, scan downstream from the termination codon, and re-initiate translation at various sites in all frames without the presence of an initiation codon . Re-initiation does not occur upstream from the termination codon nor beyond a downstream initiation signal . RRF inactivation was bacteriostatic in the growing phase and bactericidal during the transition between the stationary and growing phase, confirming the essential nature of the fourth step of protein synthesis in vivo.

J Bacteriol, 1998 Mar, 180(6), 1402 - 10
Balance between endogenous superoxide stress and antioxidant defenses; Gort AS et al.; Cells devoid of cytosolic superoxide dismutase (SOD) suffer enzyme inactivation, growth deficiencies, and DNA damage . It has been proposed that the scant superoxide (O2-) generated by aerobic metabolism harms even cells that contain abundant SOD . However, this idea has been difficult to test . To determine the amount of O2- that is needed to cause these defects, we modulated the O2- concentration inside Escherichia coli by controlling the expression of SOD . An increase in O2- of more than twofold above wild-type levels substantially diminished the activity of labile dehydratases, an increase in O2- of any more than fourfold measurably impaired growth, and a fivefold increase in O2- sensitized cells to DNA damage . These results indicate that E . coli constitutively synthesizes just enough SOD to defend biomolecules against endogenous O2- so that modest increases in O2- concentration diminish cell fitness . This conclusion is in excellent agreement with quantitative predictions based upon previously determined rates of intracellular O2- production, O2- dismutation, dehydratase inactivation, and enzyme repair . The vulnerability of bacteria to increased intracellular O2- explains the widespread use of superoxide-producing drugs as bactericidal weapons in nature . E . coli responds to such drugs by inducing the SoxRS regulon, which positively regulates synthesis of SOD and other defensive proteins . However, even toxic amounts of endogenous O2- did not activate SoxR, and SoxR activation by paraquat was not at all inhibited by excess SOD . Therefore, in responding to redox-cycling drugs, SoxR senses some signal other than O2-.

Vestn Ross Akad Med Nauk, 1998, (1), 50 - 5
{Criteria for prognosis of infectious complications of radiation injuries}; Alekseev GI et al.; The results of comprehensive study of clinical, hematological, immunological, and bacteriological parameters in irradiated rats and dogs are presented . There is a growth of the automicroflora with decreases in leukocytes by 30-40%, lysozyme, beta-lysine, phagocytosis, total serum bactericidal activity by 10-20% . The appearance of pathogenic properties and resistance of Escherichia coli to antibiotics was observed with 60-70% reductions in the activity of nonspecific antiinfectious resistance and with 75-85% falls in neutrophils . This causes clinical manifestations of infectious complications . The appearance of hemolytic properties of Escherichia coli promotes the development of hemorrhagic complications at the sites of its natural habitation . The changes found in different periods after irradiation in the hematologic, immunological, and particularly bacteriological parameters may be criteria for predicting infectious complications and using therapeutical features in time.

Klin Khir, 1997, (9-10), 42 - 3
{Characteristics of the myeloperoxidase-dependent bactericidal system of blood neutrophil granulocytes in patients with suppurative-septic diseases}; Tumasova EP; It was established on the ground of comparative analysis of the blood neutrophils oxidant activity (OA), estimated on the reason of chemoluminescence index and morphometric determination of bactericidal enzyme myeloperoxidase activity in 150 patients with septic-purulent diseases and 30 healthy persons, that frequent association of high indexes of the neutrophils OA and the low enzyme activity don't reflect the functional full value of myeloperoxidase system of the neutrophils bactericidal activity.

Lepr Rev, 1997 Dec, 68(4), 350 - 66
The impact of multiple drug therapy on leprosy disabilities; Willcox ML; In an overview of controlled trials, it is shown that bactericidal drugs increase the short-term risk of Type I reactions, but prevent the long-term development of new impairments caused by bacterial proliferation . Clinical experience suggests that the clofazimine component of multiple drug therapy (MDT) has reduced the incidence of Type II reactions or erythema nodosum leprosum (ENL) . The principal impact of MDT, compared with monotherapy, has been to reduce the duration of active disease, thus preventing the deterioration of disability scores . Reduction of population disability rates is mainly achieved by earlier detection and treatment . MDT has a number of indirect benefits such as improved compliance, decreased cost, and increased motivation and availability of leprosy workers . However, MDT must be supplemented by other measures to prevent and treat disabilities.

FEMS Microbiol Lett, 1998 Feb 15, 159(2), 261 - 6
Rapid accumulation of intracellular 2-keto-3-deoxy-6-phosphogluconate in an Entner-Doudoroff aldolase mutant results in bacteriostasis; Fuhrman LK et al.; The accumulation of 2-keto-3-deoxy-6-phosphogluconate, the key intermediate of the Entner-Doudoroff pathway, has long been thought to inhibit growth of bacteria, but careful measurements of 2-keto-3-deoxy-6-phosphogluconate accumulation by growing cells and the correlation of intracellular 2-keto-3-deoxy-6-phosphogluconate levels to growth inhibition had not been made . A system designed for this purpose was developed in Escherichia coli strains, allowing 2-keto-3-deoxy-6-phosphogluconate accumulation to be experimentally induced and measured by extraction of the cell pool . Addition of gluconate to a strain which lacked 2-keto-3-deoxy-6-phosphogluconate aldolase and overproduced 6-phosphogluconate dehydratase resulted in an increase in the intracellular concentration of 2-keto-3-deoxy-6-phosphogluconate from undetectable levels to 2.0 mM within 15 s, as measured by anion-exchange HPLC . The accumulation of 2-keto-3-deoxy-6-phosphogluconate was correlated with an immediate and significant decrease in growth; this inhibition was determined to be bacteriostatic and not bactericidal . It had been proposed that the mechanism of 2-keto-3-deoxy-6-phosphogluconate toxicity involves competitive inhibition of 6-phosphogluconate dehydrogenase and the consequent block of the pentose phosphate pathway . An experiment addressing this hypothesis failed to provide any supporting data.

J Heart Valve Dis, 1998 Jan, 7(1), 72 - 4
Application of cyanoacrylate adhesive (Krazy Glue) in critical cardiac injuries; Eastman DP et al.; BACKGROUND AND AIMS OF THE STUDY: Although small lacerations of the myocardium may be repaired easily using conventional methods, larger tears or ruptures, especially if they occur in infarcted myocardial tissue, may create formidable technical challenges . Described is a method for applying sutureless pericardial patches for control of hemorrhage . METHODS: A sutureless pericardial patch was glued to the myocardium with commercially available household cyanoacrylate (Krazy Glue) in seven patients . RESULTS: No patient in this series developed any evidence of mediastinal infection as a result of this technique . Six patients were discharged home without any long-term sequelae noted . One patient developed reinfarction and died of arrhythmia two weeks following surgery . Autopsy revealed that the laceration had healed and that the patch was closely adherent . Bacteriology studies revealed that different brands of cyanoacrylate are not only bacterium-free but also exhibit a bactericidal effect . CONCLUSIONS: Sutureless pericardial patches fastened to the myocardium with cyanoacrylate glue to control hemorrhage under critical situations were easy to apply, safe and effective in this series of patients.

Acupunct Electrother Res, 1997, 22(3-4), 191 - 206
Acupuncture enhances the efficacy of antibiotics treatment for canine otitis crises; Sanchez-Araujo M et al.; External bacterial otitis is a very frequent dog's recurrent complaint and should be useful for studying acupuncture effects in infectious disease . Conventional treatment based on antibiotics, washes and bactericidal ointments, solves most cases within 1 to 6 weeks . To establish if the well known effect of this conventional treatment could be modified by acupuncture as additional treatment, a comparative randomized placebo controlled study was performed . From a sample of 25 dogs, two groups were selected at random . In addition to a conventional treatment, each group also received real acupuncture or placebo acupuncture . A veterinarian M.D . and the animal's owner evaluated at blind, through a Visual Analogous Scale, (VAS) the clinical signs and symptoms of otitis . The results show a significant difference in the complete recovery time (p < 0.01); in the disappearance period of the pain (p < 0.01) in the disappearance period of the secretion (p < 0.001) and in the animal's owner evaluation of symptoms (p < 0.001) by 6th . day of treatment . These results suggest that acupuncture stimulates animal defensive systems increasing the conventional treatment effect, which seems to work mainly against the invading agent while acupuncture seems to act by reinforcing the host's defensive barriers . Combination of both shows a synergistic effect capable of empowering the anti-infectious therapy . A three months follow-up revealed all animals were without symptoms . Further evaluation of both groups after a one year follow up will allow us to establish if relapse incidence is also modified by acupuncture, and the potential usefulness of this complementary treatment on recurrent infectious pathology.

Anticancer Res, 1997 Nov-Dec, 17(6D), 4403 - 6
Correlation between structure and diverse biological activities of "half-mustard type" phenothiazines; Motohashi N et al.; The structure and activity relationship of fifteen "half-mustard type" phenothiazines and related compounds were investigated . These compounds did not show any direct bactericidal activity, possibly due to the lack of radical generation activity . Pretreatment with phenothiazines significantly reduced the lethality of Escherichia coli GN2411 infection, possibly due to activation of the host defense mechanism . Higher concentrations of these compounds showed cytotoxic activity against several cultured tumor cell lines . However, no clear-cut relationship was established between biological activity and two dipole moments (mu g, mu e).

Mikrobiologiia, 1997 Nov-Dec, 66(6), 785 - 9
{Detection of two new extracellular adaptogenic factors in Escherichia coli K-12}; Nikolaev IuA; The culture liquid of Escherichia coli K-12 was able to promote the adaptation of this bacterium to various stress factors . Exponentially growing E . coli cells were found to secrete an extracellular "factor of accelerated adaptation to new media," capable of shortening by several hours the lag phase of LB-grown cells transferred to minimal medium with glucose . At sublethal temperatures, E . coli cells secreted a "growth rate reducer," capable of slowing down the growth rate and enhancing the resistance of cells to bactericidal concentrations of N-ethylmaleimide . Such factors have not yet been described in the literature.

Microbiol Immunol, 1997, 41(12), 909 - 16
Growth inhibition of Helicobacter pylori by monoclonal antibody to heat-shock protein 60; Yamaguchi H et al.; The H20mAb recognizing the 60-kilodalton protein, which existed in the outer membrane and was induced by heat shock at 42 C, was established . The molecule recognized with the mAb was a heat-shock protein 60 (HSP60) of Helicobacter pylori . To understand the role of HSP60 on the cell surface of H . pylori, whether or not H20mAb affects the growth of H . pylori was investigated . When bacteria were cultured with H20mAb, growth was markedly inhibited after 24 hr, although an initial 5 hr-incubation with the mAb induced no significant inhibition of H . pylori growth . The 24- and 48 hr growth of the bacteria after washing to remove the mAb at 5 hr was also inhibited though the inhibitory effect was not strong . In electron microscopical analysis, the spots with high electron density in the cytoplasm of the bacteria treated with H20mAb were increased, depending on the length of incubation time from 5 to 24 hr . After 24 hr treatment with H20mAb, bacterial destruction was also observed, indicating bactericidal activity by H20mAb . These results suggest that the HSP60 on the cell surface of H . pylori might have an essential role in the growth of the bacteria.

J Microw Power Electromagn Energy, 1997, 32(4), 205 - 14
Modification of bactericidal effects of microwave heating and hyperthermia by hydrogen peroxide; Kuchma T; Two different approaches for studying of bactericidal effects of microwave heating and hyperthermia were introduced . Low concentration of hydrogen peroxide (0.05%) was used to modify the sensitivity of isogenous strains of Escherichia coli K-12 to microwave heating and hyperthermia with the following assessment of their combined action . This was carried out simulataneously and successively under equal conditions of temperature rise at 50 degrees C . A method of anomalous viscosity time dependencies (AVTD) was used for measurement of the changes in genome conformational state simultaneously with bacterial survival determination . Experiments were performed to study isolated effects of hyperthermia and microwave heating over a range of temperatures from 40 to 80 degrees C and hydrogen peroxide concentrations from 0.05 to 0.3% during 10-minute exposures and their combined action . No difference was found between isolated effects of microwave heating and hyperthermia when survival of E . coli AB 1157 cells was determined . It was shown by the AVTD method that microwave heating at a temperature increase of 6 degrees C per second caused greater damage to cell genome than hyperthermia . The synergistic interaction of microwave heating and low concentrations of hydrogen peroxide was found in simulataneous and successive exposures . The essential distinctions observed in recognition of the action of microwave heating and hyperthermia combined with hydrogen peroxide in various sequences on cellular and molecular levels were attributed to the different effects of microwave and conventional heating on the systems of DNA repair.

Vopr Kurortol Fizioter Lech Fiz Kult, 1997 Nov-Dec, (6), 18 - 9
{The effect of sanatorium-health resort treatment on the function of the bactericidal oxidative mechanism of the blood neutrophils in patients with different forms of chronic bronchitis}; Iarosh AM et al.; The authors' studies show that sanatorium treatment contributes to normalization of bactericidal oxidant mechanism (BOM) of blood neutrophils in patients with chronic bronchitis . In a complete remission this normalization is complete, in partial remission positive trends were seen . In chronic obstructive bronchitis the effect was minor.

Anesth Analg, 1998 Feb, 86(2), 422 - 6
Propofol and thiopental in a 1:1 volume mixture is chemically stable; Lazar ER et al.; Propofol and thiopental have been used clinically in combination for induction of anesthesia . Studies suggest that this mixture has synergistic activity, recovery characteristics similar to propofol alone, and bactericidal effects on multiple organisms . It may therefore be both clinically useful and cost-effective . In this study, we examined the chemical stability of this mixture . We used high-performance liquid chromatography to quantify the concentration of both propofol and thiopental in a given sample . This technique allows the detection of loss in total drug mass and of the appearance of breakdown products resulting from drug interaction . Ten samples of a 1:1 mixture by volume were prepared and assayed at Time 0 and Days 1, 3, and 7 . Half the samples were incubated at 23 degrees C and the rest were stored at 4 degrees C . Other mixtures were assayed before and after filtration at Time 0 and Days 1 and 7 after storage at 23 degrees C . The assay was able to measure accurately the quantity of drug present in the samples . There was no significant decrease in the quantities of either propofol or thiopental in the mixture over the 7-day period . We conclude that the 1:1 volume mixture of propofol and thiopental is chemically stable for 1 wk at room temperature . Implications: A mixture of propofol and thiopental has been used to induce anesthesia . We investigated the chemical stability of this mixture using high-performance liquid chromatography and found it to be stable for at least 24 h.

Vestn Otorinolaringol, 1997, (5), 50 - 1
{The use of xenon irradiator in otology}; Kulakova LA et al.; Xenon irradiator " " (Yakhont-F) has been used in combined treatment of chronic purulent diseases of the external and middle ear in 50 patients . The unit is supplied with flexible multifiber light guides which can operate in the sites not easily accessible for other instruments . Such radiation leads to good bactericidal, antiinflammatory and antiedema effects . In reconstructive and cleansing surgery on the middle ear the transplant retension and epidermization of the postoperative cavity took less time . The technique is proposed for wide otiatric practice.

Infect Immun, 1998 Feb, 66(2), 540 - 8
Mapping of a protective epitope of the CopB outer membrane protein of Moraxella catarrhalis; Aebi C et al.; A monoclonal antibody (MAb) (MAb 10F3) directed against the CopB outer membrane protein of Moraxella catarrhalis previously was found to enhance pulmonary clearance of M . catarrhalis in an animal model (M . Helminen, I . Maciver, J . L . Latimer, L . D . Cope, G . H . McCracken, Jr., and E . J . Hansen, Infect . Immun . 61:2003-2010, 1993) . In the present study, this same MAb was shown to exert complement-dependent bactericidal activity against this pathogen in vitro . Nucleotide sequence analysis of the copB gene from two MAb 10F3-reactive and two MAb 10F3-unreactive strains of M . catarrhalis revealed that the deduced amino acid sequences of these four CopB proteins were at least 90% identical . Comparison of the amino acid sequences of these proteins allowed localization of possible MAb 10F3 binding sites to five relatively small regions of the CopB protein from M . catarrhalis O35E . When five synthetic peptides representing these regions were tested for their ability to bind MAb 10F3 in a direct enzyme-linked immunosorbent assay system, an oligopeptide containing 26 amino acids was shown to bind this MAb . The actual binding region for MAb 10F3 was localized further through the use of overlapping decapeptides that spanned this 26-mer . A fusion protein containing the same 26-mer readily bound MAb 10F3 and was used to immunize mice . The resultant antiserum contained antibodies that reacted with the CopB protein of the homologous M . catarrhalis strain in Western blot analysis and bound to the surface of both homologous and heterologous strains of M . catarrhalis.

Infect Immun, 1998 Feb, 66(2), 486 - 91
Lactoferrin inhibits the endotoxin interaction with CD14 by competition with the lipopolysaccharide-binding protein; Elass-Rochard E et al.; Human lactoferrin (hLf), a glycoprotein released from neutrophil granules during inflammation, and the lipopolysaccharide (LPS)-binding protein (LBP), an acute-phase serum protein, are known to bind to the lipid A of LPS . The LPS-binding sites are located in the N-terminal regions of both proteins, at amino acid residues 28 to 34 of hLf and 91 to 108 of LBP . Both of these proteins modulate endotoxin activities, but they possess biologically antagonistic properties . In this study, we have investigated the competition between hLf and recombinant human LBP (rhLBP) for the binding of Escherichia coli 055:B5 LPS to the differentiated monocytic THP-1 cell line . Our studies revealed that hLf prevented the rhLBP-mediated binding of LPS to the CD14 receptor on cells . Maximal inhibition of LPS-cell interactions by hLf was raised when both hLf and rhLBP were simultaneously added to LPS or when hLf and LPS were mixed with cells 30 min prior to the incubation with rhLBP . However, when hLf was added 30 min after the interaction of rhLBP with LPS, the binding of the rhLPS-LBP complex to CD14 could not be reversed . These observations indicate that hLf competes with rhLBP for the LPS binding and therefore interferes with the interaction of LPS with CD14 . Furthermore, experiments involving competitive binding of the rhLBP-LPS complex to cells with two recombinant mutated hLfs show that in addition to residues 28 to 34, another basic cluster which contains residues 1 to 5 of hLf competes for the binding to LPS . Basic sequences homologous to residues 28 to 34 of hLf were evidenced on LPS-binding proteins such as LBP, bactericidal/permeability-increasing protein, and Limulus anti-LPS factor.

Anaesth Intensive Care, 1997 Dec, 25(6), 640 - 2
Do intensive care drug infusions support microbial growth?
Graystone S, Wells MF, Farrell DJ.
The potential for commonly infused drug solutions to support or inhibit microbial growth was explored in this study . Drugs examined were midazolam HCl, morphine sulphate, fentanyl citrate, pethidine HCl, bupivacaine HCl, atracurium besylate, vecuronium bromide, adrenaline, dopamine, dobutamine, noradrenaline, isoprenaline, glyceryl trinitrate, sodium nitroprusside and propofol . All drugs were studied in both diluted and undiluted forms . Of the 15 medications investigated, all except propofol were found to be bactericidal and to reduce fungal growth.

Antimicrob Agents Chemother, 1998 Jan, 42(1), 154 - 60
The lantibiotic mersacidin inhibits peptidoglycan synthesis by targeting lipid II; Brotz H et al.; The lantibiotic mersacidin exerts its bactericidal action by inhibition of peptidoglycan biosynthesis . It interferes with the membrane-associated transglycosylation reaction; during this step the ultimate monomeric peptidoglycan precursor, undecaprenyl-pyrophosphoryl-MurNAc-(pentapeptide)-GlcNAc (lipid II) is converted into polymeric nascent peptidoglycan . In the present study we demonstrate that the molecular basis of this inhibition is the interaction of mersacidin with lipid II . The adsorption of {14C}mersacidin to growing cells, as well as to isolated membranes capable of in vitro peptidoglycan synthesis, was strictly dependent on the availability of lipid II, and antibiotic inhibitors of lipid II formation strongly interfered with this binding . Direct evidence for the interaction was provided by studies with isolated lipid II . {14C}mersacidin associated tightly with {14C}lipid II micelles; the complex was stable even in the presence of 1% sodium dodecyl sulfate . Furthermore, the addition of isolated lipid II to the culture broth efficiently antagonized the bactericidal activity of mersacidin . In contrast to the glycopeptide antibiotics, complex formation does not involve the C-terminal D-alanyl-D-alanine moiety of the lipid intermediate . Thus, the interaction of mersacidin with lipid II apparently occurs via a binding site which is not targeted by any antibiotic currently in use.

Int J Tuberc Lung Dis, 1997 Oct, 1(5), 460 - 7
Activity of KRM 1648 or rifabutin alone or in combination with clarithromycin against Mycobacterium avium complex in human alveolar macrophages; Suzuki K et al.; SETTING: The activity of KRM 1648 (KRM), a new benzoxazinorifamycin, and rifabutin (RBT), alone or in combination with clarithromycin (CLA), was evaluated against Mycobacterium avium complex (MAC) that multiplied in human alveolar macrophages (AM) . DESIGN: AM were recovered by bronchoalveolar lavage, incubated in RPMI 1640 medium with 10% human AB serum, infected with four strains of MAC (of non-acquired immune deficiency syndrome {AIDS} origin), and then treated with each drug alone or in combination . After incubation for 7 days, colony forming units in each well were counted on 7H10 agar . RESULTS: Although concentrations between 0.2 microgram/ml and 20 micrograms/ml of both rifamycins showed clear dose-dependent activities against all MAC strains tested, only 20 micrograms/ml of each drug had modest bactericidal effect . In combination with 2.0 micrograms/ml of CLA, however, 0.2 microgram/ml of both drugs caused a bactericidal response against two of the four MAC strains examined . CONCLUSION: According to this human alveolar macrophage model of MAC infection, KRM and RBT in combination with CLA was found to be a promising candidate against human pulmonary MAC infection, and deserves clinical evaluation.

Scand J Immunol, 1997 Dec, 46(6), 609 - 18
Activated human gamma delta T lymphocytes express functional lactoferrin receptors; Mincheva-Nilsson L et al.; Lactoferrin (Lf), an iron-binding protein in milk, mucosal secretions and neutrophil granules has bactericidal properties and is a source of iron for breast-fed infants . In this paper the authors show that most in vivo activated lymphocytes, i.e . freshly isolated lymphocytes from first trimester human decidua, and most in vitro activated human blood lymphocytes, express lactoferrin receptors (Lf-R), while unstimulated blood lymphocytes do not . All major lymphocyte subsets, i.e . alpha beta T cells, gamma delta T cells, CD8+ T cells, CD4+ T cells, B cells and NK cells, express Lf-R after activation . The proportion of Lf-R expressing activated gamma delta T cells is significantly larger than that of activated alpha beta T cells . Lf-R and transferrin receptors (Tr-R/CD71) show the same kinetics of appearance on activated blood lymphocytes and are, to a large extent, expressed on the same cells . However, 35% of decidual lymphocytes and 15% of activated blood lymphocytes express Lf-R only . Addition of Lf to cultures containing an optimal concentration of Tr augments the proliferative response to polyclonal T cell activators and alloantigens, suggesting that presently used standard culture conditions for in vitro activation are suboptimal in particular for gamma delta T cells . Lf-R on decidual lymphocytes contain bound Lf, which probably is produced locally . The results suggest that Lf is a growth-supporting factor, especially important in local immune responses in the mucosa.

J Trauma, 1997 Dec, 43(6), 875 - 9
Effects of exogenous cytokines on intravascular clearance of bacteria in normal and splenectomized mice; Hebert JC et al.; BACKGROUND: Pretreatment with interleukin-1 (IL-1), granulocyte colony-stimulating factor (G-CSF), and granulocyte macrophage colony-stimulating factor (GM-CSF) can improve alveolar macrophage bactericidal activity against pneumococci . These effects vary in eusplenic and asplenic mice . Likewise, these cytokines have been shown to improve survival after an aerosol pneumococcal challenge . Mice dying in these studies had positive blood cultures and disseminated infection . The purpose of this study was to determine the effect of cytokine pretreatment on intravascular clearance of bacteria from eusplenic and asplenic mice . METHODS: Two weeks after splenectomy or sham operation, mice were pretreated for various times with IL-1, G-CSF, or GM-CSF or their corresponding vehicles . Mice then received tail-vein injections of bacteria (0.1 mL), and quantitative blood cultures were performed 15 and 30 minutes thereafter . RESULTS: Splenectomized mice had impaired clearance of both pneumococci and Pseudomonas compared with sham-operated mice (p < 0.05) . IL-1 enhanced clearance in splenectomized mice (p < 0.001) but not in sham-operated mice (p not significant) . G-CSF enhanced bacterial clearance in sham-operated mice (p < 0.01) but not in splenectomized mice (p not significant) . GM-CSF enhanced clearance in both groups (p < 0.001) . CONCLUSION: The net effects of exogenous cytokine therapy for infections depends on the state of the host defenses at the time of therapy . These agents may be useful as adjuvants for the treatment of infections, but further study is warranted.

Antimicrob Agents Chemother, 1997 Dec, 41(12), 2786 - 9
Antipneumococcal activity of BAY 12-8039, a new quinolone, compared with activities of three other quinolones and four oral beta-lactams; Visalli MA et al.; Activities of BAY 12-8039 against 205 pneumococci were tested by agar dilution . MICs (in micrograms per milliliter) at which 50 and 90% of the isolates are inhibited (MIC50s and MIC90s, respectively) were 0.125 and 0.25 (BAY 12-8039), 2.0 and 4.0 (ciprofloxacin and ofloxacin), and 0.25 and 0.5 (sparfloxacin) . Beta-lactam MIC50s and MIC90s for penicillin-susceptible, -intermediate, and -resistant strains, in that order, were 0.016 and 0.03, 0.25 and 2.0, and 2.0 and 4.0 (amoxicillin); 0.03 and 0.06, 0.25 and 4.0, and 4.0 and 8.0 (ampicillin); 0.03 and 0.06, 0.5 and 4.0, and 4.0 and 8.0 (cefuroxime); and 0.03 and 0.125, 0.25 and 2.0, and 4.0 and 8.0 (cefpodoxime) . At two times their MICs after 24 h, BAY 12-8039, ciprofloxacin, ampicillin, and cefuroxime were uniformly bactericidal (99.9% killing) against 12 strains; other compounds were bactericidal at four times their MICs.

Antimicrob Agents Chemother, 1997 Dec, 41(12), 2699 - 704
Inhibition of the multiple antibiotic resistance (mar) operon in Escherichia coli by antisense DNA analogs; White DG et al.; The multiple antibiotic resistance operon (marORAB) in Escherichia coli controls intrinsic susceptibility and resistance to multiple, structurally different antibiotics and other noxious agents . A plasmid construct with marA cloned in the antisense direction reduced LacZ expression from a constitutively expressed marA::lacZ translational fusion and inhibited the induced expression of LacZ in cells bearing the wild-type repressed fusion . The marA antisense construction also decreased the multiple antibiotic resistance of a Mar mutant . Two antisense phosphorothioate oligonucleotides, one targeted to marO and the other targeted to marA of the mar operon, introduced by heat shock or electroporation reduced LacZ expression in the strain having the marA::lacZ fusion . One antisense oligonucleotide, tested against a Mar mutant of E . coli ML308-225, increased the bactericidal activity of norfloxacin . These studies demonstrate the efficacy of exogenously delivered antisense oligonucleotides targeted to the marRAB operon in inhibiting expression of this chromosomal regulatory locus.

Optom Vis Sci, 1997 Oct, 74(10), 865 - 7
Sterility of ophthalmic drugs dispensed from spray bottles; Kim GE et al.; BACKGROUND: Spray application of cycloplegics and mydriatics is efficacious and frequently easier to use than a standard dropper bottle in the pediatric population . However, no documentation regarding the sterility of drugs dispensed from spray bottles is available . This study was conducted to determine whether contamination of ophthalmic drugs occurs with spray bottle use . METHODS: Fifteen milliliters of 1% cyclopentolate hydrochloride or 0.5% tetracaine hydrochloride were transferred to each of 15 disinfected spray bottles, stored at room temperature or refrigerated, and sprayed three times weekly for 12 weeks . Cultures were obtained from the spray bottles and drugs before transfer of the drug and from spray bottle contents at 0, 2, 4, and 6 to 12 weeks of storage . RESULTS: No cultures showed significant bacterial growth . The bactericidal action of the preservative and sterility of the drugs were maintained . CONCLUSIONS: Despite the transfer to and use of a spray bottle there appears to be minimal risk of instilling contaminated diagnostic drugs using the spray method when a single drug is stored in a spray bottle.

Int Urol Nephrol, 1997, 29(4), 473 - 8
Chemiluminescence response of whole blood in patients undergoing urological operations; Sakumoto M et al.; Polymorphonuclear leukocytes (PMNs) are one of the most important components of the defence mechanisms against bacterial infection . The functions of PMNs are believed to be impaired in patients during the perioperative period . Bactericidal function of PMNs was investigated together with the luminol-dependent chemiluminescence (CL) reaction of whole blood in 23 patients, 12 undergoing open surgery and 11 undergoing endoscopic surgery . Blood samples were collected one day before surgery (day -1) and 2 hours (day 0), 24 hours (day 1) and 7 days (day 7) after surgery . Counts of whole white blood cells (WBCs), PMNs and lymphocytes were not different between the two surgery groups . CL responses in the open surgery group were increased on days 0, 1 and 7 . In the endoscopic surgery group, CL response was increased on day 1, but not on day 0 or day 7 . These results suggest that the PMN function during the perioperative period was not impaired, but increased just after surgery, mainly due to an increasing number of WBC caused by the surgical intervention.

C R Seances Soc Biol Fil, 1997, 191(4), 617 - 25
{Modulation of respiratory activity of renal macrophages in sea bass (Dicentrarchus labrax) by chronic exposure to sublethal concentration of ammonia}; Gourdon I et al.; Sea bass (Dicentrarchus labrax) were exposed for 71 days to three different ammonia concentrations corresponding to 0, 15 and 25% of the lethal threshold concentration causing the death of half a fish population in 96 hours . The study of the respiratory burst from sea bass renal macrophages showed that the luminol dependent chemiluminescence (CL) emitted following stimulation with mezerein is higher when fishes were previously exposed to ammonia . Therefore, it seemed that chronically exposure of fishes to sublethal concentrations of ammonia primed their renal macrophages to secrete higher amounts of oxygen activated species during respiratory burst, even several days after the transfer of fishes into a standard environment . The stimulated-macrophage CL was partially inhibited by sodium azide, superoxide dismutase and a nitric oxide-synthesis inhibitor, the N5-(-1-iminoethyl)-L-ornithine monochloride, showing that hydrogen peroxide, superoxide anions and nitric oxide were released by renal macrophages from sea bass during the respiratory burst . These reactive species could react together to generate peroxynitrite, a strong bactericidal agent.

Clin Infect Dis, 1997 Nov, 25(5), 1213 - 21
The clinical use of fluoroquinolones for the treatment of mycobacterial diseases; Alangaden GJ et al.; Mycobacterial diseases often require prolonged therapy with multidrug regimens . Fluoroquinolones have excellent bactericidal activity against many mycobacteria; achieve effective serum, tissue, and intracellular levels following oral administration; and produce few adverse effects . These properties have led to the increasing use of fluoroquinolones for the treatment of mycobacterial infections . We reviewed clinical studies and reports involving the use of fluoroquinolones for mycobacterial diseases . Ofloxacin, ciprofloxacin, sparfloxacin, and pefloxacin exhibit clinical efficacy against mycobacterial diseases, especially tuberculosis and leprosy . Fluoroquinolones have generally been administered in regimens that include other agents . However, when a fluoroquinolone has been found to be the sole active agent in a multidrug regimen, the ready emergence of resistance to fluoroquinolones has been recognized, just as when they have been used as monotherapy . Therefore, to forestall the emergence of resistance to fluoroquinolones during the treatment of mycobacterial diseases, these drugs should always be used in combination with at least one other active agent, and they should be used only when effective alternative drugs are not available.

Helicobacter, 1996 Mar, 1(1), 28 - 33
Susceptibility of Helicobacter pylori to the bactericidal activity of human serum; Gonzalez-Valencia G et al.; BACKGROUND: Human serum represents an important barrier to the entry of most mucosal organisms into tissues and to the systemic circulation . If at all present, Helicobacter pylori within gastric tissue is rare, and bacteremia for this organism has been described only once . METHODS: To assess the susceptibility of H . pylori to the bactericidal activity present in normal human serum (NHS), we examined 13 H . pylori isolates . To assess the contributions of the classical and alternative complement pathways to killing, we added either C2-deficient or factor B-deficient serum, respectively, to heat-inactivated NHS . Also we assessed the ability of the strains to bind 125I-C3 . RESULTS: After incubation for 60 minutes at 37 degrees C, all 13 H . pylori strains were killed by NHS; heating to 56 degrees C for 30 minutes ablated killing, indicating complement dependence for this phenomenon . In the absence of an antibody source, there was no killing when either an alternative or classical complement pathway source was used . Adding B-deficient serum to heat-inactivated normal human serum did not restore killing, but adding C2-deficient serum permitted partial killing . All of the 13 strains bound 125I-C3 . Although the kinetics varied from strain to strain, C3 bound was significantly correlated (r = 0.61, p = 0.03) with serum susceptibility . CONCLUSIONS: H . pylori are susceptible to complement, alternative pathway activation appears critical, and C3 binding is a major locus of variability.

Kekkaku, 1997 Oct, 72(10), 579 - 85
{Basic and clinical studies on pathogenesis of pulmonary Mycobacterium avium complex disease}; Suzuki K; I have studied pathogenesis of pulmonary Mycobacterium avium complex disease (PMAC), using mouse and human alveolar macrophage (PAM) model of the infection as well as clinical evaluations . The mouse model revealed no relation between natural resistance against the bacteria and the activation of macrophages which was evaluated on the basis of releasing capacities of prostaglandin E2 and superoxide anion . The PAM model suggested that TNF-alpha and GM-CSF could activate PAM to restrict the intracellular growth of the bacteria, probably not through the superoxide anion release, but through the myeloperoxidasae-halide system . It was also found that rifamycins in combination with clarithromycin could have a good bactericidal effect in the PAM-model of the infection . Clinical evaluations suggested that defect in local pulmonary defense, such as healed pulmonary tuberculous lesions, pneumoconiosis, and COPD was more important predisposing factor than defect in systemic defense in the development of PMAC . Most patients having PMAC without predisposing factors are elderly women, the reason of which is the most important question to be answered in the future studies.

