Antimicrobial

Ann Pharmacother . 2005 Jan 11; {Epub ahead of print}
Furazolidone-Induced Pulmonary Hypersensitivity (February); Kowalski TJ et al.; OBJECTIVE: To report a case of pulmonary hypersensitivity associated with furazolidone use and review the literature on this topic . CASE SUMMARY: A 43-year-old white female presented with fever and dyspnea . She had recently completed a course of furazolidone 125 mg 4 times daily for 10 days for enteritis . Investigations revealed bibasilar interstitial infiltrates on chest X-ray, hypoxia, and 21% eosinophilia . Her fever, hypoxia, and dyspnea rapidly abated following discontinuation of furazolidone and administration of corticosteroids . DISCUSSION: Furazolidone is a bactericidal agent used to treat infectious enteropathies . It is chemically similar to nitrofurantoin, which is well known to cause pulmonary hypersensitivity reactions . Application of the Naranjo probability scale suggests that a furazolidone adverse reaction in this patient was probable . A MEDLINE search from 1966 to October 2004 revealed 2 previously reported cases suggestive of furazolidone pulmonary hypersensitivity . All published reports closely resemble each other and descriptions of nitrofurantoin-associated pulmonary hypersensitivity reactions . CONCLUSIONS: Furazolidone may induce pulmonary hypersensitivity reactions; clinicians should be aware of this potentially serious adverse effect.

Haematologica, 2005 Jan, 90(1), 38 - 44
Biosynthetic profiles of neutrophil serine proteases in a human bone marrow-derived cellular myeloid differentiation model; Garwicz D et al.; BACKGROUND AND OBJECTIVES: Human leukocyte elastase, proteinase 3 and cathepsin G are neutrophil granule proteins belonging to the hematopoietic serine protease superfamily . In addition to their established roles in inflammation, they have recently been implicated as regulators of granulopoiesis and mediators of apoptosis . We set out to characterize the individual biosynthetic profiles of these proteins in a neutrophil differentiation model . DESIGN AND METHODS: CD34+CD38+ hematopoietic progenitor cells from 21 healthy human bone marrow donors were cultured in vitro in the presence of recombinant human granulocyte colony-stimulating factor (G-CSF) . Biosynthetic radiolabeling was performed in cells from 13 subjects after various periods of differentiation induction . Following protein extraction, the proteins were specifically immunoprecipitated from cell lysates and media and run in gel electrophoresis . Biosynthetic profiles of azurophil granule proteins, in particular members of the neutrophil serine protease family, were examined during myeloid differentiation . RESULTS: The onset of synthesis of myeloperoxidase, lysozyme, leukocyte elastase, and proteinase 3 occurred early after differentiation induction with G-CSF, while synthesis of cathepsin G, azurocidin, and bactericidal/permeability-increasing protein was detected somewhat later . Cathepsin G and proteinase 3 were retained intracellularly relatively efficiently, while leukocyte elastase and lysozyme were secreted to a greater extent . Cell morphology and positive immunocytochemistry for lactoferrin as well as flow cytometric analysis of selected surface antigens confirmed neutrophil-like maturation . INTERPRETATION AND CONCLUSIONS: We demonstrate that azurophil granule proteins, including proforms of human leukocyte elastase, proteinase 3 and cathepsin G, are constitutively secreted to various degrees during in vitro myeloid differentiation of human hematopoietic progenitor cells, in addition to being stored intracellularly in active forms . These findings suggest protein-specific sorting mechanisms and may have implications for the regulation of granulopoiesis.

Aquat Toxicol, 2005 Jan 18, 71(1), 73 - 84 Epub 2004 Nov 24.
'In vivo' effects of Bisphenol A in Mytilus hemocytes: modulation of kinase-mediated signalling pathways; Canesi L et al.; Endocrine disrupting chemicals (EDCs) include a variety of natural and synthetic estrogens, as well as estrogen-mimicking chemicals . We have previously shown that in the hemocytes of the mussel Mytilus galloprovincialis Lam . both natural and environmental estrogens in vitro can rapidly affect the phosphorylation state of components of tyrosine kinase-mediated cell signalling, in particular of mitogen activated protein kinases (MAPKs) and signal transducers and activators of transcription (STAT), that are involved in mediating the hemocyte immune response . These effects were consistent with the hypothesis that 'alternative' modes of estrogen action involving kinase-mediated pathways similar to those described in mammalian systems are also present in invertebrate cells . This possibility was investigated in vivo with Bisphenol A (BPA): mussels were injected with BPA and hemocytes sampled at 6, 12, and 24h post-injection . The results show that BPA (25nM nominal concentration in the hemolymph) lead to a significant lysosomal membrane destabilisation at all times post-injection, indicating BPA-induced stress conditions in the hemocytes, whereas lower concentrations were ineffective . BPA induced significant changes in the phosphorylation state of MAPK and STAT members, as evaluated by SDS-PAGE and WB of hemocyte protein extracts with specific antibodies, although to a different degree at different exposure times . In particular, BPA induced a dramatic decrease in phosphorylation of the stress-activated p38 MAPK, whose activation is crucial in mediating the bactericidal activity . Moreover, BPA decreased the phosphorylation of a CREB-like transcription factor (cAMP-responsive element binding protein) . The results demonstrate that BPA can affect kinase-mediated cell signalling in mussel hemocytes also in vivo, and suggest that EDCs may affect gene expression in mussel cells through modulation of the activity of transcription factors secondary to cytosolic kinase cascades . Overall, these data address the importance of investigating full range responses to EDCs in ecologically relevant marine invertebrate species.

Immunobiology, 2004, 209(8), 629 - 35
Inhibition of C5 or absence of C6 protects from sepsis mortality; Buras JA et al.; Inhibiting complement anaphlytoxin C5a during sepsis may prevent sepsis mortality . Although human anti-C5 antibodies exist, their therapeutic use in microbial sepsis has been avoided because of the hypothesis that inhibiting C5b will prevent formation of the bactericidal membrane attack complex (MAC) and worsen clinical outcome . We wished to test the hypothesis that inhibition of C5 would improve outcomes in sepsis . Sepsis was induced in rats by laparotomy and cecal ligation and puncture (CLP) by an IACUC-approved protocol . Sham animals underwent laparotomy without CLP . Following CLP rats were randomized to receive a single IV dose of purified IgG ant-C5 antibody (Ab) or control IgG Ab . Anti-C5 Ab treated rats (n = 20) had significantly lower mortality vs . controls (n = 21), 20% vs . 52% (P = 0.019, log-rank) . Analysis of bacterial load by culture of spleen and liver homogenates showed a reduction in colony forming units in anti-C5 Ab treated rats vs . control IgG (P = 0.003 and 0.009, respectively) . Anti-C5 treatment reduced lung injury as measured by total MPO content of lung tissue (P = 0.024) . Finally, rats genetically deficient in C6 production, unable to form MAC but capable of producing C5a and C5b, were protected from CLP-induced sepsis mortality . Our results show that in anti-C5 antibody therapy prevents CLP sepsis-induced mortality and improves lung injury . Inhibition of the complement MAC does not increase bacterial load or mortality, therefore, the use of anti-C5 therapy may be beneficial rather than detrimental in sepsis.

Int J Tuberc Lung Dis, 2004 Dec, 8(12), 1396 - 400
The WHO/IUATLD diagnostic algorithm for tuberculosis and empiric fluoroquinolone use: potential pitfalls; Sterling TR; According to the current WHO/IUATLD diagnostic algorithm for tuberculosis, to establish the diagnosis of smear-negative pulmonary disease, patients should first demonstrate no clinical response to a course of broad-spectrum antibiotics . The fluoroquinolones have broad-spectrum activity against respiratory pathogens and are generally considered first-line therapy for the treatment of community-acquired pneumonia; they also have bactericidal activity against Mycobacterium tuberculosis . Of note, empiric fluoroquinolone monotherapy has been associated with delays in the initiation of appropriate anti-tuberculosis therapy, and also resistance in M . tuberculosis . Delays in the diagnosis and treatment of tuberculosis are associated with increased morbidity and mortality . Resistance to fluoroquinolones in M . tuberculosis could limit the use of this potentially first-line class of anti-tuberculosis agents . The WHO/IUATLD diagnostic criteria for smear-negative tuberculosis should be revised to ensure that fluoroquinolones are not used inappropriately and that the detrimental effects of empiric fluoroquinolone monotherapy in tuberculosis patients are avoided.

J Immunol, 2005 Jan 15, 174(2), 595 - 9
Cutting Edge: Macrophage Inhibition by Cyclic AMP (cAMP): Differential Roles of Protein Kinase A and Exchange Protein Directly Activated by cAMP-1; Aronoff DM et al.; cAMP has largely inhibitory effects on components of macrophage activation, yet downstream mechanisms involved in these effects remain incompletely defined . Elevation of cAMP in alveolar macrophages (AMs) suppresses FcgammaR-mediated phagocytosis . We now report that protein kinase A (PKA) inhibitors (H-89, KT-5720, and myristoylated PKA inhibitory peptide 14-22) failed to prevent this suppression in rat AMs . We identified the expression of the alternative cAMP target, exchange protein directly activated by cAMP-1 (Epac-1), in human and rat AMs . Using cAMP analogs that are highly specific for PKA (N6-benzoyladenosine-3',5'-cAMP) or Epac-1 (8-(4-chlorophenylthio)-2'-O-methyladenosine-3',5'-cAMP), we found that activation of Epac-1, but not PKA, dose-dependently suppressed phagocytosis . By contrast, activation of PKA, but not Epac-1, suppressed AM production of leukotriene B(4) and TNF-alpha, whereas stimulation of either PKA or Epac-1 inhibited AM bactericidal activity and H(2)O(2) production . These experiments now identify Epac-1 in primary macrophages, and define differential roles of Epac-1 vs PKA in the inhibitory effects of cAMP.

Ann Agric Environ Med, 2004, 11(2), 227 - 31
Selected parameters of immunological response in hop growers during the period of intensive application of pesticides; Tokarska-Rodak M et al.; The aim of the study was determination of selected parameters of immunological response among hop growers and farmers in conditions of intensive exposure to means of plant protection . Survey data was collected from 238 males aged 25-70 living in the area of Wilkow near Pulawy (Lublin Region) . Control group were males from the area of Witoszyn (Lublin Region)--53 people aged 25-70 occupied mainly with land cultivation . Based on an environmental survey conducted among hop growers and farmers, the respondents were divided into 3 age groups: 25-40, 41-55 and 56-70 . Laboratory tests covered the determination of selected morphological parameters, phagocytic test, NBT test, and myeloperoxidasis (MPO) concentration in blood serum of hop growers and farmers.A significant decrease was noted in the number of platelets in the general population of hop growers and in individual age groups, compared to the control groups of farmers . Analysis of individual sub-populations of leukocytes showed a significantly higher number statistically of basophils and lymphocytes among hop growers, compared to farmers . A detailed analysis of the degree of phagocytic and bactericidal activity of neutrophils allowed us to presume that during the period of spraying there occurred a mobilisation of the granulocytic system, manifested by the presence of over 90% of neutrophils of intensified phagocytic activity, and 20% of neutrophils of intensified bactericidal activity . The preparations prepared by the routine NBT test method were analysed with the use of LUCIA computer programme (version 4.51) . The analysis of the level of MPO in blood serum in the populations examined showed the presence of statistically significant differences . In hop growers, the MPO level was significantly higher statistically (60.0 ng/ml), compared to the control group of farmers (43.4 ng/ml).

Proc Natl Acad Sci U S A, 2005 Jan 11, 102(2), 467 - 72 Epub 2004 Dec 30.
S-nitroso proteome of Mycobacterium tuberculosis: Enzymes of intermediary metabolism and antioxidant defense; Rhee KY et al.; The immune response to Mycobacterium tuberculosis (Mtb) includes expression of nitric oxide (NO) synthase (NOS)2, whose products can kill Mtb in vitro with a molar potency greater than that of many conventional antitubercular agents . However, the targets of reactive nitrogen intermediates (RNIs) in Mtb are unknown . One major action of RNIs is protein S-nitrosylation . Here, we describe, to our knowledge, the first proteomic analysis of S-nitrosylation in a whole organism after treating Mtb with bactericidal concentrations of RNIs . The 29 S-nitroso proteins identified are all enzymes, mostly serving intermediary metabolism, lipid metabolism, and/or antioxidant defense . Many are essential or implicated in virulence, including defense against RNIs . For each of two target enzymes tested, lipoamide dehydrogenase and mycobacterial proteasome ATPase, S-nitrosylation caused enzyme inhibition . Moreover, endogenously biotinylated proteins were driven into mixed disulfide complexes . Targeting of metabolic enzymes and antioxidant defenses by means of protein S-nitrosylation and mixed disulfide bonding may contribute to the antimycobacterial actions of RNIs.

Infect Immun, 2005 Jan, 73(1), 193 - 200
Acute-phase concentrations of lipopolysaccharide (LPS)-binding protein inhibit innate immune cell activation by different LPS chemotypes via different mechanisms; Hamann L et al.; The chain length of bacterial lipopolysaccharide (LPS) is a crucial factor for host-pathogen interaction during bacterial infection . While rough (R)-type and smooth (S)-type LPSs have been shown to differ in their ability to interact with the bactericidal/permeability-increasing protein, little is known about the differential mode of interaction with the acute-phase reactant LPS-binding protein (LBP) . At lower concentrations, LBP catalyzes the binding of LPS to CD14 and enhances LPS-induced cellular activation via Toll-like receptor 4 . In humans, however, concentrations of LBP in serum increase during an acute-phase response, and these LBP concentrations exhibit inhibitory effects in terms of cellular activation . The mechanisms of inhibition of LPS effects by LBP are not completely understood . Here, we report that human high-dose LBP (hd-LBP) suppresses binding of both R-type and S-type LPS to CD14 and inhibits LPS-induced nuclear translocation of NF-kappaB, although cellular uptake of R-type LPS was found to be increased by hd-LBP . In contrast, we found that hd-LBP enhanced the binding and uptake of S-type LPS only under serum-free conditions, whereas in the presence of serum, hd-LBP inhibited cellular binding and uptake . This inhibitory effect of serum could be mimicked by the addition of purified high-density lipoprotein (HDL) to serum-free medium, indicating an LBP-mediated transfer of preferentially S-type LPS to plasma lipoproteins such as HDL . A complete understanding of the host's mechanisms to modulate the proinflammatory effects of LPS will most likely help in the understanding of inflammation and infection and may lead to novel therapeutic intervention strategies.

J Perinatol . 2004 Dec 09; {Epub ahead of print}
Neutrophil Function in Neonates Born to Gestational Diabetic Mothers; Mehta R et al.; OBJECTIVE:: To examine neutrophil functional activity in the cord blood of term neonates born to gestational diabetic mothers, in association with the type of diabetes and the development of neonatal hypoglycemia . METHODS:: Neutrophil chemotaxis, random motility, and chemiluminescence was evaluated in the cord blood of 30 healthy term neonates: 12 were born to gestational diabetic mothers who received no-insulin (GDM-NI), eight to gestational diabetic mothers who received insulin (GDM), and 10 to mothers without diabetes (neonatal controls) . In addition, the neutrophil functional activity in the peripheral venous blood of 10 healthy adults was analyzed . RESULTS:: Neutrophil functional activity in the cord blood of the neonates with and without maternal gestational diabetes was significantly lower than that in adults . As compared to neonatal controls, neonates born to both groups of GDM had decreased chemotaxis, random motility, and chemiluminescence (GDM-NI: 52.8+/-2.1 mum, p<0.001, 42.1+/-4.4 mum, p<0.001, and 140.1+/-6.9 counts per minute (cpm) x 10(3), p<0.01, respectively, and GDM: 53.0+/-1.9 mum, p<0.01, 41.8+/-4.0 mum, p<0.001, and 143.0+/-6.8 cpm x 10(3), p<0.01, respectively) . Unlike controls, a tight correlation was identified between the tested neutrophil parameters in the cord blood of neonates born to diabetic mothers (r=0.70 to 0.91) . The prevalence of hypoglycemia after birth was almost equal (50.0 to 41.7%) in the two groups of neonates born to diabetic mothers . There were differences in the neutrophil functional activity in the cord blood of the neonates with and without hypoglycemia . CONCLUSION:: Maternal gestational diabetes leads to impairment of cord blood neutrophil motility and postphagocytic bactericidal capacity independently from the insulin requirements for the maintenance of normoglycemia during pregnancy.Journal of Perinatology advance online publication, 9 December 2004; doi:10.1038/sj.jp.7211241.

Acta Dermatovenerol Croat, 2004, 12(4), 294 - 313
Antineutrophil Cytoplasmic Antibodies (ANCA): Diagnostic Utility and Potential Role in the Pathogenesis of Vasculitis; Malenica B et al.; Antineutrophil cytoplasmic antibodies (ANCA) are a heterogeneous group of circulating antibodies directed toward the cytoplasmic constituents of neutrophils and monocytes . ANCA have been described in various diseases including idiopathic systemic vasculitides, connective tissue diseases, inflammatory bowel diseases, autoimmune liver diseases, infectious diseases, and some drugs . ANCA recognize different target antigens such as proteinase 3 (PR3-ANCA), myeloperoxidase (MPO-ANCA), cathepsin G, lactoferrin, bactericidal/permeability-increasing protein (BPI), and some others . However, only PR3-ANCA and MPO-ANCA are closely associated with systemic vasculitides, in particular Wegener's granulomatosis, microscopic polyangiitis and its renal limited manifestation, and Churg-Strauss syndrome . Both in vitro and in vivo experimental data strongly support a pathogenic role for ANCA in vasculitis and glomerulonephritis.

Clin Orthop, 2004 Dec, (429), 33 - 8
Titanium surface with biologic activity against infection; Parvizi J et al.; Despite immense improvements, periprosthetic infection continues to compromise the result of otherwise successful joint arthroplasty . There are various limitations in the treatment of periprosthetic infection, the most important of which is the inability to deliver antibiotics to the local tissue without the need for intravenous administration . We have developed a novel route to covalently tether vancomycin to a metal (titanium) surface, which showed effective bactericidal activity because of a vancomycin coupling . The chemistry of tethering does not affect the biological activity of the biofactors that are attached to the metal surface . This technology holds great promise for the manufacturing of "smart" implants that can be self protective against periprosthetic infection, or can be used for the treatment of periprosthetic infections when they occur.

Appl Environ Microbiol, 2004 Dec, 70(12), 7093 - 102
Biosynthetic gene cluster of the herbicide phosphinothricin tripeptide from Streptomyces viridochromogenes Tü494; Schwartz D et al.; The antibiotic phosphinothricin tripeptide (PTT) consists of two molecules of L-alanine and one molecule of the unusual amino acid phosphinothricin (PT) which are nonribosomally combined . The bioactive compound PT has bactericidal, fungicidal, and herbicidal properties and possesses a C-P-C bond, which is very rare in natural compounds . Previously uncharacterized flanking and middle regions of the PTT biosynthetic gene cluster from Streptomyces viridochromogenes Tu494 were isolated and sequenced . The boundaries of the gene cluster were identified by gene inactivation studies . Sequence analysis and homology searches led to the completion of the gene cluster, which consists of 24 genes . Four of these were identified as undescribed genes coding for proteins that are probably involved in uncharacterized early steps of antibiotic biosynthesis or in providing precursors of PTT biosynthesis (phosphoenolpyruvate, acetyl-coenzyme A, or L-alanine) . The involvement of the genes orfM and trs and of the regulatory gene prpA in PTT biosynthesis was analyzed by gene inactivation and overexpression, respectively . Insight into the regulation of PTT was gained by determining the transcriptional start sites of the pmi and prpA genes . A previously undescribed regulatory gene involved in morphological differentiation in streptomycetes was identified outside of the left boundary of the PTT biosynthetic gene cluster.

Nippon Jinzo Gakkai Shi, 2004 Oct, 46(7), 700 - 8
{Innate immunity in hemodialysis and continuous ambulatory peritoneal dialysis patients: impaired cytokine synthetic response to ex-vivo stimuli in mononuclear cells}; Shibuya A et al.; Dialysis patients are weak in immune host defense, which is associated with their high morbidity of infection . Polymorphonuclear leukocytes (PMNLs) and mononuclear cells play a key role in innate host defense . PMNLs and monocytes have bactericidal activity through the process of phagocytosis . Monocytes and lymphocytes contribute to the development of innate immunity by their cytokine actions . We studied the intracellular cytokine syntheses in response to ex-vivo stimuli, which may reflect the potential reactivity of immune cells in cytokine syntheses when pathogens invade humans . Furthermore, phagocytic activity was assessed in granulocytes and monocytes . Twenty HD, 15 CAPD, and 10 age-matched controls were enrolled in this study . One milliliter of whole blood from each subject was incubated with lipopolysaccharides (LPS) or mitogens for 4 hours at 37 degrees C . Monoclonal antibodies to CD14+ and CD4+ were used for identifying monocytes and helper T cells, respectively . Intracellular cytokines were stained using FASTIMMUNE staining kits . Interleukin-1beta and TNF-alpha syntheses were examined in monocytes, which are the most important early-response cytokines in innate immunity . IFN-gamma and IL-4 syntheses were examined in helper T cells to observe their polarization into Th1 and Th2 cells . IFN-gamma is a key factor in establishing innate immunity . The percentage of cells that stained positive for each cytokine was analyzed using a flow cytometer . The following results were obtained: 1) In CAPD patients, IL-1beta and TNF-alpha response to LPS in monocytes were significantly reduced, as compared to other subjects . Polarization of helper T cells was reduced, resulting in a significant decrease in Th1 cells . 2) In HD patients, monokine responses were not altered, but polarization of helper T cells was skewed toward a Th1 type . Phagocytic activities were not impaired in both dialysis groups . In conclusion, mononuclear cells from CAPD patients have the potential to exhibit failure of a cytokine response to ex-vivo stimuli in terms of innate immunity.