Biochim Biophys Acta, 1997 Oct 23, 1329(2), 357 - 69
Effects on mollicutes (wall-less bacteria) of synthetic peptides comprising a signal peptide or a membrane fusion peptide, and a nuclear localization sequence (NLS) -- a comparison with melittin; Beven L et al.; In order to investigate the effect of primary amphipathic peptides on mollicutes (wall-less bacteria), we have synthesised five molecules (P1, P2, P3, JM123, and JM133) comprising a 16 to 18-residue hydrophobic sequence and the nuclear localization sequence (NLS) PKKKRKV of simian virus 40 large-T antigen, C-terminated by a cysteamide group . The hydrophobic cluster was in P1 the signal sequence of the heavy chain of Caiman crocodilus immunoglobulin G and in JM123 the fusion peptide of human immunodeficiency virus 1 glycoprotein gp41 in which phenylalanine7 was replaced by a tryptophan residue . The homologues P2, P3, and JM133 were obtained by slight alterations of these sequences . Circular dichroism spectroscopy revealed that, in liposomes, P-series peptides were mainly under the form of beta-sheets whereas JM-series peptides displayed a high proportion of turns . These peptides proved to be bactericidal for some mollicutes, notably Acholeplasma laidlawii, but were much less potent than melittin . Furthermore, their antibiotic activity was independent of the average thickness of the plasma membrane hydrophobic core whilst that of melittin was inversely related to the thickness . Melittin and the synthetic peptides abolished spiroplasma cell motility and helicity, but only melittin and P-series peptides split the cells into globular forms displaying an average diameter of ca . 1 microm . In contrast to melittin, the synthetic peptides agglutinated spiroplasmas, suggesting that their polycationic NLS was exposed on the cell surface . P-series peptides decreased, though less efficiently than melittin, A . laidlawii and Spiroplasma melliferum membrane potential (delta psi) and transmembrane pH gradient (delta pH), at concentrations much lower than their minimal inhibitory concentrations whilst JM-series peptides had no effect on delta psi and delta pH in the same conditions . Actually, the bactericidal activity of these peptides towards mollicutes was proportional to their ability to collapse the electrochemical transmembrane potential.

Cancer, 1997 Nov 15, 80(10), 1897 - 903
Vitamin C inhibits the growth of a bacterial risk factor for gastric carcinoma: Helicobacter pylori; Zhang HM et al.; BACKGROUND: Helicobacter pylori infection is a risk factor for gastric carcinogenesis . High dietary vitamin C intake appears to protect against gastric carcinoma . It has been suggested that vitamin C exerts the protective effect by scavenging free radicals that may be enhanced by H . pylori . However, vitamin C has not been investigated in relation to the direct action on H . pylori . In this study, the authors attempted to clarify this possibility both in vitro and in vivo . METHODS: Susceptibility testing of H . pylori (64 strains) was performed by the agar dilution method . Bactericidal actions were determined by a broth cultivation technique . The effect of vitamin C on in vivo H . pylori colonization was evaluated by using the Mongolian gerbil model . RESULTS: At concentrations of 2048, 512, and 128 microg/mL (minimum inhibitory concentrations {MICs}), vitamin C could inhibit the growth of 90% of the bacterial stains incubated at pH values of 7.4, 6.0, and 5.5, respectively . The broth cultures exposed to the MICs of vitamin C displayed a 1.57 approximately 2.5-log decrease in the number of viable bacteria, and the loss of viability was observed in 24 hours at concentrations 8-fold higher than the MICs . In an in vivo experiment, H . pylori colonies decreased significantly in animals treated with vitamin C after oral administration of vitamin C (10 mg/head/day) for 7 days . CONCLUSIONS: High doses of vitamin C inhibit the growth of H . pylori in vitro as well as in vivo.

J Immunol, 1997 Nov 15, 159(10), 5079 - 83
Two functionally independent pathways for lipopolysaccharide-dependent activation of mouse peritoneal macrophages; Amura CR et al.; We have investigated the effects of human LPS-binding protein (LBP) and human bactericidal/permeability-increasing protein (BPI) on LPS-dependent activation of mouse thioglycolate-elicited peritoneal macrophages in vitro, in comparison with human PBMCs . Confirming earlier published studies, BPI inhibited, and LBP enhanced, the ability of LPS to stimulate PBMC production of the cytokines TNF-alpha and IL-6 . In marked contrast to these results, under identical conditions of in vitro culture, both LBP and BPI suppressed, in a dose-dependent manner, the ability of LPS to stimulate cytokine production in mouse macrophages . Further, while human BPI also suppressed LPS-dependent NO secretion in mouse macrophages, human LBP had no inhibitory effect on NO secretion under conditions that inhibited TNF-alpha secretion . These data provide the first direct evidence that mouse macrophages may utilize two independent pathways in response to LPS, thus leading to different phenotypic responses.

Nutrition, 1997 Oct, 13(10), 863 - 9
Glutamine-enhanced bacterial killing by neutrophils from postoperative patients; Furukawa S et al.; Neutrophils play an important role in host defense by phagocytosing and destroying invading bacteria . A recent investigation revealed that glutamine (Gln) augmented the in vitro bactericidal activity of neutrophils from burn patients . However, it is unclear whether Gln enhances the function of neutrophils in postoperative patients . This study was designed to investigate the effect of Gln on the in vitro Escherichia coli-killing activity of neutrophils from postoperative patients . Nine randomly selected patients were included in this study . On the morning of the first postoperative day, blood was drawn and neutrophils were isolated . Eight healthy volunteers served as controls . E . coli was opsonized with pooled normal serum . Neutrophils (5 x 10(6)), together with opsonized E . coli (5 x 10(5)), were incubated for 2 h at 37 degrees C in Hanks' balanced salt solution supplemented with 0, 100, 500, or 1000 nmol/mL of Gln . The bactericidal function of neutrophils was determined by counting the number of viable bacteria . Tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta, IL-8, and granulocyte elastase levels in the cell culture supernatant were measured . Plasma C-reactive protein (CRP), cortisol, and amino acids were also analyzed . The plasma concentration of Gln was significantly lower in the postoperative patients than in the controls . Following culture with patient neutrophils, the number of viable E . coli decreased by 26% as the in vitro Gln concentration was increased from 500 to 1000 nmol/mL (P < 0.01) . We defined the Gln 1000/Gln 500 ratio of the number of viable bacteria as the number of viable E . coli at an in vitro Gln concentration of 1000 nmol/mL divided by the number of viable E . coli at an in vitro Gln concentration of 500 nmol/mL . A positive correlation was thus demonstrated between the plasma Gln level and the Gln 1000/Gln 500 ratio of the number of viable bacteria in the patients (r = 0.69, P = 0.04) . This finding indicated that as plasma Gln fell, there was an enhancement of neutrophil E . coli-killing activity by neutrophils in in vitro tests when the Gln concentration was increased from 500 to 1000 nmol/mL . Gln supplementation caused no appreciable changes in TNF-alpha, IL-1 beta, IL-8, or granulocyte elastase levels in cell culture supernatants . A negative correlation was recognized between the patient plasma Gln level and the Gln 1000/Gln 500 ratio of the cell culture supernatant IL-8 level (r = -0.73, P = 0.025) . In conclusion, Gln supplementation enhanced the in vitro bactericidal function of neutrophils from postoperative patients.

Microbiology, 1997 Oct, 143 ( Pt 10), 3367 - 73
The pncA gene from naturally pyrazinamide-resistant Mycobacterium avium encodes pyrazinamidase and confers pyrazinamide susceptibility to resistant M . tuberculosis complex organisms; Sun Z et al.; The antituberculosis drug pyrazinamide (PZA) needs to be converted into pyrazinoic acid (POA) by the bacterial pyrazinamidase (PZase) in order to show bactericidal activity against Mycobacterium tuberculosis . M . avium is naturally resistant to PZA . To investigate whether this natural resistance to PZA is due to inability of the M . avium PZase to convert PZA to bactericidal POA, the M . avium PZase gene (pncA) was cloned by using the M . tuberculosis pncA gene as a probe . Sequence analysis showed that the M . avium pncA gene is 561 bp long, encoding a protein with a predicted size of about 19.8 kDa; but Western blotting showed that the M . avium PZase migrated as a 24 kDa band when expressed in M . bovis BCG and Escherichia coli . Sequence comparison revealed that M . avium PZase has 67.7% and 32.8% amino acid identity with the corresponding enzymes from M . tuberculosis and E . coli, respectively . Southern blot analysis with the M . avium pncA gene as a probe showed that M . terrae, M . gastri, M . marinum, M . fortuitum, M . xenopi, M . gordonae, M . szulgai, M . celatum and M . kansasii have close pncA homologues, whereas M . chelonae and M . smegmatis did not give significant hybridization signals . Transformation with the M . avium pncA gene conferred PZA susceptibility to PZA-resistant M . tuberculosis complex organisms, indicating that the nonsusceptibility of M . avium to PZA is not due to an ineffective PZase enzyme, but appears to be related to other factors such as transport of POA.

Nephrol Dial Transplant, 1997 Sep, 12(9), 1863 - 8
Rapid screening assay for anti-GBM antibody and ANCAs; an important tool for the differential diagnosis of pulmonary renal syndromes; Westman KW et al.; BACKGROUND: Pulmonary renal syndrome is encountered in several diseases such as Goodpasture's syndrome, antineutrophil cytoplasmic antibody (ANCA) associated systemic vasculitis, systemic lupus erythematosus (SLE) and infection-associated or drug-induced glomerulonephritis . To preserve organ function it is of vital importance to make the correct diagnosis and institute adequate therapy early, in the acute phase . METHOD: An enzyme-linked immunosorbent assay (ELISA), specially designed as a rapid screening assay for antiglomerular basement membrane (anti-GBM) antibody, proteinase-3 (PR3-) ANCA and myeloperoxidase-(MPO-) ANCA were evaluated from 1060 serum samples drawn from patients with clinically suspected pulmonary renal syndrome or rapidly progressive glomerulonephritis (RPGN) . RESULTS: Of the 1060 serum samples, 142 were positive for anti-GBM antibody (n = 19), PR3-ANCA (n = 60), or MPO-ANCA (n = 73) . Of the 142 samples, 10 were double positive . Reanalysis of positive sera with a quantitative ELISA yielded results manifesting good correlation with those of the the rapid screening assay . Of 918 sera found to be negative in the rapid screening assay, 105 were also tested with IIF, 11 being found to be positive . However, these 11 sera manifested no specificity for PR3 or MPO, but some were specific for bactericidal/permeability-increasing proteins, lactoferrin or elastase ANCAs . Two of the patients whose sera yielded negative results in the rapid assay had systemic vasculitis . CONCLUSION: The ELISA thus detects the true antibodies to PR3, MPO, and GBM, whereas IIF detects additional specificities . The findings suggest the rapid assay results to be of high positive predictive value, and the assay to be of high diagnostic specificity and sensitivity and thus useful in the diagnostic workup in suspected cases of RPGN or pulmonary renal syndrome.

Anasthesiol Intensivmed Notfallmed Schmerzther, 1997 Jul, 32(7), 420 - 5
{Effects of human i.v . immunoglobulin on bacterial clearance and granulocyte function in endotoxinemia}; Koch T et al.; PURPOSE: The therapeutic impact of intravenous immunoglobulins (ivIG) in septic patients remains controversial . Until now, the mechanisms of action have not been fully elucidated . Since polymorphonuclear neutrophils (PMN) play a key role in host defence, this study focuses on the effects of ivIG on bacterial clearance and PMN respiratory burst activity during endotoxinaemia . For this purpose, it was investigated whether ivIG improves blood clearance and organ colonisation as well as PMN functions after experimentally induced bacteraemia in rabbits . METHODS: The experiments were performed in 30 anaesthetised rabbits . To determine quantification of bacterial killing in vivo, defined numbers of exogenous Escherichia (E.) coli 1.3 x 10(8) CFU) were injected intravenously in untreated animals (n = 10) or 60 min after infusion of endotoxin (LPS: 40 micrograms/kg/h) in groups without (n = 10), and after pretreatment with ivIG (Sandoglobulin, 0.5 g/kg body weight, n = 10), respectively . Parameters monitored were rates of bacterial elimination from the blood, LPS clearance, arterial pressure, blood gases and white blood cell counts, PMN burst activity was determined using a flow cytometry assay . Samples of liver, kidney, spleen and lung were collected for bacterial counts 180 min following E . coli injection . RESULTS: Compared to controls, endotoxinaemia resulted in a prolonged elimination of the injected E . coli out of the blood associated with a significantly (p < 0.01) higher colonisation of all organs . Pretreatment with ivIG improved LPS clearance and significantly reduced bacterial colonisation of lung and kidney (p < 0.01) . This was paralleled by an enhanced PMN respiratory burst activity compared to untreated animals (p < 0.05) . CONCLUSION: The reduced bacterial colonisation of lung and kidney in correlation with an increased PMN bactericidal activity in endotoxinaemia suggest an improved granulocyte-dependent bacterial killing due to ivIG application.

Vet Immunol Immunopathol, 1997 Sep, 58(2), 133 - 45
Effects of 4-ipomeanol on bovine alveolar macrophage function; Li X et al.; The objective of this study was to determine whether 4-ipomeanol toxicosis in calves impairs alveolar macrophage functions important in pulmonary defense against infectious agents . Male Holstein calves were given either 4-ipomeanol (3 mg kg-1, i.v.) or vehicle (polyethylene glycol 400) . Alveolar macrophages were recovered by pulmonary lavage 3 days later, and their capacities to phagocytose and kill E . coli, migrate toward zymosan-activated immune bovine serum, and produce interferon and interleukin-1 activity were evaluated in vitro . Alveolar macrophages recovered from 4-ipomeanol-treated calves had over a 70% decrease (p < 0.01) in chemotactic activity and over a 37% decrease (p < 0.005) in their capacity to phagocytose E . coli as compared to macrophages from control calves . Interleukin-1 activity in macrophages from 4-ipomeanol-treated calves tended to be higher than that from control calves, but the differences were not statistically significant (p = 0.06) . 4-ipomeanol did not affect macrophage bactericidal activity or production of interferon . These results indicate that 4-ipomeanol suppresses select functions of alveolar macrophages in cattle that may be important in pulmonary defense against bacterial pathogens.

Genetics, 1997 Oct, 147(2), 713 - 24
Molecular population genetics of Drosophila immune system genes; Clark AG et al.; A striking aspect of many vertebrate immune system is the exceptionally high level of polymorphism they harbor . A convincing case can be made that this polymorphism is driven by the diversity of pathogens that face selective pressures to evade attack by the host immune system . Different organisms accomplish a defense against diverse pathogens through mechanisms that differ widely in their requirements for specific recognition . It has recently been shown that innate defense mechanisms, which use proteins with broad-spectrum bactericidal properties, are common to both primitive and advanced organisms . In this study we characterize DNA sequence variation in six pathogen defense genes of Drosophila melanogaster and D . mauritiana, including Andropin; cecropin genes CecA1, CecA2, CecB, and CecC; and Diptericin . The necessity for protection against diverse pathogens, which themselves may evolve resistance to insect defenses, motivates a population-level analysis . Estimates of variation levels show that the genes are not exceptionally polymorphic, but Andropin and Diptericin have patterns of variation that differ significantly from neutrality . Patterns of interpopulation and interspecific differentiation also reveal differences among the genes in evolutionary forces.

Langenbecks Arch Chir, 1997, 382(4 Suppl 1), S14 - 7
{Therapy of peritonitis today . Surgical management and adjuvant therapy strategies}; Reith HB; Acute necrotizing pancreatitis and fylecal or diffuse purulent peritonitis are the diseases primarily responsible for mortality due to surgical infections of the abdomen . The most recent figures indicate that a mortality rate of 50%-80% in this specialized treatment group is still a reality . Without doubt, surgical sanitation of the focus is the most important therapeutic measure . A generalized inflammation reaction has been regularly observed in nearly all patients within this disease category . Local surgical therapy has the greatest effect on prognosis . If the therapeutic goal is not reached with the first intervention, adjuvant surgical therapy is necessary . The different forms are continuous peritoneal lavage (CPL), open dorsoventral lavage, and relaparotomy or scheduled reoperation ("Etappenlavage") . Adjuvant medical treatments include TNF alpha and interleukin-1 synthesis inhibitors or antibodies . Unfortunately, clinical studies with these mediators have only been partly successful in the subgroups, so that a general clinical adjuvant treatment is not considered viable . The bacterial properties of taurolidine destroy the bacterial membrane and, at the same time, lead to cross-linking of the membrane components and functional proteins (LPS), so that a bactericidal effect and endotoxin reduction take place simultaneously . Both local and intravenous routes of administration can be used.

Kansenshogaku Zasshi, 1997 Aug, 71(8), 763 - 9
{Isolation of Legionella pneumophila from 24 hr-home bath water and an eradication trial of the bacteria from the bath}; Li N et al.; Contamination of 24 hr home bath with Legionella pneumophila is recently well recognized . Eradication of the water-bath contamination from L . pneumophila and other bacteria is an important matter to prevent the infection because the 24 hr-bathing facility is widely accepted in Japanese houses . Among the 16 bathing water samples we tested, Legionella pneumophia was isolated from 6 cases (37.5%) when the bathing water was not treated with disinfectants . Number of L . pneumophila increased up to 10(3) cfu/ml and total culturable bacterial counts reached to 10(5) cfu/ml within 5 days when the water was not treated . We selected 5 water baths among 6 positive cases to study the bactericidal effect of chlorine . As a result we concluded that the growth of L . pneumophila in 24 hr-water bath could be stopped by the 2 ppm chlorination program every day.

Lancet, 1997 Sep 20, 350(9081), 855 - 9
Randomised placebo-controlled trial of granulocyte-colony stimulating factor in diabetic foot infection; Gough A et al.; BACKGROUND: Diabetic foot infections cause substantial morbidity and mortality . Neutrophil superoxide generation, a crucial part of neutrophil bactericidal activity, is impaired in diabetes . Granulocyte-colony stimulating factor (G-CSF) increases the release of neutrophils from the bone marrow and improves neutrophil function . We assessed G-CSF as adjuvant therapy for the treatment of severe foot infections in diabetic patients . METHODS: 40 diabetic patients with foot infections were enrolled in a double-blind placebo-controlled study . On admission, patients were randomly assigned G-CSF (filgrastim) therapy (n = 20) or placebo (n = 20) for 7 days . Both groups received similar antibiotic and insulin treatment . Neutrophils from the peripheral blood of these participants and from healthy controls were stimulated with opsonised zymosan, and superoxide production was measured by a spectrophotometric assay (reduction of ferricytochrome C) . FINDINGS: G-CSF therapy was associated with earlier eradication of pathogens from the infected ulcer (median 4 {range 2-10} vs 8 {2-79} days in the placebo group; p = 0.02), quicker resolution of cellulitis (7 {5-20} vs 12 {5-93} days; p = 0.03), shorter hospital stay (10 {7-31} vs 17.5 {9-100} days; p = 0.02), and a shorter duration of intravenous antibiotic treatment (8.5 {5-30} vs 14.5 {8-63} days; p = 0.02) . No G-CSF-treated patient needed surgery, whereas two placebo recipients underwent to amputation and two had extensive debridement under anaesthesia . After 7 days' treatment, neutrophil superoxide production was significantly higher in the G-CSF group than in the placebo group (16.1 {4.2-24.2} vs 7.3 {2.1-11.5} nmol per 10(6) neutrophils in 30 min; p < 0.0001) . G-CSF therapy was generally well tolerated . INTERPRETATION: G-CSF treatment was associated with improved clinical outcome of foot infection in diabetic patients . This improvement may be related to an increase in neutrophil superoxide production.

Am J Respir Crit Care Med, 1997 Sep, 156(3 Pt 1), 895 - 900
The early bactericidal activity of isoniazid related to its dose size in pulmonary tuberculosis; Donald PR et al.; Collections of sputum from 105 patients with newly diagnosed pulmonary tuberculosis were made before and at 1 and 2 d after the start of chemotherapy with isoniazid (INH) alone given to groups of patients in doses of 600 mg, 300 mg, 150 mg, 75 mg, 37.5 mg, 18.75 mg, and 9 mg daily, as well as from an untreated group . Counts of colony forming units (cfu) of Mycobacterium tuberculosis in the collections were set up on plates of selective 7H10 medium . The early bactericidal activity (EBA) of INH was defined as the decrease in log10 cfu/ml sputum/day during the first 2 d of treatment . A smooth curve relating EBA to log dose was obtained, with 600 mg INH yielding the highest mean EBA of 0.539, and 18.75 mg INH yielding the lowest EBA (0.111) that could be distinguished from the bactericidal activity of the untreated group . The ratio of the usual dose of 300 mg INH to the lowest dose, of 18.75 mg, that produced a detectable EBA, termed the therapeutic margin, was therefore 16, in contrast to the lower therapeutic margin of 4 for rifampin . The EBA was related to the INH acetylator genotype of patients treated with 600 mg or 9 mg INH.

Gerontology, 1997, 43(5), 277 - 82
Studies on haemagglutination and serum resistance status of strains of Moraxella catarrhalis isolated from the elderly; Murphy S et al.; A total of 40 strains of Moraxella catarrhalis, isolated from the sputum of elderly persons with respiratory tract infections and from nasopharyngeal swabs from healthy elderly, were examined for haemagglutination of human red blood cells and resistance to bactericidal activity in normal human serum (NHS) . 15 of 20 strains isolated from the infected elderly and 3 of 20 strains isolated from the healthy elderly showed haemagglutinating properties (p < 0.001) . Similarly, 13 of 20 strains from the infected group and 2 of 20 strains from the healthy group were found to be resistant to the bactericidal activity of NHS (p < 0.001) . Strains of M . catarrhalis which were associated with respiratory tract infections in the elderly, in contrast to strains colonizing the healthy elderly, were found to be predominantly haemagglutinating for human red blood cells and resistant to complement killing in NHS.

Int J Biochem Cell Biol, 1997 Jun, 29(6), 857 - 60
Nitric oxide; Robbins RA et al.; Nitric oxide (NO) is a gas with diverse biological activities produced from arginine by NO synthases . It is capable of interacting with a number of molecules, most notably superoxide, forming peroxynitrite, which, in turn, can mediate bactericidal or cytotoxic reactions . Nitric oxide also mediates smooth muscle relaxation, neurotransmission, and modulation of inflammation in a number of organ systems and pathophysiologic conditions . Modulation of NO by administration of inhaled NO for respiratory distress syndromes and infusion of NO synthase inhibitors in bacterial sepsis are ongoing . Levels of exhaled NO are being evaluated for their utility in assessing inflammation in respiratory disorders such as asthma.

Rinsho Kyobu Geka, 1989 Jun, 9(3), 261 - 6
Surgical reintervention in prosthetic valve endocarditis; von der Emde J et al.; Early successful conservative treatment and thereafter early surgical intervention on prosthetic-valve-endocarditis may reduced the mortality rate . Therefore, early surgical intervention in native valve endocarditis with perioperative antibiotic treatment in primary valve replacement reduced the incidence of PVE . Soaking of prostheses and suture material with bactericidal antibiotics during the first or second procedures with precise surgical sterility and short perioperative exposure time are successful points which should be taken . Mortality rate decreases from 100 to 60-90% to 10% with combined antibiotic and surgical treatment.

Arch Surg, 1997 Sep, 132(9), 991 - 6
Wound hypoxia and acidosis limit neutrophil bacterial killing mechanisms; Allen DB et al.; BACKGROUND: "Respiratory burst" activity, ie, O2- production, is dependent on PO2, temperature, pH, and glucose concentrations within the physiologic range . OBJECTIVES: To determine whether environmental conditions characteristic of wounds may limit human neutrophil respiratory burst metabolism and to clarify the degree to which bactericidal oxidant production depends on local PO2 . METHODS: Human blood and wound neutrophils were stimulated with phorbol myristate acetate . Oxygen consumption and superoxide production were measured over a range of 30 to 300 mm Hg PO2, 0 to 40 mmol/L glucose, pH 6.0 to 8.0, and 30 degrees C to 37 degrees C . The apparent Michaelis Menten constant for oxidant production with respect to PO2 was calculated . RESULTS: Oxygen consumption and O2- production were dependent on PO2 throughout the range tested . Half-maximal oxidant production occurred in the range of 45 to 80 mm Hg PO2 and maximal at PO2 higher than 300 mm Hg . These data agree with the highest previous estimates . Oxidant generation was also dependent on pH, temperature, and glucose concentration, but to a lesser extent . CONCLUSIONS: Leukocyte bacterial killing capacity as measured by oxygen consumption and superoxide production are substantially impaired at the low oxygen tensions often found in wounds . Changes in pH, temperature, and glucose concentration have lesser but nonetheless significant consequences . The data provide a plausible mechanism for the vulnerability of some wounds to infection and for the previous finding that increasing oxygen tension at wound sites enhances bactericidal function . Thus, the data serve as a basis for future studies on prevention of wound infection.

Eur J Surg, 1997 Aug, 163(8), 605 - 9
Effect of omentectomy on peritoneal defence mechanisms in rats; Agalar F et al.; OBJECTIVE: To assess the effect of omentectomy on peritoneal defence mechanisms in rats . DESIGN: Randomised study . MATERIAL: Sixty Wistar Albino rats . INTERVENTIONS: Rats were divided in three groups of 20 . All rats underwent midline laparotomy and 10 rats in each group had omentectomy . Samples were taken two hours (early period) and 30 days (late period) after omentectomy . MAIN OUTCOME MEASURES: Effect of omentectomy on bactericidal activity of peritoneal fluid (PBA), chemotactic indices (CI) of polymorphonuclear leucocytes (PMNL), phagocytic activity of PMNL in the peritoneal fluid (PHA), total cell counts, and cell types of peritoneal washing fluid . RESULTS: In the early period omentectomy reduced total cell counts from 3440 (1400-4800) x 10(6)/10 ml to 1480 (800-2080) x 10(6)/10 ml (p = 0.0022), and the CI of PMNL from 2.86 (2.32-4.02) to 1.43 (1.29-1.77) (p = 0.0002), and increased the PHA from 11.9 (8.3-17) to 17 (16-19) (p = 0.0006) . The PBA was not significantly altered . The proportion of macrophages decreased (p = 0.0001), while the proportion of lymphocytes increased in the peritoneal fluid (p = 0.0002) . In the late period total cell counts in the control and omentectomy groups were 3440 (1400-4800) x 10(6) and 3160 (1040-5120) x 10(6)/10 ml fluid, respectively (p = 0.52) . Omentectomy reduced the CI of PMNL from 2.86 (2.32-4.02) to 2.01 (1.82-1.49) (p = 0.0003) . The difference between the proportion of PHA of either group was not significant (p = 0.06) . PBA in the control and omentectomy groups was 99.03 (70-100) and 99.48 (71.5-100), respectively (p = 0.97) . Although the total cell count in the peritoneal fluid did not differ, the proportion of macrophages decreased (p = 0.0003) while the proportion of lymphocytes increased (p = 0.0002) . The proportion of PMNL did not change in either of the experimental settings . CONCLUSIONS: The omentum has an important role in local peritoneal defence mechanisms in experimental conditions . Its removal may effect some of these mechanisms adversely.

Cytometry, 1997 Sep 1, 29(1), 58 - 64
Assessment of the effects of gramicidin, formaldehyde, and surfactants on Escherichia coli by flow cytometry using nucleic acid and membrane potential dyes; Comas J et al.; Two membrane potential sensitive dyes (Rhodamine 123 and bis-oxonol) and three nucleic acid dyes (propidium iodide, SYTO-13, and SYTO-17) were used to assess the effect of surfactants on Escherichia coli . The ability of E . coli to be stained by these probes was validated at different physiological states . Propidium iodide was used to assess the integrity of cell envelopes . Two double staining methods based on propidium iodide with SYTO-13 and bis-oxonol with SYTO-17 were used to improve the discrimination between bacteria and micelles or aggregated particles generated by the presence of surfactants . A rapid (1 h contact time between cells and surfactants, and less than 5 min for staining and obtaining data) Rhodamine 123 flow cytometric assay was developed to assess the bactericidal effect of surfactants.

J Immunol Methods, 1997 Jul 14, 205(2), 127 - 33
Use of native and recombinant bactericidal/permeability-increasing proteins (BPI) as antigens for detection of BPI-ANCA; Schultz H et al.; Anti-neutrophil cytoplasmic antibodies (ANCA) against native bactericidal/permeability-increasing protein (nBPI) have gained increasing diagnostic significance in inflammatory bowel disease and cystic fibrosis . However, routine detection of BPI-ANCA requires pure antigen in large quantities . As nBPI is difficult to isolate and is very susceptible to proteolytic cleavage with subsequent epitope loss, it was the aim of this study to determine whether recombinant BPI (rBPI) can be used as an alternative to nBPI as target antigen for ANCA in diagnostic procedures . Therefore, 93 BPI-ELISA-positive sera and controls were compared in different ELISAs using nBPI, rBPI, unglycosylated rBPI and a 21-kDa amino-terminal fragment of rBPI . ELISA results were confirmed by immunoblotting and all sera were tested in indirect immunofluorescence (IFT) . There was an 88% (82/93) agreement in recognition of nBPI and rBPI by ANCA in both ELISA systems, yet the quantitation of BPI-ANCA in relative units showed a less optimal result and correlated only by 45% (p < 0.01) . Most sera recognized nBPI, rBPI and unglycosylated rBPI equally suggesting that glycosylation has no influence on antigen recognition . Only two sera were positive for the 21-kDa nBPI indicating that the binding sites for ANCA are either conformational epitopes and/or are located mainly on the carboxy-terminal part of the BPI molecule . Most BPI-ELISA-positive sera were negative in IFT (43%), but a perinuclear (pANCA, 30%), a cytoplasmic (cANCA,10%) or an atypical ANCA (aANCA, 2%) staining pattern, as well as a cytoplasmic pattern only on formaldehyde-fixed granulocytes (13%) were also observed . Overall, no characteristic pattern was seen for BPI-ELISA-positive sera in IFT . Taken together, these data suggest that rBPI offers an excellent alternative to nBPI for broad-based BPI-ANCA ELISA and will be of great value in further investigations of BPI-ANCA interactions.

J Cataract Refract Surg, 1997 Jul-Aug, 23(6), 883 - 8
Half-life of intracameral gentamicin after phacoemulsification; Lehmann OJ et al.; PURPOSE: To determine the half-life of intracameral gentamicin administered during phacoemulsification . SETTING: Southampton Eye Unit, Southampton General Hospital, England . METHODS: Thirty-one patients having scleral tunnel phacoemulsification were given intracameral gentamicin in the irrigation fluid . Samples of fluid were taken from the anterior chamber at the end of the operation and at various times postoperatively . The concentration of gentamicin in the samples was determined by fluorescence polarization immunoassay and the half-life calculated for a single compartment model using a peeling algorithm . RESULTS: The concentration of gentamicin in the anterior chamber after phacoemulsification decreased by half every 51 minutes (95% confidence interval, 42 to 66 minutes) . CONCLUSION: Intracameral gentamicin was cleared from the anterior chamber after phacoemulsification at a rate that prevents the maintainance of the bactericidal levels required for reliable antibiotic prophylaxis.

Infect Immun, 1997 Sep, 65(9), 3672 - 9
Coccoid forms of Helicobacter pylori are the morphologic manifestation of cell death; Kusters JG et al.; Helicobacter pylori can transform from its normal helical bacillary morphology to a coccoid morphology . Since this coccoid form cannot be cultured in vitro, it has been speculated that it is a dormant form potentially involved in the transmission of H . pylori and in a patient's relapse after antibiotic therapy . In this study we determined the effects of aging, temperature, aerobiosis, starvation, and antibiotics on the morphologic conversion rate and culturability of H . pylori . Aerobiosis and the addition of a bactericidal antibiotic to the culture medium resulted in the highest conversion rate . During the conversion to coccoid forms, the cultures always lost culturability at the stage where 50% of the organisms were still in bacillary form; this result indicated that culturability and coccoid morphology are two separate but related entities . Independent of the conditions used to induce the conversion into coccoids, the morphological conversion was accompanied by several marked antigenic and ultrastructural changes . Also, both the total amounts and the integrity of RNA and DNA were significantly reduced in coccoid forms . With the potential-sensitive probe diOC(5)-3, a clear loss of membrane potential in coccoid forms was observed . Inhibition of protein or RNA synthesis by the addition of bacteriostatic antibiotics did not prevent the conversion to coccoid forms but resulted in an increased conversion rate . Hence, we conclude that conversion of H . pylori from the bacillary to the coccoid form is a passive process that does not require protein synthesis . Our data suggest that the coccoid form of H . pylori is the morphologic manifestation of bacterial cell death.

Surgery, 1997 Aug, 122(2), 380 - 5
Design of a potent novel endotoxin antagonist; Uknis ME et al.; BACKGROUND: Bactericidal permeability increasing protein (BPI) binds to and neutralizes lipopolysaccharide (LPS, endotoxin) . Small synthetic peptides based on the amino acid sequence of the LPS binding domain of BPI neutralize LPS, albeit inefficiently . Although the LPS binding domain of native BPI possesses a beta-turn secondary structure, this structure is not present in small derivative peptides . The purpose of this study was to determine whether the addition of a beta-turn to a BPI-derived peptide is associated with more potent endotoxin antagonism . METHODS: We generated a hybrid peptide (BU3) on the basis of (1) a portion of the LPS binding domain from BPI and (2) amino acids known to initiate a beta-turn . BU3 folds with a beta-turn, and we tested its effects on LPS neutralization and LPS-induced tumor necrosis factor-alpha secretion, comparing it with BPI-derived peptide BG22 that lacks a beta-turn and to an irrelevant peptide (BG16) . RESULTS: Compared with BG22, BU3 demonstrated enhanced LPS neutralization and inhibition of LPS-induced tumor necrosis factor-alpha secretion in vitro and a similar diminution of endotoxemia and tumor necrosis factor-alpha secretion in a murine model of endotoxemia . CONCLUSIONS: These data demonstrate the potential for enhancing the biologic activity of a BPI-derived peptide endotoxin antagonist via manipulation of its conformational structure.

Crit Care Med, 1997 Aug, 25(8), 1283 - 8
Recombinant N-terminal fragment of bactericidal/permeability increasing protein (rBPI21) prevents shock-induced microcirculatory alterations in the liver; Bauer C et al.; OBJECTIVE: To investigate the effects of the recombinant 21-kilodalton N-terminal fragment of recombinant bactericidal and permeability increasing protein (rBPI21) on leukocyte adhesion and the hepatic microcirculation after hemorrhagic shock . DESIGN: Prospective, randomized, blinded, and placebo-controlled experimental study . SETTING: University research laboratory . SUBJECTS: Anesthetized Sprague-Dawley rats, weighing 220 to 250 g . INTERVENTIONS: Rats were subjected to 60 mins of hemorrhagic shock and subsequent resuscitation to sufficiently restore systemic circulation . The microcirculation of the liver was investigated by intravital fluorescence microscopy 5 hrs after hemorrhagic shock . Four shock groups were compared with a sham-control group . Shock groups received either rBPI21 (10 mg/kg) or placebo either before or after shock period . MEASUREMENTS AND MAIN RESULTS: No differences were observed in hemodynamic, respiratory, or metabolic parameters between the shock groups . However, the hepatic microcirculation showed severe deterioration 5 hrs after shock, indicated by significantly narrowed sinusoids in all shock groups compared with controls (8.5 +/- 0.3 microm vs . 10.0 +/- 0.4 pm) . Leukocyte adhesion was markedly increased to comparable values in both placebo groups (619 cells/mm2 and 644 cells/mm2; sham, 168 cells/mm2) . Neutralization of endotoxin by administration of rBPI21 before or after shock resulted in plain reduction of pathologic leukocyte-endothelial interaction (138 cells/mm2 and 85 cells/mm2) . CONCLUSION: The results support the hypothesis that endotoxin induces microcirculatory alterations after shock, and further suggest a potentially beneficial role of rBPI21 in the treatment of posttraumatic endotoxin-induced inflammatory reactions.

Thromb Res, 1997 Aug 1, 87(3), 323 - 9
Bactericidal/permeability-increasing protein ameliorates hypercoagulability after hemorrhagic shock; Yamashita M; We recently showed that both plasminogen activator inhibitor-1 (PAI-1) and tissue factor (TF) are induced after a massive hemorrhage . In this study, we determined if bactericidal/permeability-increasing protein (BPI) has any effects on the induction of these factors after hemorrhagic shock . Three days after cannulation, rats were bled and maintained at a mean blood pressure of 40 mmHg for 60 min, and then were resuscitated with the shed blood and an equal volume of saline over 60 min . Rats in the BPI group were given at 6 mg/kg of rBPI21 (XOMA, Berkeley, CA; a 3-mg/kg dose at the beginning of hemorrhage followed by two doses of 1.5 mg/kg at the end of shock and at the end of resuscitation) . The control group was treated similarly to the BPI group, but received control protein in the same dose as rBPI21 . Plasma endotoxin concentration, whole blood clotting time (WBCT) and plasma PAI activity were measured at times 0, 2, 4, 6, and 8 h . The time-course changes in mRNA of TF, PAI-1, tumor necrosis factor alpha (TNF alpha), interleukin-1 beta (IL-1 beta), and interleukin-6 (IL-6) were also detected in the liver by reverse transcription and polymerase chain reaction . The plasma endotoxin levels increased after hemorrhagic shock and showed a peak at 2 h in the control group . These increases were significantly neutralized by rBPI21 treatment at 2 h in the BPI group . WBCT decreased and PAI activity increased rapidly after hemorrhagic shock in the control group . These changes were significantly smaller in the BPI group at 6 and 8 h . The increases in mRNA of TF, PAI-1, TNF alpha, and IL-6 were also attenuated by rBPI21 treatment . These results show that BPI ameliorates hypercoagulability after hemorrhagic shock and suggest that endotoxin plays a role in the pathogenesis of thrombogenic responses after hemorrhagic shock.

FEMS Microbiol Lett, 1997 Aug 1, 153(1), 199 - 204
Acridine orange as an indicator of bacterial susceptibility to gentamicin; Mason DJ et al.; We have studied the response of Escherichia coli NCTC10418 to gentamicin with flow cytometry . The susceptibility of individual bacterial cells to the antibiotic was assessed by differential staining using the metachromatic dye, acridine orange . Exponential phase cultures were exposed to the minimum bactericidal concentration of gentamicin and analysed at regular intervals over 90 min . Within 60 min of exposure to the drug, two sub-populations of organisms could be distinguished in cultures by their different acridine orange-associated fluorescence emissions of < 550 nm and > 550 nm . The number of bacteria exhibiting acridine orange-associated fluorescence at > 550 nm corresponded to counts of colony forming units.

J Biol Chem, 1997 Jul 25, 272(30), 18682 - 5
Lipopolysaccharide (LPS)-binding proteins BPI and LBP form different types of complexes with LPS; Tobias PS et al.; Lipopolysaccharide (LPS)-binding protein (LBP) and bactericidal/permeability-increasing protein (BPI) are closely related LPS-binding proteins whose binding to LPS has markedly different functional consequences . To gain better insight into the possible basis of these functional differences, the physical properties of LBP-LPS and BPI-LPS complexes have been compared in this study by sedimentation, light scattering, and fluorescence analyses . These studies reveal dramatic differences in the physical properties of LPS complexed to LBP versus BPI . They suggest that of the two proteins, only LBP can disperse LPS aggegates . However, BPI can enhance both the sedimentation velocity and apparent size of LPS aggregates while inhibiting LPS-LBP binding even at very low (1:40 to 1:20) BPI:LPS molar ratios.

Biochem Biophys Res Commun, 1997 Jul 18, 236(2), 427 - 30
The genomic organization of the genes for human lipopolysaccharide binding protein (LBP) and bactericidal permeability increasing protein (BPI) is highly conserved; Hubacek JA et al.; We have determined the exon/intron organization of the human lipopolysaccharide binding protein (LBP) and bactericidal permeability increasing protein (BPI) genes . The LBP gene spans approximately 28.5 kb and is composed of 14 exons while the 31.5-kb-long BPI gene is composed of 15 exons . Comparison of the genomic organization of the LBP and BPI genes together with the genomic structures of the PLTP (phospholipid transfer protein) and CETP (cholesteryl ester transfer protein) genes, which all together constitute a gene family of functionally related proteins, revealed high homology with a remarkable conservation of exon/intron transitions . The exon/intron junctions of the LBP, BPI, and PLTP genes are almost identical, with most of the exons being of the same size . In addition, functional domains are conserved in these proteins . The C-terminal octapeptide important for CETP anchoring in lipoprotein particles is also present in LBP, BPI, and PLTP . The LPS binding motif in exons 3 and 4 has been retained in LBP and BPI . Our results indicate that the LBP, BPI, and PLTP genes, and probably the CETP gene, may have evolved from a common primordial gene and may share similar functional properties.

Rinsho Ketsueki, 1997 Jul, 38(7), 566 - 71
{Combined deficiency in neutrophil functions after bone marrow transplantation and the in vitro effect of granulocyte colony-stimulating factor}; Katoh M et al.; In order to investigate the mechanism of susceptibility to bacterial infection after bone marrow transplantation (BMT), we evaluated the neutrophil functions at 1, 3, 6, and 12 months after hematological reconstitution in a 29-year-old AML patient who received allogeneic bone marrow transplantation . Acute graft-versus-host disease (GVHD) was grade I, and chronic GVHD was not present . The patient exhibited complications of upper and lower respiratory infections several times after BMT, all of which subsided within a month . Chemotactic responses toward all three chemotactic factors, random mobility, phagocytosis, superoxide (O2-) release and bactericidal activity were severely impaired early after reconstitution . These neutrophil functions gradually improved with time after BMT, and all normalized at 12 months after reconstitution of transplanted bone marrow . Both O2- release and bactericidal activity of neutrophils were significantly enhanced at 6 and 12 months after marrow reconstitution following pretreatment of 50 ng/ml of granulocyte colony-stimulating factor (G-CSF) in vitro . These findings suggest that the combined disorders in neutrophil functions in early phase after BMT may play an important role in susceptibility to bacterial infections until one year after BMT . Administration of G-CSF, which potentiates the bactericidal activity of neutrophils, is recommended for preventing infectious complications and for treating prolonged and unmanageable infections after BMT.

Kekkaku, 1997 Jul, 72(7), 449 - 53
{Anti-Mycobacterium avium complex activities of KRM-1648, clarithromycin and levofloxacin in 7HSF medium at peak or average blood concentrations after their oral administration of clinical dosages}; Sato K et al.; We evaluated the anti-Mycobacterium avium complex (MAC) activities of KRM-1648 (KRM), clarithromycin (CAM) and levofloxacin (LVFX) in 7HSF medium at concentrations in human blood after their oral administration of clinical dosages . MAC organisms were inoculated into tubes containing the medium with or without the addition of test drugs at either 1/10 Cmax, Cmax or C0-8h (average concentrations during the first 8 h) in the blood . KRM at Cmax or C0-8h showed strong bactericidal activity against MAC, CAM showed weak or moderate degree of bactericidal activity at Cmax and C0-8h, while LVFX added at Cmax or C0-8h showed only a weak bacteriostatic effect against M . avium, but did not show any effect against M . intracellulare.

Chem Res Toxicol, 1997 Jul, 10(7), 802 - 10
Toxicity of peroxynitrite and related reactive nitrogen species toward Escherichia coli; Hurst JK et al.; The toxicity of peroxynitrite toward Escherichia coli (expressed as LD50, the concentration required to kill 50% of the bacteria) was found to be independent of bacterial cell densities over a wide experimental range, spanning 10(6)-10(10) colony-forming units/mL; the magnitude of LD50 was also pH-independent over the range pH 5.9-8.3 . This highly unusual behavior can be quantitatively reproduced by a dynamical model in which (i) ONO2H is identified as the toxic form of the oxidant and (ii) the bulk of the added peroxynitrite decays to nitrate ion under these conditions . From the model, one estimates that 10(6)-10(7) ONO2H molecules are required to kill a bacterium, indicating a very high intrinsic toxicity (cf . HOCl, for which LD50 = 10(7)-10(8) molecules/cell of E . coli) . Nearly complete protection was observed when bicarbonate ion was added to the buffer, even when concentrations of peroxynitrite exceeded 50 times the LD50 measured in the absence of bicarbonate . Consistent with previous reports, combinations of H2O2 and NO and, in weakly acidic media, H2O2 and NO2- were found to exhibit enhanced toxicities relative to the individual reactants . Protection by bicarbonate was utilized to assess the potential role of intermediary formation of ONO2H in bacterial killing in these systems . Approximately 25% protection by bicarbonate was observed for media containing H2O2 and NO2-, consistent with a minor contribution to killing by ONO2H under the experimental conditions . No protection was observed for media containing H2O2 and *NO in both anaerobic and aerobic environments, excluding extracellularly generated ONO2H as a participant in these bactericidal reactions.

J Antimicrob Chemother, 1997 Jul, 40(1), 39 - 45
Correlation between bactericidal activity and postantibiotic effect for five antibiotics with different mechanisms of action; Li RC et al.; Theoretically, if the postantibiotic effect (PAE) reflects the duration of cellular recovery, then the extent of cellular damage inflicted on bacteria by an antibiotic, as suggested by the degree of bactericidal activity, should reflect the length of PAE; this is especially true if binding of the antibiotic to bacterial receptors is irreversible . To test this hypothesis, correlation between PAEs and bactericidal rate constants measured simultaneously at various antibiotic concentrations was examined for five antibiotic-bacterium combinations . Each of the five antibiotics used, i.e . tobramycin, ciprofloxacin, dicloxacillin, trimethoprim and tetracycline, has a different mechanism of action: the first three bind irreversibly to bacterial receptors, while trimethoprim and tetracycline bind reversibly . Both PAE and bactericidal activity increased nonlinearly with concentrations in a saturable manner for all the combinations studied . Linear least-square regression analyses showed strong correlations (P < 0.01) between the two responses for individual combinations . Such a linear relationship also extended, with good correlation (P < 0.05), across the five combinations when individual maximal bactericidal rate constants and PAEs were considered separately . These observations suggest that cellular recovery from nonlethal damage following antibiotic exposure may be a major determinant of PAE.

Burns, 1997 Jun, 23(4), 338 - 40
Hypernatremic suppression of neutrophils; Kuroda T et al.; Phagocytic and bactericidal activities are important functions of the human polymorphonuclear leucocytes (PMNL) as the host defence against burn wound infection . The zone of stasis, just below the zone of coagulation, is a site of interaction between invading bacteria and PMNL . For many reasons the osmotic pressure at this site is elevated . In this study we evaluated the in vitro phagocytic and bactericidal activities of PMNL under hypernatremic conditions . At a sodium concentration of 180 mEq/l, phagocytic activity was suppressed . Although on average superoxide production was maintained within the normal range, it was suppressed in three of the ten cases studied . We conclude that the hypernatremic condition may weaken local defence against burn wound infection at the zone of stasis and may be a risk factor for burn wound sepsis.

Gut, 1997 Jun, 40(6), 731 - 8
In vitro evaluation of the role of antibodies against Helicobacter pylori in inhibiting adherence of the organism to gastric cells; Clyne M et al.; BACKGROUND: Once Helicobacter pylori infection is established, it is difficult to eradicate despite a persistent systemic and local immune response . It is not known whether immunisation can be used to prevent H pylori infection in humans . AIMS: To evaluate the effect of the human immune response on adherence of H pylori to gastric cells . METHODS: Human milk from a woman infected with H pylori and milk from a non-infected woman were each fractionated by chromatography on DEAE cellulose . Bacteria were incubated with either serum, human milk, human milk fractions, or secretory IgA before incubation with Kato III cells (cells from a gastric adenocarcinoma cell line) . Bacterial adherence to the cells was assessed using flow cytometry . RESULTS: Serum from both the H pylori infected and non-infected women killed H pylori . This resulted from the action of complement as heating the serum to 56 degrees C for 30 minutes abolished the bactericidal activity . Immunoglobulin fractions from serum of both infected and non-infected women did not inhibit H pylori adherence to Kato III cells . Human milk from the woman infected with H pylori and from the non-infected woman inhibited binding of H pylori to Kato III cells by 50 to 70% . Secretory IgA isolated from human milk had minimal inhibitory effect on adherence and this was notably less than the inhibitory effect of whole human milk . CONCLUSIONS: Human milk inhibits adherence of H pylori to Kato III cells and this inhibition is independent of whether or not the donor is infected with H pylori . Secretory IgA has minimal inhibitory effect on H pylori adherence.

Infect Immun, 1997 Jun, 65(6), 2272 - 7
Saturable CD14-dependent binding of fluorescein-labeled lipopolysaccharide to human monocytes; Troelstra A et al.; We used rough lipopolysaccharide (ReLPS) to construct a fluorescein-labeled LPS (FITC-LPS) with a very high labeling efficiency that bound to isolated human monocytes in a CD14-dependent fashion and that in this respect behaved indistinctively from native LPS . The CD14-dependent binding could be inhibited either by a 1,000-fold excess of unlabeled LPS or by polymyxin B, bactericidal/permeability-increasing protein, cationic protein 18, or soluble CD14 . Although this FITC-LPS preparation no longer possessed the ability to prime neutrophils for the production of reactive oxygen species or to stimulate human monocytes to produce tumor necrosis factor, activation of the Limulus amoebocyte lysate cascade was comparable to activation by native LPS . Binding to monocytes was enhanced by human pooled serum (HPS) or LPS-binding protein (LBP) for LPS concentrations up to 100 ng/ml and was completely CD14 dependent . For LPS concentrations exceeding 100 ng/ml, binding was still partially CD14 dependent, but not HPS or LBP dependent . CD14-dependent association of LPS with monocytes was shown to be totally saturable . In conclusion, we found an HPS- or LBP-dependent binding of FITC-LPS to monocytes that was CD14 dependent at up to 100 ng of LPS per ml, and saturation of binding was shown.

Infect Immun, 1997 Jun, 65(6), 2136 - 44
Effects of adenosine on the functions of circulating polymorphonuclear leukocytes during hyperdynamic endotoxemia; Thiel M et al.; Endotoxin-activated polymorphonuclear leukocytes (PMNL) adhere to the vascular endothelium and cause damage by the release of toxic superoxide anions (O2-) . Because adenosine is a potent inhibitor of PMNL in vitro, the present study investigates the effects of this nucleoside on the functions of circulating PMNL in a standardized porcine model of hyperdynamic endotoxemia . Ten anesthesized pigs received an intravenous (i.v.) 330-min infusion of endotoxin (5 microg/kg of body weight per h) . Another 10 pigs were also infused with endotoxin plus adenosine (150 microg/kg/min {i.v.}); this treatment was begun 30 min prior to the beginning of endotoxin treatment . Control groups (five animals per group) received either adenosine or physiological saline . Infusion of endotoxin caused severe neutropenia, shedding of L-selectin, upregulation of beta2-integrins, increased binding of C3-coated zymosan particles, and subsequent phagocytosis by PMNL . While phagocytosis-induced production of oxygen radicals appeared to decrease, extracellular release of superoxide anions was strongly enhanced . Infusion of adenosine during endotoxemia had no effect on neutropenia, expression of adhesion molecules, C3-induced adhesion, phagocytosis, or intracellular production of oxygen radicals, whereas extracellular release of O2- was strongly inhibited . Thus, i.v . infusion of adenosine during endotoxemia could be useful in protecting from O2(-)-mediated tissue injury without compromising the bactericidal mechanisms of PMNL.

Diabetes Care, 1997 Jun, 20(6), 995 - 8
Epalrestat, an aldose reductase inhibitor, improves an impaired generation of oxygen-derived free radicals by neutrophils from poorly controlled NIDDM patients; Sato N et al.; OBJECTIVE: To study the in vivo effect of epalrestat (Epa), an aldose reductase inhibitor, on the generation of oxygen-derived free radicals by neutrophils from poorly controlled NIDDM patients (HbA1c > 10%) . RESEARCH DESIGN AND METHODS: A total of 31 diabetic patients were randomly divided into two groups: an Epa(+) group of 16 patients treated with 150 mg/day epalrestat and an Epa(-) group of 15 patients treated without epalrestat . A control group of 20 age- and sex-matched normal healthy subjects also participated . HbA1c, postprandial plasma glucose (PPG), and neutrophil bactericidal function were measured before and at the end of the drug treatment period (4 weeks) . Neutrophil bactericidal function was measured as chemilu-minescence amplified by a Cypridina luciferin analog (CLA), which is dependent on O2- generation, and by luminol (L), which is highly dependent on OCl- generation, in response to formyl-methonyl-leucyl-phenylalanine (fMLP) . RESULTS: At the start of the experiment, both CLA-dependent chemiluminescence (CLA-DCL) and L-dependent chemiluminescence (L-DCL) were clearly decreased in diabetic subjects (64 and 54%, respectively; P < 0.05) compared with control subjects (2,182 +/- 144 and 3,221 +/- 173 kc.min-1.10(-6) cells, respectively) . At the end of the experiment, CLA-DCL and L-DCL in the Epa(+) group were significantly improved by 44 and 46%, respectively; however, these values were still lower than the corresponding results in the control group . HbA1c and PPG in both the Epa(+) and Epa (-) groups were significantly higher than in the control group, and treatment had no effect on either HbA1c or PPG . CONCLUSIONS: These data suggest that epalrestat may be a useful drug to prevent infection by improving the impaired O2- and OCl- generation by neutrophils from poorly controlled NIDDM patients.

J Immunol, 1997 Jun 1, 158(11), 5400 - 8
Adenosine inhibits neutrophil degranulation in activated human whole blood: involvement of adenosine A2 and A3 receptors; Bouma MG et al.; Adenosine, acting via A2 receptors, is a potent inhibitor of neutrophil oxidative burst, but its effects and mechanisms of action on neutrophil degranulation have been less well characterized . We, therefore, investigated the effects of adenosine and its receptor-specific analogues on neutrophil degranulation in stimulated human whole blood . Adenosine dose-dependently inhibited the LPS- and TNF-alpha-induced release of the azurophilic granule proteins bactericidal/permeability-increasing protein, elastase, and defensins to approximately the same extent, with a maximum inhibition of 70 to 80% and an IC50 ranging from 14 to 24 microM . The inhibitory effects of adenosine were partially blocked by the A2 receptor antagonist 3,7-dimethyl-1-propargylxanthine, the A1/A2 antagonist 8(p-sulfophenyl)theophyline, and the A1/A3 antagonist xanthine amine congener, but not by the A1 antagonist 1,3-dipropyl-8-cyclopentylxanthine . The highly selective A3 agonist N6-(3-iodobenzyl)-adenosine-5'-N-methyluronamide and the nonselective agonist 2-chloroadenosine reduced degranulation more potently than the A1 agonist N6-cyclopentyladenosine . The inhibitory effects of N6-(3-iodobenzyl)-adenosine-5'-N-methyluronamide and 2-chloroadenosine were strongly reversed by xanthine amine congener, but were not affected by 8(p-sulfophenyl)theophyline . In addition, the adenosine kinase inhibitor GP515 attenuated degranulation via an adenosine-mediated mechanism . These data indicate that adenosine acts via A2 as well as A3 receptors to inhibit neutrophil degranulation and add to the anti-inflammatory potential of adenosine and adenosine-regulating agents in neutrophil-mediated tissue injury.

Eur J Biochem, 1997 May 15, 246(1), 45 - 9
Influence of the Gloeophyllum metabolite oosponol and some synthetic analogues on protein and RNA synthesis in target cells; Sonnenbichler J et al.; Oosponol {4-(hydroxyacetyl)-8-hydroxy-1H-2-benzopyran-1-one}, a toxic metabolite of the basidiomycete Gloeophyllum abietinum, and some synthetic analogues were studied to clarify the molecular basis of their fungicidal, bactericidal, and phytotoxic effects . The antibiotic activity was due to a strong inhibition of RNA and protein synthesis in target cells.

Schweiz Med Wochenschr, 1997 May 10, 127(19), 805 - 11
{Surgical therapy of severe acute pancreatitis}; Uhl W et al.; There is some evidence that the incidence of acute pancreatitis is increasing worldwide . Improved treatment concepts, especially in the severe course of the disease, have significantly reduced formerly high mortality . According to the different clinical courses it is of the utmost importance for the therapeutic approach to this disease to differentiate between mild (morphologically characterized as edema) and severe (intra- and extrapancreatic necroses) as early as possible . In this respect, contrast-enhanced CT scanning and the determination of so-called necrosis indicating parameters (e.g . C-reactive protein) have been established as the "gold-standard" . While patients with acute edematous pancreatitis are successfully treated in a normal ward, patients with a proven necrotizing course of the disease should undergo intensive monitoring and maximum intensive care therapy in the ICU . Additionally, these latter patients should receive antibiotics which are capable of penetrating the pancreas and the pancreatic necroses in bactericidal concentrations . It seems more and more evident that only patients under this treatment regimen who develop infected pancreatic necrosis and sepsis are candidates for surgical intervention . Infected pancreatic necrosis can be easily diagnosed with a high level of safety and reliability by fine needle puncture and aspiration of pancreatic necrosis and fluid collections under imaging-guided procedures . Patients with sterile necrosis respond in most cases to intensive care therapy and in these patients the indication for surgery will be only exceptional . Surgery should be performed as late as possible to ensure sufficient demarcation of the necroses . In our experience the best surgical treatment modality for infected pancreatic necrosis is necrosectomy combined with postoperative continuous local lavage of the retroperitoneum . Mortality of severe acute pancreatitis has been reduced under this treatment concept to below 10%.

Anesteziol Reanimatol, 1997 May-Jun, (3), 18 - 20
{Use of ofloxacin in the treatment of suppurative-inflammatory complications in surgery}; Bogomolova NS et al.; Analysis of ofloxacin use in clinical surgery over the latest decade showed that despite its wide clinical application for the prevention and therapy of postoperative complications, this drug remains highly active for the majority of micro-organisms . Hence, a wide spectrum, high bactericidal activity of the drug, optimal pharmacokinetics, and a high bio-availability (95-100%) still recommend ofloxacin, a highly effective agent for the treatment of infectious complications of surgery.

Contraception, 1997 May, 55(5), 315 - 8
Nitrites and L-citrulline levels in copper intrauterine device users; Pradhan M et al.; The mechanism of copper in limiting intrauterine infections in intrauterine device (Cu IUD) users is poorly understood . Copper ions may enhance the release of reactive oxygen radicals, which in turn decrease the release of reactive nitrogen intermediates (RNI) . RNI are known to have bactericidal effect . The present study compares the levels of RNI prior to Cu-T insertion and at different post-insertion intervals up to 12 weeks . The decrease in RNI was evident by one week and continued until 12 weeks . Therefore, the bactericidal effect of copper in IUD is via reactive oxygen intermediates . The superoxide anion inactivates this active intermediate nitric oxide . Therefore, excess of superoxide radical will markedly shorten the half-life of nitric oxide but will not prevent its conversion to nitrites and nitratesPIP: Copper ions may enhance the release of reactive oxygen radicals that decrease the release of reactive nitrogen intermediates (RNIs), known to have a bactericidal effect . The capability of the Copper-T IUD to generate RNIs was investigated in women seeking IUD insertion at a family planning clinic in Chandigarh, India . A significant decrease in nitric oxide levels was recorded from pre-insertion to 1 week post-insertion . Moreover, the values at 4 weeks' post-insertion were significantly lower than those at 1 week, and the 12 weeks' post-insertion level was lower than that recorded at 4 weeks . Uterine aspirate L-citrulline levels at 1 week post-insertion were significantly lower than pre-insertion levels; again, values at 4 and 12 weeks post-insertion were significantly less than those of the receding levels . There was no significant difference in levels of nitrites or serum L-citrulline before and after IUD insertion . Although the exact mechanism of the decrease in RNI levels at the local site after IUD insertion is unknown, it appears related to the sterile intrauterine milieu . At the peak of copper release, the inflammatory cells are activated to release reactive oxygen radicals . Copper potentiates the effect of superoxide dismutase, which influences the half-life of nitric oxide, but does not prevent its conversion to nitrites and nitrates .

Baillieres Clin Rheumatol, 1997 May, 11(2), 395 - 421
Laboratory findings in the vasculitides; Kallenberg CG; The primary vasculitides are diseases of unknown aetiology . They are characterized by inflammation of blood vessel walls . Measuring non-specific laboratory markers of inflammation is useful in the monitoring of patients with vasculitis . The diagnostic specificity of these markers is, however, restricted . In the last decade, autoantibodies reacting with myeloid granule proteins have been detected in the sera from patients with Wegener's granulomatosis, microscopic polyangiitis, Churg-Strauss syndrome, and the renal limited form of these vasculitides (i.e . idiopathic rapidly progressive glomerulonephritis) . Anti-neutrophil cytoplasmic antibodies (ANCA) in the aforementioned disorders react with proteinase 3 (Pr3) or myeloperoxidase (MPO), and only incidentally to other antigens such as elastase and bactericidal-permeability increasing protein . The presence of ANCA alone, in particular perinuclear ANCA, as detected by indirect immunofluorescence, has a low specificity for those vasculitides . However, in combination with the presence of anti-Pr3 or anti-MPO antibodies as detected by enzyme-linked immunosorbent assay, sensitivity and specificity for the vasculitides is high . Several in vitro and in vivo data have suggested a pathophysiological role for anti-Pr3 and anti-MPO in the associated disorders . Measuring levels of the autoantibodies seems useful for the follow-up of patients with these vasculitides . The sensitivity and specificity of rises in ANCA levels for ensuing relapses appears somewhat lower than previously suggested . Refinement of the assays, for example, by measuring subclasses and functional characteristics of the autoantibodies, may improve their value in monitoring patients with vasculitides.

Curr Opin Hematol, 1997 May, 4(3), 183 - 90
Effects of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor on the bactericidal functions of neutrophils; Pitrak DL; The hematopoietic growth factors granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) not only regulate the numbers of circulating neutrophils but also modulate the function of mature cells . Additionally, newly developed neutrophils subsequently released from the bone marrow in response to colony-stimulating factors (CSFs) also have enhanced function . A variety of different functions are affected, including changes in adherence, movement, phagocytosis, priming and stimulation of the respiratory burst, secretion, and degranulation . These effects also can cause increased microbicidal capacity in vitro, ex vivo, and in vivo . Both G-CSF and GM-CSF have such effects on neutrophil function, but there are differences that may result in precise modulation of the immune responses and may have implications for choice of agents for immune-based therapy for different conditions.

Chem Res Toxicol, 1997 May, 10(5), 618 - 26
Inactivation of the inducible nitric oxide synthase by peroxynitrite; Huhmer AF et al.; The simultaneous production of superoxide and nitric oxide by stimulated human neutrophils leads to the formation of peroxynitrite, a physiologically important bactericidal agent . We have investigated two possible pathways for the inactivation of inducible nitric oxide synthase (NOS-II) by peroxynitrite: inactivation of NOS-II through oxidation of the tightly bound cofactor calmodulin (CaM) and direct interaction of ONOO-/ONOOH with the NOS-II protein . Studies of two model peptides indicated that the Ca2+-dependent binding to CaM of a typical high-affinity sequence, melittin, significantly prevented Met oxidation in CaM by ONOO-/ONOOH . In contrast, binding of the putative CaM-binding domain of human hepatocyte NOS-II (NOS-II509-534) to CaM only marginally prevented the oxidation of Met residues in CaM . When the native NOS-II/CaM complex was exposed to peroxynitrite, CaM was inert toward oxidation . Nevertheless, even small amounts of peroxynitrite abolished the activity of NOS-II through direct interaction with the heme . The loss of activity was paralleled by a decrease in heme absorbance and a shift of the absorbance maximum from 419 to 409 nm . The presence of the cofactor tetrahydrobiopterin during peroxynitrite exposure did not prevent inactivation of the enzyme but altered the change of the heme spectrum, i.e., a shift of lambda(max) from 419 to 420 nm rather than to 409 nm . In conclusion, peroxynitrite inactivates NOS-II through changes in the heme or its environment in NOS-II rather than via oxidation of the cofactor CaM.

Shock, 1997 May, 7(5), 339 - 44
Studies on polymorphonuclear leukocyte bactericidal function: II . The role of oxidative stress; Simms HH et al.; We have previously detailed the effects of exogenous cytokines (IL-1 beta, TNF-alpha, and IL-8) on polymorphonuclear leukocyte (PMN) bactericidal function during normoxia . In these studies, we have investigated the effects of hypoxia +/- reoxygenation independently and with IL-1 beta, TNF-alpha, and IL-8 on PMN bactericidal activity . Hypoxia in and of itself did not significantly alter PMN bactericidal activity against Escherichia coli or Staphylcoccus aureus; however, after 2 h of reoxygenation (H/R), PMN bactericidal activity against E . coli was significantly reduced compared with levels seen after 2 h of hypoxia . Similar to what was observed during normoxia, TNF-alpha, IL-1 beta, and IL-8 increased PMN bactericidal activity during hypoxia compared with buffer control PMN for S . aureus but not E . coli after 4 h of hypoxia . Following H/R, neither TNF-alpha, IL-1 beta, nor IL-8 reversed the decline in bactericidal activity induced by reoxygenation alone . Monoclonal antibodies that blocked the functional epitope of particular cytokine receptors demonstrated that during both hypoxia and H/R IL-1 beta R type I, IL-8R type A, and TNF-alpha R p60 were the predominant receptors responsible for mediating the bactericidal effect of the cytokines . During hypoxia and H/R, the addition of exogenous cytokines did prevent the fall in bactericidal activity seen as PMN: target ratios decreased . The decline in bactericidal activity following H/R was mediated in part by reduced phagocytosis of serum-opsonized bacteria following H/R . These results demonstrated that oxidative stress in the form of hypoxia +/- reoxygenation independently modulate the effect of exogenously added cytokines on PMN bactericidal capability.

J Microencapsul, 1997 May-Jun, 14(3), 335 - 48
In vitro kinetics of drug release and pulmonary retention of microencapsulated antibiotic in liposomal formulations in relation to the lipid composition; Beaulac C et al.; In previous in-vivo studies, we demonstrated that liposomal entrapment of tobramycin resulted in an increased availability of the antibiotic in the lungs without increasing bactericidal efficacy (Omri et al . 1994) . With the aim of developing liposomal formulations allowing more efficient liposome-bacteria interactions, we studied the influence of lipid composition on both drug release and pulmonary retention of encapsulated tobramycin . The phase transition temperatures of nine liposome-tobramycin formulations consisting of two synthetic phospholipids (distearoyl phosphatidylcholine (DPSC) or dipalmitoyl phosphatidylcholine (DPPC) with dimyristoyl phosphatidyl-glycerol (DPMG) or dimyristoyl phosphatidylcholine (DMPC) were determined by differential scanning calorimetry . Liposomes, varying in terms of membrane fluidity and charge were submitted to in-vitro and in-vivo kinetic studies while retention and release of tobramycin were measured by high-performance liquid chromatography (HPLC) . Five less fluid liposome formulations showed absence or very low tobramycin release in in-vitro tests and long term pulmonary retention of tobramycin . Four fluid liposome formulations showed in vitro tests modulated tobramycin release while pulmonary retention of tobramycin was dependent of the presence of charged phospholipids . Administration of charged fluid liposomes in mice showed a low level of tobramycin in the kidneys; non-charged fluid liposomes exhibited a relatively high level of tobramycin retention in the kidneys.

Antimicrob Agents Chemother, 1997 May, 41(5), 1170 - 2
Postantibiotic effect assessments for antibiotics exhibiting a wide range of bactericidal activities by using a modified total-cell-counting method; Li RC et al.; We recently described a total-cell-counting method for postantibiotic effect (PAE) assessments that performs well with weakly bactericidal antibiotics . This note presents a modified method for the study of PAE with extended capability to cover a broad range of bactericidal activities.

J Membr Biol, 1997 May 1, 157(1), 27 - 37
Transmembrane insertion of the colicin Ia hydrophobic hairpin; Kienker PK et al.; Colicin Ia is a bactericidal protein that forms voltage-dependent, ion-conducting channels, both in the inner membrane of target bacteria and in planar bilayer membranes . Its amino acid sequence is rich in charged residues, except for a hydrophobic segment of 40 residues near the carboxyl terminus . In the crystal structure of colicin Ia and related colicins, this segment forms an alpha-helical hairpin . The hydrophobic segment is thought to be involved in the initial association of the colicin with the membrane and in the formation of the channel, but various orientations of the hairpin with respect to the membrane have been proposed . To address this issue, we attached biotin to a residue at the tip of the hydrophobic hairpin, and then probed its location with the biotin-binding protein streptavidin, added to one side or the other of a planar bilayer . Streptavidin added to the same side as the colicin prevented channel opening . Prior addition of streptavidin to the opposite side protected channels from this effect, and also increased the rate of channel opening; it produced these effects even before the first opening of the channels . These results suggest a model of membrane association in which the colicin first binds with the hydrophobic hairpin parallel to the membrane; next the hairpin inserts in a transmembrane orientation; and finally the channel opens . We also used streptavidin binding to obtain a stable population of colicin molecules in the membrane, suitable for the quantitative study of voltage-dependent gating . The effective gating charge thus determined is pH-independent and relatively small, compared with previous results for wild-type colicin Ia.

Infect Immun, 1997 May, 65(5), 1908 - 15
Characterization of the physiological requirements for the bactericidal effects of a monoclonal antibody to OspB of Borrelia burgdorferi by confocal microscopy; Escudero R et al.; A confocal microscopy study was undertaken to characterize the bactericidal effects of the Fab fragments of CB2, an immunoglobulin G1kappa murine monoclonal antibody, to an epitope in the carboxy region of the outer surface protein B (OspB) of Borrelia burgdorferi . Simultaneous direct labeling of both fixed and live spirochetes with fluorochrome-labeled Fab-CB2 and 11G1, and an immunoglobulin Mkappa monoclonal antibody to OspA, showed that OspA and OspB seem to colocalize in dead spirochetes but do not appear to be physically associated when the organisms are alive . A polar bleb composed of a Fab-CB2-OspB complex, followed by incorporation of 11G1-OspA, precedes the formation of a spheroplast . The spheroplasts contain both OspA and OspB and are a terminal stage in the bactericidal process induced by Fab-CB2 . Outer membrane destabilization by Fab-CB2, but not cell wall or cytoplasmic membrane alterations, was demonstrated experimentally by the sequential treatment of spirochetes with Fab-CB2 and monoclonal antibodies to flagellin and DnaK . The action of Fab-CB2 is epitope specific, as another monoclonal antibody to an epitope in the amino terminus of OspB was not bactericidal . The bactericidal effect of Fab-CB2 is not dependent on the induction of spirochetal proteases but is dependent on the presence of Ca2+ and Mg2+ . Supplementation of Ca2(+)- and Mg2(+)-free medium with these cations restored the bactericidal effects of Fab-CB2 . The mechanism by which a Fab fragment of an antibody destroys a bacterium directly may represent a novel form of antibody-organism interaction.