Infect Immun, 2004 Dec, 72(12), 7265 - 74
Characterization of immunodominant and potentially protective epitopes of Mannheimia haemolytica serotype 1 outer membrane lipoprotein PlpE; Ayalew S et al.; Mannheimia haemolytica serotype 1 (S1) is the most common bacterial isolate found in shipping fever pneumonia in beef cattle . Currently used vaccines against M . haemolytica do not provide complete protection against the disease . Research with M . haemolytica outer membrane proteins (OMPs) has shown that antibodies to one particular OMP from S1, PlpE, may be important in immunity . In a recently published work, members of our laboratory showed that recombinant PlpE (rPlpE) is highly immunogenic when injected subcutaneously into cattle and that the acquired immunity markedly enhanced resistance to experimental challenge (A . W . Confer, S . Ayalew, R . J . Panciera, M . Montelongo, L . C . Whitworth, and J . D . Hammer, Vaccine 21:2821-2829, 2003) . The objective of this work was to identify epitopes of PlpE that are responsible for inducing the immune response . Western blot analysis of a series of rPlpE with nested deletions on both termini with bovine anti-PlpE hyperimmune sera showed that the immunodominant region is located close to the N terminus of PlpE . Fine epitope mapping, in which an array of overlapping 13-mer synthetic peptides attached to a derivatized cellulose membrane was probed with various affinity-purified anti-PlpE antibodies, identified eight highly reactive regions, of which region 2 (R2) was identified as the specific epitope . The R2 region is comprised of eight imperfect repeats of a hexapeptide (QAQNAP) and is located between residues 26 and 76 . Complement-mediated bactericidal activity of affinity-purified anti-PlpE bovine antibodies confirmed that antibodies directed against the R2 region are effective in killing M . haemolytica.

Infect Immun, 2004 Dec, 72(12), 7172 - 82
Distinct roles of reactive nitrogen and oxygen species to control infection with the facultative intracellular bacterium Francisella tularensis; Lindgren H et al.; Reactive nitrogen species (RNS) and reactive oxygen species (ROS) are important mediators of the bactericidal host response . We investigated the contribution of these two mediators to the control of infection with the facultative intracellular bacterium Francisella tularensis . When intradermally infected with the live vaccine strain F . tularensis LVS, mice deficient in production of RNS (iNOS(-/-) mice) or in production of ROS by the phagocyte oxidase (p47(phox-/-) mice) showed compromised resistance to infection . The 50% lethal dose (LD(50)) for iNOS(-/-) mice was <20 CFU, and the LD(50) for p47(phox-/-) mice was 4,400 CFU, compared to an LD(50) of >500,000 CFU for wild-type mice . The iNOS(-/-) mice survived for 26.4 +/- 1.8 days, and the p47(phox-/-) mice survived for 10.1 +/- 1.3 days . During the course of infection, the serum levels of gamma interferon (IFN-gamma) and interleukin-6 were higher in iNOS(-/-) and p47(phox-/-) mice than in wild-type mice . Histological examination of livers of iNOS(-/-) mice revealed severe liver pathology . Splenocytes obtained 5 weeks after primary infection from antibiotic-treated iNOS(-/-) mice showed an in vitro recall response that was similar in magnitude and greater secretion of IFN-gamma compared to cells obtained from wild-type mice . In summary, mice lacking expression of RNS or ROS showed extreme susceptibility to infection with F . tularensis LVS . The roles of RNS and ROS seemed to be distinct since mice deficient in production of ROS showed dissemination of infection and died during the early phase of infection, whereas RNS deficiency led to severe liver pathology and a contracted course of infection.

Rheumatology (Oxford) . 2004 Nov 16; {Epub ahead of print}
Low seroprevalence and poor specificity of antineutrophil cytoplasmic antibodies in tuberculosis; Teixeira L et al.; Objectives . Recently published findings suggested that antineutrophil cytoplasmic antibodies (ANCA), particularly those with a cytoplasmic (C-ANCA) labelling pattern and targeting proteinase 3 (anti-PR3), might be markers of tuberculosis (TB) . This is a critical issue, because C-ANCA/anti-PR3 were considered to be a highly specific hallmark of Wegener's granulomatosis or microscopic polyangiitis and because TB may clinically mimic Wegener's granulomatosis . We therefore undertook a study with the aim of investigating further the prevalence and specificity of ANCA in TB . Methods . We evaluated serum samples from 67 patients diagnosed with culture-proven TB and 10 previously untested control samples from patients known to be ANCA positive (four Wegener's granulomatosis and two microscopic polyangiitides) or negative . All 77 sera were screened for ANCA using commercially available indirect immunofluorescence (IIF) and enzyme-linked immunosorbent assay (ELISA) for anti-PR3 and antimyeloperoxidase (MPO) . IIF-positive and anti-PR3- and anti-MPO-negative sera were also tested for bactericidal/permeability-increasing protein, lactoferrin, elastase and cathepsin G specificities with commercially available ELISA . Results . IIF detected ANCA in seven (10%) of the TB sera, including three C-ANCA and four atypical perinuclear-labelling ANCA . Only one IIF-negative specimen was anti-PR3 positive in ELISA . ANCA testing of the control sera yielded IIF and ELISA results concordant with previous findings, except for one borderline ELISA . Conclusion . Our results indicate that TB is associated with low ANCA seroprevalence and poor specificity, with no test serum showing combined C-ANCA/anti-PR3 activity . In a clinical setting of Wegener's granulomatosis/TB mimicry, such combined reactivity would seem to be more suggestive of Wegener's granulomatosis.

Clin Infect Dis, 2004 Nov 15, 39(10), 1425 - 30 Epub 2004 Oct 25.
The influence of human N-acetyltransferase genotype on the early bactericidal activity of isoniazid; Donald PR et al.; The elimination of isoniazid is subject to the influence of the N-acetyltransferase 2 (NAT2) genotype, and individuals may be homozygotic slow, heterozygotic fast, or homozygotic fast acetylators of isoniazid . The early bactericidal activity (EBA) of an antituberculosis agent can be determined by quantitative culture of Mycobacterium tuberculosis in sputum samples obtained from patients with pulmonary tuberculosis during the first days of treatment . In these studies, the EBA of isoniazid during the first 2 days of treatment was determined for 97 patients with sputum smear-positive pulmonary tuberculosis following isoniazid doses of < or =37.5 mg, 75 mg, 150 mg, 300 mg, and 600 mg . The NAT2 genotype was determined in 70 patients, and the association between EBA and genotype was examined in this subgroup . Similarly, the relationship between EBA and isoniazid serum concentration was evaluated in 87 patients . The mean EBA of isoniazid increased with dose, but it levelled off between doses of 150 mg (mean EBA, 0.572) and 300 mg (mean EBA, 0.553) . Significant differences were found in the mean EBA of isoniazid between the homozygous slow acetylator group and the heterozygous fast acetylator group and between the homozygous slow acetylator group and the homozygous fast acetylator group, but not between the heterozygous fast acetylator group and the homozygous fast acetylator group . The EBA appeared to reach a maximum at a 2-h isoniazid concentration of 2-3 microg/mL . These data confirm a significant effect of NAT2 genotype on the EBA of isoniazid over a range of doses.

Immunopharmacol Immunotoxicol, 2004 Aug, 26(3), 411 - 24
Immunostimulatory role of tryptic digest of Abrus agglutinin; Tripathi S et al.; Many bioactive peptides are in therapeutic use as immunomodulators at present . The origin of these bioactive peptides is diverse . Such bioactive peptides are reported to be present in enzymatic digest of food and milk proteins . In our previous work we have reported that Abrus agglutinin retains its bioactivity even after heat denaturation but loses its haemagglutination properties . This leads to the supposition that immunostimulatory regions in the protein might be responsible for its in vivo and in vitro stimulatory properties . Thus the bioactivity of tryptic digest of Abrus agglutinin (TDA) was checked in vitro to ascertain the presence of some bioactive region in the protein, which will lead to the discovery of certain immunostimulatory peptides which might be of use in nonspecific immunotherapy . In this study it is observed that TDA stimulates macrophage increasing the phagocytic and bactericidal activity as well as hydrogen peroxide production . TDA also proliferates splenocytes leading to Th1 response and NK cell activation.

Clin Infect Dis . 2004 Oct 25;39(10) {Epub ahead of print}
The Influence of Human N-Acetyltransferase Genotype on the Early Bactericidal Activity of Isoniazid; Donald PR et al.; The elimination of isoniazid is subject to the influence of the N-acetyltransferase 2 (NAT2) genotype, and individuals may be homozygotic slow, heterozygotic fast, or homozygotic fast acetylators of isoniazid . The early bactericidal activity (EBA) of an antituberculosis agent can be determined by quantitative culture of Mycobacterium tuberculosis in sputum samples obtained from patients with pulmonary tuberculosis during the first days of treatment . In these studies, the EBA of isoniazid during the first 2 days of treatment was determined for 97 patients with sputum smear-positive pulmonary tuberculosis following isoniazid doses of </=37.5 mg, 75 mg, 150 mg, 300 mg, and 600 mg . The NAT2 genotype was determined in 70 patients, and the association between EBA and genotype was examined in this subgroup . Similarly, the relationship between EBA and isoniazid serum concentration was evaluated in 87 patients . The mean EBA of isoniazid increased with dose, but it levelled off between doses of 150 mg (mean EBA, 0.572) and 300 mg (mean EBA, 0.553) . Significant differences were found in the mean EBA of isoniazid between the homozygous slow acetylator group and the heterozygous fast acetylator group and between the homozygous slow acetylator group and the homozygous fast acetylator group, but not between the heterozygous fast acetylator group and the homozygous fast acetylator group . The EBA appeared to reach a maximum at a 2-h isoniazid concentration of 2-3 mu g/mL . These data confirm a significant effect of NAT2 genotype on the EBA of isoniazid over a range of doses.

Antimicrob Agents Chemother, 2004 Nov, 48(11), 4103 - 12
Antipneumococcal activities of two novel macrolides, GW 773546 and GW 708408, compared with those of erythromycin, azithromycin, clarithromycin, clindamycin, and telithromycin; Matic V et al.; The MICs of GW 773546, GW 708408, and telithromycin for 164 macrolide-susceptible and 161 macrolide-resistant pneumococci were low . The MICs of GW 773546, GW 708408, and telithromycin for macrolide-resistant strains were similar, irrespective of the resistance genotypes of the strains . Clindamycin was active against all macrolide-resistant strains except those with erm(B) and one strain with a 23S rRNA mutation . GW 773546, GW 708408, and telithromycin at two times their MICs were bactericidal after 24 h for 7 to 8 of 12 strains . Serial passages of 12 strains in the presence of sub-MICs yielded 54 mutants, 29 of which had changes in the L4 or L22 protein or the 23S rRNA sequence . Among the macrolide-susceptible strains, resistant mutants developed most rapidly after passage in the presence of clindamycin, GW 773546, erythromycin, azithromycin, and clarithromycin and slowest after passage in the presence of GW 708408 and telithromycin . Selection of strains for which MICs were >/=0.5 microg/ml from susceptible parents occurred only with erythromycin, azithromycin, clarithromycin, and clindamycin; 36 resistant clones from susceptible parent strains had changes in the sequences of the L4 or L22 protein or 23S rRNA . No mef(E) strains yielded resistant clones after passage in the presence of erythromycin and azithromycin . Selection with GW 773546, GW 708408, telithromycin, and clindamycin in two mef(E) strains did not raise the erythromycin, azithromycin, and clarithromycin MICs more than twofold . There were no change in the ribosomal protein (L4 or L22) or 23S rRNA sequences for 15 of 18 mutants selected for macrolide resistance; 3 mutants had changes in the L22-protein sequence . GW 773546, GW 708408, and telithromycin selected clones for which MICs were 0.03 to >2.0 microg/ml . Single-step studies showed mutation frequencies <5.0 x 10(-10) to 3.5 x 10(-7) for GW 773546, GW 708408, and telithromycin for macrolide-susceptible strains and 1.1 x 10(-7) to >4.3 x 10(-3) for resistant strains . The postantibiotic effects of GW 773546, GW 708408, and telithromycin were 2.4 to 9.8 h.

Bull Exp Biol Med, 2003 Dec, 136(6), 548 - 50
Dependence of the nonspecific resistance in cows from their physiological and clinical state; Gugushvili NN; In cows phagocytic, bactericidal, and lysozyme activity increased with increasing pregnancy term, decreased by the 2nd day after labor, and increased in the follow-up period . The nonspecific resistance decreased most significantly in the winter-spring period.

Free Radic Res, 2004 Aug, 38(8), 805 - 11
Anti-inflammatory effects of Melaleuca alternifolia essential oil on human polymorphonuclear neutrophils and monocytes; Caldefie-Chezet F et al.; OBJECTIVE AND DESIGN: The fungicidal and bactericidal actions of the essential oil (EO) of Melaleuca alternifolia seem well established, but their anti-inflammatory and anti-oxidative effects remain unclear . In this study, we investigated in vitro the possible role of whole M . alternifolia EO as a modulator of the oxidative response, i.e . reactive oxygen species (ROS) production, of leukocytes (monocytes and polymorphonuclear neutrophils (PMNs)) in humans . METHODS: Whole blood leukocytes from healthy human volunteers (n = 7), isolated from erythrocytes by haemolytic shock, were incubated for 30 min with M . alternifolia EO (0-0.1%) to determine their ROS production by flow cytometry with or without stimulation of cells . We compared the effects of 3 different stimulating agents acting differently on transductional pathways to stimulate the ROS production: a phorbol ester (PMA), formyl-methionyl-leucyl-phenylalanine (fMLP) and opsonised zymosan (OZ) . RESULTS: As attested by the Kruskall-Wallis test, M . alternifolia EO at 0.1% directly stimulated ROS production by PMNs (x 8.7 vs . 0% EO, p < 0.05) and increased the intracellular ROS produced by monocytes . Whichever the stimulating agent used (PMA, fMLP or OZ), M . alternifolia EO decreased the intracellular ROS production at the dilution of 0.1% by PMNs and monocytes, more so with PMNs . CONCLUSION: M . alternifolia EO may be both a direct active mediator of the bactericidal action of the circulating leukocytes and may be efficient in protecting the organism from an excess of ROS, through an anti-oxidant and radical scavenging activity.

Shock, 2004 Nov, 22(5), 460 - 6
Innate immune response in Th1- and Th2-dominant mouse strains; Watanabe H et al.; C57BL/6 and BALB/c mice are prototypical Th1- and Th2-type mouse strains, respectively . In the present study, we attempted to characterize the innate immune response of macrophages from these mouse strains . Macrophages from C57BL/6 mice produced higher levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-12 than those from BALB/c mice after stimulation with macrophage-activating lipopeptide-2 (MALP-2, a synthetic TLR-2 ligand) or lipopolysaccharide (LPS, a TLR-4 ligand) . The augmented IL-12 production by C57BL/6 macrophages increased interferon-gamma and, in contrast, decreased IL-13 production by CD4+ T cells . On stimulation with MALP-2 or LPS, C57BL/6 macrophages produced lysosomal enzyme and nitric oxide, effector molecules for bacterial killing, whereas BALB/c macrophages did not . Bactericidal activity of BALB/c macrophages was impaired relative to C57BL/6 macrophages when cells were infected with live bacteria in vitro . In a murine model of septic peritonitis induced by cecal ligation and puncture (CLP), BALB/c mice failed to facilitate bacterial clearance relative to C57BL/6 mice despite an augmented peritoneal leukocyte infiltration that was associated with increased peritoneal levels of cytokines/chemokines . BALB/c mice exhibited increased plasma and hepatic levels of cytokines/chemokines, resulting in an exaggerated systemic inflammation as determined by acute-phase proteins . Finally, BALB/c mice were vulnerable to CLP-induced lethality relative to C57BL/6 mice . Altogether, innate immune response of macrophages is different between these mouse strains, which may affect the development of Th1 and Th2 adaptive immunity in these strains . Reduced systemic inflammatory response in C57BL/6 mice that may result from an eminent local response appears to be beneficial during sepsis.

Shock, 2004 Nov, 22(5), 438 - 45
Effects of burn serum on myocardial inflammation and function; Horton JW et al.; Large cutaneous burns are clearly recognized to produce acute myocardial contractile dysfunction . This study used a model of burn serum challenge in either primary cardiomyocyte cultures or isolated perfused hearts to examine several aspects of burn-serum-related contractile dysfunction as well as myocardial inflammatory responses . Despite the absence of detectable LPS in burn serum, pretreating isolated cells or perfused hearts with recombinant bactericidal permeability-increasing protein (rBPI21) prevented both the inflammatory cytokine cascade and the cardiac contractile dysfunction induced by burn serum treatment of myocytes or ventricular muscle preparations . Our finding that anti-TNF strategies applied to isolated myocytes or hearts before burn serum challenge prevented myocardial inflammation and contractile dysfunction suggested that LPS or LPS-like factors may require the action of second messengers such as TNF-alpha and IL-1beta to mediate LPS-related myocardial depressant effects . Our finding that experimental approaches neutralizing circulating LPS provided cardioprotection suggested that bacterial endotoxin or LPS-like molecules contribute, in part, to burn-related myocardial contractile dysfunction.

Expert Rev Anti Infect Ther, 2003 Jun, 1(1), 141 - 55
Early bactericidal activity of antituberculosis agents; Donald PR et al.; The early bactericidal activity (EBA) of an antituberculosis agent is arbitrarily defined as the fall in log(10) colony forming units (cfu) of Mycobacterium tuberculosis per ml sputum per day during the first 2 days of treatment . Determining the EBA is an important preliminary step in the clinical evaluation of an antituberculosis agent . We review the results of eight published studies of the EBA of different antituberculosis agents, the impact of these results on our understanding of the actions of the respective agents, the clinical characteristics and sputum findings of patients included in these studies, and explore sources of variation in the EBA results . Patients in these studies had a mean age of 31-36 years, a mean weight of 50-57 kg, 67% were male and 56% had lung involvement covering an area of more than one lung, and 90% had multicavitary disease . None of these findings were related to EBA in any study . The mean log(10) cfu per ml sputum in the first specimen was 6.474 . This was related to radiological extent of disease and cavity size in one study (p < 0.001) and, in the case of isoniazid to EBA with a rise in EBA of 0.094 (95% CL 0.029-0.158) for each tenfold rise in cfu counts/ml sputum . The overall variation in EBA in these studies was 0.0303, that due to laboratory processing of specimens was 0.0011, and due to patient characteristics and sputum sampling 0.0212 . The EBA is a reproducible investigation that has contributed significantly to our knowledge of the actions and characteristics of both established and new antituberculosis agents . The greatest source of variation in EBA results appears to be that due to interpatient variation in disease characteristics and sputum sampling.

Cytokine, 2004 Nov 7, 28(3), 101 - 8
Splenectomy differentially influences immune responses in various tissue compartments of the body; Shih-Ching K et al.; Patients without spleens have an increased risk of infection . Previous studies have shown that splenectomy (Spx) influences Kupffer cells (KC), peritoneal macrophages (pMphi) and alveolar macrophages (aMphi) phagocytosis and bactericidal activity . This study examined the effect of Spx on the cytokine production by peripheral blood monocular cells (PBMC), KC, aMphi, pMphi and cells from mesenteric lymph nodes (MLN) . We also determined if Spx influences survival following sepsis induced by cecal ligation and puncture (CLP) . C57BL/6J male mice (8-10 weeks old) underwent sham operation or Spx . Two weeks after sham or Spx, KC, pMphi, aMphi, PBMC and cells from MLN were isolated and stimulated with LPS . Cell-free supernatants were analyzed for TNF-alpha, IL-6, and IL-10 . A significant increase in KC, pMphi and aMphi TNF-alpha and IL-6 release was observed following Spx . The production of IL-10 was also significantly higher in KC under those conditions . In contrast, the release of TNF-alpha, IL-6 and IL-10 was significantly decreased in PBMC after Spx . Similarly, TNF-alpha and IL-6 was also decreased in MLN after Spx . Overall survival after CLP was not different in mice subjected to either sham or Spx . However, the mean time to death was significantly decreased in mice subjected to Spx compared to sham injured mice . These findings suggest that Spx modulates immune cell functions in various tissue compartments of the body and results in early deaths following sepsis.

Endocr Res, 2004 May, 30(2), 205 - 13
The prevalence and target antigens of antithyroid drugs induced antineutrophil cytoplasmic antibodies (ANCA) in Chinese patients with hyperthyroidism; Gao Y et al.; OBJECTIVE: Antithyroid drugs such as propylthiouracil (PTU) and methimazole (MMI) are common medications in Chinese patients with hyperthyroidism and PTU-induced antineutrophil cytoplasmic antibody (ANCA) positive vasculitis has been reported . The current cross-sectional study aimed to investigate the prevalence and the target antigens of ANCA in Chinese patients with hyperthyroidism pre- and post-antithyroid medication therapy . METHODS: Sera from 216 patients with hyperthyroidism in our hospital were collected from January to July in 2002 . Patients were divided into four groups: untreated (n = 34); treated with PTU (n = 62); treated with MMI (n = 77); and treated with both PTU and MMI (n = 43) . Indirect immunofluorescence (IIF) assay was used to detect ANCA and ANA . Antigen-specific ELISAs were used to detect antigen specificities . The known antigens included myeloperoxidase (MPO), proteinase 3 (PR3), human leukocyte elastase (HLE), lactoferrin, bactericidal/permeability-increasing protein (BPI), cathepsin G and azurocidin . RESULTS: 33/216 sera were IIF positive, 20 of the 33 samples were ANCA positive, 11 samples were ANA positive, and two samples were both P-ANCA and ANA positive . The prevalence of positive ANCA in patients receiving PTU (14/62, 22.6%) was significantly higher than that of untreated patients (1/34, 2.9%) and patients treated with MMI (0/77, 0), P < 0.017 . Of the 22 IIF-ANCA positive samples, 12 (54.5%) sera recognized lactoferrin, seven (31.8%) sera recognized HLE, four sera recognized MPO and azurocidin respectively, three sera recognized PR3 and cathepsin G respectively, and one serum recognized BPI . Six of the 22 (27.3%) patients with ANCA positive had clinical evidence of vasculitis . All patients with MPO-ANCA and two of the three patients with PR3-ANCA had clinical vasculitis . CONCLUSION: PTU is associated with the production of ANCA in patients with hyperthyroidism . PTU-induced ANCA are due to polyclonal activation of B cells . Anti-MPO and anti-PR3 antibodies may associate the occurrence of clinical vasculitis.