J Chromatogr B Biomed Sci Appl, 1997 Apr 25, 692(1), 79 - 86
Fast specific separation and sensitive quantification of bactericidal and sporicidal aldehydes by high-performance liquid chromatography: example of glutaraldehyde determination; Menet MC et al.; This article describes the design and the validation of the HPLC determination of glutaraldehyde at g/l and mg/l concentrations, after derivatization by 2,4-dinitrophenylhydrazine and using the external standard method . At low concentrations, the reaction mixture needs to be heated and a weight ratio of 500 for the 2,4-dinitrophenylhydrazine reagent and the glutaraldehyde ensures a linear calibration curve . In contrast, high concentrations do not require heating of the reaction mixture and a weight ratio of 32 proved to be sufficient . The optimized HPLC method has been validated for both ranges of concentrations . Between 1.25 and 10 mg/l, the content can be determined by the external standard method, with a repeatability of 0.5% . The detection limit is 0.2 mg/l . Between 0.31 and 2.5 g/l, the content can also be determined by the external standard method, with a repeatability of 0.4% . Finally, statistical analysis has demonstrated that aqueous solutions of glutaraldehyde are stable for at least three days at 4 degrees C within the mg to g range.

Biochim Biophys Acta, 1997 Apr 4, 1338(2), 282 - 94
X-ray absorption and resonance raman spectroscopy of human myeloperoxidase at neutral and acid pH; Yue KT et al.; Myeloperoxidase (MPO), an important enzyme in the oxygen-dependent host defense system of human polymorphonuclear leukocytes, utilizes hydrogen peroxide to catalyze the production of hypochlorous acid, an oxidizing bactericidal agent . While MPO shows significant sequence homology with other peroxidases and this homology is particularly striking among the active-site residues, MPO exhibits unusual spectral features and the unique ability to catalyze the oxidation of chloride ions . We have investigated the MPO active-site with X-ray absorption (XAS) and resonance Raman (RRS) spectroscopies at neutral pH and also at the physiological acidic pH (pH approximately 3) and have compared these results with those of horseradish peroxidase (HRP) . At pH 7.5, XAS results show that the iron heme active site is 6-coordinate where the distal ligand is likely nitrogen or oxygen, but not sulfur . The heme is distorted compared to HRP, other peroxidases, and heme compounds, but at pH approximately 3, the distal ligand is lost and the heme is less distorted . RRS results under identical pH conditions show that the skeletal core-size sensitive modes and v3 are shifted to higher frequency at pH approximately 3 indicating a 6- to 5-coordination change of high spin ferric heme . In addition, a new band at 270 cm(-1) is observed at pH approximately 3 which is consistent with the loss of the sixth ligand . The higher symmetry of the heme at pH approximately 3 is reflected by a single v4 mode in the (RRS) spectrum . HRP also loses its loosely associated distal water at this pH, but little change in heme distortion is observed . This change suggests that loss of the distal ligand in MPO releases stress on the heme which may facilitate binding of chloride ion.

Am J Physiol, 1997 Apr, 272(4 Pt 1), L608 - 13
Activation of endothelial cell phospholipase D by polycations; Vepa S et al.; Naturally occurring polycations and cationic proteins are implicated in vascular disorders . It is known that activated leukocytes and platelets release polycations, such as polylysine (PLys), of varying molecular sizes into the vasculature, and some of these have been described to be bactericidal . Polycations interact with endothelial cells (ECs) and cause alterations in permeability and cellular functions . The precise mechanism(s) by which polycations bring about cellular changes is unknown . Here, we report that the polycations PLys and polyarginine (PArg) induce phospholipase D (PLD) activation in ECs . Polycation-mediated PLD activation was both time and concentration dependent, and activation of PLD was not due to cytotoxicity . PArg was more potent compared with PLys of the same molecular weight in stimulation of PLD . Treatment with bisindolylmaleimide, a specific protein kinase C (PKC) inhibitor, and heparin attenuated polycation-mediated PLD activation . Furthermore, downregulation of PKC by 12-O-tetradecanoylphorbol-13-acetate (100 nM, 18 h) also blocked polycation-mediated PLD stimulation . These data suggest that polycation-mediated PLD stimulation probably involves PKC and may represent an important cellular response to leukocyte/platelet activation in the vascular endothelium.

Am J Physiol, 1997 Apr, 272(4 Pt 1), C1352 - 64
Cytokine control of PMN phagocytosis: regulatory effects of hypoxemia and hypoxemia-reoxygenation; Knowles R et al.; We investigated the effects of hypoxemia and hypoxemia-reoxygenation (H/R) on interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha), or IL-1beta stimulation of whole blood polymorphonuclear leukocyte (PMN) phagocytosis and bactericidal activity . Whole blood PMN were rendered hypoxemic (venous PO2 < 15 mmHg), normoxic (venous PO2 60-80 mmHg), or reoxygenated after hypoxemia (H/ R = venous PO2 150-200 mmHg) and were incubated with IL-8, TNF-alpha, or IL-1beta before sequential addition of serum-opsonized fluorescent microspheres and fluorescein isothiocyanate-conjugated mouse anti-human CD64, CD32w, CD16, CD35, or CD11b/CD18 . Concomitant two-color flow cytometric analyses were then performed measuring mean channel fluorescence and the percentage of PMN positive for phagocytosis, with simultaneous subset receptor analysis on populations of PMN that exceeded control levels of phagocytosis . During hypoxemia, whole blood PMN phagocytosis in the presence of IL-8, TNF-alpha, or IL-1beta was increased compared with normoxia . Northern blot analyses revealed an increase in steady-state mRNA levels for CD32w during hypoxemia + IL-8 and CD64 during hypoxemia + IL-1beta . During reoxygenation, both whole blood PMN phagocytosis and bactericidal activity were reduced in the presence of IL-8, TNF-alpha, or IL-1beta, and in subsets of PMN with reduced phagocytosis H/R reduced CD64, CD32w, CD16, CD35, and CD11b/CD18 expression in the presence of each cytokine . Northern blot analyses revealed that H/R reduced mRNA levels for opsonic receptors primarily for IL-1beta-stimulated PMN . These results demonstrate a direct regulatory effect of hypoxemia and H/R on whole blood PMN phagocytosis, receptor expression, and steady-state mRNA levels of both Fc(gamma) and complement receptors.

Infect Immun, 1997 Apr, 65(4), 1228 - 36
Complement-mediated serum sensitivity among spirochetes that cause Lyme disease; van Dam AP et al.; Borrelia burgdorferi-related isolates were tested for their sensitivity to normal human serum (NHS) and their ability to activate complement . By dark-field microscopy, electron microscopy, and subsurface plating, it was shown that exposure of a Borrelia garinii isolate to 10% or more NHS resulted in immobilization, blebbing, and killing of the spirochetes . These effects were mediated by complement, since they were not seen after heat treatment of NHS, in the presence of EDTA, or in an agammaglobulinemic serum . All seven B . garinii type 5 or 6 and all four VS116/M19 strains were serum sensitive, whereas all eight Borrelia afzelii, five of eight B . garinii type 4, and three of seven B . burgdorferi sensu stricto isolates were serum resistant . The other isolates were partially serum sensitive . Four serum-sensitive B . garinii isolates had been isolated from human cerebrospinal fluid . Most likely, activation of both the alternative pathway and the classical pathway of complement was involved, since bactericidal activity was diminished in properdin-deficient sera as well as in a C1q-depleted serum and in a C4-deficient serum . Bactericidal activity could be restored when a serum lacking C1q or C4 was mixed with a properdin-deficient serum . Isolates with various genetic backgrounds were equally able to activate C3 as measured by enzyme-linked immunosorbent assay . In the presence of Mg-EGTA, C3 was activated by all isolates after exposure to > or = 10% NHS . This study shows that B . burgdorferi-related spirochetes can be either serum sensitive or serum resistant in vitro and that this characteristic is associated with their genetic background.

Immunol Cell Biol, 1997 Apr, 75(2), 136 - 41
Enhancement of macrophage-mediated bactericidal activity by macrophage-mannose receptor-ligand interaction; Lefkowitz DL et al.; Neutrophils represent one of the host's primary defenses against invading organisms . These cells often arrive at the site of infection prior to macrophages (M phi) . Neutrophils release myeloperoxidase (MPO) into the micro-environment during phagocytosis . Previous studies by the present investigators have shown that M phi bactericidal activity is enhanced by exposure to MPO . A recent report suggests that as much as 40% of this protein is enzymatically inactive once it is released into the micro-environment . In the present study, exposure of M phi to an enzymatically inactive form of MPO (iMPO) or another mannosylated protein, mannosylated bovine serum albumin (mBSA), can induce the same enhanced Mo-mediated bacterial cell killing observed with the active form of MPO . Furthermore, this phenomenon is limited as galactosylated BSA (gBSA) did not induce enhancement of bacterial killing . The data suggest that interaction of either enzymatically active or inactive mannosylated proteins with the M phi mannose receptor (MMR), is sufficient to enhance M phi bactericidal activity and further underscores the binding of the MMR and resultant responses as a major host defense mechanism.

Appl Environ Microbiol, 1997 Apr, 63(4), 1396 - 9
Infection of Acanthamoeba castellanii by Chlamydia pneumoniae; Essig A et al.; Chlamydia pneumoniae is an intracellular respiratory pathogen, which, similar to Legionella, might have developed mechanisms to escape the intracellular bactericidal activity of both human host cells and amoeba . We therefore investigated the intracellular growth and survival of C . pneumoniae in Acanthamoeba castellanii by using cell culture, immunofluorescence microscopy, and electron microscopy . A castellanii was incubated with purified elementary bodies of C . pneumoniae TW 183 at a concentration of 10(6) inclusion-forming units (IFU)/ml to give a ratio of approximately 1 IFU of C . pneumoniae per amoeba . Quantitative determination of chlamydial growth within A . castellanii revealed viable and infective C . pneumoniae in the range of 10(4) to 10(5) IFU/ml between days 7 and 14 postinfection . Immunofluorescence analysis and transmission electron microscopy with subsequent immunogold staining confirmed evidence of infection of the amoebae by C . Pneumoniae and additionally revealed that C . pneumoniae entered the typical growth cycle . Our results show that amoebae allow the survival of C . pneumoniae, suggesting that amoebae may serve as an additional reservoir for Chlamydia or Chlamydia-related organisms.

Radiats Biol Radioecol, 1997 Mar-Apr, 37(2), 228 - 32
{The immunocorrection with Aerosil-350 of the natural resistance of mice found under conditions of an elevated radiation background}; Ganova LA et al.; Experiments on the CBA mice that had been within 30 km of the Chernobyl NPP accident for 3 months revealed a decrease in the phagocytic and bactericidal properties of peritoneal macrophages accompanied with a disturbance in the production of interleukin-1 and a tumor necrosis factor responsible for the cell immunity . A single administration of a silicon-containing enterosorbent Aerosil-350 expedient to remove radionuclides and ensure the organism detoxication made it possible to correct and restore the affected cell and organism immunity for 7 days.

Med Microbiol Immunol (Berl), 1997 Mar, 185(4), 253 - 60
Heterogeneity in the complement-dependent bacteriolysis within the species of Borrelia burgdorferi; Breitner-Ruddock S et al.; Sixteen Borrelia burgdorferi strains, including all three species, were compared in a colorimetric bactericidal assay for their ability to escape the complement-dependent bacteriolysis on incubation in normal human serum free of specific antibodies (NHS) . The species B . afzelii was found to be serum resistant (EB1, EB3, FEM1, FEM2, Pko), whereas strains of the species B . garinii were found to be serum sensitive (1/B29, G1, G2, PSth, PBr, PTrob) . Six strains, mainly B . burgdorferi sensu stricto, were only partially sensitive (Z25, 297, B31, PKa-I, PBi) . All strains activated the complement cascade in NHS, whereas only four strains (G1, G2, PBr, PSth) could activate complement in the presence of EGTA-Mg . After complement activation, covalently bound C3 fragments (C3b, iC3b) were detected on serum-sensitive as well as serum-resistant borrelial strains . Heterogeneity, however, was observed between serum-resistant and serum-sensitive strains with respect to deposition of C6 and C9 . Whereas serum-sensitive strains were strongly positive for C6 and C9 and were, therefore, killed by the terminal complement complex (TCC), serum-resistant strains were devoid of C6 and C9 on their cell surface . The serum resistance may, therefore, be due to an absent or only transient formation of TCC on the bacterial surface.

Anticancer Res, 1997 Mar-Apr, 17(2A), 1099 - 106
Effects of conjugated dienoic derivatives of linoleic acid and beta-carotene in modulating lymphocyte and macrophage function; Chew BP et al.; The in vitro effects of conjugated dienoic derivatives of linoleic acid (CLA) in combination with beta-carotene on lymphocyte and macrophage function was studied . Porcine blood lymphocytes and murine peritoneal macrophages were incubated with 0 (control), 1.78 x 10(-5), 3.57 x 10(-5) and 7.14 x 10(-5) M CLA and 0 (control), 10(-9), 10(-8) and 10(-7) M beta-carotene . CLA alone stimulated mitogen-induced lymphocyte proliferation, lymphocyte cytotoxic activity and macrophage bactericidal activity . In contrast, CLA inhibited interleukin-2 production by lymphocytes and suppressed the phagocytic activity of macrophages . beta-Carotene alone stimulated the cytotoxicity of lymphocytes and increased superoxide production by peritoneal macrophages . When present together, CLA and beta-carotene interacted in an additive manner to further enhance lymphocyte cytotoxicity and spontaneous lymphocyte proliferation . In addition, beta-carotene was able to negate the inhibitory action of CLA on the phagocytic activity of macrophages . Also, CLA and beta-carotene together seemed to suppress mitogen-induced lymphocyte proliferation . Therefore, CLA and beta-carotene; alone and in concert, act to modulate different aspects of cellular host defense.

Lepr Rev, 1997 Mar, 68(1), 16 - 24
Release of reactive nitrogen intermediates from the peripheral blood-derived monocytes/macrophages of leprosy patients stimulated in vitro by tuftsin; Khare S et al.; The production of reactive nitrogen intermediates (RNI) by macrophages is critical to host defence, particularly for exerting the bactericidal and tumoricidal properties . Nitric oxide (NO) were measured in the peripheral blood-derived monocytes/macrophages of normal and leprosy patients (BT/TT and BL/LL) in the presence and absence of 'tuftsin' as a function of in vitro culture age (on 1, 3, 7 days) . Macrophages from both groups of leprosy patients were able to produce NO during the unstimulated state but only BL/LL macrophages could be activated by tuftsin to produce significantly high levels of NO . This increase was highest on day 1, then gradually decreased with in vitro culture age . Surprisingly, tuftsin was unable to enhance the NO production in normal macrophages above the basal level . Further, normal and BT/TT macrophages had only Cu-Zn derived superoxide dismutase (SOD) activity whereas BL/LL cultures has Cu-Zn and Mn derived SOD activity . These studies indicate that in BL/LL cultures: a, apart from tuftsin, some additional signal is required to activate nitric oxide synthase (NOS) gene for NO production; and b, Mn-SOD produced by Mycobacterium leprae is playing a defensive role against toxic-free radicals . The final outcome of this mechanism is the survival of M . leprae inside the macrophages.

Chest, 1997 Mar, 111(3), 577 - 83
Release of lipopolysaccharide toxicity-modulating proteins in patients undergoing cardiopulmonary bypass using noncoated and heparin-coated extracorporeal circuits . A clinical pilot study; Bouma M et al.; STUDY OBJECTIVE: Cardiopulmonary bypass (CPB) induces a generalized inflammatory response, including activation of leukocytes, contributing to postoperative morbidity . The inflammatory pathways leading to this systemic inflammatory response syndrome are considered identical to those involved in septic shock . Therefore, we studied the release of bactericidal/permeability-increasing protein (BPI), lipopolysaccharide binding protein (LBP), and soluble CD14 (sCD14)-all proteins that modulate the effects of lipopolysaccharide (LPS)-in patients undergoing CPB . In addition, the effect of heparin coating of the extracorporeal bypass circuit on the release of these parameters was assessed . DESIGN: Prospective, randomized clinical pilot study . SETTING: Cardiothoracic Surgery Department in a university hospital . PATIENTS: Fourteen patients undergoing elective coronary artery bypass grafting were included . Seven patients underwent CPB using a standard, noncoated extracorporeal circuit, and seven patients had CPB using a heparin-coated extracorporeal circuit (Duraflo II) . INTERVENTIONS: Blood samples were taken after induction of anesthesia, just before aortic crossclamping, and 0, 0.5, 1.5, 3, 6, 12, and 24 h after declamping . MEASUREMENTS AND RESULTS: CPB with a noncoated extracorporeal circuit induced a sharp increase in neutrophil-derived BPI, manifest directly after release of the aortic crossclamp, which was significantly attenuated using a heparin-coated system . Also, CPB induced a gradual increase of the acute-phase reactant LBP, which was identical in the noncoated and heparin-coated groups . Systemic release of sCD14 after crossclamp release was significantly higher in the noncoated group compared with the heparin-coated group, but did not rise above baseline levels . CONCLUSIONS: These data confirm that CPB-induced leukocyte activation is attenuated using a heparin-treated extracorporeal circuit and point to the possible role of LPS toxicity-modulating proteins in the systemic inflammatory response after bypass surgery.

J Antimicrob Chemother, 1997 Mar, 39(3), 331 - 7
Bactericidal activity and synergy studies of proton pump inhibitors and antibiotics against Helicobacter pylori in vitro; Midolo PD et al.; The bactericidal activity of the proton pump inhibitors omeprazole and lansoprazole alone and in combination with a beta-lactam or macrolide antibiotic were investigated in vitro . Time-kill curves against Helicobacter pylori NCTC 11637 and a recent clinical isolate revealed significant concentration-dependent killing with all drugs other than amoxycillin . Combinations of proton pump inhibitor and erythromycin showed synergic activity . In contrast, proton pump inhibitor plus amoxycillin showed additive activity against the clinical isolate only . Bactericidal investigations of anti-helicobacter drugs in vitro may suggest optimum treatment strategies for this common infectious disease.

FEMS Immunol Med Microbiol, 1997 Mar, 17(3), 187 - 99
The major outer membrane protein, CD, extracted from Moraxella (Branhamella) catarrhalis is a potential vaccine antigen that induces bactericidal antibodies; Yang YP et al.; The major outer membrane protein of Moraxella (Branhamella) catarrhalis, CD, was detergent-extracted from the bacterial cell wall and purified to homogeneity in high yields by a simple process . The purified protein appeared to exhibit immunogenic properties similar to those of native CD exposed on the surface of the bacterium . Antibodies to CD raised in mice specifically bound to intact B . catarrhalis, as determined by flow cytometry analysis . The IgG subclass distributions of anti-CD antibodies in sera from mice immunized with purified CD or with B . catarrhalis were also similar . CD was found to be antigenically conserved among a panel of B . catarrhalis isolates, as demonstrated by the consistent reactivities of mouse anti-CD antisera with a common 60 kDa protein on immunoblots . Furthermore, convalescent sera collected from patients with otitis media due to B . catarrhalis infection were found to be reactive with the CD protein by immunoblotting . Finally, the purified protein induced antibodies in guinea pigs and mice that exhibited in vitro bactericidal activity against the pathogen . Therefore, the native CD outer membrane protein represents a potentially useful antigen for inclusion in a vaccine against B . catarrhalis.

Pediatr Infect Dis J, 1997 Mar, 16(3 Suppl), S65 - 9
Acute otitis media in pediatrics: are there rational issues for empiric therapy?
Boccazzi A, Careddu P.
BACKGROUND: Acute otitis media in children is a significant clinical problem that requires a rational approach to treatment . The condition is extremely common and has important economic implications . At present there is considerable controversy over the most appropriate strategy and over the use and choice of antibiotics . OBJECTIVES: To analyze the various factors that influence therapeutic decisions and consider how these may assist in the formulation of a rational approach to therapy . DISCUSSION: Otitis media has a multifactorial etiology but it is extremely difficult to differentiate between bacterial and viral causes on clinical grounds . Culture of the middle ear fluid is rarely practicable; however, nasal swabs are relatively noninvasive and can provide useful microbiologic information, especially in excluding a bacterial cause . Published information provides little guidance on the most appropriate therapy; a rational approach to treatment is based on many considerations including the local epidemiology . The minimum criteria for the empiric choice of an antibiotic for acute otitis media are that it should be rapidly bactericidal and reach adequate concentrations in the middle ear fluid . In areas where beta-lactamase-producing strains are prevalent, a beta-lactamase-stable antibiotic should be chosen; good absorption from the gastrointestinal tract and high and consistent penetration into the middle ear are important characteristics . Compliance-enhancing factors such as fewer doses per day and good palatability are also important.

J Am Soc Nephrol, 1997 Mar, 8(3), 463 - 70
Lipopolysaccharide-binding protein and bactericidal/permeability-increasing factor during hemodialysis: clinical determinants and role of different membranes; Sundaram S et al.; The host response to the presence of lipopolysaccharide (LPS) is complex and varied . Two closely related endogenous serum proteins, LPS-binding protein (LBP) and bactericidal/permeability-increasing factor (BPI), regulate delivery of LPS to CD14 antigen on effector cell surfaces and modulate the host response to LPS . In the study presented here, plasma levels of LBP and BPI were measured, predialysis, 15 min into dialysis and postdialysis in patients dialyzed with cellulose, cellulose-tri-acetate (CTA), and polysulfone dialyzers . Further, the association between LBP levels and BPI release during hemodialysis and clinical and laboratory characteristics of patients, complement activation represented by plasma C3a levels, and monocyte cytokine production represented by interleukin-1 receptor antagonist (IL-1Ra) synthesis was also studied . Predialysis plasma levels of LBP were 14,459 +/- 544, 13,889 +/- 1362 and 12,622 +/- 6305 ng/mL, respectively, with cellulose, CTA, and polysulfone dialyzers, and postdialysis levels were 17,834 +/- 861, 20,979 +/- 8485 and 18,177 +/- 1656 ng/mL, respectively . Postdialysis plasma levels of LBP were consistently higher than predialysis levels with all three dialyzers (P < 0.05) . However, plasma LBP levels were not significantly different between the three dialyzers either predialysis (P = 0.28) or postdialysis (P = 2.8) . There were no significant differences in predialysis BPI levels between the three dialyzers (P = 0.21) . BPI levels at 15 min of dialysis with CTA (10.91 +/- 3.65 ng/mL) and polysulfone (10.73 +/- 2.24 ng/mL) dialyzers were significantly greater (P < 0.05) than that observed with cellulose (5.49 +/- 0.66 ng/mL) . Similarly, postdialysis levels with CTA and polysulfone were significantly greater (P < 0.05) than that observed with cellulose dialyzers . The percentage change in BPI levels between predialysis and 15 min was 1341 +/- 243%, 2935 +/- 1033%, and 3790 +/- 1151% for cellulose, CTA, and polysulfone dialyzers, respectively . The changes in BPI levels from predialysis to 15 min and between pre- and postdialysis samples were statistically significant for all three dialyzers (P < 0.05) . Postdialysis LBP:BPI ratios were 50 +/- 6%, 18 +/- 4%, and 22 +/- 6% of predialysis ratios for cellulose, CTA, and polysulfone dialyzers, respectively . These changes were statistically significant (P < 0.05) for all three dialyzers . There was no significant correlation between baseline clinical or laboratory characteristics and predialysis LBP levels . Similarly, the correlation between BPI levels at 15 min of dialysis with the clinical and laboratory characteristics was also poor, with the exception of serum albumin (r = 0.43, P = 0.008) . The correlation between BPI levels at 15 min of dialysis with plasma LBP levels (r = -0.29; P = 0.08), plasma C3a levels (r = -0.1; P = 0.55), peripheral blood mononuclear cells (PBMC) content of IL-1Ra (r = 0.01; P = 0.94), and IL-1Ra production by unstimulated (r = 0.13; P = 0.45), and endotoxin-stimulated PBMC (r = 0.32; P = 0.06) was not statistically significant . The results of this study demonstrate that dialysis with cellulose, CTA, and polysulfone dialyzers results in a significant increase in LBP and BPI levels . BPI release is probably mediated by non-complement factors and may be related to the nutritional status of the patient . The release of BPI during HD and consequent lowering of the LBP:BPI ratio could potentially afford some protection against endotoxin in the dialysate.

Antimicrob Agents Chemother, 1997 Mar, 41(3), 607 - 10
Which aminoglycoside or fluoroquinolone is more active against Mycobacterium tuberculosis in mice?
Lounis N, Ji B, Truffot-Pernot C, Grosset J.
To identify the most active aminoglycoside or fluoroquinolone for the treatment of tuberculosis, the in vivo activities of four different aminoglycosides and three different fluoroquinolones were compared with that of isoniazid (INH) in a murine tuberculosis model . Mice were each inoculated intravenously with 2.3 x 10(7) CFU of Mycobacterium tuberculosis H37Rv . Treatment began the next day (D1) after inoculation and continued for 4 weeks, at the frequency of six times weekly with one of the following regimens: INH, 25 mg/kg; ofloxacin, 200 mg/kg; levofloxacin, 100 or 200 mg/kg; sparfloxacin (SPFX), 50 mg/kg; and streptomycin, kanamycin, amikacin (AMIKA), and isepamicin, all at 200 mg/kg . The dosages of the treatments were presumably equivalent to their clinically tolerated dosages . The severity of infection and effectiveness of the treatment were assessed by the survival rate, spleen weights, gross lung lesions, and the numbers of CFU in the spleens . The results indicate that INH is more bactericidal than any of the aminoglycosides or fluoroquinolones tested, that AMIKA is the most active aminoglycoside, and that SPFX at 50 mg/kg is far more bactericidal than the treatment with other fluoroquinolones.

Antimicrob Agents Chemother, 1997 Mar, 41(3), 540 - 3
Characterization of pncA mutations in pyrazinamide-resistant Mycobacterium tuberculosis; Scorpio A et al.; Pyrazinamide (PZA) is a first-line drug for short-course tuberculosis therapy . Resistance to PZA is usually accompanied by loss of pyrazinamidase (PZase) activity in Mycobacterium tuberculosis . PZase converts PZA to bactericidal pyrazinoic acid, and the loss of PZase activity is associated with PZA resistance . The gene (pncA) encoding the M . tuberculosis PZase has recently been sequenced, and mutations in pncA were previously found in a small number of PZA-resistant M . tuberculosis strains . To further understand the genetic basis of PZA resistance and determine the frequency of PZA-resistant strains having pncA mutations, we analyzed a panel of PZA-resistant clinical isolates and mutants made in vitro . Thirty-three of 38 PZA-resistant clinical isolates had pncA mutations . Among the five strains that did not contain pncA mutations, four were found to be falsely resistant and one was found to be borderline resistant to PZA . The 33 PZA-resistant clinical isolates and 8 mutants made in vitro contained various mutations, including nucleotide substitutions, insertions, or deletions in the pncA gene . The identified mutations were dispersed along the pncA gene, but some degree of clustering of mutations was found at the following regions: Gly132-Thr142, Pro69-Leu85, and Ile5-Asp12 . PCR-single-strand conformation polymorphism (SSCP) analysis was shown to be useful for the rapid detection of pncA mutations in the PZA-resistant strains . We conclude that a mutation in the pncA gene is a major mechanism of PZA resistance and that direct sequencing by PCR or SSCP analysis should help to rapidly identify PZA-resistant M . tuberculosis strains.

Pharm Res, 1997 Feb, 14(2), 224 - 9
The role of liver and kidney on the pharmacokinetics of a recombinant amino terminal fragment of bactericidal/permeability-increasing protein in rats; Bauer RJ et al.; PURPOSE: The pharmacokinetics of rBPI23, a recombinant amino terminal fragment of bactericidal/permeability-increasing protein that binds to and neutralizes endotoxin, was investigated . METHODS: rBPI23 was administered to rats at doses 0.01-10 mg/kg and plasma rBPI23 levels were measured by ELISA . rBPI23 was also administered to bilaterally nephrectomized rats . In addition, rBPI23 was administered intra-hepatically via the pyloric vein to determine the first-pass effect by the liver . rBPI23 concentrations were also simultaneously measured in the right atrium and aorta to determine the removal of rBPI23 by the lungs . RESULTS: The concentration-time profile of rBPI23 was described by a 3-compartmental model with parallel first order and Michaelis-Menten (saturable) elimination . The clearance of rBPI23 was not altered by bilateral nephrectomy . Clearance of intra-hepatically administered rBPI23 was 4.5 fold lower than intra-femorally administered rBPI23 . The concentration difference of rBPI23 between aortic and right atrial blood was no greater than 11% . Clearance of rBPI23 in rats could be reduced up to 10 fold by co-administration of heparin . Uptake by liver of intra-hepatically administered rBPI23 was prevented by co-administration of heparin . CONCLUSIONS: rBPI23 is not significantly cleared by the kidneys, and no more than 11% of the rBPI23 was removed by the lungs with each pass . The liver could remove 78% of the rBPI23 from the hepatic circulation . Studies with heparin suggest rBPI23 is cleared by binding to heparan sulfate sites in the liver.

Ann Hematol, 1997 Feb, 74(2), 83 - 7
The putative role of arylsulfatase in interleukin-2-mediated cytotoxicity and interleukin-7-mediated bactericidal activity of natural killer cells; Toren A et al.; We have examined the cytolytic and bactericidal activity of resting and cytokine-stimulated natural-killer (NK) cells against K562 and Daudi cell lines and Escherichia coli, respectively . Unstimulated NK cells showed considerable cytolytic activity against K562 (64 +/- 4%) and relatively low activity against the Daudi cell lines (22 +/- 9%) . Pretreatment of NK cells with the arylsulfatase (AS) type-II-specific inhibitor NaH2PO4 reduced cytotoxicity towards K562 and Daudi 1.3- and 2.9-fold (p < 0.05; n = 12), respectively, indicating that AS participates in NK-mediated cytotoxicity . Interleukin-2 (IL-2) (200 units/ml) caused a 1.3- and 3.5-fold (p < 0.5; n = 12) enhancement of NK cytotoxic activity against K562 and Daudi, respectively . Pretreatment of these cells with the AS type-II-specific inhibitor NaH2PO4 reduced cytotoxicity 1.1-fold towards K562 (p < 0.05; n = 12) and 1.2-fold towards Daudi (p > 0.05; n = 12) indicating that AS does not participate in IL-2-mediated NK cytolytic activity against these cell lines . IL-7 (3 units/ml) did not cause any enhancement of NK cytolytic activity . Unstimulated NK cells showed considerable bactericidal activity against E . coli (23 +/- 4%) . Incubation of resting NK cells with NaH2PO4 reduced the bactericidal effect only by 1.09-fold (p > 0.05; n = 12), indicating that AS does not mediate this effect . IL-2 (200 units/ml) and IL-7 (3 units/ml) enhanced the bactericidal activity 1.5- and 2.2-fold, respectively (p < 0.05; n = 12) . This effect was not influenced by incubation of IL-2-stimulated cells with NaH2PO4, indicating that AS does not participate in the IL-2-mediated NK bactericidal effect . IL-2 seems to exert its stimulatory effect upon NK-mediated bactericidal activity by a different, non-AS-dependent mechanism . However, incubation of IL-7-stimulated NK cells with NaH2PO4 reduced the NK bactericidal effect by 1.2-fold (p < 0.05; n = 12), indicating that AS may have a role in this reaction . These data can be further confirmed by detection of AS through degranulation of NK cells, showing that IL-2 induced only mild degranulation of resting and f-MLP-stimulated NK cells (26 +/- 1% vs 22 +/- 2% and 31 +/- 2% vs 29 +/- 2, respectively) (p > 0.05; n = 8) . In contrast, IL-7 showed significant enhancement of AS release in resting or f-MLP-induced NK cells (36 +/- 3% vs 22 +/- 2% and 49 +/- 3% vs 29 +/- 2%, respectively) (p < 0.05; n = 8).

Antimicrob Agents Chemother, 1997 Feb, 41(2), 326 - 30
Experimental evaluation of possible new short-term drug regimens for treatment of multibacillary leprosy; Banerjee DK et al.; Groups of nude mice, with both hind footpads infected with 10(8) Mycobacterium leprae organisms, were treated with 4-week courses of different drug combinations . The effect treatment on each group was evaluated by subinoculating footpad homogenates from the treated mice into groups of normal and nude mice for subsequent regrowth, assessed 1 year later . A combination of rifampin (RMP) with clarithromycin (CLARI), minocycline (MINO), and ofloxacin (OFLO) resulted in the complete killing of M . leprae after 3 weeks of treatment . A combination of sparfloxacin (SPAR) and RMP also resulted in a similar bactericidal effect after 3 weeks of treatment . Other drug combinations showed variable effects . Very little or no effect was observed with any regimen if the treatment was given for less than 2 weeks . World Health Organization (WHO) multidrug therapy (MDT) given for 8 weeks was as effective as the two combinations described above . The results suggest that multidrug combinations consisting of RMP-OFLO (or SPAR)-CLARI (and/or MINO) are as effective as the WHO MDT for the treatment of experimental leprosy . Moreover, they imply that these combinations, which were found to be active in a 4-week experimental treatment protocol, could be administered as treatment to patients for a period of time shorter than the present 2-year regimen without a loss of effectiveness.

J Biol Chem, 1997 Jan 24, 272(4), 2149 - 55
Biochemical characterization of recombinant fusions of lipopolysaccharide binding protein and bactericidal/permeability-increasing protein . Implications in biological activity; Abrahamson SL et al.; The physiological response to endotoxin (lipopolysaccharide (LPS)) can be regulated by two closely related LPS-binding proteins, LPS-binding protein (LBP), which potentiates LPS' inflammatory activity via interaction with the monocytic antigen CD14, and bactericidal/permeability-increasing protein (BPI), which neutralizes LPS . Both proteins bind LPS with high affinity sites in their N-terminal domains, whereas interaction between LBP and CD14 is dependent upon the LBP C-terminal domain . We have created fusions of the N- and C-terminal domains from each protein and compared the functional activities and pharmacokinetics of these fusions, the individual N-terminal domains, and the parent proteins . The N-terminal domains of BPI and LBP bound lipid A with their characteristic apparent affinity constants, regardless of the C-terminal fusion partner . In addition, the C-terminal domain of LBP allowed transfer of LPS to CD14 in conjunction with either N-terminal LPS binding domain . Proteins containing a BPI N-terminal domain had greater heparin binding capacities in vitro and were cleared more rapidly from the plasma of whole animals . Taken together, these data better define how closely related proteins such as BPI and LBP can have opposing effects on the body's response to LPS.

C R Seances Soc Biol Fil, 1997, 191(2), 237 - 46
{Signal transduction by Rac small G proteins in phagocytes}; Dorseuil O et al.; Rac1 and Rac2 are 92% homologous cytosolic small GTPase proteins . Both Rac1 and Rac2 have been implicated with NADPH oxidase activation in vitro, however, Rac2 is largely predominant in human phagocytes . NADPH oxidase is a plasma membrane enzyme of phagocytes, generating superoxide anions which serve as bactericidal agents . Activation of this multimolecular enzyme, minimally requires assembly at the membrane with flavocytochrome b258 of cytosolic components p47phox, p67phox and Rac proteins . Using the yeast two hybrid system, we provide data demonstrating in vivo interactions between human p47phox, p67phox, and Rac proteins . Rac proteins interact with p67phox in a GTP-dependent manner, but do not interact with p47phox . Moreover, Rac effector site mutants which are known to be inactive in NADPH oxidase lose their interaction with p67phox . Finally, we observe that p67phox interacts six fold better with Rac2 than with Rac1 . We also show a strong intracellular interaction between p47phox and p67phox . These results indicate that activated Rac, and particularly Rac2, can regulate superoxide production by NADPH oxidase of phagocytic cells through direct interaction with p67phox subunit . Recently published data suggest that Rac proteins could transduce mitogenic signals in non-phagocytic cells through superoxide production by a phagocytic-related NADPH oxidase enzymatic system which remains to be determined . NADPH oxidase regulation by Rac proteins in phagocytes could then be used as a model to understand the molecular mechanisms underlying Rac functions in various cell types.

Zh Mikrobiol Epidemiol Immunobiol, 1997 Jan-Feb, (1), 3 - 6
{The role of the thrombocytic cationic protein (beta-lysin) in anti-infectious protection}; Bukharin OV et al.; The biological activity of beta-lysin, cationite protein of thrombocytic origin with bactericidal activity, isolated from human blood serum was studied . The mechanisms of its bactericidal action was determined: the inhibition of bacterial catalase and peroxidase . The study revealed that the activity of beta-lysin in anti-infectious protection was due to both its direct bactericidal action and the enhancement the phagocytic reaction of the body by the stimulation of phagocytes and the opsonization of the pathogen.

Lik Sprava, 1997 Jan-Feb, (1), 73 - 5
{The possibility of using highly disperse iron for the directed transport of thyroxin in the body}; Shvets TM et al.; It has been ascertained that high-disperse powders of iron obtained by a thermochemical method in the process of formation of particles with pre-determined physical-and-chemical and medical-and-biological properties (essentially monodisperse, harmless, capable of exerting a bactericidal action, having an as-needed specific surface, corrosion resistance, withstanding sterilization heat up to 120 degrees C, allowing the magnetic characteristics to be controlled, etc.) can be used for directed transport of medicinal substances (thyroxin), hormone control, protection of the organism from overdosing of the administered drug preparations . Feasibility is shown of prolonged administration of thyroid hormones through the gastrointestinal wall with the aid of high-disperse ferromagnetic compositions as containers-carriers, which fact may secure the organism-controlled maintenance of the thyroid status in various thyroidal pathologies.

Probl Tuberk, 1997, (1), 53 - 4
{Bactericidal effects of maxaquin on mycobacterium tuberculosis}; Kunichian AD et al.; Maxaquine is a fluoroquinolone agent that is effective against Mycobacterium tuberculosis (MBT) . The drug was tested for effects on MBT growth in the infected lung tissue in relation to its concentration and exposure time . The bactericidal effect was found to be achieved no earlier than 72 hours of its administration . The minimum bactericidal concentration of maxaquine was 2 micrograms/ml.

Gut, 1997 Jan, 40(1), 105 - 9
Antineutrophil cytoplasm autoantibodies against bactericidal/permeability-increasing protein in inflammatory bowel disease; Walmsley RS et al.; BACKGROUND: Bactericidal/permeability-increasing protein (BPI), a constituent of primary neutrophil granules, is a potent natural antibiotic and an antineutrophil cytoplasm antibody (ANCA) antigen in cases of vasculitis in which the target antigen is neither myeloperoxidase (MPO) nor proteinase-3 (PR3) . AIM: To investigate BPI as a possible target antigen for ANCAs in inflammatory bowel disease . METHODS: ANCAs were detected by routine immunofluorescence (IIF) and solid phase enzyme linked immunosorbent assay (ELISA) performed for antibodies to the purified neutrophil granule proteins; MPO, PR3, cathepsin-G, lactoferrin, and BPI in serum samples from 88 patients with inflammatory bowel disease (36 with Crohn's disease, 52 with ulcerative colitis) . Thirty patients with bacterial enteritis acted as controls . RESULTS: Significantly more patients with ulcerative colitis were ANCA positive by IIF (60%) than patients with Crohn's disease (28%) or infectious enteritis (23%) (p < 0.001) . IgG anti-BPI antibodies were present in 29% of patients with ulcerative colitis, 14% of patients with Crohn's disease, and 23% of patients with infectious enteritis, occurring in 44% of those patients with inflammatory bowel disease who were ANCA positive by IIF . Antibodies to other ANCA antigens were rare . The presence of ANCAs was not related to either disease activity or extent; presence of anti-BPI antibodies was significantly related to both a lower serum albumin concentration (p = 0.001) and a higher erythrocyte sedimentation rate (p = 0.02) in patients with ulcerative colitis, and to colonic involvement in patients with Crohn's disease (p = 0.01) . CONCLUSION: BPI is a significant minority target antigen for ANCAs in inflammatory bowel disease that seems related to colonic Crohn's disease and disease activity in ulcerative colitis . Anti-BPI antibodies occur in infectious enteritis.

Annu Rev Immunol, 1997, 15, 749 - 95
Cellular responses to interferon-gamma; Boehm U et al.; Interferons are cytokines that play a complex and central role in the resistance of mammalian hosts to pathogens . Type I interferon (IFN-alpha and IFN-beta) is secreted by virus-infected cells . Immune, type II, or gamma-interferon (IFN-gamma) is secreted by thymus-derived (T) cells under certain conditions of activation and by natural killer (NK) cells . Although originally defined as an agent with direct antiviral activity, the properties of IFN-gamma include regulation of several aspects of the immune response, stimulation of bactericidal activity of phagocytes, stimulation of antigen presentation through class I and class II major histocompatibility complex (MHC) molecules, orchestration of leukocyte-endothelium interactions, effects on cell proliferation and apoptosis, as well as the stimulation and repression of a variety of genes whose functional significance remains obscure . The implementation of such a variety of effects by a single cytokine is achieved by complex patterns of cell-specific gene regulation: Several IFN-gamma-regulated genes are themselves components of transcription factors . The IFN-gamma response is itself regulated by interaction with responses to other cytokines including IFN-alpha/beta, TNF-alpha, and IL-4 . Over 200 genes are now known to be regulated by IFN-gamma and they are listed in a World Wide Web document that accompanies this review . However, much of the cellular response to IFN-gamma can be described in terms of a set of integrated molecular programs underlying well-defined physiological systems, for example the induction of efficient antigen processing for MHC-mediated antigen presentation, which play clearly defined roles in pathogen resistance . A promising approach to the complexity of the IFN-gamma response is to extend the analysis of the less understood IFN-gamma-regulated genes in terms of molecular programs functional in pathogen resistance.

J Burn Care Rehabil, 1997 Jan-Feb, 18(1 Pt 1), 17 - 21
Effects of recombinant bactericidal, permeability-increasing protein on bacterial translocation and pulmonary neutrophil sequestration in burned mice; Rennekampff OH et al.; Burn injury induces bacterial translocation (BT) from the gut in multiple animal models . Etiologic factors contributing to BT may be an ischemia-reperfusion injury to the gut, the release of inflammatory cytokines, oxygen metabolites and other mediators, and cytotoxic effects mediated by endotoxin (lipopolysaccharide) . Bactericidal, permeability-increasing protein is a neutrophil granule protein with potent bactericidal and lipopolysaccharide-neutralizing activities . The use of this protein has not been previously reported in a burn-injury model . The purpose of this study was to determine whether recombinant bactericidal, permeability-increasing protein (rBPI23) affects the incidence of BT and myeloperoxidase content in lung tissue (a measure of leukocyte sequestration) in a burn-injury model . Mice received a 32% total body surface area, full-thickness, scald burn, and 10 mg/kg body weight rBPI23 in saline solution was given by intraperitoneal injection at 0, 3, and 6 hours after the burn . Control animals received intraperitoneal saline solution only . All animals received a total of 1 ml saline solution intraperitoneally immediately after burn injury for fluid resuscitation . At 24 hours after burn injury, mesenteric lymph nodes (MLN) were harvested, homogenized, and plated . Lung tissue was harvested and assayed for myeloperoxidase . Burned mice treated with rBPI23 had significantly (p = 0.005, Fisher's Exact Test, two-tailed) decreased incidence of BT, compared to burned mouse controls . Leukosequestration into lung tissues was not affected by rBPI23 . Postburn administration of rBPI23 reduces but does not abolish the incidence of BT after burn injury in mice, perhaps by reducing intestinal injury during burn shock and the ischemia-reperfusion period by inhibiting the effects of lipopolysaccharide . An alternate explanation may be that rBPI23 could increase clearance and killing of bacteria by host defenses.

Nephron, 1997, 75(1), 41 - 7
Polymorphonuclear cells in chronic hemodialysis patients have intact phagocytotic and impaired bactericidal activities; Iida T et al.; Although it has been well documented that chronic hemodialysis (HD) patients are highly susceptible to infectious diseases, the reasons for this have yet to be clarified . The present study was thus designed in order to better define this issue . Fifty-eight stable chronic HD patients without any evidence of infection were selected for the study . Blood samples were collected before and after HD from the same patient to determine the effect of HD . Reactive oxygen species (ROS) production in polymorphonuclear cells (PMNC) was measured by chemiluminescence using luminol . When the PMNC collected after HD were stimulated in vitro with a calcium ionophore (A23187), they produced a larger amount of ROS than that obtained from healthy volunteers {mean 7.4 x 10(5) photon counts (n = 58) vs . 3.0 x 10(5) photon counts (n = 17); p < 0.01} . A higher production of ROS after HD was seen in patients using membranes such as cellulose triacetate, polymethylmetacrylate and cellulose diacetate, whereas cuprophane did not seem to augment ROS production at all . On the other hand, when the PMNC after HD were stimulated with phorbol myristate acetate, their photon counts (mean 4.3 x 10(7)) were comparable to those before HD (mean 3.5 x 10(7)), and to those of PBMC obtained from healthy volunteers (mean 4.1 x 10(7)) . It was thus suggested that the enhanced ROS production of PMNC was related to some stimuli, possibly even to the assay used to measure ROS . The phagocytotic activity and bactericidal effect of PBMC were measured by coculturing 1 x 10(5) PMNC with 1 x 10(5) CFU of Escherichia coli . Similar phagocytotic activities were noted in the PMNC from healthy volunteers and chronic HD patients before and after HD: the mean number of phagocytosed bacteria (log10 CFU) was 3.3, 3.3, and 3.3, respectively . However, in the case of a bactericidal effect, only the PMNC from healthy volunteers, but not the PMNC from HD patients, could effectively kill the bacteria, since the number of bacteria in PMNC decreased from 10(3.3) to 10(2.1) . The PMNC from HD patients could not kill the bacteria-regardless of the characteristics of the membranes . It was thus concluded that the PMNC of chronic HD patients possess an intact phagocytotic activity which impaired bacterial killing, and was probably due to an abnormality occurring in the ROS production pathway.

J Infect Dis, 1997 Jan, 175(1), 108 - 17
Bactericidal/permeability-increasing protein release in whole blood ex vivo: strong induction by lipopolysaccharide and tumor necrosis factor-alpha; Dentener MA et al.; In this study, the release of bactericidal/permeability-increasing protein (BPI), which is stored in polymorphonuclear leukocytes (PMNL), was analyzed in a whole blood ex vivo system . Of the microbial products tested, lipopolysaccharide (LPS) most potently induced BPI release; FMLP, serum-treated zymosan (STZ), and lipoteichoic acid (LTA) also induced BPI release . In addition, the inflammatory mediator tumor necrosis factor (TNF)-alpha potently activated PMNL in whole blood, via TNF receptor p55, to release BPI, whereas interleukin (IL)-1, IL-8, platelet activating factor, and C5a were poor inducers of BPI release . STZ and phorbol myristate acetate, but not LPS, FMLP, or LTA, stimulated isolated PMNL to release BPI . BPI was released in comparable magnitude with the azurophilic granule protein elastase . Furthermore, both proteins were released with similar kinetics, which started within 30 min after onset of stimulation and lasted 1-4 h.

Antimicrob Agents Chemother, 1997 Jan, 41(1), 49 - 53
Once-daily gentamicin therapy for experimental Escherichia coli meningitis; Ahmed A et al.; In vitro and in vivo studies have demonstrated that the bacteriologic efficacy of once-daily aminoglycoside therapy is equivalent to that achieved with conventional multiple daily dosing . The impact of once-daily dosing for meningitis has not been studied . Using the well-characterized rabbit meningitis model, we compared two regimens of the same daily dosage of gentamicin given either once or in three divided doses for 24 or 72 h . The initial 1 h mean cerebrospinal fluid (CSF) gentamicin concentration for animals receiving a single dose (2.9 +/- 1.7 micrograms/ml) was threefold higher than that for the animals receiving multiple doses . The rate of bacterial killing in the first 8 h of treatment was significantly greater for the animals with higher concentrations in their CSF (-0.21 +/- 0.19 versus -0.03 +/- 0.22 log10 CFU/ml/h), suggesting concentration-dependent killing . By 24h, the mean reduction in bacterial titers was similar for the two regimens . In animals treated for 72 h, no differences in bactericidal activity was noted for 24, 48, or 72 h . Gentamicin at two different dosages was administered intracisternally to a separate set of animals to achieve considerably higher CSF gentamicin concentrations . In these animals, the rate of bacterial clearance in the first 8 h (0.52 +/- 0.15 and 0.58 +/- 0.15 log10 CFU/ml/h for the lower and higher dosages, respectively) was significantly greater than that in animals treated intravenously . In conclusion, there is evidence of concentration-dependent killing with gentamicin early in treatment for experimental E . coli meningitis, and once-daily dosing therapy appears to be at least as effective as multiple-dose therapy in reducing bacterial counts in CSF.

Recenti Prog Med, 1996 Dec, 87(12), 623 - 7
{Pulmonary complications in diabetes mellitus}; Marvisi M et al.; The lung is not considered a target organ in diabetes mellitus . In English language literature there are many papers showing the opposite . Many studies demonstrated a thickened alveolar epithelial and pulmonary capillary basal lamina and a reduced lung elasticity, others showed that these histopathological alterations developed into functional abnormalities: reduced lung volumes, reduced pulmonary diffusion capacity and elastic recoil . The pathogenesis is currently thought to involve the nonenzymatic glycosylation (NEG) of tissue proteins inducing an alteration in connective tissue . In patients with diabetic autonomic neuropathy there is an abnormal basal airway tone due to an alteration in vagal pathways: these patients have a reduced bronchial reactivity and bronchodilatation . Diabetic patients have an increased propensity to acquire infections, in particular tuberculosis and pulmonary fungal diseases (coccidioidomycosis, aspergillosis and mucormycosis) . The frequency of occurrence of tuberculosis is reported to be four times than in non diabetics, there is a predilection for the lower lobes and the disease is more aggressive in poorly controlled diabetes mellitus . Pulmonary mucormycosis is an infection caused by Phycomycetes, the fungus has the propensity to invade vascular structures giving hemoptysis and leading to a high mortality unless diagnosed promptly . The mechanism for the increased susceptibility to infection is due to an alteration in chemotactic, phagocytic and bactericidal activity of polymorphonuclear leukocytes.

Tuber Lung Dis, 1996 Dec, 77(6), 516 - 23
Immune activation, allergic drug toxicity and mortality in HIV-positive tuberculosis; Wallis RS et al.; SETTING: Tuberculosis Treatment Center, Kampala, Uganda . OBJECTIVE: HIV-1 affects outcome in pulmonary tuberculosis (TB) . Immune mechanisms triggered by Mycobacterium tuberculosis may lead to increased HIV expression and accelerated disease progression . This study was conducted to correlate serum levels of markers of immune activation with mortality and drug toxicity in HIV + TB . DESIGN: Substudy of a randomized clinical trial of streptomycin-thiacetazone-isoniazid (STH) vs . rifampin-isoniazid-pyrazinamide (RHZ) in HIV + TB . RESULTS: Neopterin > or = 14 ng/ml, TNF-alpha receptors > or = 6.5 ng/ml, and negative skin test were independently associated with increased mortality (P < 0.01) . Among STH-treated subjects, dermatologic toxicity and mortality were respectively 13- and 6.3-fold more likely to occur in subjects with elevated neopterin (P < 0.05), although these two adverse events occurred independently . Activation markers increased from baseline after 2 months of therapy with the less rapidly bactericidal STH regimen, whereas they declined in those treated with RHZ, suggesting a relationship with continued mycobacterial replication . CONCLUSIONS: Immune activation in HIV + TB is associated with shortened survival and increased risk of drug toxicity . HIV + TB patients with elevated serum neopterin should be treated with a rapidly-bactericidal drug regimen which does not include thiacetazone.

J Antimicrob Chemother, 1996 Dec, 38(6), 1055 - 60
Bactericidal activity, post antibiotic effect and modified controlled effective regrowth time of meropenem at high concentrations; Bowker KE et al.; The effect of increasing meropenem concentrations up to 250 mg/L, as might occur if 3 g was given as a single daily intravenous dose, was investigated in terms of bactericidal activity, post antibiotic effect (PAE) and modified controlled effective regrowth time (mCERT) . Increasing the meropenem concentration above 50 mg/L did not result in increased bacterial killing, while concentrations over 75 mg/L did not result in longer PAE or mCERT.

J Am Soc Nephrol, 1996 Dec, 7(12), 2664 - 9
Vascular reactivity during combined ultrafiltration-hemodialysis: influence of dialysate-derived contaminants; van Kuijk WH et al.; It has been suggested that hemodynamic instability and impaired vascular reactivity during combined ultrafiltration-hemodialysis are related to bioincompatibility factors such as dialysate-derived contaminants or the dialyzer . The study presented here investigated whether vascular reactivity could be improved by the use of sterile dialysate . Forearm vascular resistance and venous tone (measured by strain-gauge plethysmography) as well as arterial blood pressure (by Dinamap) and heart rate (by electrocardiogram) were measured in ten stable dialysis patients (age range, 28 to 71 yr) during 2 h of combined ultrafiltration-hemodialysis (bicarbonate; ultrafiltration rate 1.0 L/h) . In addition, a dialysate sample was obtained for culture and limulus amebocyte lysate testing while blood was withdrawn for the estimation of plasma bactericidal/permeability increasing factor (measured by ELISA) and the soluble tumor necrosis factor receptor p75 (measured by ELISA) . Patients served as their own control, comparing dialysis with nonsterile and sterile dialysate . No bacterial growth was observed in sterile dialysate, whereas all samples were positive for Pseudomonas in culture in nonsterile dialysis . All limulus amebocyte lysate tests were negative . Bactericidal/permeability increasing factor tended to increase during nonsterile dialysis (P = 0.063) and remained unchanged during sterile dialysis . In both treatments, tumor necrosis factor receptor p75 increased significantly (P < 0.01) . There were no significant differences in hemodynamic parameters between the treatment modalities . Despite use of sterile dialysate, forearm vascular resistance remained unchanged whereas venous tone decreased significantly . These results indicate that vascular reactivity during combined ultrafiltration-hemodialysis is not improved by the use of sterile dialysate.

Graefes Arch Clin Exp Ophthalmol, 1996 Dec, 234(12), 765 - 9
Efficacy of iontophoresis in the rat cornea; Frucht-Pery J et al.; BACKGROUND: Iontophoresis can enhance penetration of drugs into tissues . We examined the extent of penetration of gentamicin into the cornea of rats during iontophoresis and the effect of varying the concentrations of gentamicin, the duration of iontophoresis and the current densities during iontophoresis . METHODS: Eight groups of rats underwent corneal iontophoresis using gentamicin dissolved in agar . Low and high concentrations of gentamicin were used, as well as low and high current densities and long and short durations of iontophoresis . Control groups received topical or subconjunctival gentamicin, topical saline solution and mock iontophoresis with the agar-gentamicin mixture . The Mann-Whitney test was used for statistical evaluation . RESULTS: Highly bactericidal concentrations of gentamicin were obtained in all the iontophoresis-treated corneas . The high concentration compared to the low concentration of gentamicin in agar significantly increased the concentration of gentamicin in the corneas, as did the longer duration of iontophoresis . However, higher current intensity did not significantly enhance the drug concentration in the cornea . CONCLUSION: Iontophoresis with a concentrated gentamicin-agar mixture may provide a rapid increase of gentamicin levels in the cornea.

Microb Pathog, 1996 Dec, 21(6), 421 - 34
Characterization of HEp-2 cell projection formation induced by diffusely adherent Escherichia coli; Cookson ST et al.; Diffusely adherent Escherichia coli (DAEC) are diarrheagenic E . coli whose pathogenetic mechanisms are largely unknown . DAEC have been shown to induce an unusual phenotype upon adherence to HEp-2 cells in culture characterized by the induction of long thin membrane processes extending from the cell surface . In addition, DAEC have been shown to be protected from the bactericidal effects of gentamicin when incubated with HEp-2 cells . In our studies, we found that three DAEC strains induced formation of eukaryotic cell processes and were protected from gentamicin killing after a 3 h incubation . Preincubation of HEp-2 cells with colchicine or cytochalasin D prior to infection with DAEC strain C1845 resulted in decreased projection formation, suggesting that the effect was dependent upon microfilament and microtubule rearrangement . When the standard gentamicin protection assay was extended for an additional 3 h incubation in the presence of gentamicin, a greater number of DAEC survived gentamicin treatment, more eukaryotic projections were seen in association with the bacteria and the bacteria were actually observed to be "embedded' within these projections . Projection formation was not observed when the bacteria were separated from the cells by a permeable membrane or when the inoculum was inactivated by ultraviolet irradiation . Transposon TnphoA mutants of C1845 were screened for decreased gentamicin protection . All three mutants which were deficient in gentamicin protection demonstrated less projection formation . Insertion mutations affecting gentamicin protection were localized to both the chromosome (two) and a plasmid (one) . Eukaryotic projections are a novel interaction of DAEC with epithelial cells, may play a role of the survival of the bacteria against host defenses and may contribute to DAEC pathogenesis . The effect is dependent upon epithelial cell contact and requires multiple bacterial genes.

J Trauma, 1996 Dec, 41(6), 1013 - 7
Granulocyte colony-stimulating factor enhances killing of translocated bacteria but does not affect barrier function in a burn mouse model; Eaves-Pyles T et al.; BACKGROUND: Granulocyte colony-stimulating factor drives the proliferation and differentiation of granulocytes and also enhances their bactericidal and phagocytic activity . The present study was undertaken to investigate the effects of murine granulocyte colony-stimulating factor (mG-CSF) on bacterial translocation and gut-derived sepsis after burn injury . METHODS: In experiment I, BALB/c mice were randomized into two treatment groups, which received 1 microgram/mouse of mG-CSF subcutaneously for either 1 (n = 16) or 2 days (n = 15) . Controls received saline (n = 16) . After treatment, all animals were gavaged with 10(10) 111In Escherichia coli and then given a 20% burn . All groups were observed 10 days for survival . In experiment II, three additional groups (n = 6/group) received the same treatment as above but were killed 4 hours after burn injury . Mesenteric lymph nodes, liver, and spleen were harvested to measure radionuclide counts (disintegrations per minute per gram of tissue) and colony-forming units (CFU/g of tissue) and to calculate the percentage of viable bacteria (% alive) . RESULTS: Experiment I: 10-day survival was significantly higher in groups treated with mG-CSF for 1 or 2 days (75% and 73%, respectively), compared with controls (43.7%), p = 0.001 . Experiment II: no differences in translocation to the tissues were observed among any of the groups, according to radionuclide counts . However, quantitative colony counts and calculated percentage of viable bacteria showed that killing was enhanced in the mesenteric lymph nodes and liver of animals that received mG-CSF, but this was significant only in the liver for both treatment times (1 day, p = 0.021 and 2 day, p = 0.009) . CONCLUSION: These data suggest that treatment with mG-CSF does not improve gut barrier function, but does enhance the host's ability to kill translocated organisms and improve survival in a gut-derived sepsis model.

Biopharm Drug Dispos, 1996 Dec, 17(9), 761 - 74
Alteration of the pharmacokinetics of small proteins by iodination; Bauer RJ et al.; The pharmacokinetics of several proteins were investigated using two different assays . A 23 kDa recombinant protein fragment of bactericidal/permeability-increasing protein (rBPI23) was radiolabeled with 125I using Iodo-beads and administered rats . Plasma samples were collected and assayed for 125I-rBPI23 by radioactivity . In a separate experiment, rBPI23 was administered to rats and plasma samples were assayed for rBPI23 by ELISA . The clearance determined from plasma concentrations of 125I-rBPI23 measured by radioactivity was about 2.5-fold lower than that of rBPI23 determined by ELISA . In addition, the steady state volumes of distribution and mean residence times of 125I-rBPI23 measured by radioactivity were four-fold and 10-fold greater, respectively, compared to those measured by the ELISA method . By studying several proteins with a range of molecular weights, we found that the pharmacokinetics of proteins below about 60 kDa were different when assayed by radioactivity or ELISA, but those of proteins with molecular weights of at least 80 kDA revealed only minor differences . To determine which assay method yielded the correct plasma pharmacokinetic profile, rBPI23 was metabolically labeled with 35S-methionine and administered to rats, and plasma samples were assayed by radioactivity . The concentration-time profile assessed by this method was very close to that determined by ELISA . Exposing rBPI23 to chloramine-T (the oxidant used in the iodination process) and measuring its plasma concentration by ELISA revealed pharmacokinetics similar to those of the iodinated protein measured by radioactivity . In contrast, radiolabeling rBPI23 using iodinated Bolton-Hunter reagent (which avoids exposing the protein to oxidant), and measuring 125I-rBPI23 by radioactivity, yielded pharmacokinetics that were similar, although not identical, to the pharmacokinetics of rBPI23 measured by ELISA . Thus, our data suggest that directly iodinating low-molecular-weight proteins by oxidation procedures alters their clearance from the blood, preventing reliable determination of pharmacokinetic parameters.

Ann Thorac Surg, 1996 Dec, 62(6), 1783 - 9
Toward further reducing wound infections in cardiac operations; Brown IW Jr et al.; BACKGROUND: Serious wound infections such as mediastinitis still occur at a rate of 0.8% to 2.0%, according to the most recently published cardiac operative series . METHODS: Data from careful surveillance for infection have been collected prospectively during a 4.5-year period on 1,717 patients who underwent cardiac operations performed under direct ultraviolet C radiation . RESULTS: The rate for mediastinitis was 0.23%, and for deep incisional infection without mediastinitis, 0.12%; these rates are significantly lower than those for eight of nine of the most recently published cardiac series . When our infection rates were stratified using the National Nosocomial Infection Surveillance risk index, they were also significantly lower in the most important risk categories than the corresponding stratified rates collected from the participating hospitals of the Centers for Disease Control and Prevention National Nosocomial Infection Surveillance system . CONCLUSIONS: Though we lack the proof that only a large, randomized study might provide, certainly, one possible explanation for our lower wound infection rate was the use of bactericidal ultraviolet C radiation during operation . This is a simple and effective means of minimizing operating room airborne bacteria as one possible source of these infections.

Curr Microbiol, 1996 Dec, 33(6), 383 - 9
Role of colanic acid polysaccharide in serum resistance in vivo and in adherence; Lopez-Torres AJ et al.; The production of an extracellular layer of polysaccharide, termed the capsule, is a common feature of many bacteria . Capsules play a vital role in permitting evasion of the host immune specific and nonspecific defenses as well as helping in adhesion for colonization of host tissue . Colanic acid capsule is usually produced in low quantities and is a common feature of several enteric bacteria . The role of the colanic acid capsule in aiding adhesion and virulence was investigated . Encapsulated and unencapsulated cells were injected into granuloma pouches that were formed on the backs of rats . During the first 50 h, the viability of the matched encapsulated and unencapsulated cells decreased . These studies showed that colanic acid capsule does not confer resistance to the bactericidal activity of serum or to phagocytosis in vivo, since there was no significant difference in the survival rates of both strains over time . Adherence studies were conducted in monolayers of human carcinoma intestinal cells (T84) with the same matched strains . After incubating radioactively labeled bacteria with the colon cells, the level of adherence was determined by measuring the radioactivity remaining in the tissue culture wells . The results of these experiments indicated that the unencapsulated cells adhered more readily to the intestinal cells, suggesting that colanic acid capsule interferes with adherence in this model system.

Med Clin (Barc), 1996 Nov 30, 107(19), 726 - 9
{In vitro effect of heroin on neutrophil polymorphonuclear leukocytes from peripheral blood}; Sala R et al.; BACKGROUND: Infections are the most common medical complications in drug addicts . Some studies suggest that heroin itself could facilitate them by altering the polymorphonuclear leukocyte (PMNL) function of these patients . The aim of this study was to analyze the heroin effect on the chemotaxis, the phagocytosis and the bactericidal oxidative metabolic activity on PMNL from 10 healthy adults . MATERIAL AND METHODS: Three samples of 20 ml of blood were obtained from each donor, separating the leukocytes later . The first sample was used as control (A group); heroin was added to the blood of the second sample before PMNL separation (1 mg of heroin into 20 ml of blood)(B group) and to the third sample after PMNL separation (0.05 mg of heroin in 1 ml of PMNL suspension)(C group) . The concentration of heroin used was 50 microliters/ml of blood (this concentration was higher than the lethal concentration found in the blood of drug addicts who die from heroin overdose) . The PMNL functions studied in vitro were the chemotaxis of PMNL applying the under agarosa gel method, and for the phagocytosis and the intracellular oxidative metabolic activity the following two tests were used: the ingestion of bacto-latex particles combined with nitroblue tetrazolium (NBT) reduction test and the chemoluminiscence method . The statistical analysis was done using parametric and non-parametric tests . RESULTS: There were no differences between the three groups studied (A, B or C) regarding chemotaxis, the ingestion of bacto-latex particles and the NBT reduction test . Concerning chemoluminiscence, it was inferior in the C group (with PMNL directly incubated with heroin) compared with A group (control) and B group (with PMNL from blood with heroin)(p < 0.05) . However, there were no statistically significant differences between A and B groups . CONCLUSIONS: In this study, heroin did not have any in vitro significative effect of chemotaxis, phagocytosis and oxidative metabolic activity on the human PMNL.

Kekkaku, 1996 Nov, 71(11), 607 - 14
{Differential growth inhibition of mycobacteria by interferon-gamma-or tumor necrosis factor-alpha-treated murine peritoneal macrophages}; Sato K et al.; Growth inhibition of the intracellular mycobacteria such as Mycobacterium tuberculosis, M . bovis, M . kansasii, M . avium, M . intracellulare, M . fortuitum, and M . chelonae subsp . abscessus by interferon-gamma (IFN-gamma)- or tumor necrosis factor-alpha (TNF-alpha)-treated murine peritoneal macrophages elicited by proteose peptone was studied in vitro . Macrophages were infected with slowly growing mycobacteria and the extracellular mycobacteria were washed out . Then, macrophages were treated with IFN-gamma or TNF-alpha at a concentration of 10 to 1000 U/ml for 2 days . In another experiment, macrophages were pretreated with these cytokines for 1 day then infected with rapidly growing mycobacteria as before . Macrophages were cultured with or without IFN-gamma or TNF-alpha for additional day . Mycobacterial growth was assessed by determination of colony-forming units on 7H11 agar plates after destruction of the macrophages . Stimulation of macrophages with IFN-gamma reduced the growth of mycobacteria . However, except for M . tuberculosis and M . bovis, growth was not inhibited by macrophages treated with TNF-alpha . IFN-gamma seems to be an important cytokine for the activation of mycobactericidal mechanisms in murine macrophages . Stimulation with IFN-gamma or TNF-alpha and subsequent phagocytosis of M . tuberculosis or M . intracellulare increased O2- production, which was assayed by the method of cytochrome C reduction by murine peritoneal macrophages . Phorbol myristate acetate-triggered-O2- production was also elevated by the cytokine pretreatment of the macrophages, suggesting that mycobacterial growth inhibition did not parallel the production of reactive oxygen intermediates in TNF alpha-activated murine peritoneal macrophages . These data suggest that bactericidal mechanisms of murine macrophages against nontuberculous mycobacteria may not depend on reactive oxygen intermediates.

Photochem Photobiol, 1996 Nov, 64(5), 814 - 6
Significant reduction of UVB caused by smoke from biomass burning in Brazil; Mims FM 3rd; Solar UVB radiation, total ozone and the aerosol optical thickness (AOT) of the atmosphere were measured at various sites in Brazil during season of 1995 . Smoke from biomass burning caused very significant AOT and up to an 81% reduction in UVB at Cuiaba, hundreds of kilometers from the most widespread burning . Little or no smoke was apparent upwind of the major burning regions at and near Manaus, and UVB was close to the expected values . There is an increased incidence of respiratory, cardiopulmonary and other diseases associated with severe air pollution, but the responsible biological mechanisms are unknown . The bactericidal effects of solar UVB are well known, and significantly reduced UVB resulting from severe air pollution in regions where UVB levels are ordinarily high might enhance the survivability of pathogenic organisms in air and water and on surfaces exposed to sunlight.

J Hosp Infect, 1996 Nov, 34(3), 217 - 22
An in-vitro evaluation of the activity of povidone-iodine against nosocomial bacterial strains; Traore O et al.; Two povidone-iodine (PVP-I) preparations, one, an antiseptic handwash and one, a skin disinfectant, were tested against 504 bacterial strains isolated from nosocomial infections in 12 French hospitals . In vitro bactericidal activity was determined by a micromethod, using specific interfering substances over a range of dilutions, after 1, 3 and 5 min exposure times . A 5 log10 reduction of the challenge inoculum was considered as the criterion of efficacy . Any resistant strains were tested with the French Standard (T72300) . When the micromethod was carried out at 20 degrees C, 10.7% (54/504) of the strains were resistant to the PVP-I skin disinfectant (dilution 1:10) and 1.6% (8/504) were resistant to the handwashing formulations (dilution 1:3) after 1 min exposure . By increasing the temperature to 32 degrees C, the resistance rate to the skin disinfectant fell to 1.9% (10/504) . All of the 18 strains found resistant with the micromethod were sensitive using the French standard.

J Med Microbiol, 1996 Nov, 45(5), 383 - 7
Identification and characterisation of a superoxide dismutase and catalase from Mycobacterium ulcerans; Roberts B et al.; Previous investigations have demonstrated the presence of both superoxide dismutase and catalase enzymes in several intracellular pathogens, including a number of mycobacterial species . These enzymes are believed to be involved in the protection of the pathogen from the bactericidal products of oxidative metabolism . Superoxide dismutase and catalase were identified in crude extracts of Mycobacterium ulcerans by polyacrylamide gel electrophoresis . Inhibition experiments showed that the superoxide dismutase probably contained manganese as the metal cofactor . Other mycobacterial species examined for comparison produced bands of superoxide dismutase activity with a different mobility to that of M . ulcerans, suggesting possible structural differences between the enzymes.