Biophys J, 2004 Dec, 87(6), 3901 - 11 Epub 2004 Dec.
Colicin occlusion of OmpF and TolC channels: outer membrane translocons for colicin import; Zakharov SD et al.; The interaction of colicins with target cells is a paradigm for protein import . To enter cells, bactericidal colicins parasitize Escherichia coli outer membrane receptors whose physiological purpose is the import of essential metabolites . Colicins E1 and E3 initially bind to the BtuB receptor, whose beta-barrel pore is occluded by an N-terminal globular "plug" . The x-ray structure of a complex of BtuB with the coiled-coil BtuB-binding domain of colicin E3 did not reveal displacement of the BtuB plug that would allow passage of the colicin (Kurisu, G., S . D . Zakharov, M . V . Zhalnina, S . Bano, V . Y . Eroukova, T . I . Rokitskaya, Y . N . Antonenko, M . C . Wiener, and W . A . Cramer . 2003 . Nat . Struct . Biol . 10:948-954) . This correlates with the inability of BtuB to form ion channels in planar bilayers, shown in this work, suggesting that an additional outer membrane protein(s) is required for colicin import across the outer membrane . The identity and interaction properties of this OMP were analyzed in planar bilayer experiments.OmpF and TolC channels in planar bilayers were occluded by colicins E3 and E1, respectively, from the trans-side of the membrane . Occlusion was dependent upon a cis-negative transmembrane potential . A positive potential reversibly opened OmpF and TolC channels . Colicin N, which uses only OmpF for entry, occludes OmpF in planar bilayers with the same orientation constraints as colicins E1 and E3 . The OmpF recognition sites of colicins E3 and N, and the TolC recognition site of colicin E1, were found to reside in the N-terminal translocation domains . These data are considered in the context of a two-receptor translocon model for colicin entry into cells.

Biotechnol Bioeng, 2004 Nov 20, 88(4), 502 - 11
Mineralization of LCFA associated with anaerobic sludge: Kinetics, enhancement of methanogenic activity, and effect of VFA; Pereira MA et al.; Long-chain fatty acids (LCFA) associated with anaerobic sludge by mechanisms of precipitation, adsorption, or entrapment can be biodegraded to methane . The mineralization kinetics of biomass-associated LCFA were established according to an inhibition model based on Haldane's enzymatic inhibition kinetics . A value around 1,000 mg COD-LCFA..g VSS(-1) was obtained for the optimal specific LCFA content that allowed the maximal mineralization rate . For sludge with specific LCFA contents of 2,838 +/- 63 and 4,571 +/- 257 mg COD-LCFA..g VSS(-1), the specific methanogenic activities in the presence of acetate, butyrate, and H(2)/CO(2) were significantly enhanced after the mineralization of the biomass-associated LCFA . For sludge with a specific LCFA content near the optimal value defined by the kinetic model, the effect of adding VFA to the medium was studied during the mineralization of the biomass-associated LCFA . Different patterns were obtained for each individual substrate . Acetate and butyrate were preferentially consumed by the consortium, but in the case of propionate no evidence of a sequential consumption pattern could be withdrawn . It was concluded that LCFA do not exert a bactericidal neither a permanent toxic effect toward the anaerobic consortia . A discussion is addressed to the relative roles of a reversible inhibitory effect and a transport limitation effect imposed by the LCFA surrounding the cells . (c) 2004 Wiley Periodicals, Inc

Prikl Biokhim Mikrobiol, 2004 Jul-Aug, 40(4), 387 - 97
{Extracellular factors of bacterial adaptation to unfavorable environmental conditions}; Nikolaev IuA; Data on extracellular compounds of bacteria involved in their adaptation to unfavorable environmental conditions are reviewed, including high or low temperatures, growth-inhibiting or bactericidal concentrations of toxic substances (oxidants, phenols, and heavy metals) and antibiotics, deviation of pH values from optimum levels, and salinity of the medium . Chemically, the compounds identified belong to diverse types (proteins, hydrocarbons, organic acids, nucleotides, amino acids, lipopeptides, volatile substances, etc.) . Most of them remain unidentified, and their properties are studied using biological testing . It is proposed to view extracellular adaptation factors (EAFs) as a new group of biologically active substances . EAFs may be divided into several subgroups by the mechanism of action . These subgroups include protectors (stabilizers), signaling molecules inducing defense responses, regulators (e.g., adhesion regulators) not acting as inducers, and antidotes (neutralizers) . The fields of EAF study include screening (search for new compounds, using biological tests), identification, and research into mechanisms of action . EAFs may find utility in biotechnology, medicine, agriculture, and environmental protection.

Acta Crystallogr D Biol Crystallogr, 2004 Oct, 60(Pt 10), 1897 - 9 Epub 2004 Sep 23.
Crystallization and preliminary crystallographic analysis of 3'-aminoglycoside kinase type IIIa complexed with a eukaryotic protein kinase inhibitor, CKI-7; Fong DH et al.; 3'-Aminoglycoside kinase type IIIa {APH(3')-IIIa} catalyzes the transfer of gamma-phosphate from ATP to the 3'-hydroxyl of many aminoglycoside antibiotics, abolishing their bactericidal effects . Despite very low sequence identity, APH(3')-IIIa and eukaryotic protein kinases share structural and functional similarities, including a sensitivity to isoquinolinsulfonamide-type inhibitors . APH(3')-IIIa has been cocrystallized with CKI-7, a casein kinase 1 inhibitor . These crystals were grown using PEG 3000 as precipitant and required consecutive cycles of microseeding . Data were collected to 2.5 A . The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 49.84, b = 91.90, c = 131.2 A.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 3884 - 91
Novel concentration-killing curve method for estimation of bactericidal potency of antibiotics in an in vitro dynamic model; Liu YQ et al.; The bactericidal pharmacodynamics of antibiotics against Escherichia coli were analyzed by a concentration-killing curve (CKC) approach, and the novel parameters median bactericidal concentration (BC(50)) and bactericidal intensity (r) for bactericidal potency were proposed . By using the agar plate method, about 500 E . coli cells were inoculated onto Luria-Bertani plates containing a series of antibiotic concentrations, and after 24 h of incubation at 37 degrees C, all the viable colonies were enumerated . This resulted in a sigmoidal CKC that could be perfectly fitted (R(2) > 0.9) with the function N = N(0)/{1 + e(r(x - BC(50)))}, where N is number of colonies surviving on each plate with an x series of concentrations of an antibiotic, and N(0) represents the meaningful inoculum size . Construction of the CKC method was based on the bactericidal effect of each antibiotic against the bacterial strain versus the concentration in two dimensions and may be a more valid, accurate, and reproducible method for estimating the bactericidal effect than the endpoint minimum bactericidal concentration (MBC) method . Mathematically, the CKC approach was point symmetrical toward its inflexion (BC(50), N(0)/2); thus, 2BC(50) could replace MBC . The parameter BC(1) can be defined as BC(50) + {ln(N(0) - 1)/r}, which is the drug concentration at which only one colony survived and which is the least critical value of MBC in the CKC . The variate r, which determined the tangent slope on inflexion when N(0) was limited, could estimate the bactericidal intensity of an antibiotic . This verified that the CKC approach may be useful in studies with other classes of antibiotics and has considerable value as a tool for the accurate and proper administration of antibiotics.

J Immunol, 2004 Oct 1, 173(7), 4598 - 606
The emerging pathogen Moraxella catarrhalis interacts with complement inhibitor C4b binding protein through ubiquitous surface proteins A1 and A2; Nordstrom T et al.; Moraxella catarrhalis ubiquitous surface protein A2 (UspA2) mediates resistance to the bactericidal activity of normal human serum . In this study, an interaction between the complement fluid phase regulator of the classical pathway, C4b binding protein (C4BP), and M . catarrhalis mutants lacking UspA1 and/or UspA2 was analyzed by flow cytometry and a RIA . Two clinical isolates of M . catarrhalis expressed UspA2 at a higher density than UspA1 . The UspA1 mutants showed a decreased C4BP binding (37.6% reduction), whereas the UspA2-deficient Moraxella mutants displayed a strongly reduced (94.6%) C4BP binding compared with the wild type . In addition, experiments with recombinantly expressed UspA1(50-770) and UspA2(30-539) showed that C4BP (range, 1-1000 nM) bound to the two proteins in a dose-dependent manner . The equilibrium constants (K(D)) for the UspA1(50-770) and UspA2(30-539) interactions with a single subunit of C4BP were 13 microM and 1.1 microM, respectively . The main isoform of C4BP contains seven identical alpha-chains and one beta-chain linked together with disulfide bridges, and the alpha-chains contain eight complement control protein (CCP) modules . The UspA1 and A2 bound to the alpha-chain of C4BP, and experiments with C4BP lacking CCP2, CCP5, or CCP7 showed that these three CCPs were important for the Usp binding . Importantly, C4BP bound to the surface of M . catarrhalis retained its cofactor activity as determined by analysis of C4b degradation . Taken together, M . catarrhalis interferes with the classical complement activation pathway by binding C4BP to UspA1 and UspA2.

ANZ J Surg, 2004 Sep, 74(9), 769 - 72
In-use efficacy of a chlorhexidine in alcohol surgical rub: a comparative study; Grabsch EA et al.; BACKGROUND: Although full surgical scrubs are performed prior to each case on an operating list, optimum regimens for hand cleaning have yet to be determined, and in-use efficacy evaluations are very limited . METHODS: A crossover study was undertaken comparing a chlorhexidine in detergent/alcohol regimen with povidine-iodine detergent scrub, within an orthopaedic operating environment . Depending on the skin asepsis regimen used, five surgical team members scrubbed or rubbed prior to each case for a complete operating list . Bactericidal efficacy was measured using the 'glove-juice' technique before and after hand asepsis, and at the completion of each case . RESULTS: The chlorhexidine regimen caused substantial and sustained reductions in hand bacterial counts (>50-fold prior to case 1) during surgical cases . Application of alcoholic chlorhexidine prior to each subsequent case reduced bacterial counts to the same level as the original scrub . In contrast, the povidine-iodine scrub reduced counts <3-fold prior to the first case and <2-fold in subsequent cases . The chlorhexidine regimen also resulted in persistent bactericidal effects between cases, as counts prior to application of cases 2 and higher were significantly lower than prior to case 1 (>7-fold for case 2 vs case 1) . CONCLUSIONS: The chlorhexidine regimen demonstrated excellent bactericidal efficacy throughout an operating list, and was superior to povidine-iodine scrubbing in all aspects . The alcoholic chlorhexidine regimen is simpler and should have wide surgical application.

J Infect Dis, 2004 Oct 15, 190(8), 1476 - 80 Epub 2004 Sep 15.
Role of human neutrophil peptide-1 as a possible adjunct to antituberculosis chemotherapy; Kalita A et al.; We report the role of human neutrophil peptide (HNP)-1 as an adjunct to antituberculosis (anti-TB) drugs . The combination of HNP-1, isoniazid, and rifampicin was evaluated against Mycobacterium tuberculosis H(37)Rv in vitro, ex vivo, and in vivo, and synergism was observed on the basis of reductions in minimum inhibitory concentrations (MICs) of these agents . In vitro results revealed >1-log unit reductions even when HNP-1 and anti-TB drugs were used at 1/16 MICs . This combination was also found to be bactericidal against intracellular mycobacteria even at 1/8 MICs of HNP-1 and drugs . HNP-1 used in conjunction with anti-TB drugs resulted in significant clearance of bacterial load from lungs, liver, and spleen of infected, compared with control animals . The effective therapeutic dosage of drugs could be reduced to half by supplementing HNP-1 in the therapeutic schedule . These results clearly suggest that HNP-1 can be used as adjunct chemotherapy with conventional drugs against TB .

Mol Pharmacol, 2004 Dec, 66(6), 1599 - 606 Epub 2004 Dec.
Suppression of pathogenicity of Porphyromonas gingivalis by newly developed gingipain inhibitors; Kadowaki T et al.; Arg-gingipain (Rgp) and Lys-gingipain (Kgp) are cysteine proteinases produced by Porphyromonas gingivalis, a major etiological bacterium of periodontal diseases . Here we show a series of small peptide analogs able to inhibit either Rgp or Kgp, which are synthesized on the basis of the cleavage site specificity of human salivary histatins by each enzyme . Among this series of compounds, carbobenzoxy-Lys-Arg-CO-Lys-N-(CH2)2 (KYT-1) and carbobenzoxy-Glu(NHN(CH3)Ph)-Lys-CO-NHCH2Ph (KYT-36) were found to be the most potent inhibitors of Rgp and Kgp, respectively, with Ki values of 10(-11) to 10(-10) M order . Both inhibitors exhibited slight or no inhibition on mammalian proteinases such as trypsin and cathepsins B, L, and H . All of the virulence induced by the culture supernatant of P . gingivalis tested, including the degradation of various host proteins such as human type I collagen, immunoglobulins, fibronectin, and fibrinogen, disruption of the bactericidal activity of polymorphonuclear leukocytes, and enhancement of the vascular permeability, were strongly inhibited by a combined action of both inhibitors . The functions essential for the bacterium to grow and survive in the periodontal pocket, such as coaggregation and acquisition of amino acids, were also strongly inhibited by the combined action of both inhibitors . The disruption of the adhesion and viability of human fibroblasts and hemagglutination by the organism were strongly suppressed by a single use of KYT-1 . These results thus indicate that the newly developed KYT-1 and KYT-36 both should provide a broader application in studies of this important class of enzymes and facilitate the development of new approaches to periodontal diseases.

J Med Microbiol, 2004 Oct, 53(Pt 10), 953 - 8
Factors affecting the escape of Francisella tularensis from the phagolysosome; Lindgren H et al.; The highly virulent bacterium Francisella tularensis is well adapted to the intracellular habitat but the mechanisms behind its intracellular survival have been elusive . Recently, it was shown that the bacterium is capable of escaping from the phagosome of human and mouse monocytic cells . Here it is shown that this escape is affected by gamma interferon (IFN-gamma) treatment of mouse peritoneal exudate cells since in treated cells the proportion that escaped was significantly lower (80%) than in untreated cells (97%) as determined by transmission electron microscopy . By contrast, < 1% of mutant bacteria lacking expression of a 23 kDa protein denoted IglC were able to escape from the phagosome . Infection with the DeltaiglC strain complemented with the iglC gene resulted in 60% of the bacteria escaping from the phagosome . Whereas IFN-gamma treatment conferred a static effect on intracellular wild-type bacteria, the treatment had a bactericidal effect on the DeltaiglC strain . The results show that the activation status of infected cells affects the escape of F . tularensis from the phagosome . An even more profound effect on this escape is related to expression of IglC by F . tularensis . Its absence rendered the mutant bacteria incapable of escaping from the phagosome and of multiplying intracellularly.

Front Biosci, 2004 Sep 01, 9, 2605 - 17
Beta-lactams and their potential use as novel anticancer chemotherapeutics drugs; Kuhn D et al.; The discovery of natural and synthetic antibiotics is one of the most important medical breakthroughs in human history . Many diseases, such as bacterial meningitis, pneumonia, and septicemia, are now curable with the use of antibiotics . Antibiotics are efficacious, generally well tolerated in patients, and have a low toxicity level . It is for these reasons antibiotics remain an attractive target for drug discovery . Traditional beta-lactam antibiotics (e.g . penicillins, penems, cephalosporins) have a bicyclic ring structure that is conformationally rigid and functions to inhibit bacterial cell wall synthesis . In addition to the bactericidal action of antibiotics, it has been discovered that many antibiotics are capable of inhibiting tumor cell growth . There are currently many antitumor antibiotics approved for cancer therapy, which work to inhibit tumor cell growth by DNA intercalation . The use of beta-lactams as prodrugs has also met with success by aiding delivery of the chemotherapeutic directly to tumor sites . Recently, a novel class of N-thiolated monobactams, so termed because they possess a monocyclic ring instead of the bicyclic ring, has been found to induce apoptosis potently and specifically in many tumor cell lines but not in normal, non-transformed cell lines . Other beta-lactams, such as the polyaromatics, have been found to slow or inhibit tumor cell growth, and the 4-alkylidene beta-lactams are capable of inhibiting matrix metalloproteinases and leukocyte elactase activity . These data indicate that synthesis and evaluation of beta-lactams are a promising area for further development in anticancer research.

J Altern Complement Med, 2004 Aug, 10(4), 681 - 3
Effect of qi training on neutrophil function in young and elderly males; Lee MS et al.; OBJECTIVES: To examine the effect of qi training on neutrophil bactericidal function (superoxide generation and adhesion) . METHODS: We studied the effects of one session of qi training on superoxide generation and adhesion of neutrophils immediately after (Post I), and 2 hours after qi training (Post II), in nine young and nine elderly male subjects . RESULTS: The qi training significantly enhanced the superoxide generation and adhesion of neutrophils, and there were significant differences at Post I compared to before qi training (Pre) in both groups . CONCLUSION: Our current observations show that qi training enhances superoxide generation and adhesion of neutrophils . It is supposed that qi training may increase the resistance of trained individuals against common infection and inflammation.

J Hosp Infect, 2004 Sep, 58(1), 78 - 80
Alcoholic fixation of blood to surgical instruments-a possible factor in the surgical transmission of CJD?
Prior F, Fernie K, Renfrew A, Heneaghan G.
While developing a new protein removal test for the quality control of surgical instrument cleaning, it was noted that alcohol firmly binds blood to stainless steel . Creutzfeldt-Jacob disease is one of the transmissible spongiform encephalopathies (TSE) that has been transmitted between humans and chimpanzees by electroencephalogram electrodes, previously 'sterilized' using ethanol and formaldehyde . Although ethanol has a bactericidal action, it also binds protein to metal . Prion proteins found in TSE are thought to be the causal agents of spongiform disease and it is likely that these proteins are also bound to the stainless steel of surgical instruments by alcohols . Where spongiform disease is a possibility, alcohol, and probably formaldehyde, should not be used to decontaminate neurosurgical instruments.

J Immunol Methods, 2004 Sep, 292(1-2), 187 - 93
High-throughput imaging of bacterial colonies grown on filter plates with application to serum bactericidal assays; Liu X et al.; The ability to accurately enumerate viable bacteria has applications in antibiotic screening assays, toxicology testing, and serological assays for functional antibodies . An impediment to high-throughput bacterial assays is the requirement to grow bacteria as individual colonies on semisolid media containing agar . We have now developed a method for growth, staining, and counting of bacterial colonies in 96-well filter plates . A unique feature of the method is that colony size is inversely proportional to the number of colonies in each well, presumably due to nutrient depletion . As a result, as many as 300 colony-forming units (cfu) can be detected as discrete colonies within a single assay well . The resulting colonies can be counted automatically using an imaging system originally developed for ELISPOT assays . The method has been applied to the measurement of serum bactericidal activity (SBA) in human sera.

J Leukoc Biol, 2004 Dec, 76(6), 1104 - 10 Epub 2004 Dec.
In vivo evidences that insulin regulates human polymorphonuclear neutrophil functions; Walrand S et al.; Polymorphonuclear neutrophils (PMN) are able to destroy invasive mircoorganisms by a wide variety of functions . Whereas insulin does not stimulate hexose transport in PMN, previous reports have clearly shown that this hormone regulates glucose metabolism inside this cell, raising the question of insulin action on PMN functions in humans . It is interesting that in vitro studies established a strong relationship between specific binding of insulin to its PMN membrane receptor and the activation of the main PMN functions . Therefore, investigation in healthy subjects under strict euglycemia and physiological insulinemia was performed to understand the in vivo-specific action of insulin on PMN functions without hyperglycemia interferences . We determined numerous PMN functions before and after hyperinsulinemia (0.5 mU/kg/min) and euglycemia (0.9 g/l) clamp for 4 h in eight adult healthy volunteers (24+/-6 years) . The total number of PMN and the number of PMN expressing CD11b, CD15, CD62L, and CD89 were significantly increased over baseline (P<0.001), whereas the density of these receptors was down-regulated (P<0.01) by insulin . PMN chemotaxis (+117%, P<0.05), phagocytosis (+29%, P<0.001), and bactericidal (+17-25%, P<0.001) capacities were increased during the insulin clamp (P<0.05) . Therefore, insulin treatment may modulate PMN functions not only by attainment of a better metabolic control, as suggested by in vivo studies in diabetic patients, but also through a direct effect of insulin.

Antibiot Khimioter, 2004, 49(3), 3 - 5
{Maintenance of bactericidal activity of pectin stabilized aqueous solutions}; Potievskii EG et al.; Maintenance of the bactericidal activity and organoleptic properties of 3% and 5% stabilized aqueous solutions of pectin (Pepidol PEG, 3% and Pepidol PEG, 5%) stored for prolonged periods and at various temperatures was studied . Pepidol PEG was shown to preserve its bactericidal activity and organoleptic properties for 28 months when stored at rather wide ranges of the environmental temperature . The drug did not lose its properties after freezing and subsequent melting . The results of the study allowed to increase the period of Pepidol PEG use up to 18 months and the storage temperature ranges up to -30 degrees to +30 degrees C.

Bioorg Med Chem, 2004 Sep 15, 12(18), 4809 - 13
Quinones as antimycobacterial agents; Tran T et al.; Mycobacterium tuberculosis is a serious worldwide health threat, killing almost 3 million people per year . Other mycobacterial species, especially Mycobacterium avium, are emerging pathogens in the immunocompromised population, most notably AIDS patients . These nontuberculous mycobacteria (NTM) are ubiquitous in the environment, and naturally resistant to many disinfection procedures . Treatment options are limited, and no new antibiotics have been developed against mycobacteria since the 1970s . There is a desperate need for new biocides and antibiotics to prevent and treat mycobacterial infections . A small aromatic compound library has been screened for effectiveness in growth inhibition or killing of mycobacteria . Four species, representing the M . tuberculosis complex, the slow-growing NTM, and the rapid-growing NTM were used . Active compounds had minimal inhibitory concentrations as low as 12.5 microg/mL, with the active component being a quinone . The primarily bactericidal activity observed represents a unique mechanism of action . A fluorescent assay involving M . smegmatis expressing gfp was analyzed as a rapid assay for predicting inhibitory activity, but failed to predict activity well . Our compounds may have significant utility as soluble biocides against mycobacteria and other hardy nosocomial pathogens.

Antimicrob Agents Chemother, 2004 Sep, 48(9), 3556 - 8
SRI-286, a thiosemicarbazole, in combination with mefloquine and moxifloxacin for treatment of murine Mycobacterium avium complex disease; Bermudez LE et al.; Treatment of Mycobacterium avium disease remains challenging when macrolide resistance develops . We infected C57 beige mice and treated them with mefloquine, SRI-286, and moxifloxacin . SRI-286 (80 mg/kg) was bactericidal in the liver . Mefloquine plus moxifloxacin or mefloquine plus SRI-286 were better than mefloquine alone.