Clin Exp Immunol, 1996 Nov, 106(2), 273 - 9
IL-1 beta production by human polymorphonuclear leucocytes stimulated by anti-neutrophil cytoplasmic autoantibodies: relevance to systemic vasculitis; Brooks CJ et al.; Neutrophils accumulate in the acute blood vessel lesions of patients with autoimmune systemic vasculitis . They have been shown previously to produce the cytokine IL-1 beta in response to stimulation with TNF . This study demonstrates that neutrophils can be stimulated by anti-neutrophil cytoplasmic antibodies (ANCA), which are present in patients with systemic vasculitis, to express mRNA and protein for IL-1 beta . Both human ANCA and MoAbs to a variety of autoantigens recognized by ANCA, including proteinase 3, myeloperoxidase, bactericidal/permeability increasing protein and elastase, are effective . This response can be inhibited by actinomycin and cycloheximide, suggesting a requirement for de novo protein synthesis . IL-1 beta production can be inhibited by pooled human intravenous immunoglobulins but not by FK506 or cyclosporin A . These data suggest that ANCA in patients with active vasculitis may stimulate neutrophils to produce cytokines . It is hypothesized that cytokine production from neutrophils that accumulate in significant numbers in vasculitic lesions contribute to and augment the local inflammatory response by the activation of vascular endothelial cells and infiltrating leucocytes.

J Immunol Methods, 1996 Oct 16, 197(1-2), 121 - 30
A comprehensive method to purify three major ANCA antigens: proteinase 3, myeloperoxidase and bactericidal/permeability-increasing protein from human neutrophil granule acid extract; Zhao MH et al.; Indirect immunofluorescence (IIF) techniques have shown that anti-neutrophil cytoplasm autoantibodies (ANCA) are useful serological markers for certain small vessel vasculitides and the non-vasculitic inflammatory disorders . ELISA procedures, using purified molecules as solid phase ligands, helped to identify proteinase 3 (PR3) and myeloperoxidase (MPO) as two major ANCA antigens; and recently we characterised bactericidal/permeability-increasing protein (BPI) as another important ANCA antigen . ANCA against these three antigens are associated with different clinical disorders . Therefore purified antigens are needed to determine these different autoantibody specificities in order to help diagnosis and guide treatment . Here we describe a method using Orange-A dye ligand chromatography and cation exchange chromatography for the sequential purification of PR3, MPO and BPI, from the same starting material, an acid extract of normal human neutrophil granules . After separation the three antigens were free of contamination by each other and no traces were found of other known minor ANCA antigens.

Eur J Ophthalmol, 1996 Oct-Dec, 6(4), 361 - 7
Comparative evaluation of the short-term bactericidal potential of a steroid-antibiotic combination versus steroid in the treatment of chronic bacterial blepharitis and conjunctivitis; Shulman DG et al.; The effects of four days' treatment with topical Maxitrol (neomycin sulphate 3500 IU/mL, polymyxin-B sulphate 6000 IU/mL with dexamethasone 0.1%) were compared with those of Maxidex (dexamethasone 0.1% alone) in a double-masked study in 111 patients with bacterial blepharitis or conjunctivitis, 95 of whom were evaluable for efficacy . The majority of patients (N = 80) had chronic blepharitis . Maxitrol treatment resulted in a significantly greater reduction (90%) in bacterial counts and bacterial eradication (50%) compared with Maxidex (34% and 17% respectively) . Maxitrol treatment also produced a significantly greater reduction in conjunctival discharge than did Maxidex, while the treatments were equally effective in alleviating other ocular signs and symptoms . It was concluded that use of a fixed dose combination steroid-antibiotic product was more effective for bacterial control and therapeutic efficacy in the treatment of chronic blepharitis and conjunctivitis patients than treatment with steroid alone . However, in the long-term treatment of chronic blepharitis the well-known toxic problems of neomycin sulphate have to be taken into account.

Am J Infect Control, 1996 Oct, 24(5), 396 - 401
Transmission of Helicobacter pylori infection via flexible fiberoptic endoscopy; Akamatsu T et al.; BACKGROUND: Public concern has been raised with regard to the possibility of transmission of instrument mediated Helicobacter pylori infection after upper gastrointestinal endoscopy . METHODS: Disinfection procedures for gastrointestinal endoscopes were surveyed in 20 Japanese institutions, and in vitro bactericidal activities of seven disinfectants against H . pylori were determined . RESULTS: Screening tests for infection before endoscopy were not consistently performed; only 11 institutions always screened for hepatitis B virus, nine for hepatitis C virus, and two for tuberculosis . All 20 institutions used the same flexible fiberoptic endoscope on more than one patient in succession, with minimal cleanings only . Only two used glutaraldehyde for disinfection . Most used ethyl alcohol, benzalkonium chloride, or alkyldiaminoethylglycine hydrochloride as an external wipe . Bactericidal testing of nine strains of H . pylori against disinfectants revealed that ethyl alcohol (80%) and glutaraldehyde (0.5%) killed all nine strains within 15 seconds, whereas chlorhexidine gluconate (0.05%, 0.1%), benzalkonium chloride (0.025%, 0.1%), alkyldiaminoethylglycine hydrochloride (0.1%), povidone-iodine (0.1%), and sodium hypochlorite (150 ppm) killed all nine strains within 30 seconds . CONCLUSION: H . pylori is readily killed by many common disinfectants and antiseptics . However, practices for disinfection of flexible fiberoptic endoscope were not appropriate.

Antimicrob Agents Chemother, 1996 Oct, 40(10), 2306 - 10
Simultaneous pharmacodynamic analysis of the lag and bactericidal phases exhibited by beta-lactams against Escherichia coli; Li RC; Antibiotic-bacterium interactions are complex in nature . In many cases, bacterial killing does not commence immediately after the addition of an antibiotic, and a lag period is observed . Antibiotic permeation and/or the intermediate steps that exist between antibiotic-receptor binding and expression of cell death are two major possible causes for such lag period . This study was primarily designed to determine the relationship, if any, between antibiotic concentrations and the lag periods by a modeling approach . Short-term time-kill studies were conducted for amoxicillin, ampicillin, penicillin-G, oxacillin, and dicloxacillin against Escherichia coli . In conjunction with the use of a saturable rate model to describe the concentration-dependent killing process, a first-order induction (initiation) rate constant was used to characterize the delay in bacterial killing during the lag period . For all of the beta-lactams tested, parameters describing the bactericidal effect suggest that amoxicillin and ampicillin were much more potent than oxacillin and dicloxacillin . The induction rate constant estimates for both ampicillin and amoxicillin were found to relate linearly to concentrations . Nevertheless, these induction rate constant estimates were lower for penicillin-G, oxacillin, and dicloxacillin and increased nonlinearly with concentrations until an apparent plateau was observed . These findings support the hypothesis that the permeation process is potentially a rate-limiting step for the rapid bactericidal beta-lactams such as ampicillin and amoxicillin . However, as suggested by previous observations of the various morphological changes induced by beta-lactams, the contribution of the steps following antibiotic-receptor complex formation to the lag period might be significant for the less bactericidal antibiotics such as oxacillin and dicloxacillin . Findings from the present modeling approach can potentially be used to guide future bench experimentation.

J Trauma, 1996 Oct, 41(4), 663 - 6
Granulocyte-macrophage colony-stimulating factor (GM-CSF) enhances pulmonary defenses against pneumococcal infections after splenectomy; Hebert JC et al.; BACKGROUND: Splenectomized individuals are at risk for pneumococcal sepsis . Alveolar macrophage bactericidal function is depressed after splenectomy . Granulocyte-macrophage colony-stimulating factor (GM-CSF) has pronounced effects on the number and function of macrophages . We hypothesized that GM-CSF treatment could improve alveolar macrophage bactericidal activity against pneumococci, and improve survival with pneumococcal infection . METHODS: Two weeks after splenectomy or sham operation, mice were treated with GM-CSF or saline twice daily for varying times . Alveolar macrophages were obtained by bronchopulmonary lavage, and bactericidal activity was measured . Survival was assessed after pneumococcal aerosol challenge . RESULTS: Alveolar macrophage bactericidal activity was improved with GM-CSF treatment in both eusplenic and asplenic mice (p < 0.001) . GM-CSF treatment improved survival in both groups (p < 0.001) . CONCLUSIONS: GM-CSF can augment alveolar macrophage function and provide protection against pneumococcal infections . It may be a useful adjuvant therapy for normal and splenectomized individuals.

Pathol Biol (Paris), 1996 Sep, 44(7), 675 - 80
{Kinetics of in vitro bactericidal activity of the antiseptic biseptine combining 3 active principles}; Reverdy ME et al.; Biseptine, is an association of chlorhexidine digluconate, benzalkonium chloride and benzylic alcohol . Little is known, in literature, on this antiseptic, used for cutaneous antisepsis . We studied killing kinetic of Biseptine, at different concentrations, on 4 AFNOR bacterial strains . Killing curves were studied at antiseptic concentration of 90 to 0.1% in 10 ml of distilled water . Bacterial counts were determined after neutralization in liquid medium . Synergy of chlorhexidine and benzalkonium chloride in Biseptine, allowed to obtain similar bactericidal activity than Hibitane champ with chlorhexidine concentrations 2 fold less . At 90, 50, 25, 10 and 5% concentrations, bactericidal activity (5 log10 reduction of the initial bacterial count) was effective in one minute . After 5 to 15 minutes, activity persisted at 1 and 0.5% concentrations . The 0.1% solution was inefficacious . This report disclosed an important security margin in antiseptic activity.

Kekkaku, 1996 Sep, 71(9), 527 - 31
{Chemotherapy of pulmonary Mycobacterium kansasii infection}; Mizutani S; A very favorable outcome after chemotherapy of 122 cases of M . kansasii lung disease was reported by Dr . Mizutani, who emphasized RFP as the "Key drug", and concluded that three-drug combination (not two-drug), including RFP (RFP.INH.EB or SM) for 1 year, could be a standard regimen for M . kansasii lung disease at the time of the moment . In addition, the following itemes were discussed . (1) In cases resistant to RFP, one could possibly replace RFP by TH, one of new quinolones (NQ), or the new macrolide (NM) (clarithromycin, CAM) . (2) In low grade resistant cases to INH (0.1 microgram /ml) or EB (2.5 micrograms/ml), the replacement of the drugs may not be necessary, however, in higher-grade resistance to INH or EB, many cases were looked for the change of drugs according the results of the questionnaire done by the author . The present status of basic preclinical evaluations of new drugs were presented by Dr . Tomioka, who summarized in vitro and in vivo antimycobacterial activities of NMs and NQs . The most potent activity among NMs was demonstrated in CAM, which is probably the candidate for M . kansasii and possibly for M . avium complex (MAC) disease, followed by roxithromycin (RXM) and azithromycin (AZM) in sequence . NQs including the ones under development were generally potent against Mycobacterium tuberculosis, M . kansasii and M . fortuitum . NQs were not potent enough for MAC . In addition, the author discussed more suitable in vitro techniques which should reflect in vivo evaluations, and proposed the observation of in vitro bactericidal activity using both Cmax (maximal in vivo concentration) and C (0-8h) (the average concentration during 8 hours after administration) of drugs, and also the assessment of bactericidal activities of drugs in macrophages as better choices . As additional comments, the results of in vitro activities of NQs and NMs against MAC were supplemented by two authors, Dr . Tsuyuguchi and Dr . Kawahara . The assessment using 7 H 9 liquid medium by the former author demonstrated the potent activities of both CS-940* and sparfloxacin (SPFX), followed by AM-1155*, ciprofloxacin (CPFX), levofloxacin (LVFX), OPC-17116*, NM-394* in sequence . The author gave attention also to a high Cmax in CS-940* . In vitro activities with 7 H 11 agar medium reported by Dr . Kawahara demonstrated generally higher activities against M . avium than M . intracellulare, and reported potent activities of CPFX, SPFX, LVFX, grepafloxacin (GPFX), AM-1155*, and DU-6859 a* among 14 NQs tested . The author reported a rather potent activity of CAM against MAC followed by RXM and AZM in sequence . There was an impression that the MICs in both liquid and agar medium were comparable . (* : under development) . The present status in the treatment of MAC lung disease was precisely reported by Dr . Harada, who summarized the results of survey both in 13 National Chest Hospitals (by questionnaire) and in the author's Hospital . The former survey demonstrated that 73% of the cases with the initial chemotherapy became consecutively negative for 6 months in the span of 9 months observation, which clearly showed the early response of MAC disease was rather favorable, in spite of very few cases with 4 drug-or more combinations . However, longer the follow up, the percentage of negative cases went down, which suggested bacteriological relapse occurred in relatively high percentage of the early converted cases . The evaluation of 117 cases of pulmonary MAC disease in the author's Hospital disclosed 2 drug-combination, RFP and INH, was clearly less potent than 3-drug combination, RFP.IHN.SM or EB, and 1 year after the begining of initial chemotherapy, around 60% of cases were negative, while only a little more than 40% of cases were negative in retreatments . The author suggested that around 50% of MAC lung disease may progress with episodes of "relapse" . Death occurred 20% in the cases

J Pharm Pharmacol, 1996 Sep, 48(9), 985 - 7
Transient relaxation of plasmid DNA in Escherichia coli by fluoroquinolones; Matsuo M et al.; We examined the influence of fluoroquinolones (norfloxacin, enoxacin, ofloxacin, levofloxacin, and sparfloxacin) on DNA supercoiling of plasmids in Escherichia coli cells by analysis with agarose gel electrophoresis in the presence of chloroquine . All the fluoroquinolones tested immediately induced DNA relaxation . The relaxed DNA was re-supercoiled, and the process was sensitive to chloramphenicol, suggesting that newly synthesized proteins participate in the reaction . The concentrations of fluoroquinolones required for DNA relaxation were much higher than those required for cell killing . The bactericidal effect of fluoroquinolones is apparently related to mechanisms other than DNA relaxation.

J Antimicrob Chemother, 1996 Sep, 38(3), 387 - 97
Bactericidal effect of plaunotol, a cytoprotective antiulcer agent, against Helicobacter pylori; Koga T et al.; In order to investigate the bactericidal effect of plaunotol, an oily antiulcer agent, against Helicobacter pylori, comparative studies were conducted using its derivatives, M-4, M-5, and M-6, whose hydrophobicity decreased in the order of plaunotol > M-6 > M-5 > M-4 by log P determination . Plaunotol rapidly reduced the viability of H . pylori in vitro, and cell death was associated with cell lysis . In addition, plaunotol showed eightfold stronger bactericidal activity against H . pylori than M-6 and M-5, while the compound with the lowest hydrophobicity, M-4, showed no bactericidal activity . The bactericidal activities of plaunotol and its derivatives were related to the hydrophobicity of these compounds . To investigate a possible interaction between these compounds and the cell membrane of H . pylori, their effects on liposomal membranes prepared from phosphatidylethanolamine and cardiolipin, which are known to be present in the membrane of H . pylori, were determined by detection of glucose release from the liposomes . Plaunotol showed eight-fold higher activity than M-6 and M-5, while M-4 showed no activity . The effects of plaunotol and its derivatives on liposomal membrane were therefore related to their bactericidal activities . In addition, it was confirmed that the bactericidal effect of plaunotol against H . pylori was neutralized by the liposomal membrane, and that plaunotol led to an increase in permeability of the membrane, as evidenced by measurement of the leakage of 260 nm absorbing-material from H . pylori . These results suggest that the bactericidal effect of plaunotol against H . pylori is due to the interaction between this compound and the bacterial cell membrane.

Antimicrob Agents Chemother, 1996 Sep, 40(9), 2137 - 41
Bactericidal activity of single dose of clarithromycin plus minocycline, with or without ofloxacin, against Mycobacterium leprae in patients; Ji B et al.; Fifty patients with newly diagnosed lepromatous leprosy were allocated randomly to one of five groups and treated with either a month-long standard regimen of multidrug therapy (MDT) for multibacillary leprosy, a single dose of 600 mg of rifampin, a month-long regimen with the dapsone (DDS) and clofazimine (CLO) components of the standard MDT, or a single dose of 2,000 mg of clarithromycin (CLARI) plus 200 mg of minocycline (MINO), with or without the addition of 800 mg of ofloxacin (OFLO) . At the end of 1 month, clinical improvement accompanied by significant decreases of morphological indexes in skin smears was observed in about half of the patients of each group . A significant bactericidal effect was demonstrated in the great majority of patients in all five groups by inoculating the footpads of mice with organisms recovered from biopsy samples obtained before and after treatment . Rifampin proved to be a bactericidal drug against Mycobacterium leprae more potent than any combination of the other drugs . A single dose of CLARI-MINO, with or without OFLO, displayed a degree of bactericidal activity similar to that of a regimen daily of doses of DDS-CLO for 1 month, suggesting that it may be possible to replace the DDS and CLO components of the MDT with a monthly dose of CLARI-MINO, with or without OFLO . However, gastrointestinal adverse events were quite frequent among patients treated with CLARI-MINO, with or without OFLO, and may be attributed to the higher dosage of CLARI or MINO or to the combination of CLARI-MINO plus OFLO . In future trials, therefore, we propose to reduce the dosages of the drugs to 1,000 mg of CLARI, 100 mg of MINO, and 400 mg of OFLO.

J Trauma, 1996 Sep, 41(3), 465 - 70
Fluconazole increases bactericidal activity of neutrophils through non--cytokine-mediated pathway; Zervos EE et al.; BACKGROUND: Polymorphonuclear neutrophils (PMNs) preincubated with fluconazole (FCZ) demonstrate enhanced bactericidal activity in vitro . OBJECTIVE: This study was undertaken to determine the role of cytokines in FCZ-induced augmentation of PMN function . METHODS: PMNs were preincubated with PBS or FCZ and exposed to Escherichia coli . Cell culture supernatants and mRNA were isolated after preincubation and again after exposure to E . coli . Tumor necrosis factor-alpha, interleukin (IL)-1 beta, and IL-8 protein and mRNA levels were determined using enzyme-linked immunosorbent assay and polymerase chain reaction, respectively . Results were compared using the Student's t test . RESULTS: Preincubation of PMNs with FCZ resulted in enhanced killing but no difference in cytokine protein or mRNA levels when compared to control . After exposure to E . coli, PMNs significantly up-regulate IL-8 and tumor necrosis factor-alpha independent of the solution with which they were preincubated . CONCLUSIONS: Up-regulation of the cytokine cascade plays a minor role, at most, in the mechanism through which FCZ augments the bactericidal activity of PMNs.

Appl Environ Microbiol, 1996 Sep, 62(9), 3094 - 100
Mechanisms of acid resistance in enterohemorrhagic Escherichia coli; Lin J et al.; Enterohemorrhagic strains of Escherichia coli must pass through the acidic gastric barrier to cause gastrointestinal disease . Taking into account the apparent low infectious dose of enterohemorrhagic E . coli, 11 O157:H7 strains and 4 commensal strains of E . coli were tested for their abilities to survive extreme acid exposures (pH 3) . Three previously characterized acid resistance systems were tested . These included an acid-induced oxidative system, an acid-induced arginine-dependent system, and a glutamate-dependent system . When challenged at pH 2.0, the arginine-dependent system provided more protection in the EHEC strains than in commensal strains . However, the glutamate-dependent system provided better protection than the arginine system and appeared equally effective in all strains . Because E . coli must also endure acid stress imposed by the presence of weak acids in intestinal contents at a pH less acidic than that of the stomach, the ability of specific acid resistance systems to protect against weak acids was examined . The arginine- and glutamate-dependent systems were both effective in protecting E . coli against the bactericidal effects of a variety of weak acids . The acids tested include benzoic acid (20 mM; pH 4.0) and a volatile fatty acid cocktail composed of acetic, propionic, and butyric acids at levels approximating those present in the intestine . The oxidative system was much less effective . Several genetic aspects of E . coli acid resistance were also characterized . The alternate sigma factor RpoS was shown to be required for oxidative acid resistance but was only partially involved with the arginine- and glutamate-dependent acid resistance systems . The arginine decarboxylase system (including adi and its regulators cysB and adiY) was responsible for arginine-dependent acid resistance . The results suggest that several acid resistance systems potentially contribute to the survival of pathogenic E . coli in the different acid stress environments of the stomach (pH 1 to 3) and the intestine (pH 4.5 to 7 with high concentrations of volatile fatty acids) . Of particular importance to the food industry was the finding that once induced, the acid resistance systems will remain active for prolonged periods of cold storage at 4 degrees C.

J Infect Dis, 1996 Sep, 174(3), 636 - 8
Moraxella catarrhalis in acute laryngitis: infection or colonization?
Hol C, Schalen C, Verduin CM, Van Dijke EE, Verhoef J, Fleer A, Van Dijk H.
The complement phenotypes of Moraxella catarrhalis isolates obtained from adult patients with acute laryngitis were investigated using a microliter serum bactericidal assay and compared with those of other donor groups . Laryngitis isolates had a higher proportion (57%) of complement-resistant strains than did carrier strains from healthy 8- to 13-year-old schoolchildren (16%) . The difference between these groups was statistically significant (chi2 {3 x 2 table} = 21.55; P < .001) . The relatively frequent occurrence of the complement-resistant (virulence-associated) phenotype in adults with acute laryngitis supports the theory of an active role of M . catarrhalis in the pathogenesis of acute laryngitis.

Pediatr Res, 1996 Aug, 40(2), 257 - 62
Lactoferrin or a fragment thereof inhibits the endotoxin-induced interleukin-6 response in human monocytic cells; Mattsby-Baltzer I et al.; Human milk is in several ways anti-inflammatory . This study investigates whether or not human milk lactoferrin (LF) in comparison with bovine LF can affect the IL-6 release from human cells . Human, as well as bovine, LF and a bactericidal pepsin-derived fragment of bovine LF (lactoferricin B) were found to suppress the IL-6 response in a monocytic cell line (THP-1) when stimulated by lipopolysaccharide (LPS) . The suppression of bovine LF was similar to or higher than that of human LF . Lactoferricin B was the strongest inhibitor of the LPS-induced IL-6 response . A time-dependence regarding the inhibitory capacity of LF was found . For human LF, the strongest inhibition was observed when added 15-30 min after the addition of LPS . Addition of LF before the LPS induced an approximately 45% reduction of the IL-6 response . The results suggest an anti-inflammatory activity of both human and bovine LF, and of the LF fragment lactoferricin B through their suppressive effects on the cytokine release.

Protein Expr Purif, 1996 Aug, 8(1), 28 - 40
Expression and characterization of cysteine-modified variants of an amino-terminal fragment of bactericidal/permeability-increasing protein; Horwitz AH et al.; rBPI23 is a biologically active, recombinant N-terminal fragment of human bactericidal/permeability-increasing protein (BPI) . While rBPI23 is readily purified from culture supernatants of Chinese hamster ovary (CHO)-K1 transfectants, it is heterogeneous, consisting of monomer and disulfide-linked dimer, characteristics due presumably to the presence of three cysteines within the molecule . We have examined the role of these cysteines in rBPI23 expression, function, and dimer formation by mutating their codons to alanine (C132A), serine (C135S), or alanine (C175A) and expressing analogues of N-terminal fragments ("variants") lacking one, two, or all three cysteines in permanently transfected CHO-K1 cells . We also expressed a variant in which serine 18 was changed to cysteine (S18C), as found in both bovine and rabbit BPI . The C132A variant was readily secreted and purified as a homogeneous, stable monomeric protein species . The C135S and S18C variants were produced as mixtures of monomer and dimer; the C135S variant was poorly secreted, difficult to purify, and unstable on storage . In contrast, the C175A variant and those lacking any two or all three cysteines were expressed but not secreted . Purified rBPI23 and the C132A and S18C variants had comparable bactericidal and lipopolysaccharide (LPS) binding activities and were similarly effective at neutralizing LPS-induced tumor necrosis factor synthesis by THP-1 cells; the purified C135S variant lacked all activities . From these studies with CHO-K1 transfectants, we conclude that (i) cysteines 135 and 175 are both necessary for efficient secretion of a biologically active N-terminal BPI fragment, presumably through the formation of a disulfide bond, (ii) cysteine 132 is responsible for dimer formation, and (iii) only the C132A modification yields a stable, biologically active, N-terminal BPI fragment (designated rBPI21) that is free of dimeric species.

Biochem Mol Med, 1996 Aug, 58(2), 184 - 91
Complement-mediated killing of porphyromonas gingivalis 381 by the immunoglobulin G induced by recombinant 40-kDa outer membrane protein; Saito S et al.; Porphyromonas gingivalis has been implicated as an important pathogen in severe adult periodontitis . We have previously cloned a 40-kDa outer membrane protein from P . gingivalis 381 and succeeded in producing sufficient quantities of the recombinant protein (r40-kDa OMP) . r40-kDa OMP has been the subject of considerable interest to us as a possible vaccine candidate . To understand the role of anti-r40-kDa OMP antibody in the host defense mechanisms against P . gingivalis, we examined the involvement of a rabbit antibody against r40-kDa OMP (r40-kDa OMP Ab) to an in vitro complement-mediated bactericidal assay for P . gingivalis 381 . By measuring the absorbance values in order to assay the surviving bacteria, we found significant anti-P . gingivalis activity of r40-kDa OMP Ab when guinea pig complement was present . Using affinity-purified immunoglobulin G of r40-kDa OMP Ab (IgG-r40-kDa OMP), we demonstrated that the IgG contributed to anti-P . gingivalis activity in the antibody-complement system . This was effected by measuring the incorporation of tritiated thymidine into newly synthesized nucleic acids . Finally, we confirmed the cell lysis of P . gingivalis 381 exposed to IgG-r40-kDa OMP in the presence of complement sources in a radioactive bactericidal assay using bacteria labeled with {14C}sodium acetate . Assembling the data from experiments using component-deficient complements, we concluded that IgG-r40-kDa OMP was related to the killing of P . gingivalis 381 by mediation in the complement activated through both the classical and the alternative pathways.

Nervenarzt, 1996 Aug, 67(8), 630 - 8
{Classical and current aspects of purulent meningitis}; Ring A et al.; Even today acute purulent meningitis is a life-threatening disease requiring immediate diagnosis and therapy . Despite the availability of bactericidal antibiotics, the mortality has not changed within the last 20 years . A therapeutic approach interfering specifically with the pathogenesis of inflammation is necessary in order to prevent long-term neurological sequelae and death associated with bacterial meningitis . In this paper we discuss the present level of understanding of antibiotics and adjunctive therapy, especially with regard to the molecular pathophysiological events occurring during a CNS infection.

J Anat, 1996 Aug, 189 ( Pt 1), 193 - 7
Immunocytochemical demonstration that human duodenal Brunner's glands may participate in intestinal defence; Coutinho HB et al.; The immunocytochemical demonstration of IgA and IgM in some secretory units of human Brunner's glands, associated with the presence of secretory component in all secretory cells, indicates the possibility that these glands assist the function of the intestinal crypts in transporting immunoglobulins into the gut lumen . In addition, the presence of muramidase (lysozyme) in the cells of the secretory units suggests that Brunner's glands continuously secrete bactericidal enzyme, thus reinforcing the function of the Paneth cells as contributors to nonspecific defence (innate immunity) in the intestinal tract.

South Med J, 1996 Aug, 89(8), 775 - 8
Efficacy of ciprofloxacin in the eradication of Helicobacter pylori; Dresner D et al.; In small preliminary trials, ciprofloxacin has failed to eradicate Helicobacter pylori . Since fluoroquinolones have a marked reduction in bactericidal activity at acidic pH, we altered the gastric pH using omeprazole and investigated the efficacy of ciprofloxacin in eradicating H pylori . Forty-four consecutive patients infected with H pylori were prospectively studied in a randomized, double-blind, controlled trial comparing ciprofloxacin with a placebo for 2 weeks . Both treatment groups received bismuth and omeprazole . In 36 patients, follow-up endoscopy was done 4 weeks after the cessation of all study drugs . The H pylori infection cleared in 13 of 17 patients (76%) in the ciprofloxacin group versus 5 of 19 (26%) in the placebo group . Concurrent administration of omeprazole with ciprofloxacin resulted in increased bactericidal activity against H pylori . Ciprofloxacin when combined with omeprazole and bismuth is efficacious for eradication of H pylori.

Cell Immunol, 1996 Jul 10, 171(1), 48 - 54
Increased lung cell cytotoxic but not bactericidal or phagocytic activity in Mycobacterium avium complex-infected mice; Saunders BM et al.; Following intranasal infection of mice with Mycobacterium avium complex (MAC) organisms, bacterial growth plateaued at the fourth week postinfection and then remained relatively constant thereafter . Inflammatory cell numbers in the lungs increased 10-fold by 4 weeks postinfection, and lung cell cytotoxicity and the production of NO, H202, and 02- by lung cell cultures had all increased significantly by this time and remained elevated throughout the 15-week experimental study . Although these parameters are generally associated with increased bactericidal activity, there appeared to be a defect in phagocytosis by lung cells, so that bactericidal activity could not be demonstrated in either in vitro or in vivo experiments . This study suggests that following intranasal infection with MAC, inflammatory cells are activated, sufficient to prevent further bacterial growth in vivo but not sufficient to clear the infection . We suggest that the deficiency may lie in the phagocytic activity of the cells.

J Endod, 1996 Jul, 22(7), 362 - 4
Intracanal pH changes of calcium hydroxide pastes exposed to carbon dioxide in vitro; Fuss Z et al.; It has been suggested that the main benefit of using calcium hydroxide as an intracanal medicament lies in its bactericidal effect, provided that the pH of the paste is above 12.5 . The purpose of this study was to measure changes in the pH of several calcium hydroxide pastes sealed in root canals for 30 days . Sixty-two extracted, single-rooted human teeth were endodontically prepared using K-files up to size 60 . The teeth were separated at random into six equal groups to be filled with either Calxyl, Hydrocalcium, or a paste made by mixing calcium hydroxide powder with either distilled water, camphorated p-monochlorophenol, local anesthetic solution, or Solvidont . Cavidentin was used to seal the coronal orifice of the teeth that were placed individually in vials containing 10 mL distilled water . Five vials of each group were exposed to air at room temperature, whereas the other five vials were exposed to carbon dioxide in a closed container . The pH of the paste in the root canal was measured after 30 days . There was no significant (p > 0.01) change in the pH (mean 13.11) of the pastes placed in teeth before and after exposure to air, whereas the pH of the pastes in teeth exposed to carbon dioxide was significantly (p < 0.01) reduced (mean 12.54) . There was no significant difference in pH between the six preparations . After 30 days of exposure to carbon dioxide, they still maintained a purportedly bactericidal pH within the root canal.

Clin Microbiol Rev, 1996 Jul, 9(3), 273 - 92
Bartonella (Rochalimaea) quintana infections; Maurin M et al.; Bartonella (formerly Rochalimaea) quintana is the etiological agent of trench fever, a disease extensively reported during the World Wars . Recent molecular biology approaches have allowed dramatic extension of the spectrum of Bartonella infections . B . quintana is now also recognized as an etiological agent of fever and bacteremia, endocarditis, bacillary angiomatosis, and chronic lymphadenopathy . Human immunodeficiency virus-infected patients and/or homeless people are the most vulnerable to infection . Poverty and louse infestation were the main epidemiological factors associated with B . quintana infections during wartime . Although poverty and chronic alcoholism have been associated with modern cases of trench fever and bacteremia due to B . quintana in Europe and the United States, vectors for B . quintana have not been clearly identified and B . quintana has not been isolated from modern-day lice . Microscopic bacillary angiomatosis lesions are characterized by tumor-like capillary lobules, with proliferating endothelial cells . In vitro experiments have shown that B . quintana survives within endothelial cells and stimulates cell proliferation . These observations, together with the finding that lesions may regress when antibiotic therapy is administered, strongly suggest that B . quintana itself stimulates angiogenesis . Bartonella infections are characterized by a high frequency of relapses after brief courses of antibiotic therapy . It is to be noted that in vitro, although Bartonella species are highly susceptible to antibiotics, only the aminoglycosides have proved to be bactericidal . However, the most effective antibiotic regimen for Bartonella infections remains to be established.

Antimicrob Agents Chemother, 1996 Jul, 40(7), 1722 - 5
Early bactericidal activity of rifabutin versus that of placebo in treatment of disseminated Mycobacterium avium complex bacteremia in AIDS patients; Dautzenberg B et al.; Rifabutin, 600 mg/day, was compared with a placebo in the early treatment of culture-proven Mycobacterium avium bacteremia in patients with AIDS . Following 14 days' treatment, bacteriological success, defined as a negative culture or a reduction in the number of CFU of M . avium organisms per milliliter of blood by a factor of > or = 0.5 log from the baseline, was observed in 7 of 10 (70%) evaluable rifabutin patients and in 1 of 13 (8%) evaluable placebo patients (P = 0.002) . Rifabutin is active against M . avium as a single agent and can make a significant contribution to combination regimens for the treatment of disseminated M . avium infection in AIDS patients.

Drugs, 1996 Jul, 52(1), 45 - 59
Optimum treatment of intracellular infection; Maurin M et al.; The intracellular location of some micro-organisms has been early recognised as a critical point to explain failure of antibiotic therapy to eradicate such pathogens from infected hosts . Most often parasites invade 'professional' phagocytic cells, including neutrophils, monocytes and macrophages, by resisting the intracellular bactericidal phagolysosomal pathway . Alternatively, they may invade 'non-professional' phagocytic cells (cells with fewer phagocytic and bactericidal abilities) such as endothelial cells, or even cells without lysosomes such as erythrocytes . The intracellular activity of an antibiotic depends on several factors including its ability to reach the eukaryotic cell membrane, its subcellular localisation as compared to that of the parasite, the possibility that the intracellular milieu may partially inactivate its activity, and the susceptibility of the intracellular form of the parasite . In vitro and animal models have been developed to investigate antibiotic activity against intracellular pathogens . However, it should be emphasised that only data obtained from patients give reliable information to define the optimum antibiotic regimen.

Infect Immun, 1996 Jul, 64(7), 2425 - 30
Determinants of activation by complement of group II phospholipase A2 acting against Escherichia coli; Madsen LM et al.; Prompt killing of many strains of Escherichia coli during phagocytosis in vitro by isolated polymorphonuclear leukocytes (PMN) requires the presence of nonlethal doses of nonimmune serum (B . A . Mannion, J . Weiss, and P . Elsbach, J . Clin . Invest . 86:631-641, 1990) . Because this requirement is bypassed in a phospholipase A (PLA)-rich mutant (pldA ) of E . coli, we have examined the effect of serum on bacteria} phospholipid (PL) degradation during phagocytosis of wild-type (pldA+) and PLA-deficient (pldA) E . coli . In parallel with increased killing, nonlethal doses of serum increased the degradation of prelabeled bacterial PL during phagocytosis by two- to fivefold, to nearly the same levels (ca . 50 to 60%) as those produced during phagocytosis of E . coli pldA in the absence of serum . The effects on the E . coli pldA mutant imply that there is a serum-mediated enhancement of granule-associated group II PMN PLA2 activity . At the same doses, serum promoted action against E . coli in the presence of purified rabbit and human group II PLA2 but did not activate bacterial PLA . Related PLA2s that lack specific structural determinants needed for optimal activity against E . coli treated with the bactericidal/permeability-increasing protein (BPI) of PMN are also less active than wild-type group II PLA2 against serum-treated E . coli . Treatment of E . coli with C7- or C9-depleted serum did not enhance bacterial killing or PL degradation during phagocytosis or the action of purified PLA2 . In summary, these findings suggest that (i) nonlethal assemblies of the membrane attack complex promote intracellular killing and destruction of E . coli ingested by PMN, in part by promoting the action of granule-associated PLA2 against ingested bacteria, and (ii) structural determinants first implicated in PLA2 action against BPI-treated E . coli are also important in PLA2 action in concert with other host defense systems, such as complement.

Proc Natl Acad Sci U S A, 1996 Jun 25, 93(13), 6437 - 42
The crystal structure of the immunity protein of colicin E7 suggests a possible colicin-interacting surface; Chak KF et al.; The immunity protein of colicin E7 (ImmE7) can bind specifically to the DNase-type colicin E7 and inhibit its bactericidal activity . Here we report the 1.8-angstrom crystal structure of the ImmE7 protein . This is the first x-ray structure determined in the superfamily of colicin immunity proteins . The ImmE7 protein consists of four antiparallel alpha-helices, folded in a topology similar to the architecture of a four-helix bundle structure . A region rich in acidic residues is identified . This negatively charged area has the greatest variability within the family of DNase-type immunity proteins; thus, it seems likely that this area is involved in specific binding to colicin . Based on structural, genetic, and kinetic data, we suggest that all the DNase-type immunity proteins, as well as colicins, share a "homologous-structural framework" and that specific interaction between a colicin and its cognate immunity protein relies upon how well these two proteins' charged residues match on the interaction surface, thus leading to specific immunity of the colicin.

J Am Soc Nephrol, 1996 Jun, 7(6), 861 - 70
Impact of single use versus reuse of cellulose dialyzers on clinical parameters and indices of biocompatibility; Pereira BJ et al.; Hemodialysis with reprocessed dialyzers has been associated with an increased mortality in patients on chronic dialysis, but the causes for this increased mortality have not been identified thus far . The aim of this study was to compare the qualitative and/or quantitative differences in activation of cellular and plasma elements, intradialytic signs and symptoms, adequacy of dialysis, and serum biochemistry and hematology in patients dialyzed with new or reprocessed cellulose dialyzers . This study measured the plasma levels and production of interleukin-1 receptor antagonist (IL-1Ra) by peripheral blood mononuclear cells (PBMC), indices of cytokine synthesis; plasma C3a levels, an index of complement activation; plasma levels of lipopolysaccharide binding protein (LBP), an acute phase reactant; and plasma levels of bactericidal-permeability increasing factor (BPI), a neutrophil primary granule protein, in 37 patients on chronic hemodialysis with glutaraldehyde and bleach-reprocessed cellulose dialyzers after random assignment to 12 wk of dialysis with new (single use) or reprocessed (reuse) cellulose dialyzers . These indices were studied before dialysis, 15 min after the start of dialysis, and at the conclusion of dialysis in both groups . Intradialytic clinical symptoms and signs, urea reduction ratios, monthly blood chemistry, and hematology were also studied during the 12-wk period . Before randomization, clinical and laboratory characteristics and IL-1Ra production by PBMC were similar in the two groups . During the 12-wk study, the mean number of dialyzer reuses was 7 +/- 1 in the reuse group and there were no breaks in protocol in the single-use group . At the end of the study, plasma levels of IL-1Ra, cell content and production of IL-1Ra by unstimulated, endotoxin-stimulated, and lgG-stimulated PBMC among patients assigned to reuse were not significantly different from those in the single-use group either before dialysis, at 15 min, or at the conclusion of dialysis . Similarly, plasma levels of C3a, LBP, and BPl were not significantly different between groups at any of the three time points . During the 12-wk study, none of the patients in either arm of the study experienced chills, rigors, or fever, and there were no differences in the number of episodes of symptomatic hypotension in patients on reused dialyzers (11 +/- 3) compared with patients on single-use dialyzers (8 +/- 2) . The mean monthly urea reduction ratio during the 3 months of the study was 63 +/- 2% and 65 +/- 2% for reuse and single-use dialyzers, respectively (not significant) . Similarly, the hematocrit, white blood cell count, serum calcium, phosphorus, cholesterol, triglycerides, total protein, and albumin levels were also not significantly different between the two groups at the end of the 12-wk study period . These results suggest that the reprocessing of cellulose dialyzers with glutaraldehyde and bleach does not affect indices of blocompatibility, intradialytic symptoms and signs, adequacy of dialysis, or serum biochemistry and hematology.

Aliment Pharmacol Ther, 1996 Jun, 10(3), 241 - 50
Ranitidine bismuth citrate: a novel anti-ulcer agent with different physico-chemical characteristics and improved biological activity to a bismuth citrate-ranitidine admixture; McColm AA et al.; BACKGROUND: Ranitidine bismuth citrate (RBC) is a new chemical entity for the treatment of peptic ulcer disease . RESULTS: RBC is freely soluble in water (more than 600 mg/mL at pH 4.6), whereas an equimolar admixture of its component molecules, bismuth citrate and ranitidine, formed an almost totally insoluble suspension . Even at very low pH values (around 2.0), the solubility of bismuth in ranitidine bismuth citrate was at least two-fold better than in the admixture . Comparison of several physico-chemical characteristics indicated that RBC possessed significantly different melting point properties, X-ray powder diffraction patterns, infra-red spectra and 13C-NMR solid-state spectra to the admixture . Ranitidine bismuth citrate inhibited human pepsin isoenzymes 1, 2, 3 and 5 but the admixture was inactive . RBC showed approximately two-fold greater anti-Helicobacter pylori activity in vitro than the admixture (geometric mean minimum inhibitory concentrations of 12.5 and 25.7 mg/L, respectively) and was more rapidly bactericidal . In a mouse model of gastric H . pylori colonization, 200 mg/kg of bismuth, given as RBC, eradicated the organism from all mice while only 10% of infections were eradicated by equivalent levels of bismuth in admixture form . CONCLUSION: It is believed that the significantly greater solubility of RBC, especially at lower pH values, is highly relevant to its better antipepsin and anti-H . pylori action compared to the insoluble admixture of bismuth citrate and ranitidine.

Antimicrob Agents Chemother, 1996 Jun, 40(6), 1482 - 5
Bacteriostatic and bactericidal activities of benzoxazinorifamycin KRM-1648 against Mycobacterium tuberculosis and Mycobacterium avium in human macrophages; Mor N et al.; Inhibitory and bactericidal activities of KRM-1648 were determined against Mycobacterium tuberculosis and M . avium residing in human monocyte-derived macrophages and extracellular M . tuberculosis and M . avium . MICs and MBCs of KRM-1648 against intracellular and extracellular bacteria were substantially lower than those of rifampin . The MICs and MBCs of either drug against the intracellular bacteria were only twofold lower than or equal to the values found for extracellular bacteria . The prolonged effect of KRM-1648 found in this study is probably associated with high ratios of intracellular accumulation, which were 50- to 100-fold higher than that found for rifampin . Further studies on intracellular distribution of KRM-1648 and on the sites of actual interaction between the drug and bacteria residing in macrophages are necessary, as well as evaluation of combined effects of KRM-1648 with other drugs in long-term macrophage culture experiments.

Infect Immun, 1996 Jun, 64(6), 1900 - 5
Evaluation of purified UspA from Moraxella catarrhalis as a vaccine in a murine model after active immunization; Chen D et al.; Moraxella catarrhalis causes otitis media, laryngitis, and respiratory infections in humans . A high-molecular-weight outer membrane protein from this bacterium named ubiquitous surface protein A (UspA) is present on all isolates . A monoclonal antibody (MAb) to UspA that recognizes a conserved epitope of this protein has been shown to promote pulmonary clearance of bacteria in passively immunized mice . In the present study, M . catarrhalis heterologous isolates were screened by dot blot with a panel of four additional MAbs specific for surface-exposed epitopes of UspA from M . catarrhalis isolate 035E . Three of the MAbs were specific for 035E, and the fourth reacted with 17 (74%) of the 23 isolates tested . Thus, UspA contains highly conserved, semiconserved, and variable surface-exposed epitopes . The UspA was purified from the 035E isolate by ion-exchange and size-exclusion chromatography, formulated with the adjuvant QS-21, and used to immunize BALB/c mice . Upon pulmonary challenge with either 035E or the heterologous isolate TTA24, significantly fewer bacteria were recovered from the lungs of immunized mice 6 h postchallenge than from control mice . The immune sera from mice or guinea pigs contained high titers of antibodies to the homologous isolate and heterologous isolates in a whole-bacterial-cell enzyme-linked immunosorbent assay . Sera against UspA, whether prepared in mice or guinea pigs, had complement-dependent bactericidal activity toward homologous and 11 heterologous M . catarrhalis isolates . These results indicate that the conserved epitopes of the UspA are highly immunogenic and elicit broadly reactive and biologically functional antibodies . UspA may offer protection against M . catarrhalis infections and is being further evaluated as a vaccine candidate.

J Trauma, 1996 Jun, 40(6), 886 - 92; discussion 892-3
Effects of recombinant bactericidal/permeability-increasing protein (rBPI23) on neutrophil activity in burned rats; Hansbrough J et al.; Bactericidal/permeability-increasing protein (BPI) is a neutrophil granule protein with potent bactericidal and lipopolysaccharide (LPS)-neutralizing activities . The purpose of this study was to determine if a human recombinant BPI product, rBPI23, would influence neutrophil (PMN) sequestration into various tissues in a rat burn injury model . Leukosequestration may produce local tissue injury from proteases and high-energy oxygen species released from PMNs . Rats received tracheostomy and venous cannulation, then received 17 to 20% total body surface area full-thickness contact burns and resuscitation with 20 ml, of intraperitoneal saline . Ten mg/kg body weight rBPI23 in saline was given by intravenous injection immediately after burn injury, followed by intravenous doses of 2 mg/kg at 2 and 4 hours . Control animals received intravenous saline only . PMN retention in lung, liver, spleen, gut, skin, muscle, kidney, and brain tissues was determined by removing (before burn injury) and differentially radiolabeling PMNs (111In) and erythrocytes (51Cr), reinfusing cells 4.5 hours after burn injury, and measuring tissue radioactivity 30 minutes later . Edema was estimated by measuring extravasated 125I-labeled albumin in the various tissues, 30 minutes after injection . Peripheral blood PMNS were analyzed for intracellular H2O2 content by flow cytometry using a fluorescent dye that reacts with H2O2 . Radioisotope studies demonstrated significant (p < 0.05) leukosequestration into lung, liver, gut, kidney, and skin tissues at 5 hours after burn injury . Tissue edema, manifested by radiolabeled albumin retention, was not observed in any tissues . Postburn PMN deposition in lungs and skin was decreased (p < 0.05) by the immediate administration of rBPI23 after burn injury . Flow cytometry showed increased intracellular H2O2 content in peripheral blood PMNs 5 hours after burn injury (p < 0.05), which was unaffected by administration of rBPI23 . Since sequestration of metabolically active PMNs may induce tissue injury, therapies that block leukosequestration after burn injury may improve clinical outcomes by limiting remote tissue injury.

J Mol Biol, 1996 May 17, 258(4), 627 - 37
DNA gyrase and topoisomerase IV on the bacterial chromosome: quinolone-induced DNA cleavage; Chen CR et al.; DNA gyrase, the bacterial enzyme that supercoils DNA, is trapped on chromosomal DNA by the 4-quinolone compounds, as drug-gyrase complexes that contain DNA breaks . Examination of chromosomal DNA extracted from Escherichia coli indicated that bacteriostatic concentrations of oxolinic acid trap gyrase and block DNA synthesis without releasing broken DNA from gyrase-DNA complexes . Release, detected as free rotation of DNA in the presence of an intercalating dye, occurred only at high, bactericidal oxolinic acid concentrations . Release of DNA breaks and cell death were both blocked by chloramphenicol, an inhibitor of protein synthesis, suggesting that synthesis of additional protein activity is required to free the DNA ends . Ciprofloxacin, a more potent quinolone, released DNA breaks and killed cells even in the presence of chloramphenicol . It is proposed that this second, chloramphenicol-insensitive mode for release of DNA breaks and cell killing arises from dissociation of gyrase subunits . Ciprofloxacin also killed a gyrase (gyrA) mutant resistant to the prototype of quinolone, nalidixic acid, and created complexes on DNA detected by DNA fragmentation . This lethal effect of ciprofloxacin was eliminated by additional mutations mapping in parC, one of the two genes encoding topoisomerase IV . Thus, the fluoroquinolone compounds have two intracellular targets . In the absence of the gyrA mutation, the parC (CipR) allele did not by itself confer resistance to ciprofloxacin, indicating that gyrase is the major quinolone target in E . coli . These findings provide a molecular explanation for quinolone action in bacteria and a new way to study topoisomerase IV-chromosome interactions.

Eksp Klin Farmakol, 1996 May-Jun, 59(3), 53 - 6
{The characteristics of oxolinic acid transport in the blood of dogs and monkeys when administered intramuscularly and perorally}; Tarkil NZ et al.; A possibility of testing oxolynic acid (OA) transport in blood of animals by its bactericidal effect was studied experimentally on macaques and non-pedigree dogs (intramuscular and peroral administration) . Kinetics of accumulation and elimination of OA and its derivatives exhibiting bactericidal effect makes it possible to predict optimum scheme of using these drugs in prevention and treatment of radiation disease.

Vet Immunol Immunopathol, 1996 May, 51(1-2), 55 - 65
Casein effects on the myeloperoxidase-mediated oxygen-dependent bactericidal activity of bovine neutrophils; Cooray R; Effects of casein on the respiratory burst and bactericidal activity of bovine neutrophils were studied in vitro using chemiluminescence measurements of neutrophils and the myeloperoxidase (MPO)/peroxide/chloride bactericidal system in neutrophil-free cultures . At physiological concentrations, casein inhibited the respiratory burst responses of unstimulated bovine neutrophils and those stimulated with opsonized zymosan or phorbol-12-myristate-13-acetate (PMA) . The inhibition did not result from impaired activation of neutrophils by the phagocytic stimuli, reduced light transmission or cellular toxicity . Neutrophil respiratory burst activity and the bactericidal activity associated with this phenomenon, i.e . the MPO/peroxide/chloride bactericidal system, were inhibited by casein through the modification of the oxygen radical generating pathway or oxygen radical scavenging . It has long been known that milk inhibits the bactericidal activity of neutrophils . However, the previously undocumented mechanism of action of the major milk protein requires further examination in order to determine whether the neutrophil oxygen-dependent bactericidal pathway plays an important role in defending the lactating bovine udder against bacteria causing mastitis.

J Pathol, 1996 May, 179(1), 100 - 5
Endotoxin- and cytokine-mediated effects on liver cell proliferation and lipid metabolism after partial hepatectomy: a study with recombinant N-terminal bactericidal/permeability-increasing protein and interleukin-1 receptor antagonist; Straatsburg IH et al.; This study was performed to clarify the mechanisms underlying post-resection changes in liver cell proliferation and metabolism . To assess the role of gut-derived endotoxaemia and endogenous cytokines in these changes, the effects of peri-operative treatment with either the lipopolysaccharide-neutralizing bactericidal/permeability-increasing protein or interleukin-1 receptor antagonist were investigated at 24 h after two-thirds hepatectomy in rats . Peri-operative treatment with either agent caused enhanced expression of proliferating cell nuclear antigen (PCNA) and reduced lipid accumulation . Activity of the hexose monophosphate shunt was significantly decreased after partial hepatectomy and restored by interleukin-1 receptor antagonist only . After partial hepatectomy, bile canalicular alkaline phosphatase activity was significantly increased in pericentral zones and redistributed to both bile canalicular and sinusoidal membranes of hepatocytes . These effects were not significantly influenced by either treatment . It is concluded that endotoxin restricts liver cell proliferation and leads to lipid accumulation following partial hepatectomy, and that interleukin-1 is a principal mediator in these processes . Furthermore, interleukin-1 mediates a repression of the pentose phosphate pathway . These changes may be of significance with respect to liver function, at least in the early phase after partial hepatectomy.

J Trauma, 1996 May, 40(5), 718 - 25; discussion 725-6
Changes in granulocyte colony-stimulating factor concentration in patients with trauma and sepsis; Tanaka H et al.; OBJECTIVE: To better understand the role of granulocyte colony-stimulating factor (G-CSF) after the inflammatory response . DESIGN: Serum G-CSF concentrations were measured serially in 19 trauma and 15 sepsis patients . Changes in G-CSF concentration were compared with those in the neutrophil ratio, phagocytic and bactericidal activities, and other cytokines . MEASUREMENTS AND MAIN RESULTS: G-CSF concentrations in trauma patients were elevated on day 1, but quickly decreased within 7 days . G-CSF reached its maximum 3 hours after injury, parallel with peaks of interleukin-6 (IL-6) and IL-8, but not of tumor necrosis factor-alpha (TNF-alpha) . In sepsis patients, G-CSF as well as TNF-alpha, IL-6, and IL-8 concentrations were markedly elevated at diagnosis and remained high during the course of the illness . These levels decreased significantly in the 11 survivors . Up to 3 days after the trauma, nonsegmented neutrophil ratios were higher than those thereafter . Neutrophil phagocytic and bactericidal activities remained normal during the course of disease in both conditions . CONCLUSIONS: These results suggest that G-CSF plays an important role in the maturation and maintenance of function of neutrophils during the inflammatory response to trauma and sepsis.

Cell, 1996 Apr 19, 85(2), 229 - 36
Cystic fibrosis airway epithelia fail to kill bacteria because of abnormal airway surface fluid; Smith JJ et al.; Despite an increased understanding of the cellular and molecular biology of the CFTR Cl- channel, it is not known how defective Cl- transport across airway epithelia causes chronic bacterial infections in cystic fibrosis (CF) airways . Here, we show that common CF pathogens were killed when added to the apical surface of normal airway epithelia . In contrast, these bacteria multiplied on CF epithelia . We found that bactericidal activity was present in airway surface fluid of both normal and CF epithelia . However, because bacterial killing required a low NaCl concentration and because CF surface fluid has a high NaCl concentration, CF epithelia failed to kill bacteria . This defect was corrected by reducing the NaCl concentration on CF epithelia . These data explain how the loss of CFTR Cl- channels may lead to lung disease and suggest new approaches to therapy.

QJM, 1996 Apr, 89(4), 259 - 65
Autoantibodies against bactericidal/permeability-increasing protein in patients with cystic fibrosis; Zhao MH et al.; Cystic fibrosis (CF), a genetic disorder, is characterized by chronic pulmonary infection/inflammation which leads to respiratory failure . The presence of anti-neutrophil cytoplasmic autoantibodies (ANCA) has previously been observed in the sera of patients with CF . In view of the known relationship of ANCA with primary vasculitis and of their putative pathogenetic role in these disorders, we studied the presence, specificity and isotype of ANCA and their clinical associations in 66 adult CF patients . None of the 66 CF samples had autoantibodies to the major ANCA antigens, proteinase 3 or myeloperoxidase . However, 60/66 (91%) CF samples contained IgG, and 55/66 (83%) IgA, autoantibodies to bactericidal/permeability-increasing protein (BPI), a recently-characterized ANCA specificity . All the IgA anti-BPI-positive samples were also IgG anti-BPI-positive . The autoantibody specificity was confirmed by inhibition assay and immunoblotting of CF sera against a neutrophil granule preparation . Furthermore, in this cross-sectional study, anti-BPI levels were inversely correlated with the observed reductions in FEV1 and FVC (IgA anti-BPI & FEV1: r = -0.508, p < 0.0001), and both IgG and IgA anti-BPI levels were higher in CF patients with secondary vasculitis (n = 6) than in those without (p < 0.05) . ANCA with specificity for BPI were present in the majority of CF sera in this study and autoimmune processes may be associated with the development of pulmonary injury in CF.

Thorax, 1996 Apr, 51(4), 369 - 73
Minimisation of aminoglycoside toxicity in patients with cystic fibrosis; Wood PJ et al.; BACKGROUND: There is evidence that administration of higher doses of aminoglycosides given less frequently improves the bactericidal effect and reduces the potential to cause side effects . To investigate this, a prospectively randomised open label therapeutic trial was undertaken in stratified groups of patients with cystic fibrosis to examine the efficacy and toxic potential of an aminoglycoside dosing regimen designed to generate high peak drug concentrations at 12 hourly intervals compared with conventional dosing at eight hourly intervals . METHODS: Patients in group A received tobramycin eight hourly using a dose aimed at generating a peak concentration of 10 mg/l with trough concentrations below 2 mg/l, and those in group B received the total daily dose required to achieve eight hourly target concentrations administered as two equal 12 hourly doses . Clinical outcomes measured and assessed included vestibular symptoms, hearing and renal function, length of hospital stay, readmission rate, and mortality . RESULTS: Twenty nine patients were recruited during a six month period, 20 to group A and nine to group B . The average peak tobramycin level was higher in group B (12.5 (2.2) mg/l) than in group A (7.9 (1.9) mg/l), whilst the average trough level was higher in group A (0.8 (0.3) mg/l) than in group B (0.5 (0.2) mg/l) . There was a difference in the number of ototoxic events between patients in group A (seven of 18, 38.9%) and group B (none of eight), but no difference was found in other outcome measures assessed . CONCLUSIONS: These results suggest that 12 hourly high peak aminoglycoside dosing may be less toxic than equivalent eight hourly dosing, without any apparent difference in efficacy.

Chest, 1996 Apr, 109(4), 1078 - 85
Cross-shift changes in blood inflammatory markers occur in the absence of airway obstruction in workers exposed to grain dust; Borm PJ et al.; Grain dust is well known to cause both acute and chronic respiratory disorders, and endotoxins are considered key components in this . Since endotoxins are known to elicit proinflammatory mediators, we investigated cytokine (tumor necrosis factor {TNF}, interleukin-6, interleukin-8) release and a number of proinflammatory and anti-inflammatory proteins (soluble TNF receptors, lipopolysaccharide (LPS) binding protein, bactericidal permeability increasing protein (BPI), C-reactive protein) in plasma of workers exposed to grain dust . In two surveys during 1 week, lung function was measured daily before and after the shift, using flow-volume curves and/or forced oscillation measurements . On Monday and Friday, blood samples (30 mL) were drawn and cytokine release was determined by enzyme-linked immunosorbent assay in supernatant of isolated monocytes or whole blood culture, either unstimulated or on the ex vivo stimulation with 3 ng/mL or 1,000 ng/mL endotoxin . Individual exposures were determined from stationary dust measurements at every workplace combined with personal task analysis during all shifts . In both surveys, no cross-week change in lung function parameters was observed . In the first survey (average exposure: 20.2 mg/m3), monocyte spontaneous TNF release was increased sevenfold cross week (p<0.001) and was significantly related both to individual dust exposure (r=0.62) of that week and the increase in soluble TNF receptor 75 kD (r=0.85) . In the second survey, where average exposure was much lower (3.67 mg/m3), impedance parameters indicated a significant improvement of airway function, and cross-week changes in inflammatory markers were minimal . Therefore, we conclude that inflammatory events can be used to monitor adverse respiratory effects of moderate grain dust exposure.

Clin Exp Immunol, 1996 Apr, 104(1), 54 - 9
Anti-neutrophil cytoplasmic antibodies (ANCA) directed against bactericidal/permeability increasing protein (BPI): a new seromarker for inflammatory bowel disease and associated disorders; Stoffel MP et al.; It was our purpose to determine the immunodiagnostic value of ANCA directed against BPI in diseases known to be associated with ANCA, such as ANCA-associated vasculitides, inflammatory bowel disease (IBD) and the associated condition primary sclerosing cholangitis . The immunoreactivity of recombinant PBI (rBPI) was established in order to develop an ELISA specific for rBPI . By means of this assay, BPI-ANCA were assessed in sera of 178 patients with IBD or the associated disorder primary sclerosing cholangitis, 112 patients with ANCA-associated vasculitides, and in sera of 182 disease and 140 health controls . BPI-ANCA were found to be closely associated with IBD and primary sclerosing cholangitis (34% and 44% of ANCA-positive sera, respectively) . By contrast, BPI-ANCA positivity was low (<10%) in the double-negative sera of patients with ANCA-associated vasculitides and in disease and health controls . BPI-ANCA appear to constitute an important marker for IBD and primary sclerosing cholangitis, but not for the ANCA-associated vasculitides.

Arch Intern Med, 1996 Mar 11, 156(5), 577 - 83
Malacoplakia . Two case reports and a comparison of treatment modalities based on a literature review; van der Voort HJ et al.; Malacoplakia is a rare infectious disease that has been almost exclusi vely reported in urology and pathology journals . We studied two cases of malacoplakia that were primarily referred to the department of internal medicine because of fever and abdominal masses . In one patient, malacoplakia was diagnosed in the unusual ovarian location, while in the other patient a large renal mass was found and ciprofloxacin therapy failed because of bacterial resistance . The clinical and radiologic appearance of malacoplakia often mimics that of a malignant tumor . The principal disorder is probably a monocytic-macrophagic bactericidal defect . A definitive diagnosis depends on microscopic detection of Michaelis-Gutmann bodies by means of von Kossa stain . We outlined treatment strategies on the basis of a review of the literature since 1981, which included 140 cases . If possible, immunosuppressive drugs should be stopped . Quinolone antibiotic treatment and surgical excision or incision and drainage lead to the highest cure rates (90% and 81%, respectively) . Specific intracellular penetration of quinolone antibiotics is a possible reason for the higher cure rate achieved with these antibiotics . Bethanechol has been suggested to correct the supposed fundamental disturbance by increasing the intrecellular cyclic guanosine monophosphate concentration, but there is still no convincing evidence of its clinical efficacy.

Antibiot Khimioter, 1996 Mar, 41(3), 25 - 8
{Effect of lomefloxacin on the functional activity of the phagocytic system}; Mozhokina GN et al.; The in vitro effect of lomefloxacin on the absorption activity of macrophages with respect to Mycobacterium tuberculosis and chemiluminescence of the phagocytizing cells was studied with the use of a wide range of the antibiotic concentrations . Lomefloxacin was shown to have no inhibitory effect on the phagocytic activity of the pulmonary macrophages and on the formation of active oxygen in the phagocytizing cells for the realization of the bactericidal action on M . tuberculosis.

FEMS Immunol Med Microbiol, 1996 Mar, 13(3), 249 - 52
Francisella tularensis resistance to bactericidal action of normal human serum; Sorokin VM et al.; Lipopolysaccharide and outer membranes from the three virulent encapsulated (Cap(+)) strains of three subspecies of Francisella tularensis and their isogenic avirulent capsule-deficient (Cap(-)) mutants were isolated . It was shown that the Cap cells and their outer membranes almost completely consumed the available complement of normal human serum whereas Cap(-) LPS (R-LPS), Cap(+) cells and their components activated the complement less effectively . Absorption of normal human serum with Cap(-) strain dramatically reduced the complement consumption for homologous strain and its surface structures . This reduction reflected the loss of bactericidal antibodies . Addition of antibodies to whole cells of F . tularensis completely restored complement activity . The cross-absorbing experiments demonstrated that Cap(-) cells more effectively deplete bactericidal antibodies than homologous virulent strain . From these results it can be concluded that normal human serum is bactericidal for serum-sensitive Cap(-) F . tularensis strains through the action of complement initiated by the classical complement pathway and serum resistance of virulent strains is not due to absence of targets for bactericidal antibodies, but is due to their low accessibility because of O-side chains of lipopolysaccharide.

Antimicrob Agents Chemother, 1996 Mar, 40(3), 743 - 9
In vitro activities of rifabutin, azithromycin, ciprofloxacin, clarithromycin, clofazimine, ethambutol, and amikacin in combinations of two, three, and four drugs against Mycobacterium avium; Yajko DM et al.; Multidrug therapy is recommended for treatment of Mycobacterium avium complex (MAC) bacteremia in patients with AIDS . Azithromycin, clarithromycin, rifabutin, ciprofloxacin, ethambutol, clofazimine, and amikacin have all been suggested for use in treating MAC bacteremia, but the most active combinations of these drugs have not been identified, nor has the minimum number of drugs needed for effective therapy been determined . To address the former, the in vitro bactericidal activities of all two-, three-, and four-drug combinations of these seven agents was determined by using 10 blood-derived strains of MAC isolated from patients with AIDS . The activities of the 132 drug combinations were compared by statistical analysis of survival means (analysis of variance) and further evaluated by determining the percentage of strains considered susceptible to each combination . When susceptibility was defined as a decrease in CFU of > or = 2 log10, no two- or three-drug combination and only two four-drug combinations were active against all 10 MAC strains . When a less stringent definition was applied (> or = 1 log10 decrease in CFU), 1 two-drug combinations, 9 three-drug combinations, and 31 four-drug combinations showed activity against all 10 strains . Eighteen selected drug combinations were also tested for intracellular activity in MAC-infected J774 cells . Combinations which contained amikacin as a component were considerably less active against intracellular MAC organisms than against organisms in broth . The opposite result was obtained for the combination of clarithromycin plus clofazimine.

Antimicrob Agents Chemother, 1996 Mar, 40(3), 546 - 51
Activities of bay Y 3118, levofloxacin, and ofloxacin alone or in combination with ethambutol against Mycobacterium avium complex in vitro, in human macrophages, and in beige mice; Bermudez LE et al.; Levofloxacin, ofloxacin, and Bay Y 3118 are new fluoroquinolones with variable in vitro bacteriostatic and bactericidal activities against the Mycobacterium avium complex (MAC) . The potential therapeutic activities of these agents both alone and combined with ethambutol were evaluated in a human macrophage test system and in the beige mouse animal test system with MAC strain 101 . Bay Y 3118 at a human-equivalent dose of 30 mg/kg/day for 4 weeks caused a significant reduction in mortality compared with that in untreated controls (P = 0.02) . Bay Y 3118 also caused significant reductions in the number of MAC organisms in the blood, liver tissue, and spleen tissue compared with those in untreated controls . Levofloxacin at a human-equivalent dose of 200 mg/kg/day was associated with a significant reduction in mortality (10 versus 39%); however, treatment with either levofloxacin or ofloxacin (200 mg/kg/day) did not result in significant reductions in the numbers of MAC organisms in blood, liver, and spleen compared with those in untreated controls . When Bay Y 3118 was combined with ethambutol, there was no enhancement in therapeutic activity except in the spleen in terms of CFU per gram (reductions of 89% compared with the untreated control, 63% compared with Bay Y 3118 alone, and 72.5% compared with ethambutol alone) . Levofloxacin in combination with ethambutol was more active than either drug alone in the reduction of organisms in blood, liver, and spleen . Bay Y 3118 was the most active fluoroquinolone for monotherapy of MAC infection in beige mice, and the combination of ethambutol plus either levofloxacin or ofloxacin was at least additive . In summary, this study demonstrates that quinolones, although active, are inhibitory against MAC in vivo and that there is little correlation between the activity of quinolones in vitro and the activity in mice.

J Am Soc Nephrol, 1996 Mar, 7(3), 479 - 87
Plasma lipopolysaccharide binding protein and bactericidal/permeability increasing factor in CRF and HD patients; Pereira BJ et al.; The recent characterization of lipopolysaccharide binding protein (LBP) and bactericidal/permeability increasing factor (BPI) have provided the opportunity to examine the natural factors that regulate cytokine production in response to endotoxin in patients on hemodialysis (HD) . Whole blood was collected in EDTA from 28 undialyzed patients with chronic renal failure (undialyzed CRF), 36 patients on chronic HD (HD) and 15 healthy controls, and plasma levels of LBP and BPI were measured by a sandwich ELISA . Plasma LBP levels in undialyzed patients with CRF (P = 0.04) and patients on HD (P = 0.01) were significantly higher than those in healthy controls, but not significantly different from each other . Plasma BPI levels in undialyzed patients with CRF and patients on HD were not significantly different from those in healthy controls . There was no correlation between serum creatinine and plasma levels of either LBP or BPI . Peripheral blood mononuclear cells (PBMC) were harvested from healthy volunteers by FLcoll-Hypaque separation, and 0.125 mL of 10 x 10(6)/mL suspensions were incubated with 0.125 mL of test plasma (containing different LBP/BPI ratios) and 0.25 mL of RPMI, containing 1 ng/mL of endotoxin, for 24 h at 37 degrees C . Samples were subjected to three freeze-thaw cycles, and total interleukin-1 receptor antagonist (IL-1Ra) or interleukin-1 alpha (IL-1 alpha) production was measured by a specific non-crossreactive RIA . The results of this study showed: (1) IL-1Ra production by endotoxin-stimulated PBMC incubated with pooled plasma from HD patients with LBP/BPI ratios of 11 x 10(2), 167 x 10(2), 379 x 10(2), and 778 x 10(2), respectively was 1466 +/- 195 pg, 3105 +/- 462 pg, 8179 +/- 1020 pg, and 4770 +/- 1185 pg (P < 0.001); (2) Paired plasma collected before dialysis (predialysis) and at 15 min after the start of dialysis (15 minute) with cellulose membranes showed a negligible change in plasma LBP levels (-3 +/- 5%), but a 6681 +/- 1788% increase in plasma BPI levels . Consequently, compared with predialysis plasma, there was a 35 +/- 6% decrease in endotoxin-stimulated IL-1 alpha production by PBMC incubated with plasma drawn at 15 min (P = 0.001); (3) Compared with the PBMC incubated with predialysis plasma from HD patients, there was a 39 +/- 5%, 53 +/- 5%, and 60 +/- 5% decrease in endotoxin-stimulated IL-1 alpha production in the presence of 1 ng/ mL, 10 ng/mL, or 1 microgram/mL of recombinant BPI, respectively (P < 0.003) . These results suggest that the plasma LBP:BPI ratio could influence cytokine production in response to bacterial endotoxin; the high LBP:BPI ratios observed in patients with chronic renal failure probably imparts an increased susceptibility to endotoxin-stimulated cytokine production; and natural or pharmacological increases in plasma BPI levels and the consequent decrease in LBP:BPI ratios could attenuate this susceptibility to endotoxin-stimulated cytokine production.

Minerva Med, 1996 Mar, 87(3), 89 - 92
{Decubitus ulcer in the calcaneus region: rapid development, difficult recovery}; Fugazza G et al.; Heel pressure sores frequently arise in patients kept in bed for a long time independently of their primary disease . In account of this event the authors completed a study concerning possible mutual relations between heel pressure sores and primary disease of the patients; to validate the pharmacological treatment in less severe sores and the surgical resolution in more serious cutaneous lesions . In the last 3 years (1992-1995) at the Rehabilitation Centre of Montescano the authors have treated 39 patients suffering from 63 different severe cutaneous lesions: from phlycten to deep necrosis . The therapeutic plane utilized pharmacological treatment for 1st, 2nd, 3rd degree pressure sores, and surgical treatment for 4th degree . Pharmacological treatment included: enzymatic drugs, bactericidal and bacteriostatic medicines and cicatrizing substances . Different healing times were related to different pressure sore severity . Surgical treatment consisted of transposition of flap into wound defect . This system caused considerable reduction in resolution times . The authors noticed how easily pressure sores arise in the heel region, and how difficultly they heal . This is probably connected with particular anatomical and vascular characteristics of this region.