Vet Microbiol, 2004 Sep 8, 102(3-4), 203 - 13
Immunogenicity of recombinant Omp31 from Brucella melitensis in rams and serum bactericidal activity against B . ovis; Estein SM et al.; Detergent-extracted recombinant Omp31 (rOmp31 extract) from Brucella melitensis produced in Escherichia coli was previously identified as a protective immunogen against B . ovis in mice . In this study, we evaluated the immunogenicity of rOmp31extract in rams . This immunogen was emulsified in an oil adjuvant and administered three times with 4 and 8 weeks intervals . Antibody response was measured in serum by whole B . ovis ELISA . Specific antibodies to purified rOmp31 (pET-Omp31) were detected by Western blotting and indirect ELISA . In addition, isotype specific antibodies were measured in tears . Serum bactericidal activity against B . ovis in the presence of complement was measured in vitro . Cellular immune response was explored by intradermal testing with purified rOmp31 . Immunization with rOmp31 extract induced IgG specific antibodies in serum able to bind to whole B . ovis cells . Furthermore, strong inhibition in a competitive ELISA (with an Omp31-specific monoclonal antibody) suggested that a proportion of Omp31-specific antibodies were directed against a loop containing a protective epitope . Serum antibodies killed efficiently B . ovis in vitro in the presence of either guinea pig or ovine serum . Tears had both IgG and IgA antibodies to equivalent titers . Finally, immunized rams showed skin reactivity to Omp31 . These data demonstrate that B . melitensis Omp31, a protective antigen identified in the mouse model, induces antibody and cellular immune mechanisms in sheep.

Trends Immunol, 2004 Sep, 25(9), 483 - 8
IL-4 in tuberculosis: implications for vaccine design; Rook GA et al.; Current attempts to find a vaccine for tuberculosis (TB) are based on the assumption that it must drive a Th1 response . We review the evidence that progressive disease might not be due to absence of Th1, but rather to the subversive effect of an unusual Th2-like response, involving interleukin-4 (IL-4) and IL-4delta2 . This Th2-like response can impair bactericidal function and lead to toxicity of tumour necrosis factor-alpha (TNF-alpha) and to pulmonary fibrosis . If this is important, effective vaccines will need to suppress pre-existing Th2-like activity . Such vaccines are feasible and are active therapeutically in mouse TB.

Infect Immun, 2004 Sep, 72(9), 5150 - 8
Mycobacterium tuberculosis triggers apoptosis in peripheral neutrophils involving toll-like receptor 2 and p38 mitogen protein kinase in tuberculosis patients; Aleman M et al.; Polymorphonuclear neutrophils (PMN) exposed to Mycobacterium tuberculosis display bactericidal responses and produce inflammatory proteins . This PMN-mediated inflammatory response is regulated by an activation of the apoptotic program, which collaborates to avoid tissue injury . In vitro, circulating PMN from patients with tuberculosis (TB) show an increased spontaneous apoptosis, and M . tuberculosis-induced activation accelerates the PMN apoptosis . In this study, we evaluated the mechanisms involved in spontaneous and M . tuberculosis-induced apoptosis . We demonstrate that apoptosis of PMN is not induced by lipoarabinomannan or by a whole-cell lysate of M . tuberculosis and that neither tumor necrosis factor alpha nor CD11b, CD14, and Fcgamma receptors are involved . Apoptosis of PMN from patients with active TB (TB-PMN) is induced by the interaction with the whole M . tuberculosis via Toll-like receptor 2 (TLR2), and, in contrast to spontaneous apoptosis, it involves the p38 mitogen-activated protein kinase (MAPK) pathway . These results correlate with a high expression of phosphorylated p38 (p-p38) in circulating TB-PMN and with the ability of M . tuberculosis to induce in vitro the expression of p-p38 in PMN . Therefore, when the bacterial burden is low, TB-PMN could be detecting nonopsonized M . tuberculosis via TLR2, leading to the activation of the p38 MAPK pathway, which in turn would induce PMN activation and apoptosis . This mechanism needs further confirmation at the site of infection.

Clin Exp Immunol, 2004 Sep, 137(3), 566 - 9
CCTTT-repeat polymorphism of the inducible nitric oxide synthase is not associated with HIV pathogenesis; Hersberger M et al.; Nitric oxide (NO) produced by the inducible form of nitric oxide synthase (iNOS) has bactericidal and virocidal effects . Although NO synthesis and iNOS expression in macrophages affect several aspects of human immunodeficiency virus (HIV) type-1 pathogenesis, their role in HIV disease remains largely unknown . In humans, the expression of iNOS is influenced by a functional CCTTT-repeat polymorphism in the promoter region of the gene . We investigated the association of this polymorphism with HIV pathogenesis in naive HIV-infected patients before the initiation of antiretroviral therapy . The allele frequencies of the iNOS CCTTT-repeat polymorphism were assessed by PCR in 857 patients from the Swiss HIV Cohort Study, including rapid progressors and long-term nonprogressors, and in 240 healthy volunteers . In HIV-infected patients, the initial viral load and the decline in total CD4 cells was calculated to estimate disease progression . Allele frequencies of the iNOS CCTTT-repeat polymorphism were similar between the HIV-infected and noninfected blood donors . In treatment-naive HIV-positive patients, there was no association of the iNOS polymorphism with viral load or with the course of CD4 cells . Regulation of iNOS expression by the functional CCTTT-polymorphism does not modify HIV pathogenesis.

Chemotherapy, 2004, 50 Suppl 1, 16 - 21
Clinical applications of levofloxacin for severe infections; Graninger W et al.; New fluoroquinolones, as exemplified by levofloxacin, possess broad spectrum activity against many common pathogens, including the majority responsible for respiratory tract infections (RTIs), atypical pathogens and those resistant to other therapeutic regimens . Following administration, levofloxacin attains high intracellular and tissue levels . This, coupled with an exceptional pharmacodynamic profile, allows levofloxacin to be administered once daily . However, in certain circumstances, such as seriously ill patients or those with difficult-to-treat pathogens, higher doses may be required . Since the bactericidal effect of levofloxacin is concentration-dependent, it is possible to increase peak concentration by increasing the dose, resulting in even better tissue concentration (and a possible reduction in the development of resistance) . High-dose levofloxacin is able to exploit these pharmacokinetic features to provide an effective treatment for severe infections . Data is now available confirming the efficacy of high-dose levofloxacin in a wide range of infections, including nosocomial pneumonia, meningitis and complicated skin and skin structure infections (CSSSIs) . Not only is this regimen effective, it is also well tolerated and provides the physician with an additional therapeutic option to manage critically ill patients.

Surgery, 2004 Aug, 136(2), 253 - 60
Structure/function studies of an endotoxin-neutralizing peptide derived from bactericidal/permeability-increasing protein; Wasiluk KR et al.; BACKGROUND: Bactericidal/permeability-increasing protein, BPI, has a beta-turn with alternating cationic and hydrophobic residues in its lipopolysaccharide (endotoxin, LPS)-binding domain . A peptide, betapep25, was designed with 9 residues of the LPS-binding domain of BPI flanked by beta-turn-inducing elements . Thereafter, we sought to use single amino acid substitutions to identify residues that are important for the biological activities of betapep25 . METHODS: Single alanine or norleucine replacement "walkthrough" peptides based on betapep25 were generated and tested for their ability to kill P aeruginosa and to neutralize endotoxin . RESULTS: Substitution of all lysines inhibited bactericidal activity . Inhibition of LPS-neutralizing activity was seen in 9 peptides in which an alanine or norleucine was substituted for each of 4 of the basic residues and 1 hydrophobic residue from the LPS-binding region of BPI and 4 hydrophobic residues from the beta-turn-inducing regions flanking the LPS-binding region on the carboxy-terminal side . Intriguingly, these last 4 substitutions resulted in peptides that exhibited increased bactericidal activity compared to betapep25 . CONCLUSIONS: These results demonstrate the importance of both cationic and hydrophobic amino acid residues to bactericidal and endotoxin-neutralizing activities . These perturbations of biological activity should be considered in the design of synthetic peptide endotoxin antagonists .

Vaccine, 2004 Aug 13, 22(23-24), 3008 - 13
PorA-specific differences in antibody avidity after vaccination with a hexavalent Men B outer membrane vesicle vaccine in toddlers and school children; Vermont CL et al.; A clinical phase II trial with an experimental hexavalent outer membrane vesicle (OMV) vaccine (HexaMen) containing six different porin A (PorAs) was carried out in toddlers (2-3 years) and schoolchildren (7-8 years) in The Netherlands . HexaMen exists of two OMVs each containing three different PorA types . The serum bactericidal activity (SBA) after vaccination against the six PorAs was significantly different and was higher in toddlers than in schoolchildren . After vaccination the SBA against P1.5-2,10 was 4-6 times higher than against P1.7-2,4 . The aim of this study was to test whether the differences in SBA could be explained by a difference in subtype-specific antibody avidity maturation . The avidity index (AI) of antibodies against three subtypes (PorA types P1.5-2,10; P1.12-1,13 and P1.7-2,4) was measured by ELISA and evaluated in relation to SBA . A significant avidity maturation for the 3 PorA subtypes was found . This maturation was most pronounced for P1.5-2,10 (mean AI = 72%), correlating with the highest SBA titres . Generally, the avidity titre correlated best with SBA . No differences in avidity indices against the three tested PorAs were found between toddlers and school children indicating that avidity maturation induced by this vaccine is not age-dependent.

J Bacteriol, 2004 Aug, 186(16), 5427 - 31
Requirements for nitric oxide generation from isoniazid activation in vitro and inhibition of mycobacterial respiration in vivo; Timmins GS et al.; Isoniazid (INH), a front-line antituberculosis agent, is activated by mycobacterial catalase-peroxidase KatG, converting INH into bactericidal reactive species . Here we investigated the requirements and the pathway of nitric oxide (NO*) generation during oxidative activation of INH by Mycobacterium tuberculosis KatG in vitro . We also provide in vivo evidence that INH-derived NO* can inhibit key mycobacterial respiratory enzymes, which may contribute to the overall antimycobacterial action of INH.

J Oral Sci, 2004 Jun, 46(2), 107 - 11
Influence of different vehicles on the pH of calcium hydroxide pastes; Pacios MG et al.; The main known benefit of calcium hydroxide as an intracanal medicament lies in the bactericidal effect conferred by its pH . The objective of this work was to determine the influence of the vehicle on the pH of calcium hydroxide pastes after usage in patients and in vitro . The incisor root canals of 180 patients were instrumented and filled with calcium hydroxide pastes containing distilled water, chlorhexidine, propylene glycol, anesthetic solution, camphorated p-monochlorophenol and camphorated p-monochlorophenol-propylene glycol . The pH of the paste in the patients' root canals was measured at 7, 14 and 21 days . Similarly, pH was measured in vitro up to 21 days . The pH of all the pastes remained constant throughout the time periods assessed . The calcium hydroxide-water combination showed significantly higher pH values than the other pastes in clinical use . Comparative analysis showed that the pH values of the anesthetic solution, camphorated p-monochlorophenol and camphorated p-monochlorophenol-propylene glycol were significantly higher in vitro . The type of vehicle was shown to influence the final pH of the pastes . However, the alkalinity of all pastes was maintained over time under the experimental conditions.

Am J Vet Res, 2004 Jul, 65(7), 957 - 63
Characterization of biological activities of feline eosinophil granule proteins; Fondati A et al.; OBJECTIVE:To characterize eosinophil granule-derived proteins in cats . SAMPLE POPULATION: Eosinophils collected via peritoneal lavage from 2 cats . PROCEDURE: The cats were infested orally with Toxocara canis eggs and subsequently challenge-exposed with T . canis antigen injected IP to induce peritoneal eosinophilia; eosinophils were collected via peritoneal lavage . Eosinophil granule proteins were acid-extracted, separated by gel-filtration chromatography, and examined for their peroxidase, ribonuclease, and bactericidal activities; the N-terminal sequence of some of these proteins was determined and compared with homologue proteins from other species . RESULTS: 3 protein peaks were separated in the chromatogram . The first peak had both peroxidase and bactericidal activities . The second peak had ribonuclease and bactericidal activities, and the N-terminal sequence of the major protein was homologous with that of proteins of the ribonuclease A superfamily, including eosinophil ribonucleases from humans and other animal species . The third protein peak had bactericidal activity, and the N-terminal sequence of the major protein was homologous with that of human and murine major basic proteins . CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that feline eosinophil granules contain major basic protein and eosinophil-associated ribonuclease and the granule proteins have peroxidase, ribonuclease, and bactericidal activities . In cats, characterization of eosinophil granule proteins may be useful in elucidation of the mechanism of tissue damage in eosinophil-associated diseases and development of new treatment options for those diseases . In addition, the identification of conserved structure and function of eosinophil granule proteins in cats is relevant from an evolutionary viewpoint.

Bull Exp Biol Med, 2004 Feb, 137(2), 206 - 10
Morphology of reparative regeneration in organs and tissues during treatment with new generation Sulfacrylate; Marchenko VT et al.; We studied morphogenesis of reparative processes in parenchymatous and hollow organs after surgery with the use of Sulfacrylate glue composition characterized by good adhesive and bactericidal properties . The glue rapidly and effectively arrested parenchymatous bleeding, reduced the volume of necrotic tissue, and promoted wound healing without suppuration . Cicatrix was completely formed 1 month after surgery and the glue completely resorbed at this term . The bioglue reliably connected and hermetically sealed sutures when used for gluing intestinal loops and for creation of intestinal anastomoses . The use of biological glue promoted the formation of elastic fibrous tissue not distorting the intestinal lumen.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 3165 - 8
In vitro activities of the newer quinolones garenoxacin, gatifloxacin, and gemifloxacin against human mycoplasmas; Pereyre S et al.; The activities of garenoxacin, gatifloxacin, and gemifloxacin were compared with those of four fluoroquinolones against human mycoplasmas and ureaplasmas, including fluoroquinolone-resistant genetically characterized strains . Garenoxacin exhibited the highest activity, followed by gemifloxacin, moxifloxacin, and gatifloxacin . The minimal bactericidal activities of these three compounds were lower than those of the four fluoroquinolones.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 3006 - 9
Nitric oxide generated from isoniazid activation by KatG: source of nitric oxide and activity against Mycobacterium tuberculosis; Timmins GS et al.; Isonicotinic acid hydrazide (INH) is a frontline antituberculosis agent . Once taken up by Mycobacterium tuberculosis, INH requires activation by the catalase-peroxidase KatG, converting INH from its prodrug form into a range of bactericidal reactive species . Here we used 15N-labeled INH together with electron paramagnetic resonance spin trapping techniques to demonstrate that nitric oxide (NO*) is generated from oxidation at the hydrazide nitrogens during the activation of INH by M . tuberculosis KatG . We also observed that a specific scavenger of NO* provided protection against the antimycobacterial activity of INH in bacterial culture . No significant increases in mycobacterial protein nitration were detected, suggesting that NOdot; and not peroxynitrite, a nitrating metabolite of NO*, is involved in antimycobacterial action . In conclusion, INH-derived NO* has biological activity, which directly contributes to the antimycobacterial action of INH.

J Immunol, 2004 Aug 1, 173(3), 1959 - 65
Bordetella pertussis lipopolysaccharide resists the bactericidal effects of pulmonary surfactant protein A; Schaeffer LM et al.; Surfactant protein A (SP-A) plays an important role in the innate immune defense of the respiratory tract . SP-A binds to lipid A of bacterial LPS, induces aggregation, destabilizes bacterial membranes, and promotes phagocytosis by neutrophils and macrophages . In this study, SP-A interaction with wild-type and mutant LPS of Bordetella pertussis, the causative agent of whooping cough, was examined . B . pertussis LPS has a branched core structure with a nonrepeating trisaccharide, rather than a long-chain repeating O-Ag . SP-A did not bind, aggregate, nor permeabilize wild-type B . pertussis . LPS mutants lacking even one of the sugars in the terminal trisaccharide were bound and aggregated by SP-A . SP-A enhanced phagocytosis by human monocytes of LPS mutants that were able to bind SP-A, but not wild-type bacteria . SP-A enhanced phagocytosis by human neutrophils of LPS-mutant strains, but only in the absence of functional adenylate cyclase toxin, a B . pertussis toxin that has been shown to depress neutrophil activity . We conclude that the LPS of wild-type B . pertussis shields the bacteria from SP-A-mediated clearance, possibly by sterically limiting access to the lipid A region.

Ann Dermatol Venereol, 2004 Apr, 131(4), 375 - 8
{Late onset of a chronic septic granulomatous disease}; Kharfi M et al.; INTRODUCTION: Chronic septic granulomatosis is a disease characterized by an impaired bactericidal potential of the neutrophilic polynuclear . The cutaneous manifestations rarely reveal the disease, but are of considerable interest in the diagnosis, notably during the late onset forms . We report such a case . CASE REPORT: A 15 year-old girl, born of consanguine parents, had a history of visceral leishmaniasis and hepatic hydatidosis . For the past 3 years she had developed dermatitis lesion on the face and skin folds, chronic folliculitis and suppurating axillary and inguinal lymphoadenitis . The absence of a reduction in tetrazolium nitro blue led to the diagnosis of chronic septic granulomatosis . Prophylactic treatment stabilized the cutaneous lesions . DISCUSSION: Chronic septic granulomatosis regroups various severe and recurrent manifestations . Its transmission is usually X-linked recessive or, on rare occasions, autosomal recessive . The clinical manifestations leading to the diagnosis are often of very early onset . They are principally pneumonia due to apergillus fumigatus and lymphoadenitis . Cutaneous involvement, although less common, must not be neglected because it can lead to the diagnosis of late onset forms, as in our patient.

Neoplasma, 2004, 51(4), 265 - 8
Bactericidal capacity of platelets in gastric cancer patients; Kamocki Z et al.; The aim of this study was to evaluate bactericidal capacity of platelets in patients suffering from gastric cancer . Number of platelets and their bactericidal activity were measured in 32 cancer patients (divided into 2 groups: I--resectable cancer, II--non-resectable one) and 32 normal donors . In group I the number of platelets was 259.136+/-84.459 x 103/microl . It was increased comparing to the normal donors 193.219+/-55.493 x 103/microl . After the surgery increase in platelet number was observed (472.05+/-111.772 x 103/microl) . In group II an increased number of platelets was observed (265.1+/-81.813 x 103/microl) and it was maintained in a post-operative period: 234.2+/-54.141 x 103/microl . In group I bactericidal capacity of platelets was 2.25+/-7.33%, whereas it increased significantly after the surgery--4.7+/-7.46% . In group II, it was 8.6+/-17.61% before and 4.72+/-4.76% after the surgery . In normal donors this ability was 21.66+/-16.66 . In gastric cancer patients increased platelet number was observed . Significant increase in platelets number occurred after a radical tumor removal . Decreased bactericidal activity of platelets was noticed in gastric cancer patients . After surgical removal of the tumor, platelets partly reclaimed bactericidal capacity . In patients presenting disseminated gastric cancer, bactericidal capacity of platelets could be permanently impaired.

Int Microbiol, 2004 Jun, 7(2), 139 - 42
An in vitro study of the susceptibility of mobile and cystic forms of Borrelia burgdorferi to tinidazole; Brorson O et al.; The susceptibility of mobile and cystic forms of Borrelia burgdorferi to tinidazole (TZ) was examined . The minimal bactericidal concentration (MBC) of TZ against the mobile spirochetes was >128 microg/ml at 37 degrees C in micro-oxic atmosphere when incubated for 14 days . TZ significantly reduced the conversion of mobile spirochetes to cystic forms during incubation . The MBC for older (10-months-old) cysts at 37 degrees C in a micro-oxic atmosphere was >0.5 microg/ml, but >0.125 microg/ml for young (1-day-old) cysts . Acridine orange staining, dark-field microscopy and transmission electron microscopy revealed that, when the concentration of TZ was > or = MBC, the contents of the cysts were partly degraded, core structures did not develop inside the young cysts, and the amount of RNA in these cysts decreased significantly . When cysts were exposed to TZ, both the spirochetal structures and core structures inside the cysts dissolved, and the production of blebs was significantly reduced . These observations may be valuable in the treatment of resistant infections caused by B . burgdorferi, and suggest that a combination of TZ and a macrolide antibiotic could eradicate both cystic and mobile forms of B . burgdorferi.

Environ Health Perspect, 2004 Jul, 112(10), 1058 - 62
Manufactured nanomaterials (fullerenes, C60) induce oxidative stress in the brain of juvenile largemouth bass; Oberdorster E; Although nanotechnology has vast potential in uses such as fuel cells, microreactors, drug delivery devices, and personal care products, it is prudent to determine possible toxicity of nanotechnology-derived products before widespread use . It is likely that nanomaterials can affect wildlife if they are accidentally released into the environment . The fullerenes are one type of manufactured nanoparticle that is being produced by tons each year, and initially uncoated fullerenes can be modified with biocompatible coatings . Fullerenes are lipophilic and localize into lipid-rich regions such as cell membranes in vitro, and they are redox active . Other nano-sized particles and soluble metals have been shown to selectively translocate into the brain via the olfactory bulb in mammals and fish . Fullerenes (C60) can form aqueous suspended colloids (nC60); the question arises of whether a redox-active, lipophilic molecule could cause oxidative damage in an aquatic species . The goal of this study was to investigate oxyradical-induced lipid and protein damage, as well as impacts on total glutathione (GSH) levels, in largemouth bass exposed to nC60 . Significant lipid peroxidation was found in brains of largemouth bass after 48 hr of exposure to 0.5 ppm uncoated nC60 . GSH was also marginally depleted in gills of fish, and nC60 increased water clarity, possibly due to bactericidal activity . This is the first study showing that uncoated fullerenes can cause oxidative damage and depletion of GSH in vivo in an aquatic species . Further research needs to be done to evaluate the potential toxicity of manufactured nanomaterials, especially with respect to translocation into the brain.

Comp Biochem Physiol C Toxicol Pharmacol, 2004 Apr, 137(4), 299 - 305
The effects of vitamin E on antiacid stress ability in juvenile soft-shelled turtles (Pelodiscus sinensis); Zhou X et al.; We determined the effect of dietary supplementation with vitamin E (0-, 50-, 250-, 500-, 1000- and 5000-mg/kg diet for 4 weeks) on antistress ability in juvenile soft-shelled turtle (Pelodiscus sinensis) . Half of the turtles per dose group were treated by acid stress for 24 h . The results showed that phagocytosis of blood cells in the control group significantly decreased after acid stress while the other five groups had no significant changes compared with those of before stress . Serum bacteriolytic activity in the control group and the group supplemented with 50-mg vitamin E/kg diet significantly decreased after acid stress . The other four groups showed no significant differences compared with those before stress . Serum bactericidal activities in all groups notably decreased after acid stress, but the difference of serum bactericidal activity in before and after stress had a decreased tendency from the control group to the highest dose group . Serum cortisol levels in the control group were significantly increased while the other five groups had no notable increases after acid stress . Liver vitamin E levels in all groups had no notable changes compared with those before stress but there was a tendency to decrease after acid stress . These results suggest that acid stress depress immune function and increase serum cortisol levels in turtles while vitamin E alleviate the adverse effects caused by acid stress.