Clin Exp Immunol, 1996 Mar, 103(3), 397 - 402
Azurocidin is a novel antigen for anti-neutrophil cytoplasmic autoantibodies (ANCA) in systemic vasculitis; Zhao MH et al.; We isolated a 27-kD protein using cation exchange chromatography from an acid extract of neutrophil granules . N-terminal amino acid sequence analysis of the first 10 residues showed that this protein is azurocidin, a member of the family of neutral serine proteinase found in the neutrophil, which shares amino acid sequence homology with the three other neutral serine proteinases, elastase, proteinase 3 (PR3) and cathepsin G, but unlike them is without proteolytic activity . To test whether, in addition to these proteases, azurocidin might be a target for the humoral autoimmune responses associated with human vasculitis, 185 indirect immunofluorescence (IIF)-positive ANCA sera, made up of four groups of sera with specificities for PR3 (n=37), myeloperoxidase (MPO; n=50), bactericidal/permeability-increasing protein (BPI; n=41) and sera that recognized none of them (triple negative, n=57), and 46 normal sera were screened for IgG anti-azurocidin antibodies using an ELISA incorporating purified azurocidin . Twenty of the 185 IIF-positive sera and 2/46 normal sera displayed reactivity with azurocidin . Positive sera could blot the 27-kD band by Western blot analysis . Further study of the 20 positive sera revealed that: (i) 10 also had autoreactivity for MPO, of which six additionally recognized lactoferrin; (ii) two had reactivity with BPI; (iii) the remaining eight sera were positive only for azurocidin . All 20 sera were from patients with systemic vasculitis, and four of the six sera with triple reactivity (for azurocidin, MPO and lactoferrin) were from patients with hydralazine-induced vasculitis . We concluded that: (i) azurocidin is a novel ANCA antigen; (ii) anti-azurocidin antibodies from a subgroup of patients might represent the consequence of a drug-induced multi-clone activation.

Nippon Rinsho, 1996 Feb, 54(2), 369 - 76
{Transfer of endothelial nitric oxide synthase gene in the purpose of gene therapy for pulmonary arterial hypertension}; Setoguchi Y et al.; Nitric oxide (NO) is a messenger molecule involved in diverse processes in many tissues . For example, NO is responsible for the bactericidal activities of macrophages, and in blood vessels it accounts for endothelium-derived relaxing factor activity . Recently, inhalation of NO gas was reported to improve the acute pulmonary arterial hypertension . Based on this knowledge, recombinant expression of endothelial nitric oxide synthase (eNOS) in lung may have profound effects on pulmonary vasomotor function and pulmonary arterial smooth muscle proliferation and platelet adhesion . On the basis of this concept, we evaluate the feasibility of gene therapy for chronic pulmonary arterial hypertension using hypoxia regulatable adenoviral vector coding eNOS cDNA.

Antimicrob Agents Chemother, 1996 Feb, 40(2), 426 - 8
In vitro bactericidal and in vivo therapeutic activities of a new rifamycin derivative, KRM-1648, against Mycobacterium tuberculosis; Yamamoto T et al.; The in vitro and in vivo activities of a new rifamycin derivative, KRM-1648, against Mycobacterium tuberculosis H37Rv were compared with those of rifampin . Bactericidal activity was evaluated by using a silicone-coated slide culture method . The MBC of KRM-1648 was 0.15 to 0.3 microgram/ml for 24 h of exposure, while that of rifampin was > 160 microgram/ml under the same conditions . Against experimental murine tuberculosis, KRM-1648 exhibited significant therapeutic effects, in terms of prolonged survival times for mice compared with those with rifampin treatment, even at lower doses, such as 1 and 3 mg/kg . At a dose of 3 mg/kg, KRM-1648 was at least as effective as rifampin at 10 mg/kg . The combination of KRM-1648 (3 mg/kg) plus isoniazid (3 mg/kg) plus ethambutol (10 mg/kg) exhibited much more activity than did rifampin (10 mg/kg) plus isoniazid (3 mg/kg) plus ethambutol (10 mg/kg) . These findings suggest that KRM-1648 is a promising candidate for the treatment of tuberculosis.

Antimicrob Agents Chemother, 1996 Feb, 40(2), 393 - 9
Bactericidal activities of combinations of new drugs against Mycobacterium leprae in nude mice; Ji B et al.; The bactericidal activities of 12 regimens with various combinations of new drugs (clarithromycin {CLARI}, minocycline {MINO}, and ofloxacin {OFLO}) and the standard antileprosy drugs, especially rifampin (RMP), were compared in nude mice with established Mycobacterium leprae infection . The longest duration of treatment was 24 weeks for intermittent (once every 4 weeks) therapy and 8 weeks for daily therapy . Bactericidal effects were monitored by titrating the proportion of viable M . leprae isolates by subinoculating the organisms into the footpads of immunocompetent and nude mice . The results indicate that RMP was more bactericidal than any combination of the new drugs . A single dose of CLARI-MINO, with or without OFLO, displayed bactericidal activity as great as that of 4 weeks of daily treatment with dapsone (DDS) plus clofazimine (CLO); thus, intermittent CLARI-MINO, with or without OFLO, may replace DDS and CLO of the standard multidrug regimen, and these will become regimens that can be administered monthly and under full supervision . Additional evidence that this may be the case is provided by the finding that intermittent RMP-CLARI-MINO or RMP-CLARI-MINO-OFLO administered for 12 or 24 weeks was as active as the standard multidrug regimen . While the intermittent treatment always displayed significantly greater bactericidal activity than the same number of doses of daily treatment, daily treatment with CLARI-MINO and CLARI-MINO-OFLO were more active than the drugs given as intermittent treatment for the same duration; therefore, unless these combinations are to be administered together with intermittent RMP, they should be given daily, especially for the treatment of RMP-resistant cases of infection . Finally, 12 weeks of daily treatment with DDS-CLO was more bactericidal than had been expected, suggesting that it may not be necessary to administer the standard multidrug regimen for multibacillary leprosy for as long as 24 months in order to minimize the risk of developing RMP resistance.

Acad Radiol, 1996 Feb, 3(2), 97 - 102
Effects of magnetic resonance imaging on polymorphonuclear neutrophil functions; Minczykowski A et al.; RATIONALE AND OBJECTIVES: Limited research has been performed on the effects of magnetic resonance (MR) imaging on the immune system . To our knowledge, there are no reported studies of MR imaging effects on the polymorphonuclear neutrophil (PMN) system . Therefore, we evaluated the influence of MR imaging exposure on PMNs . METHODS: In vivo and in vitro studies were performed on 36 patients undergoing MR imaging . The following were estimated in blood samples: leukocyte and PMN count, PMN phagocytosis and bactericidal capacity, percentage of cells with expression of surface receptor for the Fc fragment of immunoglobulin G (IgG), PMN superoxide, hydrogen peroxide production, and plasma lysozyme activity . Another sample of patients was used to eliminate temperature as an influence on changes in PMN functions . RESULTS: Both in vitro and in vivo MR imaging led to a decrease in PMNs and an increase in PMN phagocytosis, bactericidal capacity, hydrogen peroxide production, and percentage of cells with expression of surface receptor for Fc IgG . Superoxide anion production did not change significantly . Elevated temperature, stress, and anxiety were excluded as influences on our results . CONCLUSION: The PMN system is affected seriously by MR imaging.

Arch Surg, 1996 Feb, 131(2), 200 - 5; discussion 206
Mononuclear cell line THP-1 internalizes bactericidal/permeability-increasing protein by a non-receptor-mediated mechanism consistent with pinocytosis; Burnett RJ et al.; BACKGROUND: Bactericidal/permeability-increasing protein (BPI) binds lipopolysaccharide and neutralizes its toxic effects in vitro and in endotoxemic animals . Our recent work identified physiologically significant interactions between BPI, lipopolysaccharide, and mononuclear cells . OBJECTIVE: To determine whether the interaction between BPI and mononuclear cells is receptor mediated . DESIGN: Labeled BPI was incubated with THP-1 cells in the presence of up to 100-fold excess of unlabeled BPI . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting were performed to evaluate competitive binding and total uptake of BPI . Crosslinking was performed to determine whether BPI binds to a single protein entity . Acid washing experiments and flow cytometric analysis were performed to determine whether BPI remains on the cellular surface . Finally, flow cytometry analysis was used to determine whether BPI incubation with THP-1 cells affects the surface expression of the lipopolysaccharide-binding protein-lipopolysaccharide receptor CD14 . RESULTS: Labeled BPI uptake was not inhibited by the presence of 100-fold excess of unlabeled BPI at 37 degrees C or 4 degrees C in the presence of azide . Uptake was not saturable under either condition with incubation concentrations up to 10 microgram/mL . Cross-linking did not show BPI bound to a single entity . Acid washing and flow cytometry experiments disclosed rapid internalization of BPI . Finally, BPI uptake by THP-1 cells had no effect on the surface expression of CD14 . CONCLUSIONS: Bactericidal/permeability-increasing protein is rapidly internalized by mononuclear cells in a nonspecific fashion not saturable at very high doses, which is consistent with pinocytosis . This process may represent a disposal mechanism for lipopolysaccharide in closed-space infections and may be partially responsible for the rapid clearance of BPI from the peripheral circulation.

Schweiz Med Wochenschr, 1996 Jan 27, 126(4), 124 - 7
{Treatment of osteomyelitis with oral bactericidal antibiotics-- description of 9 cases}; Major S et al.; The efficacy of oral bactericidal antibiotics in the treatment of osteomyelitis is analyzed retrospectively by a review of 9 cases histories . The first patient was treated in 1982 . Clinical and laboratory controls were reviewed up to 1.1.1995 . In 6 patients S . aureus was isolated, while in 3 patients the organism could not be cultured but gram + cocci were almost likely . 5 cases of osteomyelitis were acute, 4 were chronic and in one case there was an infection of an artificial knee prothesis . The isolated S . aureus were sensitive to the antibiotics prescribed . 8 patients received a combination of flucloxacillin (2-4 g/day) and rifampicin (600 mg/day); one patient was treated with the combination amoxicillin/clavulanic acid (1500/425 mg/day) followed by ofloxacin (400 mg/day) and finally co-trimoxazole (320/1600 mg/day) . 3 patients received all antibiotics orally; in 6 patients oral administration followed a short intravenous course of the antibiotic combination . The duration of treatment varied between 6 and 23 weeks . Follow-up extended over 9 months (n = 1), 14 months (n = 3), 4-7 years (n = 3), and 10-12 years (n = 2) . In 8 cases the osteomyelitis was cured (89%) . In our view, a combination of bactericidal oral antibiotics with good penetration into bone tissue can be prescribed in selected cases of osteomyelitis to shorten or even avoid the standard intravenous therapy of acute or chronic diseases.

J Immunol Methods, 1996 Jan 16, 189(1), 37 - 45
Isolation of the C9b fragment of human complement component C9 using urea in the absence of detergents; Gu X et al.; The bactericidal activity of the C5b-9 complex of complement is dependent upon the terminal complement component C9 . The precursor C5b-8 complex is not harmful to bacterial cells until C9 is added to complete the C5b-9 complex . The C9 molecule can be proteolytically cleaved by thrombin to yield an intact, nicked molecule that remains fully functional when added to either bacterial cells or erythrocytes bearing pre-formed C5b-8 complexes . In investigating the membranolytic function of C9 in the C5b-9 complex, the carboxyl-terminal portion of the nicked molecule (C9b) has been shown to be membranolytic when added to erythrocytes, liposomes, or bacterial inner membranes in the absence of any other complement components . The isolation of C9b from nicked C9 has been accomplished by preparative gel electrophoresis using detergents, however the study of the activity of C9b in membrane systems may be complicated by the possible presence of residual detergent . To address this concern, we have used 4 M urea in conjunction with hydroxyapatite chromatography and a phosphate elution procedure to separate the domains of nicked C9 . The isolated C9b domain, free of detergents and in the absence of any other complement components, was found to be membranolytic . C9b isolated in this manner was capable of lysing erythrocytes and inhibiting the growth of bacterial spheroplasts.

J Biol Chem, 1996 Jan 5, 271(1), 83 - 8
The Rac target NADPH oxidase p67phox interacts preferentially with Rac2 rather than Rac1; Dorseuil O et al.; NADPH oxidase is a plasma membrane enzyme of phagocytes generating superoxide anions which serve as bactericidal agents . Activation of this multimolecular enzyme minimally requires assembly at the membrane with flavocytochrome b558 of cytosolic components p47phox, p67phox, and Rac proteins . Rac1 and Rac2 are 92% homologous cytosolic small GTPase proteins . Both Rac1 and Rac2 have been implicated with NADPH oxidase activation in vitro; however, Rac2 is largely predominant in human phagocytes . Here, using the yeast two-hybrid system, we provide data demonstrating in vivo interactions between human p47phox, p67phox, and Rac proteins . Rac proteins interact with p67phox in a GTP-dependent manner, but do not interact with p47phox . Moreover, Rac effector site mutants, which are known to be inactive in NADPH oxidase, lose their interaction with p67phox; Rac2L61 mutant, which has an increased NADPH oxidase affinity, shows an increased affinity for p67phox . Finally, we observe that p67phox interacts 6-fold better with Rac2 than with Rac1 . We also show a strong intracellular interaction between p47phox and p67phox . These results indicate that activated Rac can regulate NADPH oxidase by interacting with p67phox and that Rac2 is the main p67phox-interacting GTPase in human cells.

Folia Microbiol (Praha), 1996, 41(2), 159 - 64
Preservative-mediated changes of the surface properties of Escherichia coli; Kaul M et al.; The effectiveness of two commonly used preservatives, sodium benzoate and potassium disulfite, was evaluated in terms of their bactericidal activity and capacity to induce changes in the surface properties of Escherichia coli isolated from commercial food preserves . Preservative treatment over a five-week test period resulted in controlling the multiplication of these organisms and causing a decline in cell-surface hydrophobicity, hemagglutinating ability and adherence capacity to rat intestinal cells of E . coli isolates . A loss in motility was also exhibited.

Acta Otolaryngol Suppl, 1996, 523, 120 - 3
Superoxide dismutases in human palatine tonsils; Lai MT et al.; In order to investigate the protective system of human palatine tonsils against the cytotoxic superoxide radicals (O(-)(2)) generated from the oxygen-related bactericidal system, immunohistochemistry and electron spin resonance (ESR) spectrometry were used to detect the distribution and activities of superoxide dismutases (SODs) in tonsils of different related systemic diseases . Immunohistochemistry showed that SODs distribute in extrafollicular lymphatic tissue and crypt epithelium . No distribution difference could be found between tonsils of different related systemic diseases . ESR revealed no significant difference between SODs activities in tonsils of different related systemic diseases . However, the mitochondrial SOD activity was found to constitute approximately 50%-60% of the total tonsillar cellular SODs activity . The results suggest: i)tonsils possess the ability to control cytotoxic O(-)(2), ii) crypt epithelium and extrafollicular lymphatic tissue may encounter more O(-)(2) threat, iii) SODs may be important in protecting germinal centers from O(-)(2) injury, and iv) systemic diseases are less related to the local expression of tonsillar SODs.

Probl Tuberk, 1996, (6), 19 - 22
{Molecular mechanisms of cell-to-cell interactions during the progress and healing of tuberculosis}; Kaminskaia GO et al.; The role of some protein systems of humoral regulation and of some lipid mediators in the development, course, and outcomes of tuberculous inflammation has been studied . The authors discuss regularities in the changes of the kallikrein-kinin system, blood clotting and fibrinolysis systems, proteinase-inhibitor balance, pool and functional activity of fibronectin in the blood and various parts of the respiratory systems, shifts in the content and ratios of prostaglandins E and F2 alpha in pulmonary tissue and bronchoalveolar space, in the level of platelet activation factor in liquid biological media and various types of phagocytizing cells . Shifts in all these systems are uniform, starting from acute-phase mobilization and eventuating in loss of autoregulation and depletion . Changes in all the above-listed systems of humoral regulation are due to the bactericidal activity of phagocytes, specific features of the cytogram of bronchoalveolar lavage, vascular permeability, morphological characteristics, and course and outcomes of the tuberculous process in the lungs.

J Clin Immunol, 1996 Jan, 16(1), 31 - 40
Mercury-specific lymphocytes: an indication of mercury allergy in man; Stejskal VD et al.; In this study, 18 patients with oral lichen planus (OLP), adjacent to amalgam fillings, were tested in vitro with an optimized lymphocyte proliferation test, MELISA (memory lymphocyte immunostimulation assay) and with a patch test . Twenty subjects with amalgam fillings but without oral discomfort and 12 amalgam-free subjects served as controls . The results show that patients with OLP have significantly higher lymphocyte reactivity to inorganic mercury, a corrosion product of amalgam, compared to control groups . Removal of amalgam fillings resulted in the disappearance of oral mucosal changes, thus indicating a causal relationship . Positive responses to phenylmercury (phenyl-Hg), a bactericidal agent in root fillings and in pharmaceutical preparations, were also noted in the oral lichen group but not in the control groups . Thus, low-grade chronic exposure to mercury may induce a state of systemic sensitization as verified by Hg-specific lymphocyte reactivity in vitro.

Med Law, 1996, 15(2), 329 - 35
Adipocere--problems in estimating the length of time since death; Rothschild MA et al.; Establishing the length of time since death is particularly difficult in corpses showing advanced autolysis . One sign of advanced decay is the formation of adipocere . In this state bodies show evidence of a partly wax-like and partly pasty condition . Continued storage ultimately results, among other things, in further decomposition due to the action of micro-organisms from the surrounding area-even if this is chronologically delayed . An exception is provided by the formation of adipocere under air-tight conditions . Initially, autolysis and heterolysis also occur, involving the release of fatty acids . As a result of the subsequent hydrogenation of the fats under the influence of bacterial enzymes, the unsaturated fatty acids are partially converted into saturated fatty acids . As the fatty acids clearly have a bactericidal effect, further bacterial decomposition is stopped at this early adipocere stage . Additional micro-organisms from outside can no longer penetrate when this hermetic seal is in place . In addition, the lack of calcium (e.g . from water or moist earth) can be a reason for the fact that further adipocere development, leading to wax-like hardening of the fat, is arrested . Thus the condition of the body can remain constantly preserved over many years and it no longer allows a reliable estimate to be made of the period of time since death.

Microbios, 1996, 86(349), 213 - 23
Effect of culture conditions on the susceptibility of enteroaggregative Escherichia coli to human serum and polymorphonuclear leucocytes; Haque MA et al.; Recent work has indicated that the pathogenicity of enteroaggregative Escherichia coli (EAggEC) is strictly regulated by environmental conditions . The growth conditions, susceptibility to the bactericidal activity of serum, and polymorphonuclear leucocytes (PMN) using strains of EAggEC grown in two different culture conditions were examined . Strains of EAggEC grown in casamino acids-yeast extract (CYE) broth at 37 degrees C for 20 h showed resistance to killing by human serum (86.8 +/- 6.9% survival by 2 h) and PMN (67 +/- 5.1% survival by 2 h) . In contrast, these strains grown in nutrient broth (NB) showed sensitivity to serum (98.9 +/- 0.5% killing by 2 h) and PMN (98.9 +/- 0.5% killing by 2 h) . EAggEC strains cultured in NB became resistant to killing by serum when mixed with extracellular material extracted from the bacteria grown in CYE broth . These results indicate that the growth condition may modulate the sensitivity of EAggEC, and the promotion of extracellular material formation in CYE broth is responsible for the resistance to killing by serum and PMN.

Rheumatol Int, 1996, 16(3), 109 - 14
Autoantibodies and target antigens in antineutrophil cytoplasmic antibody (ANCA)-associated vasculitides; Savige JA et al.; In this study of antineutrophil cytoplasmic antibody (ANCA)-associated diseases, we determined the prevalence of other autoantibodies and the antigen specificities of ANCA . ANA were common, occurring in 7 of 36 (19%) patients with Wegener's granulomatosis, in 16 of 34 (47%) patients with microscopic polyarteritis, in 6 of 11 (55%) patients with segmental necrotising glomerulonephritis and in 8 of 18 (44%) of those with ANCA-associated systemic vasculitis without renal involvement . ANA were associated more often with pANCA and microscopic polyarteritis than with cANCA (P < 0.05) . Patterns were speckled (n = 23), homogeneous (n = 10) or nucleolar (n = 4) . Anticardiolipin antibodies were also common, occurring in 10 of 25 (40%) patients with Wegener's granulomatosis, in 8 of 14 (57%) patients with microscopic polyarteritis and in 6 of 18 (33%) of those with a systemic vasculitis . However, anticardiolipin antibodies did not correlate with the presence of ANCA in any of the disease groups . Anti-GBM antibodies were demonstrated in only 2 of 25 (8%) patients with Wegener's granulomatosis, in 1 patient with microscopic polyarteritis (1/14, 7%) and in 1 with segmental necrotising glomerulonephritis (1/11, 9%) . All four patients with anti-GBM antibodies had either cANCA or pANCA . In the second part of the study, the target antigens of ANCA were determined in Wegener's granulomatosis, microscopic polyarteritis, systemic vasculitis, inflammatory bowel disease, rheumatoid arthritis and systemic lupus erythematosus (SLE) . Of the 19 sera with cANCA, 13 (68%) were directed against proteinase 3; other antigens were myeloperoxidase (1/19, 5%), elastase and lactoferrin together (1/19, 5%), lysozyme (1/19, 5%) or unknown (3/19, 16%) . Of the 12 (58%) sera from patients with Wegener's granulomatosis who had cANCA, 7 bound to proteinase 3 . Antimyeloperoxidase antibodies were present in 14 of 45 (31%) sera with pANCA; other antigens were proteinase 3 (5/45, 11%), elastase (3/45, 78%), lactoferrin (1/45, 2%), cathepsin G (5/45, 11%) or unknown (17/45, 38%) . Antimyeloperoxidase antibodies were common in microscopic polyarteritis (6/14, 43%) and systemic vasculitis (5/16, 31%) . However, the majority of target antigens in systemic vasculitis and rheumatoid arthritis with pANCA were not determined . "Atypical" ANCA were present in four patients, one with inflammatory bowel disease (1/8, 13%) and three with SLE (3/15, 20%) . The specificities were cathepsin G, cathepsin G plus lactoferrin, or unknown in two sera . A recent report has suggested that bactericidal/permeability-increasing protein may be the target in patients with inflammatory bowel disease.

Free Radic Res, 1996 Jan, 24(1), 61 - 8
Potential role of the peroxidase-dependent metabolism of serotonin in lowering the polymorphonuclear leukocyte bactericidal function; Salman-Tabcheh S et al.; Serotonin (5-hydroxytryptamine, 5-HT) significantly and dose-dependently suppressed the luminol-enhanced chemiluminescence (CL) signal generated by polymorphonuclear leukocytes (PMN) activated with phorbol myristate acetate (PMA), but did not modify either lucigenin-enhanced CL or the reduction of superoxide dismutase-inhibitable cytochrome c . Moreover, stimulation of PMNs previously incubated with 5-HT resulted in a threefold increase in 5-HT equivalents bound to the proteins of PMN . The addition of catalase or sodium azide substantially reduced this binding . The present results suggest that 5-HT metabolism is mediated by H2O2 and myeloperoxidase (MPO) released by activated PMNs . Hence 5-HT could lower the bactericidal function of these cells by competition with hypochlorite formation from halides and MPO/H2O2.

Drugs, 1996, 51 Suppl 1, 20 - 30
Mechanism of action of streptogramins and macrolides; Vannuffel P et al.; Protein synthesis is catalysed by ribosomes and cytoplasmic factors . Bacterial ribosomes (70S) are made up of 2 subunits (50S and 30S) containing ribosomal RNA (rRNA) and ribosomal proteins: the 30S binds messenger RNA and begins the ribosomal cycle (initiation), whereas 50S binds transfer RNA (tRNA) derivatives and controls elongation . The key reaction, peptide bond formation, is promoted by the catalytic centre of 50S (the peptidyl transferase centre), and the growing peptide chain (peptidyl-tRNA) attached at the donor P site undergoes peptide linkage with an aminoacyl-tRNA at the acceptor A site . This reaction is inhibited by several antibiotics, the best known being chloramphenicol, and the macrolide-lincosamide-streptogramin (MLS) group . These inhibitors have a reversible action, except for streptogramins that are composed of A and B components, which are bacteriostatic alone, but bactericidal when combined . The peptidyl transferase centre has been identified at the 50S surface, and the binding sites of inhibitors have been mapped within this domain: some of these sites overlap (e.g . those of macrolides, and type B streptogramins, which compete for binding to ribosomes) . Chloramphenicol blocks the catalytic portion, and A streptogramins the substrate sites of the peptidyl transferase centre . Macrolides and type B streptogramins interfere with the formation of long polypeptides and cause a premature detachment of incomplete peptide chains . The synergism between types A and B streptogramins is due to induction by type A streptogramins of an increased ribosome affinity for type B streptogramins . Microbial resistance to antibiotics mainly involves inactivation of inhibitors and modification of targets (mutations of ribosomal proteins or rRNA genes) . Alterations of rRNA bases can induce resistance to a single inhibitor or to a group of antibiotics (e.g . MLSB) . The impact of resistance in chemotherapy is less important for streptogramins than for other inhibitors, because the synergistic effect of A and B streptogramins also applies to strains resistant to the MLSB group . It is proposed that mutations and modifications of rRNA bases induce conformational ribosomal changes that prevent antibiotics binding to the target . Conformational changes are also triggered by type A streptogramins: they are responsible for their synergism with type B streptogramins.

J R Soc Med, 1996 Jan, 89(1), 47P - 8P
ANCA associated with Behçet's disease; Burrows NP et al.; Behcet's disease is a multisystem disorder affecting primarily mucocutaneous and ocular sites although the gastrointestinal, cardiovascular, central nervous and respiratory systems may also be involved . Hulusi Behcet, a Turkish dermatologist, first described Behcet's disease in 1937 and suggested a possible infectious aetiology . The pathogenesis of this condition still remains unclear although it is likely that infection acts as a trigger in genetically susceptible individuals . We report a patient with unusual cutaneous manifestations of Behcet's disease and antineutrophil cytoplasmic antibodies (ANCA) directed against the cytotoxic protein, bactericidal/permeability-increasing protein (BPI) . This is the first report of Behcet's disease associated with this autoantibody.

Life Sci, 1996, 58(25), 2337 - 43
50-mile walking race suppresses neutrophil bactericidal function by inducing increases in cortisol and ketone bodies; Fukatsu A et al.; To examine the effect of intensive aerobic exercise on the interaction between endocrine and immune systems, we studied in ten normal healthy male subjects the effect of a 50-mile walking race on blood concentration of hormones (insulin, GH, ACTH, cortisol, adrenaline, noradrenaline, and dopamine), ketone bodies, specific immunological functions (IgG, IgM, and PHA/Con A-induced lymphocyte blastformation test), and nonspecific immune (CH50, and neutrophil bactericidal functions) . Neutrophil bactericidal activity was measured as chemiluminescences amplified by luciferin analog (CLA-DCL) and luminol (L-DCL) . The race increased cortisol and ketone bodies, and decreased insulin, CLA-DCL, and L-DCL (all parameters; P < 0.01) . However, other parameters were not significantly changed . There were significant negative correlations between changes of ketone bodies/cortisol and CLA/L-DCL (P < 0.05), however there was no significant correlations between changes of insulin and CLA/L-DCL . These data indicate that extensive aerobic exercise causes impaired neutrophil bactericidal function, probably due to the induced increases in both cortisol and ketone bodies . This impaired neutrophil function may cause the susceptibility to infection after an extensive exercise.

Can J Microbiol, 1996 Jan, 42(1), 60 - 5
Influence of peracetic acid on Escherichia coli H10407 strain in laboratory microcosms; Jolivet-Gougeon A et al.; The bactericidal properties of peracetic acid (PAA) were tested using Escherichia coli H10407, in sterilized artificial seawater, sewage effluent water, and distilled water microcosms . No LT enterotoxin synthesis was detected by GM1 enzyme-linked immunosorbent assay of the water supernatants, but a specific fragment of the eltB gene was always amplified by polymerase chain reaction for 21 days after PAA treatment . The resuscitation capacity of starved cells was assayed in rich medium and their inability to overcome the effects of PAA stress was observed, despite the emergence of viable but nonculturable cells in microcosms 24 or 48 h after treatment . Moreover, no obvious differences in response were obtained, concerning enterotoxigenesis, between bacteria subjected to osmotic and (or) nutrient starvation-induced stress with or without PAA treatment.

South Med J, 1996 Jan, 89(1), 68 - 70
Organism isolation and serum bactericidal titers in oral antibiotic therapy for pediatric osteomyelitis; Marshall GS et al.; Isolation of the etiologic organism and monitoring of serum bactericidal titers are considered necessary in oral antibiotic therapy for osteomyelitis . Over a 5-year period, 26 pediatric patients with hematogenous osteomyelitis were treated with sequential parenteraloral therapy . No organism was isolated in 9 cases, and bactericidal titers were determined in only 7 cases . All patients did well, raising questions about the conventional prerequisites invoked for oral treatment of skeletal infections.

J Infect Dis, 1996 Jan, 173(1), 252 - 5
Bactericidal/permeability-increasing protein, a lipopolysaccharide-specific protein on the surface of human peripheral blood monocytes; Dentener MA et al.; Bactericidal/permeability-increasing protein (BPI), a cationic protein present in the azurophilic granule and on the surface of polymorphonuclear leukocytes, specifically interacts with lipopolysaccharide (LPS) . This study demonstrates for the first time, using flow cytometry with specific anti-BPI monoclonal antibody (MAb), that human peripheral blood monocytes express BPI on their cell surface . The monocyte cell surface BPI was shown to bind to LPS, because binding of anti-BPI MAb 4E3 (which is known not to react with BPI to which LPS is bound) to cell surface BPI was strongly reduced after preincubation of cells with LPS . However, cell surface BPI did not quantitatively contribute to the interaction of LPS with the monocyte cell membrane, since preincubation of cells with 4E3 did not block binding of LPS-fluorescein isothiocyanate to monocytes . The origin of the monocyte cell surface BPI remains to be further elucidated.

Biochem Biophys Res Commun, 1995 Dec 26, 217(3), 972 - 8
Chlorogenic acid as a natural scavenger for hypochlorous acid; Kono Y et al.; The effect of chlorogenic acid (CGA) on the killing of Escherichia coli by hypochlorous acid (HOCI) was examined . CGA prevented E . coli from bactericidal action of HOCI in a concentration-dependent manner . HOCI reacted rapidly with CGA . By comparison with the one- and two-electron oxidation products produced by peroxidase and phenolase reactions, respectively, the main product was identified as o-quinone of CGA via the formation of o-semiquinone of CGA as an intermediate The quinone form of CGA reacted further with HOCI to yield unstable product(s).

Southeast Asian J Trop Med Public Health, 1995 Dec, 26(4), 639 - 41
Use of serum bactericidal activity in assessing previous antibiotic therapy in acute lower respiratory tract infection in children; Vasudevan G et al.; Bactericidal activity in sera of children with acute lower respiratory tract infection was assayed to determine its effect on the outcome of blood culture . Parental reporting of prior antibiotic therapy was also determined . 14.4% of samples without serum bactericidal activity yielded pathogens from blood culture, whereas only 2.4% of samples with serum bactericidal activity yielded pathogens . A statistically significant correlation was found between isolation of pathogens by blood culture and serum bactericidal activity . Parental reporting could not be relied upon as there was no positive correlation.

Int J Immunopharmacol, 1995 Dec, 17(12), 1027 - 31
Superoxide anion (O2-) production and bactericidal capacity of polymorphonuclear neutrophils obtained from patients subjected to glucagon test; Szczepanik A et al.; Superoxide anion (O2-) production and bactericidal capacity of morphologically mature bone marrow polymorphonuclear neutrophils (PMN) were evaluated in 30 haematologically normal individuals . These same parameters of peripheral PMNs were estimated in 15 healthy volunteers before and after glucagon-induced marrow granulocyte reserve mobilization . Bone marrow PMN in comparison with cells obtained from peripheral blood manifested impaired superoxide anion production and diminished bactericidal capacity . The admixture of bone marrow PMN released into the circulation by the use of glucagon administration significantly lowered both estimated PMN functions.

Immunol Lett, 1995 Dec, 48(3), 187 - 91
Monoclonal antibodies recognize 2300 years aged alkaline phosphatase; Etspuler H et al.; It was attempted to monitor the immunological response of monoclonal antibodies directed to human alkaline phosphatase in ancient Egyptian bones from the ptolemeic period . The intactness of the respective epitopes of the bone enzyme was successfully demonstrated in an ELISA . Fortunately, the mummified bone was not contaminated by fungi and bacteria due to the fungicidal and bactericidal reactivity of the ancient pretreatment employing resins of pistachio for mummification . The enzyme was enriched using gel chromatography, anion exchange and affinity chromatography to yield 310 +/- 7 mU/mg . The enzymically active fractions of the wheat-germ lectin affinity chromatography were subjected to ELISA . The best binding affinity was detected using the monoclonal antibody BAP A while the reactions of all the other four antibodies BAP B, BAP G, BAP 4A5 and BAP 5D4 were substantially diminished.

Aliment Pharmacol Ther, 1995 Dec, 9(6), 705 - 9
The influence of isolated doses of drugs, feeding and colonic bacterial ureolysis on urea breath test results; Perri F et al.; BACKGROUND: Aim of this work was to improve standardization of the 13C-urea breath test (13C-UBT) by evaluating the effect of drug administration, feeding and colonic ureolysis on the UBT results . METHODS: Three different studies were performed . First, a 13C-UBT was performed in 41 Helicobacter pylori infected subjects before and after 1 day of therapy with sucralfate (5 g), colloidal bismuth (600 mg), amoxycillin (2.5 g) or omeprazole (40 mg) . Second, a 13C-UBT was performed in 10 H . pylori-positive patients after an overnight fast and repeated 2 h after lunch . Finally, a 13C-UBT was carried out in seven healthy volunteers, with breath sampling prolonged to 6 h to investigate colonic bacterial ureolysis . RESULTS: Even a short course of drugs specific for H . pylori may result in a falsely negative 13C-UBT . Feeding does not interfere with the 13C-UBT in infected subjects . No significant 13C-urea breakdown by colonic bacteria is observed during the 13C-UBT when it is prolonged to 6 h . CONCLUSION: The 13C-urea breath test is a sensitive clinical tool for the non-invasive diagnosis of H . pylori infection . It is unaffected by feeding or colonic ureolysis . However, false negative results are likely even after 1 day of therapy with bactericidal, 'cytoprotective' or antisecretory drugs.

Proteins, 1995 Dec, 23(4), 588 - 90
Crystallization and preliminary X-ray crystallographic analysis of ImmE7 protein of colicin E7; Ku WY et al.; The ImmE7 protein, which can bind specifically to the DNase colicin E7 and neutralize its bactericidal activity, has been purified and crystallized in two different crystal forms by vapor diffusion method . The orthorhombic crystals belong to space group I222 or I2(1)2(1)2(1) and have unit cell dimensions a = 75.1 A, b = 50.5 A, and c = 45.4 A . The second form is monoclinic space group P2(1) with cell dimensions a = 29.3 A, b = 102.7 A, c = 53.0 A, and beta = 91.5 degrees . The orthorhombic crystals diffract to 1.8 A resolution, and are suitable for high-resolution X-ray analysis.

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