Toxicon, 2004 Jul, 44(1), 91 - 101
Structural and functional characterization of myotoxin I, a Lys49 phospholipase A2 homologue from the venom of the snake Bothrops atrox; Nunez V et al.; A new myotoxin was isolated from the venom of Bothrops atrox from Colombia . B . atrox myotoxin I is a homodimer, with a subunit molecular mass of 13,826, and a pI of 8.9 . Its complete nucleotide sequence was obtained by cDNA cloning, indicating a mature product of 122 residues that belongs to the family of Lys49 phospholipase A(2) (PLA(2)) homologues, a subgroup of catalytically inactive proteins within the group IIA . Accordingly, the toxin was devoid of phospholipase and anticoagulant activities, in vitro . In mice, it induced conspicuous local myonecrosis, edema, and a systemic interleukin-6 response . In vitro, it was cytolytic upon myoblasts, and weakly bactericidal . The toxin showed highest homology with other Lys49 PLA(2)s, both in its primary and three-dimensional modeled structure, although with an evident difference in the C-terminal region . Unlike Lys49 proteins of American crotalids having 121 residues, this toxin presents an insertion (Asn) between positions 118 and 119 . Despite several substitutions within the C-terminal region 115-129 between B . atrox myotoxin I and B . asper myotoxin II, antibodies against synthetic peptide 115-129 of the latter were strongly cross-reactive to the former, indicating the antigenic conservation of this site, known to be critical for the membrane-damaging activities of Lys49 myotoxins.

Schmerz, 2004 Jun, 18(3), 203 - 10
{Cannabinoids and the immune system . Of men, mice and cells}; Kraft B et al.; The medical use of cannabis or cannabinoid compounds is controversial . Cannabinoids like the Delta(9)-THC (tetrahydrocannabinol) or the synthetic derivative Nabilone are available against cancer- and HIV-associated cachexia, nausea and vomiting . Over the last 20 years, the cannabinoid receptors CB(1) and CB(2) and their endogenous ligands have been found . The involvement of this endogenous cannabinoid signalling system in feeding, appetite, pain perception and immunomodulation could be demonstrated using animal and in vitro studies . Thus, the concern about immunosuppressive effects in humans using medical cannabinoid preparations grew . However, up to now most human studies have failed to demonstrate a well-defined and reproducible immunosuppressive cannabinoid-effect . Only the smoking of marijuana showed a significant local immunosuppression of the bactericidal activity of human alveolar macrophages.In animal studies, cannabinoids were identified as potent modulators of cytokine production, causing a shift from Th1 to Th2 cytokines . In consequence, a compromised cellular immunity was observed in these animals, resulting in enhanced tumor growth and reduced immunity to viral infections . In vitro, immunosuppressive effects were shown in all immune cells, but only at high micromolar cannabinoid concentrations not reached under normal clinical conditions . In conclusion, there is no evidence that cannabinoids induce a serious, relevant immunosuppression in humans, with the exception of marijuana-smoking which may affect local broncho-alveolar immunity.

Tsitologiia, 2003, 45(8), 832 - 8
Some routine and novel approaches to chemical dosimetry of far-ultraviolet light emitted by bactericidal tubes; Zherebtsov SV; Detailed protocols are presented of two improved chemical procedures, based upon a photochemical decomposition of uranyl oxalate or potassium ferrioxalate, enabling a reliable measurement of the far-ultraviolet light emitted by a low-pressure mercury-vapour lamp . Besides, an original semi-quantitative method of UV dosimetry is presented and discussed, employing optical detection of a chromophore formed in the photo-oxidized glutathione . In addition to exemplary computations of UV light dose rate, a simple formula is proposed for recalculating its value while varying the distance from the lamp.

J Infect Dis, 2004 Jul 15, 190(2), 341 - 51 Epub 2004 Jun 18.
Role of cellular activation and tumor necrosis factor-alpha in the early expression of Mycobacterium tuberculosis 85B mRNA in human alveolar macrophages; Islam N et al.; BACKGROUND: Infection of alveolar macrophages (AMs), which constitute the first line of defense against Mycobacterium tuberculosis, initiates an intense interaction between the host's innate immune response and mycobacteria that may assist in the successful intracellular parasitism of M . tuberculosis . METHODS: Expression of tumor necrosis factor (TNF)- alpha and M . tuberculosis 85B mRNA was studied in M . tuberculosis-infected AMs, to better delineate the role of macrophages in the early events in initiation of infection . RESULTS: Both TNF- alpha mRNA and M . tuberculosis 85B were induced in AMs; at 24 h, the time point of maximum TNF- alpha induction, the mRNA levels for TNF- alpha and M . tuberculosis 85B correlated with one another, and induction of either gene correlated strongly with their protein levels . Inhibition of endogenous TNF- alpha by soluble (s) TNF receptor (R) I and sTNFRII reduced expression of both TNF- alpha and M . tuberculosis 85B . The activation of nuclear factor- kappa B was found to underlie expression of both TNF- alpha and M . tuberculosis 85B . Exogenous TNF- alpha was slightly more potent than interleukin (IL)-6 and granulocyte-macrophage colony-stimulating factor and was significantly stronger than IL-1 in inducing expression of M . tuberculosis 85B . Interestingly, inhibition of bactericidal mediators, reactive oxygen intermediates (ROIs) and reactive nitrogen intermediates (RNIs), reduced expression of TNF- alpha and M . tuberculosis 85B genes in M . tuberculosis-infected AMs . CONCLUSION: Activation of AMs by M . tuberculosis initiates a cascade of events whereby TNF- alpha, ROI, and RNI enhance the expression of the M . tuberculosis 85B gene.

J Biochem (Tokyo), 2004 Jun, 135(6), 713 - 9
Low pH facilitates uptake of proteins by cells through a non-endocytic pathway; Motizuki M et al.; We previously noted that bovine apolipoprotein A-II (apoA-II) had a bactericidal effect causing morphological changes in the cytoplasm . To determine whether and how apoA-II and apoA-I, which have acidic isoelectric points (pIs), enter cells, we determined the rates of uptake of FITC-labeled proteins by fibroblast cells and found that they entered cells more easily at low pH than at neutral pH under conditions where endocytosis was inhibited . The enhanced uptake of proteins at low pH was also observed for other proteins examined regardless of the molecular weight (M(r)) or pI in a time-dependent manner, although the efficiency of uptake varied among the proteins . Furthermore, a pH gradient was shown to be the main driving force for the translocation . As cells were viable above pH 4 for 2 h at 4 degrees C and internalized beta-galactosidase was active under these conditions, we suggest that this procedure is applicable to the injection of proteins into cells without the use of an apparatus such as a microinjector.

Rev Mal Respir, 2004 Apr, 21(2 Pt 1), 261 - 71
{Treatment of exacerbations of chronic obstructive pulmonary disease with pristinamycin}; Leophonte P et al.; BACKGROUND: Pristinamycin is a bactericidal antibiotic whose spectrum covers the main respiratory pathogens including S . pneumoniae poorly sensitive to penicillin . It has not yet been evaluated in short course treatment of acute exacerbations of chronic obstructive bronchitis (AECB) . METHODS: 476 patients suffering from an AECB were randomised to either a short course of pristinamycin, 3 G daily for 4 days, or conventional treatment with co-amoxiclav (AAC) 2G daily for 8 days . The duration of follow-up was 6 months . RESULTS: The clinical success rate at 21 days was the same in both groups at 87.2% and 87.9%, CI95% {-7.0%, 6.0%}, in the protocol population (FEV1<80%) . Among the 120 patients in whom a bacterial pathogen was isolated at the time of inclusion a satisfactory bacteriological response was obtained in 84.6% of the PRI patients against 78.2% of the AAC patients . The time to relapse was comparable with a relapse rate of 25% reached in 128 days in the PRI group and 125 days in the AAC group . Treatment related side effects occurred in 9.2% of the PRI group and in 10.6% of the AAC group . CONCLUSION: Pristinamycin 3 G daily for 4 days is as effective and well tolerated as co-amoxiclav 2G daily for 8 days in the treatment of AECB.

Clin Immunol, 2004 Jul, 112(1), 85 - 91
Polymorphisms of the lipopolysaccharide-signaling complex in inflammatory bowel disease: association of a mutation in the Toll-like receptor 4 gene with ulcerative colitis; Torok HP et al.; Genes encoding for receptors of the innate immune system are potential candidates for susceptibility to inflammatory bowel disease, e.g., mutations in the cytosolic receptor NOD2/CARD15 were associated with Crohn's disease . Herein, two mutations of the Toll-like receptor (TLR)-4 gene (Asp299Gly and Thr399Ile) resulting in impaired lipopolysaccharide signaling, the -159C/T promotor polymorphism of the CD14 gene, polymorphisms of the lipopolysaccharide binding protein gene and the bactericidal permeability increasing protein gene were evaluated in 102 patients with Crohn's disease, 98 patients with ulcerative colitis and 145 healthy controls . The allele and carrier frequencies for the Thr399Ile mutation in TLR4 gene were significantly increased in ulcerative colitis when compared to the controls (P = 0.014 and P = 0.018, respectively) . None of the other five polymorphisms was associated with inflammatory bowel disease . In conclusion, a novel association between a functional polymorphism in TLR4 and ulcerative colitis is reported . This observation underscores the importance of impaired innate immunity in inflammatory bowel disease.

J Periodontal Res, 2004 Aug, 39(4), 275 - 85
Potential applications of Erbium:YAG laser in periodontics; Ishikawa I et al.; OBJECTIVES: Since lasers were introduced for the treatment of oral diseases, there has been considerable advancement in technology . As a result, numerous laser systems are currently available for oral use . Neodymium:Yttrium-Aluminum:Garnet (Nd:YAG), carbon dioxide (CO(2)) laser and the semiconductor Diode lasers have already been approved by the US Food and Drug Administration for soft tissue treatment in oral cavity . The Erbium:YAG (Er:YAG) laser was approved in 1997 for hard tissue treatment in dentistry and recent studies have reported positive results . This suggests that the Er:YAG laser system is a promising apparatus, which will be able to revolutionize and improve dental practice, in particular periodontal treatment . In this mini-review, we would like to describe the positive characteristics of the Er:YAG laser which indicate its potential as a new treatment modality in periodontics . MATERIALS AND METHODS: Recent findings are summarized briefly to evaluate the potential of the Er:YAG laser for clinical application in periodontics . RESULTS: The Er:YAG laser possesses suitable characteristics for oral soft and hard tissue ablation . Recently, it has been applied for effective elimination of granulation tissue, gingival melanin pigmentation and gingival discoloration . Contouring and cutting of bone with minimal damage and even or faster healing can also be performed with this laser . In addition, irradiation with the Er:YAG laser has a bactericidal effect with reduction of lipopolysaccharide, high ability of plaque and calculus removal, with the effect limited to a very thin layer of the surface and is effective for implant maintenance . CONCLUSION: The Er:YAG laser seems to be an effective tool for periodontal therapy, however, further clinical and basic investigations are required to confirm its clinical application . Copyright Blackwell Munksgaard, 2004

Biochimie, 2004 Apr-May, 86(4-5), 283 - 6
Putative membrane assembly of EtpM-colicin V chimeras; Gerard F et al.; EtpM of the enterohemorrhagic E . coli O157:H7 is a bitopic membrane protein of the type II protein secretion apparatus . There is a twin-arginine (RR) motif in front of its signal anchor, suggesting a Tat-dependent membrane targeting of EtpM . By exploiting the periplasmic bactericidal activity of colicin V (ColV), we constructed EtpM-ColV fusions and studied the EtpM-mediated translocation of ColV . The wild type strain and the DeltatatC mutant were killed by the expressed fusions and were fully protected from the killing effect by the ColV-specific immunity protein . In contrast, cold-inactivation of YidC, which is generally required for integral membrane protein assembly, significantly attenuated the killing effect in the cold-sensitive yidC mutant . These results confirmed the predicted N(in)-C(out) EtpM topology, and suggests an EtpM-mediated, Tat-independent and YidC-dependent translocation of ColV.

J Dairy Res, 2004 May, 71(2), 154 - 61
Effect of recombinant cytokines on leucocytes and physiological changes in bovine mammary glands during early involution; Wedlock DN et al.; We examined the effects of administering recombinant bovine cytokines to non-lactating dairy cows and measured mammary gland leucocytes and the involution process . After the final milking, groups of cows were given an intramammary infusion of cytokine in two quarters . These cytokines were recombinant bovine interleukin-2 (rbolL-2) (2 x 10(5) units, n = 6), recombinant bovine granulocyte-macrophage colony stimulating factor (rboGM-CSF) (500 microg, n = 4) and recombinant bovine interleukin-1beta (rbolL-1beta) (10 microg, n = 10) . Each animal also received an infusion of phosphate-buffered saline (PBS) in the other two quarters as controls . The rbolL-2 and rboGM-CSF were produced in a yeast expression system, while rbolL-1beta was produced in Escherichia coli . Leucocyte numbers, bactericidal activity of leucocytes, and concentrations of citrate and lactoferrin in quarter secretion samples were monitored after infusion of cytokine or PBS . Infusion of rbolL-2 had minimal effect on leucocyte numbers and concentrations of citrate and lactoferrin . Both rboGM-CSF and rbolL-1beta induced a rapid increase in the number of neutrophils and macrophages compared with control PBS quarters . Concentrations of lactoferrin in secretions were increased by rboGM-CSF and rbolL-1beta compared with control PBS quarters . In addition, infusion of glands with rbolL-1beta lowered the citrate:lactoferrin molar ratio compared with PBS control quarters . The results indicate that intramammary infusion of either rboGM-CSF or rbolL-1beta at cessation of milking immediately increased the number of phagocytic cells in the gland . These cytokines, in particular rbolL-1beta, also increased the rate of mammary gland involution during the early dry period.

Biochemistry, 2004 Jun 8, 43(22), 7046 - 53
Toward better antibiotics: crystallographic studies of a novel class of DD-peptidase/beta-lactamase inhibitors; Silvaggi NR et al.; Beta-lactam antibiotics are vital weapons in the treatment of bacterial infections, but their future is under increasing threat from beta-lactamases . These bacterial enzymes hydrolyze and inactivate beta-lactam antibiotics, rendering the host cell resistant to the bactericidal effects of the drugs . Nevertheless, the bacterial D-alanyl-D-alanine transpeptidases (DD-peptidases), the killing targets of beta-lactams, remain attractive targets for antibiotic compounds . Cyclic acyl phosph(on)ates have been developed and investigated as potential inhibitors of both transpeptidases and beta-lactamases . The X-ray crystal structures of the complexes of the Streptomyces strain R61 DD-peptidase inhibited by a bicyclic {1-hydroxy-4,5-benzo-2,6-dioxaphosphorinanone(3)-1-oxide} and a monocyclic {1-hydroxy-4-phenyl-2,6-dioxaphosphorinanone(3)-1-oxide} acyl phosphate were determined to investigate the mode of action of these novel inhibitors . The structures show, first, that these inhibitors form covalent bonds with the active site serine residue of the enzyme and that the refractory complexes thus formed are phosphoryl-enzyme species rather than acyl enzymes . The complexes are long-lived largely because, after ring opening, the ligands adopt conformations that cannot directly recyclize, the latter a phenomenon previously observed with cyclic acyl phosph(on)ates . While the two inhibitors bind in nearly identical conformations, the phosphoryl-enzyme complex formed from the monocyclic compound is significantly less mobile than that formed from the bicyclic compound . Despite this difference, the complex with the bicyclic compound breaks down to regenerate free enzyme somewhat more slowly than that of the monocyclic . This may be because of steric problems associated with the reorientation of the larger bicyclic ligand required for reactivation . The structures are strikingly different in the orientation of the phosphoryl moiety from those generated using more specific phosph(on)ates . Models of the noncovalent complexes of the monocyclic compound with the R61 DD-peptidase and a structurally very similar class C beta-lactamase suggest reasons why the former enzyme is phosphorylated by this compound, while the latter is acylated . Finally, this paper provides information that will help in the design of additional DD-peptidase inhibitors with the potential to serve as leads in the development of novel antibiotics.

Am J Kidney Dis, 2004 Jun, 43(6), 1030 - 9
Defective regulation of iron transporters leading to iron excess in the polymorphonuclear leukocytes of patients on maintenance hemodialysis; Otaki Y et al.; BACKGROUND: Although hemodialysis (HD) patients are suspected of having defectively regulated iron metabolism, intracellular iron status has never been investigated thoroughly . To clarify the iron metabolism of HD patients, proteins involved in iron import (transferrin receptor {TfR}), as well as export (ferroportin 1), were investigated in polymorphonuclear leukocytes (PMNLs) . Relations between iron status and several PMNL functions also were tested . METHODS: Seventeen HD patients and 17 controls were recruited . Relative quantitative polymerase chain reaction was used to measure ferroportin 1 and TfR messenger RNA (mRNA), and ferroportin 1 and TfR expression were semiquantified by means of Western blot analysis or immunohistochemistry . PMNL functions also were examined . RESULTS: Serum iron levels were significantly lower in HD patients than controls, and serum ferritin levels, as well as PMNL ferritin and iron content, were elevated in HD patients . Ferroportin 1 mRNA levels were substantially lower in PMNLs from HD patients, whereas TfR mRNA levels were higher . Western blot analysis and immunohistochemistry confirmed that expression of the corresponding proteins paralleled those of the mRNAs . PMNL phagocytic and bactericidal activity did not differ between HD patients and controls . Chemotactic peptide f-Met-Leu-Phe-stimulated degranulation activity of lactoferrin (Lf) was decreased significantly in HD patients, whereas those of myeloperoxidase and elastase were accelerated . Lf release correlated negatively with intracellular ferritin level . CONCLUSION: We show for the first time that increased iron levels in PMNLs of HD patients were associated with downregulation of ferroportin 1 and upregulation of TfR, which might be linked to hypercytokinemia.

J Antimicrob Chemother, 2004 Jul, 54(1), 79 - 85 Epub 2004 May 26.
Pre-exposure of infected human endometrial epithelial cells to penicillin in vitro renders Chlamydia trachomatis refractory to azithromycin; Wyrick PB et al.; OBJECTIVE: The clinical significance of the potential for persistent human chlamydial infections in vivo is being actively reassessed because of the increased frequency of recurrent infection with the same serovar despite compliance with an effective antibiotic regimen . The ability to extend the length of time of in vitro cultivation of polarized human endometrial epithelial cells (HEC-1B) provided the opportunity to establish a model system to determine if a persistent form of Chlamydia trachomatis had the same susceptibility as the actively growing form to a cidal concentration of azithromycin . METHODS: Polarized HEC-1B cells cultivated on extracellular matrix were infected with C . trachomatis serovar E and exposed to penicillin at 24 h post-infection (hpi) to induce a persistent infection characterized by slowly metabolizing but non-dividing, ultrastructurally aberrant reticulate bodies within the chlamydial inclusion; at 48 hpi, infected cultures were exposed to a bactericidal concentration of azithromycin for 72 h . RESULTS: Persistent chlamydiae were phenotypically resistant to azithromycin; the number of chlamydial inclusions on subpassage of progeny from persistent chlamydiae following removal of penicillin and recovery was essentially the same as from progeny from persistent chlamydiae following removal of penicillin and azithromycin and recovery . Neutrophils were attracted in vitro to persistently infected HEC-1B cells that had been exposed to penicillin and azithromycin . CONCLUSIONS: Thus, this study provides evidence at the cellular microbiology level in vitro for mechanisms that could exist in vivo to create sustained, but perhaps clinically inapparent inflammation, which might eventually lead to conditions such as silent pelvic inflammatory disease.

Immunol Lett, 2004 May 15, 93(2-3), 143 - 9
Down-regulatory effect of N-chlorotaurine on tryptophan degradation and neopterin production in human PBMC; Wirleitner B et al.; N-Chlorotaurine (NCT) plays an important role in the human defense system as a main component of long-lived oxidants, and shows bactericidal, fungicidal, and virucidal activity . Besides this role, NCT seems to act regulatory on immunocompetent cells by altering cytokine production . NCT inhibited nitric oxide, TNF-alpha, and prostaglandin E(2) (PGE(2)) production in activated rodent macrophages, and suppressed superoxide anion, IL-6, and IL-8 formation in human polymorphonuclear leukocytes . In this study, the influence of NCT on the production of neopterin and the activation of the enzyme indoleamine-2,3 dioxygenase (IDO) was investigated in human peripheral blood mononuclear cells (PBMC) . Both events are well established to be triggered by IFN-gamma and therefore related to Th1-type immune activation . Mitogen-induced neopterin production as well as tryptophan degradation were drastically reduced upon addition of NCT . Results fit in the concept of a reduction of pro-inflammatory cytokines by this compound . In contrast to earlier results, where NCT was suggested to act primarily down-regulatory on Th2 cells, we propose also a strong suppressive effect of NCT on Th1-type immunity.

Eur J Biochem, 2004 Jun, 271(11), 2117 - 26
A Kazal prolyl endopeptidase inhibitor isolated from the skin of Phyllomedusa sauvagii; Gebhard LG et al.; Searching for bioactive peptides, we analyzed acidic extracts of Phyllomedusa sauvagii skin and found two new proteins, PSKP-1 and PSKP-2, of 6.7 and 6.6 kDa, respectively, which, by sequence homology, belong to the Kazal family of serine protease inhibitors . PSKP-1 and PSKP-2 exhibit the unprecedented feature of having proline at P(1) and P(2) positions . A gene encoding PSKP-1 was synthesized and expressed in Escherichia coli . Recombinant PSKP-1 was purified from inclusion bodies, oxidatively refolded to the native state, and characterized by chemical, hydrodynamic and optical studies . PSKP-1 shows inhibitory activity against a serum prolyl endopeptidase, but is unable to inhibit trypsin, chymotrypsin, V8 protease, or proteinase K . In addition, PSKP-1 can be rendered active against trypsin by active-site site-specific mutagenesis, has bactericidal activity, and induces agglutination of red cells at micromolar concentrations . PSKP-1 might protect P . sauvagii teguments from microbial invasion, by acting as an inhibitor of an as-yet unidentified prolyl endopeptidase or directly as a microbicidal compound.

FEMS Immunol Med Microbiol, 2004 Jun 1, 41(2), 109 - 15
Exploration of Moraxella catarrhalis outer membrane proteins, CD and UspA, as new carriers for lipooligosaccharide-based conjugates; Hu WG et al.; Moraxella catarrhalis outer membrane proteins, CD and ubiquitous surface protein A (UspA), were used as carriers for M . catarrhalis detoxified lipooligosaccharide (dLOS)-based conjugates . Our study was designed to investigate the feasibility of CD and UspA as protein carriers for dLOS-based conjugates and their possible synergic effects on protection from both anti-LOS and anti-CD or anti-UspA antibody responses . Female Balb/c mice were immunized subcutaneously three times with dLOS-CD or dLOS-UspA conjugate in Ribi adjuvant . Antisera elicited by the conjugates showed high titers of specific anti-LOS antibodies with complement-dependent bactericidal activity towards M . catarrhalis strain 25238 . In a mouse aerosol challenge model, mice immunized with both conjugates showed a significant enhancement of the clearance of strain 25238 from lungs as compared with the control mice . Although both conjugates elicited reduced (relative to unconjugated CD or UspA) but significant levels of anti-CD or UspA antibodies, they did not show synergetic effects with anti-LOS antibodies on the bactericidal activity or the pulmonary bacterial clearance . Nevertheless, CD and UspA are safe and effective new carriers for dLOS-based or other potential carbohydrate-based conjugate vaccines to help thymus-independent carbohydrate antigens for production of anti-carbohydrate antibodies against target pathogens.

Cell Mol Life Sci, 2004 May, 61(10), 1229 - 37
Periplasmic lysozyme inhibitor contributes to lysozyme resistance in Escherichia coli; Deckers D et al.; The product of the Escherichia coli ORFan gene ykfE was recently shown to be a strong inhibitor of C-type lysozyme in vitro . The gene was correspondingly renamed ivy (inhibitor of vertebrate lysozyme), but its biological function in E . coli remains unknown . In this work, we investigated the role of Ivy in the resistance of E . coli to the bactericidal effect of lysozyme in the presence of outer-membrane-permeabilizing treatments . Both in the presence of lactoferrin (3.0 mg/ml) and under high hydrostatic pressure (250 MPa), the lysozyme resistance of E . coli MG1655 was decreased by knock-out of Ivy, and increased by overexpression of Ivy . However, knock-out of Ivy did not increase the lysozyme sensitivity of an E . coli MG1655 mutant previously described to be resistant to lysozyme under high pressure . These results indicate that Ivy is one of several factors that affect lysozyme resistance in E . coli, and suggest a possible function for Ivy as a host interaction factor in commensal and pathogenic E . coli.

Dig Dis Sci, 2004 Mar, 49(3), 370 - 8
Bismuth subsalicylate increases intracellular Ca2+, MAP-kinase activity, and cell proliferation in normal human gastric mucous epithelial cells; Gilster J et al.; Clinical and laboratory studies have shown that bismuth subsalicylate (BSS) is helpful in the healing of gastric ulcers because of the bactericidal effects of bismuth (Bi3+) on H . pylori . Bismuth or BSS has also been reported to possess other nonbactericidal or "gastroprotective" effects in the stomach . It is known in other cell types that the effects of extracellular divalent or trivalent cations (e.g., Ca2+) can activate a plasma membrane-bound calcium-sensing receptor (CaSR) . In a previous study, we found the existence of a CaSR which was activated by extracellular Ca2+ and found to increase intracellular Ca2+ {Ca2+}i, MAP-kinase activity, and gastric epithelial cell proliferation . In the present study, we were interested in determining whether the effects of the trivalent cation Bi3+ (in the form of BSS) on {Ca2+}i, MAP-kinase activity, and proliferation of gastric cells . We found that BSS dose dependently increased {Ca2+}i, p44/p42 and p38 MAP-kinase activites, and gastric mucous epithelial cell growth . The addition of BAPTA to chelate intracellular Ca2+ blocked BSS-induced p44/p42 MAP-kinase activities but not p38 MAP-kinase activity . The p44/p42 MAP-kinase inhibitor PD98059 and the p38 MAP-kinase inhibitor SB203580 dose dependently decreased gastric mucous cell growth over a 24 hr . All of the BSS-induced changes in {Ca2+}i, MAP-kinase activity, and gastric cell proliferation could be reproduced with the CaSR-agonist gadolinium (Gd3+) . Our data suggest that BSS may possess additional novel effects by increasing gastric mucous epithelial cell growth through a Ca2+/MAP-kinase-dependent pathway.

Clin Diagn Lab Immunol, 2004 May, 11(3), 559 - 62
Detection rate and antigenic specificities of antineutrophil cytoplasmic antibodies in chinese patients with clinically suspected vasculitis; Xin G et al.; The detection rate of antineutrophil cytoplasmic antibodies (ANCA) in Chinese patients with clinically suspected small vessel vasculitis was investigated, and their antigen specificity and demographic features were analyzed . A number of sera (n = 5,604) sent to our referral laboratory for ANCA screening were tested by indirect immunofluorescence (IIF), enzyme-linked immunosorbent assays (ELISAs) for myeloperoxidase (MPO)- and proteinase 3 (PR3)-ANCA . Then the IIF-ANCA-positive sera that were negative for MPO- and PR3-ANCA were further tested by antigen-specific ELISA by using other five highly purified known ANCA antigens as solid-phase ligands . The known antigens included bactericidal/permeability-increasing protein (BPI), human leukocyte elastase (HLE), lactoferrin, cathepsin G, and azurocidins . Of the 5,604 sera, 267 (4.76%) sera were IIF-ANCA positive and 390 (7%) were antinuclear antibody (ANA) positive in the IIF assay . Of the IIF-positive samples, 213 were anti-MPO positive, 32 were anti-PR3 positive, and five cases were positive for both . Of the 48 sera positive for IIF-ANCA but negative for MPO- and PR3-ANCA, 13 sera (27%) recognized other target antigens, 7 sera recognized BPI, 5 recognized HLE, 1 recognize cathepsin G, and 1 recognized azurocidin . None of the sera recognized lactoferrin, and one serum sample recognized both BPI and HLE . The majority of ANCA-positive patients presented in summer or winter . There was no difference in gender (male/female ratio, 1:1.12) in ANCA-positive patients with a mean age of 53.1 years . The male/female ratio was 1.17:1 for patients over 60 years of age; however, it was 1:4 for patients under 20 years of age . We conclude that ANCA-related diseases are not rare in China, and the major antigens are MPO and PR3 . When the IIF technique is used to detect ANCA, ANA should be carefully distinguished.

Mol Cell Probes, 2004 Jun, 18(3), 167 - 70
A pncA polymorphism to differentiate between Mycobacterium bovis and Mycobacterium tuberculosis; Barouni AS et al.; The pyrazinamidase gene coding for the enzyme that activates the bactericidal drug pyrazinamide contains a polymorphic site that is preserved in Mycobacterium bovis . We synthesized two sets of primers, one encompassing a 180 bp fragment, and the second spanning a 726 bp fragment including the full pncA gene . Following PCR of Mycobacterium tuberculosis and M . bovis samples, it is possible to discriminate by this polymorphism between these species by digestion with Eco065 I . Digestion of the 180 bp fragment results in two fragments of 101 and 79 bp, specific for M . tuberculosis . Alternatively, digestion of the 726 bp fragment yields three fragments of 452, 165 and 109 bp for M . tuberculosis, but only two fragments of 561 and 165 bp for M . bovis.

J Am Anim Hosp Assoc, 2004 May-Jun, 40(3), 224 - 9
Bone marrow hypoplasia associated with fenbendazole administration in a dog; Gary AT et al.; A 1.5-year-old Doberman pinscher was presented with sudden-onset of fever and malaise . Twelve days prior to presentation, fenbendazole therapy was initiated for a suspected lungworm infection . Results of a complete blood count on presentation showed pancytopenia, while histopathological evaluation of a bone marrow core sample revealed bone marrow hypoplasia of undetermined etiology . Bactericidal antibiotics and fluid therapy, as well as discontinuation of fenbendazole administration, led to a complete resolution of clinical and hematological abnormalities within 15 days . An idiosyncratic reaction to fenbendazole was suspected based on the absence of infectious, neoplastic, autoimmune, and toxic etiologies, as well as resolution of clinical signs and pancytopenia upon drug withdrawal.

J Immunol, 2004 May 15, 172(10), 6272 - 80
Mycobacterium tuberculosis inhibits macrophage responses to IFN-gamma through myeloid differentiation factor 88-dependent and -independent mechanisms; Fortune SM et al.; Mycobacterium tuberculosis overcomes macrophage bactericidal activities and persists intracellularly . One mechanism by which M . tuberculosis avoids macrophage killing might be through inhibition of IFN-gamma-mediated signaling . In this study we provide evidence that at least two distinct components of M . tuberculosis, the 19-kDa lipoprotein and cell wall peptidoglycan (contained in the mycolylarabinogalactan peptidoglycan (mAGP) complex), inhibit macrophage responses to IFN-gamma at a transcriptional level . Moreover, these components engage distinct proximal signaling pathways to inhibit responses to IFN-gamma: the 19-kDa lipoprotein inhibits IFN-gamma signaling in a Toll-like receptor (TLR)2-dependent and myeloid differentiation factor 88-dependent fashion whereas mAGP inhibits independently of TLR2, TLR4, and myeloid differentiation factor 88 . In addition to inhibiting the induction of specific IFN-gamma responsive genes, the 19-kDa lipoprotein and mAGP inhibit the ability of IFN-gamma to activate murine macrophages to kill virulent M . tuberculosis without inhibiting production of NO . These results imply that inhibition of macrophage responses to IFN-gamma may contribute to the inability of an apparently effective immune response to eradicate M . tuberculosis.

Biomaterials, 2004 Aug, 25(19), 4555 - 62
Synthesis of metal incorporated low molecular weight polyurethanes from novel aromatic diols, their characterization and bactericidal properties; Acharya V et al.; Low molecular weight polyurethanes with sites for metal complexation were synthesized . The -SO(2) and the -COOH groups were incorporated into the polyurethane chain by the reactions of tri-functional monomers, 4,4'-bis(hydroxyphenyl)sulfone and 4,4'-bis(hydroxyphenyl) valeric acid with hexa-methylene di-isocyanate (HMDI) and 2,4 toluene di-isocyanate (TDI), respectively . The reaction was monitored from the disappearance of the -OH group and results show that the -NCO groups of the isocyanate reacted with the -OH group preferentially when compared to the other reactive groups . The pristine tri-functional monomers themselves formed stable complexes with the metals and so were chosen to be incorporated into the polyurethane chain . These polymers were characterized by IR and NMR spectroscopy . The free -SO(2) and the -COOH groups were used for metal complexation using non-toxic metals like zinc and silver . The anti-bacterial studies conducted on the six polymers in film form showed interesting results as elaborated in the paper.

Cell Biol Toxicol, 2004 Feb, 20(1), 41 - 54
The effect of enrofloxacin on cell proliferation and proteoglycans in horse tendon cells; Yoon JH et al.; Fluoroquinolone antibiotics have been used widely in humans and domestic animals, including horses, because of their broad-spectrum bactericidal activity, and relative safety . The use of fluoroquinolones, however, is not without risk . Tendonitis and spontaneous tendon rupture have been reported in people during or following therapy with fluoroquinolones . We have studied the effects of enrofloxacin, a fluoroquinolone antibiotic used commonly in domestic animals, on tendon cell cultures established from equine superficial digital flexor tendons . Effects on cell proliferation and morphology were studied using cell counting and scanning electron microscopy . Monosaccharide content and composition was determined by gas chromatography-mass spectrometry analysis . Western and Northern blot analyses were utilized to evaluate the synthesis and expression of two proteoglycans, biglycan and decorin . Our data demonstrate that enrofloxacin inhibits cell proliferation, induces morphological changes, decreases total monosacharide content and alters small proteoglycan synthesis at the glycosylation level in equine tendon cell cultures . These effects are more pronounced in juvenile tendon cells than in adult equine tendon cells . We hypothesize that morphological changes and inhibition of cell proliferation are a result of impaired production of biglycan and decorin, proteoglycans involved in fibrillogenesis of collagen, the most important structural component of the tendon of enrofloxacin-treated tendon cells . Our findings suggest that fluoroquinolones should be used with caution in horses, especially in foals.

Jpn J Infect Dis, 2004 Apr, 57(2), 52 - 4
Bactericidal effects of acidic electrolyzed water on the dental unit waterline; Kohno S et al.; Many studies have been conducted in the United States regarding the microbial contamination of dental unit waterline, but not in Japan . Recently, acidic electrolyzed water has been used in the medical and dental fields . In this study, we investigated the bactericidal effects of the temporary inflow of acidic electrolyzed water on microbial contamination of the dental unit waterline . First, in order to observe the daily bacterial contamination of the dental unit waterline, water samples were collected at the end of handpieces and three-way syringes before the inflow of acidic electrolyzed water . They were cultured to detect viable bacteria . Later, the inflow of acidic electrolyzed water was conducted through the piping box of the dental unit . Before starting operation on next day, water samples were collected and cultured, as described above . The mean viable bacteria count was 910 -/+ 190 CFU/ml at the end of handpieces, and 521 -/+ 116 CFU/ml at the end of three-way syringes before the inflow of acidic electrolyzed water . However, bacteria were detected in only small numbers at the end of handpieces and three-way syringes on the next day . These results indicated that acidic electrolyzed water could be applied as an appropriate measure against bacterial contamination of the dental unit waterline.

J Leukoc Biol, 2004 Jul, 76(1), 254 - 62 Epub 2004 Apr 23.
Disturbed granulocyte macrophage-colony stimulating factor priming of phosphatidylinositol 3,4,5-trisphosphate accumulation and Rac activation in fMLP-stimulated neutrophils from patients with myelodysplasia; Fuhler GM et al.; The production of reactive oxygen species (ROS) by human neutrophils is imperative for their bactericidal activity . Proinflammatory agents such as granulocyte macrophage-colony stimulating factor (GM-CSF) can prime ROS production in response to chemoattractants such as N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) . In neutrophils from patients suffering from Myelodysplastic syndromes (MDS), a clonal, hematological disorder characterized by recurrent bacterial infections, this GM-CSF priming is severely impaired . In this study, we set out to further delineate the defects in neutrophils from MDS patients . We examined the effect of GM-CSF priming on fMLP-triggered activation of Rac, a small GTPase implicated in neutrophil ROS production . In contrast to healthy neutrophils, activation of Rac in response to fMLP was not enhanced by GM-CSF pretreatment in MDS neutrophils . Furthermore, activation of Rac was attenuated by pretreatment of neutrophils with the phosphatidylinositol 3-kinase (PI-3K) inhibitor LY294002 . Unlike healthy neutrophils, fMLP-induced accumulation of the PI-3K lipid product PI(3,4,5)trisphosphate was not increased by GM-CSF pretreatment in MDS neutrophils . The disturbed Rac and PI-3K activation observed in MDS neutrophils did not appear to reflect a general GM-CSF or fMLP receptor-signaling defect, as fMLP-triggered Ras activation could be primed by GM-CSF in MDS and healthy neutrophils . Moreover, fMLP-induced activation of the GTPase Ral was also normal in neutrophils from MDS patients . Taken together, our data suggest that in neutrophils from MDS patients, a defect in priming of the PI-3K-Rac signaling pathway, located at the level of PI-3K, results in a decreased GM-CSF priming of ROS production.

J Korean Med Sci, 2004 Apr, 19(2), 289 - 90
Anaphylaxis caused by benzalkonium in a nebulizer solution; Kim SH et al.; Benzalkonium chloride (BAC) is commonly used as a bactericidal preservative in nebulizer solutions, and can cause paradoxical onchoconstriction following nebulizing therapy in some asthmatics . We describe a case of anaphylactic shock in a 23-yr-old asthmatic woman following an intradermal skin test with a salbutamol solution containing BAC . Since she complained of cough and dyspnea after inhalation therapy with a nebulizer solution, we conducted an intradermal skin test using the same solution, which contained BAC . About 10 min later, the patient reported dizziness, palpitations, and dyspnea . On examination, tachycardia, tachypnea, and hypotension were found . She was resuscitated with a subcutaneous injection of epinephrine and an infusion of saline . One month later, we conducted a bronchial provocation test with BAC, and she showed a positive response.

Ultrason Sonochem, 2004 May, 11(3-4), 167 - 72
Inactivation effect of sonication and chlorination on Saccharomyces cerevisiae . Calorimetric analysis; Tsukamoto I et al.; The inactivation effects of ultrasonic irradiation at 27.5 kHz and chlorination using sodium hypochlorite solution (NaOCl) on the growth of Saccharomyces cerevisiae (yeast cells) were investigated . In order to evaluate the effect of ultrasound on the growth of the yeast cells, calorimetric analysis was carried out in addition to colony counting . The heat evolution produced by the growth of yeast cells detected by calorimetry showed completely different patterns between sonication and chlorination . In case of sonication, the yeast cells were inactivated almost like a bactericidal effect, i.e . a quantitative change in cell number, at the beginning of sonication . It was similar to patterns obtained on simple dilution of yeast cells . In contrast, longer sonication increased the bacteriostatic effect, i.e . qualitative damage of the cell growth activity, together with the bactericidal effect . These results suggest that the cavitation caused by ultrasonic irradiation initially disrupted the cells located near the cavitation bubble which caused immediate cell death and the growth activity of the surviving cells was gradually damaged by further sonication . On the other hand, only a bacteriostatic effect was observed when the yeast cells were inactivated by chlorination.

Am J Vet Res, 2004 Apr, 65(4), 473 - 9
Pharmacokinetics of a high dose of amikacin administered at extended intervals to neonatal foals; Magdesian KG et al.; OBJECTIVE: To determine disposition kinetics of amikacin in neonatal foals administered high doses at extended intervals . ANIMALS: 7 neonatal foals . PROCEDURE: Amikacin was administered (21 mg/kg, i.v., q 24 h) for 10 days . On days 1, 5, and 10, serial plasma samples were obtained for measurement of amikacin concentrations and determination of pharmacokinetics . RESULTS: Mean +/- SD peak plasma concentrations of amikacin extrapolated to time 0 were 103.1 +/- 23.4, 102.9 +/- 9.8, and 120.7 +/- 17.9 microg/mL on days 1, 5, and 10, respectively . Plasma concentrations at 1 hour were 37.5 +/- 6.7, 32.9 +/- 2.6, and 30.6 +/- 3.5 microg/mL; area under the curve (AUC) was 293.0 +/- 61.0, 202.3 +/- 40.4, and 180.9 +/- 31.2 (microg x h)/mL; elimination half-life (t(1/2)beta) was 5.33, 4.08, and 3.85 hours; and clearance was 1.3 +/- 0.3, 1.8 +/- 0.4, and 2.0 +/- 0.3 mL/(min x kg), respectively . There were significant increases in clearance and decreases in t(1/2)beta, AUC, mean residence time, and plasma concentrations of amikacin at 1, 4, 8, 12, and 24 hours as foals matured . CONCLUSIONS AND CLINICAL RELEVANCE: Once-daily administration of high doses of amikacin to foals resulted in high peak plasma amikacin concentrations, high 1-hour peak concentrations, and large values for AUC, consistent with potentially enhanced bactericidal activity . Age-related findings suggested maturation of renal function during the first 10 days after birth, reflected in enhanced clearance of amikacin . High-dose, extended-interval dosing regimens of amikacin in neonatal foals appear rational, although clinical use remains to be confirmed.

Med Electron Microsc, 2004 Mar, 37(1), 16 - 28
Differentiation and function of Kupffer cells; Naito M et al.; Kupffer cells are the largest population of tissue macrophages . They are predominantly distributed in the lumen of hepatic sinusoids and exhibit endocytic activity against blood-borne materials entering the liver . Macrophage colony-stimulating factor and other growth factors regulate Kupffer cell differentiation in the fetal and adult period . Because of the unique attributes of tissue, Kupffer cells play essential roles not only in host defense but also in the homeostatic responses of tissue . Macrophage scavenger receptors and heme oxygenase are expressed in Kupffer cells from an early stage of ontogeny . Scavenger receptors are involved not only in the lipid metabolism but also in the bactericidal mechanism . Heme oxygenase in Kupffer cells is essential to the production of bilirubin . In this review, the developmental mechanism and functional activities of Kupffer cells are described . Evidence suggests that Kupffer cells represent a distinct cell population with unique differentiation mechanisms, metabolic functions, and responsiveness to inflammatory agents.

J Am Chem Soc, 2004 Apr 7, 126(13), 4432 - 6
Hydrogen isotope effects and mechanism of aqueous ozone and peroxone decompositions; Lesko TM et al.; Hydrogen peroxide exalts the reactivity of aqueous ozone by reasons that remain obscure . Should H2O2 enhance free radical production, as it is generally believed, a chain mechanism propagated by (.OH/.O2-) species would account for O3 decomposition rates in neat H2O, HR-O3, and in peroxone (O3 + H2O2) solutions, HPR-O3 . We found, however, that: (1) the radical mechanism correctly predicts HR-O3 but vastly overestimates HPR-O3, (2) solvent deuteration experiments preclude radical products from the (O3 + HO2-) reaction . The modest kinetic isotope effect (KIE) we measure in H2O/D2O: HR-O3/DR-O3 = 1.5 +/- 0.3, is compatible with a chain process driven by electron- and/or O-atom transfer processes . But the large KIE found in peroxone: HPR-O3/DPR-O3 = 19.6 +/- 4.0, is due to an elementary (O3 + HO2-) reaction involving H-O2- bond cleavage . Since the KIE for the hypothetical H-atom transfer: O3 + HO2- HO3 . +.O2-, would emerge as a KIE1/2 factor in the rates of the ensuing radical chain, the magnitude of the observed KIE must be associated with the hydride transfer reaction that yields a diamagnetic species: O3 + HO2- HO3- + O2 . HO3-/H2O3 may be the bactericidal trioxide recently identified in the antibody-catalyzed addition of O2(1Deltag) to H2O.

Ann Ig, 2003 Nov-Dec, 15(6), 885 - 94
{Evaluation of the bactericidal activity of a disinfectant containing sodium hypochlorite (Amiclor)}; Bagordo F et al.; We evaluated the stability and the bactericidal activity of a disinfectant containing sodium hypochlorite (Amiclor), which associates the buffer effect of the system sodium carbonate/sodium tetraborate to the stabilizing action of sodium chloride . The stability was determined evaluating the trend of the active chlorine title in various conservation conditions and comparing it with a product having only sodium chloride as stabilizing component . The bactericidal activity, instead, was evaluated by suspension and surface tests against Escherichia coli (ATCC 10536) and in relation to several variables, as product concentration, contact time, presence of interfering substances and water hardness . From the obtained data it is possible to affirm that Amiclor owns a greater stability as regards the traditional products stabilized only with sodium chloride . The activity tests have pointed out that the practical conditions affect the choice of the combination "product concentration/contact time" necessary to obtain an effective bactericidal activity . The activity of Amiclor is conditioned by the presence of proteins in the mixture or on the surface of reaction . This influence is not very clear in clean conditions while it considerably increases in dirty conditions . A decrease of bactericidal activity was observed when the product was diluted in hard water, mainly for contact times of 5 minutes or low product concentrations.

Izv Akad Nauk Ser Biol, 2004 Jan-Feb, (1), 86 - 91
{Immunogenic properties of the colloidal gold}; Dykman LA et al.; We studied the capacity of colloidal gold for enhancing specific and nonspecific immune response in laboratory animals (rabbits, rats, and mice) immunized with antigens of various nature . The antibody titers obtained with colloidal gold as a carrier were higher as compared to the standard immunization techniques (free antigen or Freund's adjuvant) . Application of colloidal gold increased nonspecific immune responses as well: lysozyme concentration in the blood, activity of the complement system proteins, as well as phagocytic and bactericidal activities . The obtained antibodies were tested by immunodot assay using gold markers . Immunization of the animals with colloidal gold conjugates with haptens as well as complete antigens was shown to induce formation of highly active antibodies without using other antigens such as complete Freund's adjuvant . In addition, antigen quantities for animal immunization with colloidal gold was by one order of magnitude lower as compared to the complete Freund's adjuvant immunization . This fact can point to direct adjuvant activity of colloidal gold.

Am J Kidney Dis, 2004 Apr, 43(4), e7 - 10
Azurocidin-specific-ANCA-related idiopathic necrotizing crescentic glomerulonephritis; Kimura R et al.; An 80-year-old woman who had rapidly progressive glomerulonephritis unaccompanied by systemic vasculitis is described . On renal biopsy, she showed necrotizing crescentic glomerulonephritis by light microscopy and pauci-immune glomerular lesions by immunofluorescent study . No dense deposits were present on electronmicroscopic study . On serum examination, indirect immunofluorescent study showed perinuclear pattern antineutrophil cytoplasmic antibody (ANCA), but myeloperoxidase-ANCA and proteinase 3-ANCA were both negative . Her serum reacted only to azurocidin excluding other ANCA antigens: bactericidal permeability-increasing protein, cathepsin G, elastase, lactoferrin, or lysozyme . Serum creatinine level decreased, and C-reactive protein turned negative after steroid therapy . Azurocidin-ANCA also turned negative . It is suggested that azurocidin-ANCA might have been related to the inflammatory process of pauci-immune necrotizing crescentic glomerulonephritis in this patient.

Infect Immun, 2004 Apr, 72(4), 2035 - 44
Whole-genome DNA array analysis of the response of Borrelia burgdorferi to a bactericidal monoclonal antibody; Anderton JM et al.; Identification and characterization of genes that contribute to infection with Borrelia burgdorferi and, of those, genes that are targets of host responses is important for understanding the pathogenesis of Lyme disease . The complement-independent bactericidal monoclonal antibody (MAb) CB2 recognizes a carboxy-terminal, hydrophilic epitope of the outer surface protein B (OspB) . CB2 kills B . burgdorferi by an unknown bactericidal mechanism . Upon binding of CB2 to OspB, differentially expressed gene products may be responsible for, or associated with, the death of the organism . A time course of the response of B . burgdorferi to CB2 was completed to analyze the differential gene expression in the bacteria over a period of visual morphological changes . Bacteria were treated with a sublethal concentration in which spirochetes were visibly distressed by the antibody but not lysed . Preliminary whole-genome DNA arrays at various time points within 1 h of incubation of B . burgdorferi with the antibody showed that most significant changes occurred at 25 min . Circular plasmid 32 (cp32)-encoded genes were active in this period of time, including the blyA homologs, phage holin system genes . DNA array data show that three blyA homologs were upregulated significantly, >/==" BORDER="0">2 standard deviations from the mean of the log ratios, and a P value of </=0.01 . Quantitative real-time PCR analysis verified blyA and blyB upregulation over an 18- to 35-min time course . The hypothesis to test is whether the killing mechanism of CB2 is through uncontrolled expression of the blyA and blyB phage holin system.

Infect Immun, 2004 Apr, 72(4), 1974 - 82
Effects of gamma interferon, interleukin-10, and transforming growth factor beta on the survival of Mycobacterium avium subsp . paratuberculosis in monocyte-derived macrophages from naturally infected cattle; Khalifeh MS et al.; Gamma interferon (IFN-gamma) plays a significant role in the control of mycobacterial infections, including Mycobacterium avium subsp . paratuberculosis . However, the contribution of other immunoregulatory cytokines, such as interleukin-10 (IL-10) and transforming growth factor beta (TGF-beta), in Johne's disease has not been investigated as yet . In this study, we examined the effects of in vivo and in vitro infection with M . avium subsp . paratuberculosis on the production of IFN-gamma, IL-10, and TGF-beta by peripheral blood mononuclear cells (PBMC) . We also examined the effects of exogenous IFN-gamma, IL-10, and TGF-beta on M . avium subsp . paratuberculosis survival in the cell cultures . PBMC obtained from naturally infected cows, regardless of their disease status, specifically upregulated IL-10 and TGF-beta in culture supernatants in response to stimulation with live M . avium subsp . paratuberculosis . Nonstimulated PBMC recovered from subclinically infected animals secreted the lowest levels of TGF-beta, but after stimulation with live M . avium subsp . paratuberculosis, TGF-beta levels in the culture supernatants increased to levels similar to that produced by PBMC from healthy animals . The numbers of viable M . avium subsp . paratuberculosis recovered from cultures from naturally infected animals were higher than those from healthy cows after in vitro infection with M . avium subsp . paratuberculosis . The addition of exogenous IL-10 and TGF-beta to PBMC isolated from healthy cows inhibited the bactericidal activity of these cells as evidenced by the increased number of viable M . avium subsp . paratuberculosis recovered from these cultures compared to cell cultures containing medium alone . These data suggest important immune regulatory roles for IL-10 and TGF-beta during infection with M . avium subsp . paratuberculosis that may be directly related to their effects on macrophage activation and killing of M . avium subsp . paratuberculosis.

Curr Opin Microbiol, 2004 Feb, 7(1), 71 - 7
A tale of two lipids: Mycobacterium tuberculosis phagosome maturation arrest; Chua J et al.; Mycobacterium tuberculosis persistence in human populations relies on its ability to inhibit phagosomal maturation . M . tuberculosis resides in a pathogen-friendly phagosome escaping lysosomal bactericidal mechanisms and efficient antigen presentation in the host phagocytic cell . M . tuberculosis phagosome maturation arrest includes the action of mycobacterial lipid products, which mimic mammalian phosphatidylinositols, targeting host cell membrane trafficking processes . These products interfere with membrane trafficking and organelle biogenesis processes initiated by Ca(2+) fluxes, and ending with host cell Rab GTP-binding proteins and their effectors . The block includes phosphatidylinositol 3-kinase and membrane tethering molecules that prepare phagosomes for fusion with other organelles . Understanding these processes could provide new targets for pharmacological intervention in tuberculosis.

Biochem Biophys Res Commun, 2004 Apr 9, 316(3), 720 - 30
Phosphoinositide 3-kinase regulates the phosphorylation of NADPH oxidase component p47(phox) by controlling cPKC/PKCdelta but not Akt; Yamamori T et al.; Superoxide production by NADPH oxidase is essential for the bactericidal properties of phagocytes . Phosphorylation of p47(phox), one of the cytosolic components of NADPH oxidase, is a crucial step of the oxidase activation . Some evidences suggest that phosphoinositide 3-kinase (PI3K) is involved in p47(phox) phosphorylation, but it has not been fully understood how PI3K regulates it . The aim of this study was to examine the mechanism underlying the PI3K regulation of p47(phox) phosphorylation . Pharmacological inhibition of PI3K attenuated both fMLP-stimulated p47(phox) phosphorylation and NADPH oxidase activity in HL-60 cells differentiated to a neutrophil-like phenotype . Although fMLP elicited Akt activation in a PI3K-dependent manner, an Akt inhibitor had no effect on the oxidase activity triggered by fMLP . In vitro kinase assay revealed that Akt was unable to catalyze p47(phox) phosphorylation . Interestingly, the activation of cPKC and PKCdelta after fMLP stimulation was dependent on PI3K . Furthermore, PI3K inhibitors reduced the activation of phospholipase Cgamma2 without affecting tyrosine phosphorylation on it . These results suggest that PI3K regulates the phosphorylation of NADPH oxidase component p47(phox) by controlling diacylglycerol-dependent PKCs but not Akt.

Toxicon, 2003 Dec 15, 42(8), 855 - 68
Chemical modifications of phospholipases A2 from snake venoms: effects on catalytic and pharmacological properties; Soares AM et al.; Phospholipases A2 (PLA2s) constitute major components of snake venoms and have been extensively investigated not only because they are very abundant in these venoms but mainly because they display a wide range of biological effects, including neurotoxic, myotoxic, cytotoxic, edema-inducing, artificial membrane disrupting, anti-coagulant, platelet aggregation inhibiting, hypotensive, bactericidal, anti-HIV, anti-tumoral, anti-malarial and anti-parasitic . Due to this functional diversity, these structurally similar proteins aroused the interest of many researchers as molecular models for study of structure-function relationships . One of the main experimental strategies used for the study of myotoxic PLA2s is the traditional chemical modification of specific amino acid residues (His, Met, Lys, Tyr, Trp and others) and examination of the consequent effects upon the enzymatic, toxic and pharmacological activities . This line of research has provided useful insights into the structural determinants of the action of these enzymes and, together with additional strategies, supports the concept of the presence of 'pharmacological sites' distinct from the catalytic site in snake venom myotoxic PLA2s.

Peptides, 2003 Nov, 24(11), 1795 - 805
Quantitative interactions between cryptdin-4 amino terminal variants and membranes; Satchell DP et al.; Paneth cells secrete alpha-defensins into the lumen from the base of small intestinal crypts, and cryptdin-4 (Crp4) is the most potent mouse alpha-defensin in vitro . Purified recombinant Crp4 and Crp4 variants with (des-Gly)-, (Gly1Val)-, (Gly1Asp)-, and (Gly1Arg)-substitutions were all bactericidal with Crp4 and (Gly1Arg)-Crp4 being slightly more active than other variants . Bactericidal activities correlated directly with permeabilization of live Escherichia coli, with equilibrium binding to E . coli membrane phospholipid bilayers and vesicles, and with induced graded fluorophore leakage from phospholipid vesicles . The Crp4 peptide N-terminus affects bactericidal activity modestly, apparently by influencing peptide binding to phospholipid bilayers and subsequent permeabilization of target cell membranes.

Appl Environ Microbiol, 2004 Mar, 70(3), 1795 - 803
Sensitivity of Escherichia coli O157:H7 to commercially available alkaline cleaners and subsequent resistance to heat and sanitizers; Sharma M et al.; The effects of seven commercially available alkaline cleaners used in the food processing industry, 0.025 M NaOH, and 0.025 M KOH on viability of wild-type (EDL 933) and rpoS-deficient (FRIK 816-3) strains of Escherichia coli O157:H7 in logarithmic and stationary phases of growth were determined . Cells were treated at 4 or 23 degrees C for 2, 10, or 30 min . Cleaners 2, 4, 6, and 7, which contained hypochlorite and <11% NaOH and/or KOH (pH 11.2 to 11.7), killed significantly higher numbers of cells than treatment with cleaner 3, containing sodium metasilicate (pH 11.4) and <10% KOH, and cleaner 5, containing ethylene glycol monobutyl ether (pH 10.4) . There were no differences in the sensitivities of logarithmic and stationary-phase cells to the alkaline cleaners . Treatment with KOH or NaOH (pH 12.2) was not as effective as four out of seven commercial cleaners in killing E . coli O157:H7, indicating that chlorine and other cleaner components have bactericidal activity at high pH . Stationary-phase cells of strain EDL 933 that had been exposed to cleaner 7 at 4 or 23 degrees C and strain FRIK 816-3 exposed to cleaner 7 at 23 degrees C had significantly higher D(55 degrees C) (decimal reduction time, minutes at 55 degrees C) values than control cells or cells exposed to cleaner 5, indicating that exposure to cleaner 7 confers cross-protection to heat . Cells of EDL 933 treated with cleaner 7 at 12 degrees C showed significantly higher D(55 degrees C) values than cells of FRIK 816-3, indicating that rpoS may play a role in cross-protection . Stationary-phase cells treated with cleaner 5 or cleaner 7 at 4 or 12 degrees C were not cross-protected against subsequent exposure to sanitizers containing quaternary ammonium compounds or sodium hypochlorite, or to cetylpyridinium chloride and benzalkonium chloride.

Reprod Nutr Dev, 2003 Sep-Oct, 43(5), 439 - 57
Mobilization of neutrophils and defense of the bovine mammary gland; Rainard P et al.; The leucocytes present in normal milk are not very efficient in preventing infection, because very small numbers of bacteria are able to induce infection experimentally . The mobilization of phagocytes from the blood to milk appears crucial in coping with the expansion of the bacterial population in the mammary gland . Important parameters for the outcome of mammary infections are the bactericidal efficiency of neutrophils and the antiphagocytic and cytotoxic properties of the invading bacteria, but several studies have shown that the promptness and the magnitude of the initial recruitment of neutrophils by the infected mammary gland have a profound influence on the severity and the outcome of mastitis . This is an incentive for studying the mechanisms behind the mobilization of neutrophils to the mammary gland . Although milk macrophages may play a role in the triggering of the inflammatory response, studies on several responses to infections at various epithelium sites strongly suggest that epithelial cells are capable of responding to bacterial intrusion and play a major part in the initiation of inflammation . A better knowledge of the effector cells and of the mediators involved in the mobilization of neutrophils could help in devising strategies to modulate this important determinant of milk quality and udder defense.

Dent Mater J, 2003 Dec, 22(4), 482 - 93
Corrosion behavior of dental alloys in various types of electrolyzed water; Dong H et al.; The corrosion behavior of dental alloys was examined in electrolyzed strong acid water, weak acid water and neutral water using a 7-day immersion test . The precious metal alloys, gold alloy . Au-Ag-Pd alloy and silver alloy showed the greatest surface color change and dissolution of constituents in the strong acid water and the smallest in the neutral water . The release of Au from gold alloy was especially marked in the strong acid water . Co-Cr alloy showed greater corrosion and tarnish resistance in the strong acid water rather than in the weak acid water and the neutral water . X-ray microanalysis revealed that the corrosion products on the precious metal alloys were silver chloride crystals and the thin brown products on Co-Cr alloy were cobalt and chromium oxides . Ti was sound in all three types of electrolyzed water . The neutral water appeared the least corrosive to metals among the three types showing equivalent bactericidal activity.

Biol Pharm Bull, 2004 Mar, 27(3), 277 - 81
Role of hydrogen peroxide in bactericidal action of catechin; Arakawa H et al.; Catechin (epicatechin (EC), epicatechin gallate (ECg), epigallocatechin (EGC) and epigallocatechin gallate (EGCg)), which occur in green tea and black tea, possess strong bactericidal action . We observed a reactive oxygen species that was generated from the catechins as the active mechanism: and this reactive oxygen was identified . EGCg reacted with the dissolved oxygen in aqueous solution, resulting in the generation of hydrogen peroxide . Hydrogen peroxide production derived from EGCg rose with increasing pH . EGCg (0.22 mmol/l) in neutral solution (0.1 mol/l phosphate buffer pH 7.0: PBS) quantitatively generated 0.2 mmol/l hydrogen peroxide after 60 min incubation . The bactericidal effect of EGCg is dependent on hydrogen peroxide levels produced by EGCg; moreover, EGCg action was inhibited by treatment with catalase . Both bactericidal effects correlated closely when the effects of EGCg and hydrogen peroxide for the bacterium (9 of 10 kinds of bacterial strains) were examined . Therefore, hydrogen peroxide, which is generated by EGCg, appears to be involved in the bactericidal action of EGCg.

Antimicrob Agents Chemother, 2004 Mar, 48(3), 780 - 2
Early bactericidal activity of moxifloxacin in treatment of pulmonary tuberculosis: a prospective, randomized study; Pletz MW et al.; Moxifloxacin is the most active fluoroquinolone against Mycobacterium tuberculosis in vitro . However, data about the efficacy in patients are not available . We enrolled 17 patients with tuberculosis in a prospective, randomized study . After 5 days of monotherapy with either moxifloxacin or isoniazid, we detected significant decreases in mean CFU per milliliter in sputum in both groups . The calculated early bactericidal activities for isoniazid and moxifloxacin were 0.209 and 0.273 log(10) CFU per ml of sputum per day, respectively . According to the data from our study, moxifloxacin exhibits an early bactericidal activity that is comparable to that of isoniazid.

Gen Comp Endocrinol, 2004 Mar, 136(1), 58 - 71
Rapid effects of 17beta-estradiol on cell signaling and function of Mytilus hemocytes; Canesi L et al.; Estrogens affect the functioning of several non-reproductive tissues, the immune system in particular . In mammalian immunocytes, 17beta-estradiol (E2) has both dose- and cell-type specific effects and the responses to E2 seem to be mediated by rapid, non-genomic mechanisms; these may be initiated at either membrane or cytosolic locations, and can result in both direct local effects, such as modification of ion fluxes, and regulation of gene transcription secondary to activation of different kinase cascades, including mitogen activated protein kinases (MAPKs) . In this work, the short-term effects of E(2) and the possible mechanisms of estrogen-mediated cell signaling were investigated in the hemocytes, the immune cells of the bivalve mollusc, the mussel Mytilus galloprovincialis Lam . The results show that E2 (25nM) caused a rapid and significant increase in hemocyte cytosolic {Ca2+}; lower concentrations (5 nM) showed a smaller, not significant effect . Both E2 concentrations affected the phosphorylation state of the components of tyrosine kinase-mediated signal transduction MAPK- and STAT- (signal transducers and activators of transcription) like proteins within 5-15 min from E2 addition . A greater effect and clearer time course were observed with 25 nM E2: in particular, E2 induced a transient increase in p-ERK2 MAPK and a persistent increase in p-p38 MAPK . Moreover, both STAT3 and STAT5 were tyrosine phosphorylated in response to E2 . E2 (5 nM) induced both morphological (as evaluated by SEM) and functional changes (such as extracellular release of hydrolytic enzymes, lysosomal membrane destabilisation, and stimulation of the bactericidal activity) within 10-30 min from addition . Lysosomal membrane destabilisation induced by both E2 concentrations was abolished by hemocyte preincubation with the p38 MAPK inhibitor SB203580, and significantly reduced by PD98059 and Wortmannin (inhibitors of ERK MAPK and PI3-K, respectively), this suggesting that rapid activation of kinase cascades is involved in mediating the effects of E2 in mussel hemocytes . The antiestrogen Tamoxifen prevented or strongly reduced most, but not all, the effects of E2 . Western blotting with heterologous anti-ERalpha-anti-ERbeta-antibodies revealed the presence of immunoreactive ERalpha- and ERbeta-like proteins in hemocyte protein extracts . Overall, our data support the hypothesis that the rapid effects and mechanisms of action of 17beta-estradiol are extremely conserved and that they may play a crucial role in endocrine-immune interactions in invertebrates.

Med Hypotheses, 2004, 62(3), 367 - 74
Bactericidal cationic peptides can also function as bacteriolysis-inducing agents mimicking beta-lactam antibiotics?; it is enigmatic why this concept is consistently disregarded; Ginsburg I; Although there is a general consensus that highly cationic peptides kill bacteria primarily by injuring their membranes, an additional hypothesis is proposed suggesting that a large variety of cationic peptides might also render bacteria non viable by activating their autolytic wall enzymes - muramidases (a "Trojan Horse" phenomenon), resulting in bacteriolysis . This group of cationic peptides includes: lysozyme, lactoferrin, neutrophil-derived permeability increasing peptides, defensins, elastase, cathepsin G, and secretory phopholipase A2 . In this respect, cationic peptides mimic the bactericidal/bacteriolytic effects exerted by of beta-lactam antibiotics . Bacteriolysis results in a massive release of the pro-inflammatory cell-wall components, endotoxin (LPS), lipoteichoic acid (LTA) and peptidoglycan (PPG), which if not effectively controlled, can trigger the coagulation and complement cascades, the release from phagocytes of inflammatory cytokines, reactive oxygen and nitrogen species, and proteinases . Synergism (a "cross-talk") among such agonists released following bacteriolysis, is probably the main cause for septic shock and multiple organ failure . It is proposed that a use of bacteriolysis-inducing antibiotics should be avoided in bacteremic patients and particularly in those patients already suspected of developing shock symptoms as these might further enhance bacteriolysis and the release of LPS, LTA and PPG . Furthermore, in additonal to the supportive regimen exercised in intensive care settings, a use of non bacteriolysis-inducing antibiotics when combined with highly sulfated compounds (e.g . heparin, and other clinically certified polysufates) should be considered instead, as these might prevent the activation of the microbial own autolytic systems induced either by highly cationic peptides released by activated phagocytes or by the highly bacteriolytic beta-lactams . Polysulfates might also depress the deleterious effects of the complement cascade and the use of combinations among anti-oxidants ( N-acetyl cysteine), proteinase inhibitors and phospholipids might prove effective to depress the synergistic cytotoxic effects induced by inflammatory agonists . Also, a use of gamma globulin enriched either in anti-LPS or in anti-LTA activities might serve to prevent the binding of these toxins to receptors upon macrophage which upon activation generate inflammatory cytokines . Thus, a use of "cocktails" of anti-inflammatory agents might replace the unsuccessful use of single antagonists proven in scores of clinical trials of sepsis to by ineffective in prolonging the lives of patients . It is enigmatic why the concept, and the publications which support a role for cationic peptides also as potent inducers of bacteriolysis, an arch evil and a deleterious phenomenon which undoubtedly plays a pivotal role in the pathophysiology of post-infectious sequelae, has been consistently disregarded.

Pediatr Res, 2004 May, 55(5), 807 - 13 Epub 2004 Feb 18.
Diurnal fluctuation of leukocyte G6PD activity . A possible explanation for the normal neutrophil bactericidal activity and the low incidence of pyogenic infections in patients with severe G6PD deficiency in Israel; Wolach B et al.; Acute hemolytic anemia associated with red blood cell (RBC) glucose-6-phosphate dehydrogenase (G6PD) deficiency is commonly encountered in the Mediterranean basin . Nevertheless, concomitant clinical evidence of white blood cell G6PD deficiency is extremely rare in Israel . This study sought to assess simultaneously levels of G6PD activity in polymorphonuclear leukocytes (PMN) and in red blood cells (RBC) of patients with G6PD deficiency, including full-term newborn infants . In PMN, the correlation between G6PD activity, hexose monophosphate shunt activity, and superoxide anion release was evaluated . In G6PD-deficient patients, a parallel and significantly decreased G6PD activity was found in neutrophils (range of activity 0-4.5 IU/10(6) PMN) and erythrocytes (range of activity 0-1.8 IU/g Hb), compared with healthy controls (5-23 IU/10(6) PMN and 2.4-6.4 IU/g Hb, respectively) . A positive correlation was found in PMN between the levels of G6PD activity, hexose monophosphate (HMP) shunt activity, and superoxide anion release (p < 0.01) . Nevertheless, all patients' bactericidal activity of neutrophils remained in the range of healthy controls . Although many episodes of acute hemolytic anemia were recorded, no increased incidence of pyogenic infections was observed in any group of patients investigated . Neutrophil and erythrocyte G6PD levels were re-assessed in some of these patients several times a day . A significant diurnal fluctuation of the enzyme activity was found . It is speculated that the patients produce fluctuating daily quantities of NADPH, sufficient to initiate the neutrophil respiratory burst and to achieve normal bactericidal activity, necessary to prevent the development of microbial infections.

J Histochem Cytochem, 2004 Mar, 52(3), 361 - 70
Immunocytochemical localization of histatins in human salivary glands; Ahmad M et al.; Histatins are a family of salivary proteins with bactericidal and fungicidal activities that contribute to the innate defense of the oral cavity . Histatins are present in the serous granules of the parotid and submandibular glands . The important role of histatins in saliva, and the limited information on their cellular and subcellular distribution, prompted us to further define the localization of histatins in the major salivary glands . Immunogold-silver staining of 1- micro m sections of plastic-embedded tissue with anti-histatin antibody revealed histatin immunoreactivity in the serous acinar cells of the parotid and submandibular glands, the serous demilune cells of the submandibular and sublingual glands, and in occasional intercalated duct cells . No reactivity was seen in mucous cells or in striated or excretory duct cells . Electron microscopic observations of thin sections labeled with anti-histatin and gold-labeled secondary antibodies revealed immunoreactivity associated with the rough endoplasmic reticulum and Golgi complex and in secretory granules of serous acinar and demilune cells . The granules of parotid acinar cells exhibited relatively uniform labeling of their content, whereas the granules of serous cells in the submandibular and sublingual glands showed variable labeling of the dense and light regions of their content . A few intercalated duct cells adjacent to the acinar cells also exhibited labeled granules . These results suggest that the serous cells of the major glands are the main source of histatins in human saliva . They are also consistent with several previous studies demonstrating the variable distribution of different proteins within the granule content.

Pharmazie, 2004 Jan, 59(1), 10 - 4
Mechanism and kinetics of synthesis of allicin; Nikolic V et al.; Allicin, allyl-thiosulfinate, a pharmacologically active compound with considerable fungicidal, bactericidal, antioxidant and anticarcinogenic effects, was obtained by oxidizing allyl disulfide with acid hydrogen peroxide . The synthesis mechanism was studied by the ESR spin trap method . The kinetics of allicin synthesis was ascertained by determination of the concentration of the limiting reactant during the synthesis using HPLC and it was found that the allicin synthesis reaction was of zero order . The allicin obtained was determined using UV, FT-IR, MS, 1H and 13C NMR analysis.

Xenotransplantation, 2004 Mar, 11(2), 141 - 8
Immunoglobulin M-enriched intravenous immunoglobulin inhibits classical pathway complement activation, but not bactericidal activity of human serum; Walpen AJ et al.; Acute or even hyperacute humoral graft rejection, mediated by classical pathway complement activation, occurs in allo- and xenotransplantation due to preformed anti-graft antibodies . Intravenous immunoglobulin (IVIg) preparations can prevent complement-mediated tissue injury and delay hyperacute xenograft rejection . It is known that IgM-enriched IVIg (IVIgM) has a higher capacity to block complement than IVIgG . Different IVIgs were therefore tested for specificity of complement inhibition and effect on anti-bacterial activity of human serum . IVIgM-I (Pentaglobin), 12% IgM), IVIgM-II (IgM-fraction of IVIgM-I, 60% IgM), and three different IVIgG (all >95% IgG) were used . The known complement inhibitor dextran sulfate was used as control . Hemolytic assays were performed to analyze pathway-specificity of complement inhibition . Effects of IVIg on complement deposition on pig cells and Escherichia coli were assessed by flow cytometry and cytotoxicity as well as bactericidal assays . Complement inhibition by IVIgM was specific for the classical pathway, with IC50 values of 0.8 mg/ml for IVIgM-II and 1.7 mg/ml for IVIgM-I in the CH50 assay . Only minimal inhibition of the lectin pathway was seen with IVIgM-II (IC50 15.5 mg/ml); no alternative pathway inhibition was observed . IVIgG did not inhibit complement in any hemolytic assay . Classical pathway complement inhibition by IVIgM was confirmed in an in vitro xenotransplantation model with PK15 cells . In contrast, IVIgM did not inhibit (mainly alternative pathway mediated) killing of E . coli by human serum . In conclusion, IgM-enriched IVIg is a specific inhibitor of the classical complement pathway, leaving the alternative pathway intact, which is an important natural anti-bacterial defense, especially for immunosuppressed patients.

Appl Microbiol Biotechnol, 2004 May, 64(4), 455 - 64 Epub 2004 Feb 05.
Lincomycin, clindamycin and their applications; Spizek J et al.; Lincomycin and clindamycin are lincosamide antibiotics used in clinical practice . Both antibiotics are bacteriostatic and inhibit protein synthesis in sensitive bacteria . They may even be bactericidal at the higher concentrations that can be reached in vivo . Clindamycin is usually more active than lincomycin in the treatment of bacterial infections, in particular those caused by anaerobic species; and it can also be used for the treatment of important protozoal diseases, e.g . malaria, most effectively in combination with primaquine . Resistance to lincomycin and clindamycin may be caused by methylation of 23S ribosomal RNA, modification of the antibiotics by specific enzymes or active efflux from the periplasmic space .

Ann Pharm Fr, 2004 Jan, 62(1), 43 - 8
{Immunosuppressive effects of a new phenothiazine derivative}; Ghiciuc CM et al.; An original phenothiazine, CPTZ, was tested for its effects on the mouse immune system . Serum opsonic capacity, phagocyte and bactericidal activity of peritoneal macrophages, counts of splenic cells forming hemolysis plaques, and the number of survivors after experimental infection were recorded . The effects observed were compared with those produced by levamisole (a non-selective immunomodulator) and indometacin (an antiinflammatory drug with selective immunomodulator properties) . The effects of CPTZ might be useful for the development of a new class of immunosuppressor drugs.

Vaccine, 2004 Jan 26, 22(5-6), 724 - 34
Characterization of M . Tuberculosis-derived IL-12-inducing material by alveolar macrophages; Higuchi K et al.; We have investigated the substance derived from Mycobacterium tuberculosis (Mtb) that induces interleukin (IL)-12 production by alveolar macrophages (AMs) in vitro . The cytosol fraction of live Mtb H37Rv induced IL-12 production by AMs in a dose-dependent manner . The addition of interferon-gamma (IFN-gamma) augmented IL-12 production . IL-12-inducing activity by AMs (termed as surely active keeping rescue antigen, SAKRA) was purified by gel filtration and ion exchange column chromatography, and the molecular weight of SAKRA was estimated by gel filtration to be more than 700 kDa . SDS-polyacrylamide gel electrophoresis (PAGE) and Western blotting of SAKRA using rabbit anti-SAKRA antibody suggested that SAKRA is composed with several low molecular weight proteins . Amino acids sequence analysis of several bands after SDS-PAGE suggested that SAKRA is a part of ribosomes . RT-PCR showed that SAKRA induced not only expression of IL-12 p40 mRNA, but expression of tumor necrosis factor (TNF)-alpha and inducible nitric oxide synthase (iNOS) mRNA at least 6 h after stimulation, suggesting that SAKRA activates the bactericidal activity of macrophages . To investigate the potential use of SAKRA as a vaccine against tuberculosis, SAKRA was administered to BALB/c mouse that had been immunized with BCG for 18 months, and mouse were infected with Mtb H37Rv via a respiratory route . Replication of Mtb in lungs and spleens was examined 6 weeks after infection . Administration of SAKRA to BCG-vaccinated mice significantly reduced the numbers of Mtb in lungs and spleens as compared with BCG-vaccinated control mice . Taken together, these results suggest that SAKRA is one of the Mtb-derived immunomodulatory substances which induce IL-12 production during infection and also increases mycobactericidal activities of macrophages, and that SAKRA may be a promising new vaccine candidate against tuberculosis.

Clin Exp Rheumatol, 2003 Nov-Dec, 21(6 Suppl 32), S117 - 20
ANCA against bactericidal/permeability-increasing protein, azurocidin, calprotectin and defensins in rheumatic and infectious diseases: prevalence and clinical associations; Schultz H et al.; OBJECTIVE: To determine the prevalence and clinical associations of ANCA against the antibiotic proteins and peptides: Bactericidal/permeability-increasing protein (BPI), Azurocidin (AZ), Calprotectin (CP) and beta-Defensin-1 and -2 (DF) . METHODS: Patients with ANCA-associated vasculitides (n = 99), other vasculitides and rheumatic connective tissue diseases (n = 303), HIV-infection (n = 66), other infectious diseases (n = 134) Crohn's disease (n = 12) and ulcerative colitis (n = 12) were tested for BPI-, AZ-, CP-, DF-, PR3-, and MPO-ANCA in indirect immunofluorescence technique (IFT) and ELISA . RESULTS: In ANCA associated vasculitides BPI-ANCA were detected in 6% of patients . In HIV infection, BPI was the main target antigen of ANCA-IFT positive sera (74%) . BPI-ANCA was associated with higher inflammatory activity . In Crohn's disease and ulcerative colitis BPI-ANCA was prominent (34% of patients) . AZ-ANCA were found in 5% of patients . No ANCA were detected against defensin and calprotectin . CONCLUSION: BPI-ANCA is the main autoantibody in HIV and is associated with higher inflammatory activity . In inflammatory bowel diseases BPI-ANCA is predominant, AZ-ANCA are also present to a lesser extend . Both were not useful characterize clinical subgroups . No ANCA were detected against calprotectin or defensins.

Clin Exp Rheumatol, 2003 Nov-Dec, 21(6 Suppl 32), S89 - 94
Prevalence of ANCA in mixed cryoglobulinemia and chronic hepatitis C virus infection; Lamprecht P et al.; OBJECTIVE: To determine the prevalence, target antigens and clinical associations of antineutrophil cytoplasmic antibodies (ANCA) in chronic hepatitis C without extrahepatic manifestations and in chronic hepatitis C virus (HCV)-associated mixed cryoglobulinemia (MC) in two European centers . METHODS: 50 sera from patients with chronic hepatitis C and 116 sera from HCV-associated MC were tested for cytoplasmic or perinuclear pattern (C-ANCA/P-ANCA) by indirect immunofluorescence test (IFT) . ANCA target antigens were determined by enzyme-linked immunosorbent assay (ELISA) . RESULTS: Clinical characteristics of the patients were not different between the two centers . Cryoglobulinemic vasculitis (CV) was biopsy-proven in about 90% of the MC patients . Two patients with HCV-associated MC and 1 patient with chronic hepatitis C had a P-ANCA . A C-ANCA was detected in 1 patient with HCV-associated MC . Eight patients with a HCV-associated MC and 5 patients with chronic hepatitis C had an ANCA either directed against bactericidal/permeability increasing protein (BPI) or cathepsin G (CG) . BPI- or CG-ANCA positivity was not associated with a more severe disease course . The C-ANCA titer followed disease activity in one C-ANCA positive HCV-associated MC patient . The subspecificity of the C-ANCA was not determinable in that patient . CONCLUSION: Two new target antigens of ANCA have been identified in HCV-associated MC and chronic hepatitis C in this study . BPI-ANCA and GC-ANCA were present in about 10% of patients with HCV-associated MC or chronic hepatitis C . ELISA proved to be more sensitive in the detection of ANCA than IFT . The present study on chronic HCV infection adds to various reports on the induction of CG- and BPI-ANCA in chronic infections.

Protein Sci, 2004 Feb, 13(2), 422 - 30
Phylogenetic and evolutionary analysis of the PLUNC gene family; Bingle CD et al.; The PLUNC family of human proteins are candidate host defense proteins expressed in the upper airways . The family subdivides into short (SPLUNC) and long (LPLUNC) proteins, which contain domains predicted to be structurally similar to one or both of the domains of bactericidal/permeability-increasing protein (BPI), respectively . In this article we use analysis of the human, mouse, and rat genomes and other sequence data to examine the relationships between the PLUNC family proteins from humans and other species, and between these proteins and members of the BPI family . We show that PLUNC family clusters exist in the mouse and rat, with the most significant diversification in the locus occurring for the short PLUNC family proteins . Clear orthologous relationships are established for the majority of the proteins, and ambiguities are identified . Completion of the prediction of the LPLUNC4 proteins reveals that these proteins contain approximately a 150-residue insertion encoded by an additional exon . This insertion, which is predicted to be largely unstructured, replaces the structure homologous to the 40s hairpin of BPI . We show that the exon encoding this region is anomalously variable in size across the LPLUNC proteins, suggesting that this region is key to functional specificity . We further show that the mouse and human PLUNC family orthologs are evolving rapidly, which supports the hypothesis that these proteins are involved in host defense . Intriguingly, this rapid evolution between the human and mouse sequences is replaced by intense purifying selection in a large portion of the N-terminal domain of LPLUNC4 . Our data provide a basis for future functional studies of this novel protein family.

Arterioscler Thromb Vasc Biol, 2004 Mar, 24(3), 483 - 8 Epub 2004 Jan 22.
Inhibition of plasmin activity by tranexamic acid does not influence inflammatory pathways during human endotoxemia; Renckens R et al.; OBJECTIVE: Plasmin activates several proinflammatory pathways at the cellular level in vitro . Lipopolysaccharide (LPS) administration to healthy humans results in a rapid generation of plasmin activity, accompanied by activation of a number of inflammatory systems . METHODS AND RESULTS: To determine the role of early plasmin activity in LPS-induced inflammation in vivo, 16 healthy males received an intravenous bolus injection with LPS (from Escherichia coli, 4 ng/kg) directly preceded by a 30-minute intravenous infusion of tranexamic acid (2 g, n=8), a plasmin activation inhibitor, or placebo (n=8) . LPS injection induced marked increases in the plasma levels of D-dimer and plasmin-alpha2-antiplasmin complexes, indicative of plasmin activation and generation, respectively, which were strongly attenuated by tranexamic acid (both P<0.01 versus placebo) . However, tranexamic acid did not influence LPS-induced coagulation activation, granulocytosis, neutrophil activation (expression of CD11b, CD66b, and L-selectin) or degranulation (plasma concentrations of elastase-alpha1-antitrypsin and bactericidal permeability-increasing protein), endothelial cell activation (plasma levels of von Willebrand factor and soluble E-selectin), or cytokine release . CONCLUSIONS: These data argue against a role of early plasmin generation in the subsequent activation of other inflammatory pathways during human endotoxemia.

Nat Med, 2004 Feb, 10(2), 161 - 7 Epub 2004 Jan 11.
Therapeutic effects of lysophosphatidylcholine in experimental sepsis; Yan JJ et al.; Sepsis represents a major cause of death in intensive care units . Here we show that administration of lysophosphatidylcholine (LPC), an endogenous lysophospholipid, protected mice against lethality after cecal ligation and puncture (CLP) or intraperitoneal injection of Escherichia coli . In vivo treatment with LPC markedly enhanced clearance of intraperitoneal bacteria and blocked CLP-induced deactivation of neutrophils . In vitro, LPC increased bactericidal activity of neutrophils, but not macrophages, by enhancing H(2)O(2) production in neutrophils that ingested E . coli . Incubation with an antibody to the LPC receptor, G2A, inhibited LPC-induced protection from CLP lethality and inhibited the effects of LPC in neutrophils . G2A-specific antibody also blocked the inhibitory effects of LPC on certain actions of lipopolysaccharides (LPS), including lethality and the release of tumor necrosis factor-alpha (TNF-alpha) from neutrophils . These results suggest that LPC can effectively prevent and treat sepsis and microbial infections.

J Immunol, 2004 Jan 15, 172(2), 1191 - 7
Generation of a complement-independent bactericidal IgM against a relapsing fever Borrelia; Connolly SE et al.; The spirochetemia of relapsing fever in mice is cleared by a complement-independent, polyclonal IgM response with reactivity to two prominent Ags of 20 and 35 kDa . In this study, we have dissected the polyclonal IgM Ab response against a relapsing fever spirochete to determine the specificity of its complement-independent bactericidal properties . Our experimental approach selectively generated an IgM murine mAb from the early specific immune response to a variable outer membrane protein . This IgM is bactericidal in the absence of complement and is part of the polyclonal Ab response that mediates the clearance of this bacterium from the blood . Purified monoclonal IgM caused direct structural damage to the outer membrane of the spirochete, in the absence of complement, and protected both B cell- and C5-deficient mice from challenge when administered passively . The direct, complement-independent, bactericidal activity of Abs is a critical mechanism of host defense against infection.

Pol J Vet Sci, 2003, 6(4), 279 - 96
Pharmacological and toxicological aspects of combination of beta-lactam and aminoglycoside antibiotic, prednisolone and procaine hydrochloride on the example of Vetramycin; Kania BF et al.; Vetramycin is an injectable veterinary compound for animal use only . In veterinary medicine, it has been used for a long time as a bactericidal beta-lactam and aminglycoside antibiotics combination, extending the bactericidal spectrum of these substances . This compound, in addition to bactericidal procaine penicillin and dihydrostreptomycin (DHS), contains also prednisolone acetate and procaine hydrochloride, two biologically active substances . Prednisolone, a glucocorticoide, has an antiinflammatory, antiallergic, antiitchical and analgesic effect . Procaine hydrochloride, in turn, has a local anaesthetic effect and attenuates pain caused by irritable properties of antibiotics at the injection sites . The average dosage of, respectively, procaine benzylpenicillin (I.U./kg(-1) b.w.), DHS (microg/kg(-1) b.w.), prednisolone acetate (microg/kg(-1) b.w.) and procaine hydrochloride (mg/kg(-1) b.w.) in horses, cattle, pigs is 6000-15000, 10-11, 0.24-0.6 and 1.2-3.0; s.i.d., in sheep, foals, calves, piglets is 20000-40000, 10, 0.8-1.6 and 4-8; s.i.d., in dogs and cats is 30000-200000, 10, 0.8-1.6 and 4-8; s.i.d. . Intramammary injection dose (Vetramycin antimastitis ointment in syringe) in cows is 1000000 I.U . of procaine benzylpenicillin + 1000000 I.U . of dihydrostreptycin sulphate per quarter of udder, s.i.d., during 3 successive days.

Rev Prat, 2003 Oct 31, 53(16), 1772 - 6
{Cystitis}; Fourcade RO et al.; Bacterial cystitis is a common condition characterised by a high incidence, an easy diagnosis and a simple treatment . Three subsets should be distinguished: simple cystitis, occurring in young women, requires no bacteriologic work-up and is best treated by a single dose or a three-day regimen consisting of a common antibiotic, complicated cystitis, occurring in patients with diabetes, an urologic history or immunosuppression; it requires a seven-day treatment course, the choice of which is guided by antibiotic sensitivity tests; recurrent cystitis is defined by, at least, four yearly flares; such recurrences warrant thorough vulvo-vaginal examination seeking a local lesion; if none is found, protracted treatment using an inhibitory rather than a bactericidal antibiotic dosage is an efficient prophylactic regimen.

J Biol Chem, 2004 Mar 19, 279(12), 11976 - 83 Epub 2003 Dec 31.
Structure-activity determinants in paneth cell alpha-defensins: loss-of-function in mouse cryptdin-4 by charge-reversal at arginine residue positions; Tanabe H et al.; Paneth cells secrete microbicidal enteric alpha-defensins into the small intestinal lumen, and cryptdin-4 (Crp4) is the most bactericidal of the mouse alpha-defensin peptides in vitro . Here, site-directed Arg to Asp mutations in Crp4 have been shown to attenuate or eliminate microbicidal activity against all of the bacterial species tested regardless of the Arg residue position . R31D/R32D charge-reversal mutagenesis at the C terminus and mutations at R16D/R18D, R16D/R24D, and R18D/R24D in the Crp4 polypeptide chain eliminated in vitro bactericidal activity, blocked peptide-membrane interactions, as well as Crp4-mediated membrane vesicle disruption . Lys for Arg charge-neutral substitutions in (R16K/R18K)-Crp4 did not alter the bactericidal activity relative to Crp4, showing that bactericidal activity appears not to require the guanidinium side chain of Arg at those two positions . Partial restoration of (R31D/R32D)-Crp4 bactericidal activity occurred when an electropositive Arg for Gly substitution was introduced at the peptide N terminus and the (G1R/R31D/R32D)-Crp4 peptide exhibited intermediate membrane binding capability . Also, the loss of peptide bactericidal activity in (G1D/R31D/R32D)-Crp4 and (R16D/R24D)-Crp4 mutants corresponded with diminished phospholipid vesicle disruptive activity . Fluorophore leakage from anionic phospholipid vesicles induced by the charge-reversal variants was negligible relative to Crp4 and lower than that induced by pro-Crp4, the inactive Crp4 precursor . Thus, Arg residues function as determinants of Crp4 bactericidal activity by facilitating or enabling target cell membrane disruption . The role of the Arg residues, however, was surprisingly independent of their position in the polypeptide chain.

Vet Immunol Immunopathol, 2004 Jan, 97(1-2), 105 - 14
Repeatability of flow cytometric and classical measurement of phagocytosis and respiratory burst in bovine polymorphonuclear leukocytes; Kampen AH et al.; Five methods for measurement of phagocytosis and respiratory burst activity of bovine blood polymorphonuclear leukocytes (PMNs) were evaluated . Eight cows were repeatedly sampled over a two week period and parallel samples tested in all five assays to assess the repeatability and stability of the methods . In the flow cytometric phagocytosis assay, ingestion of fluorescein labeled bacteria was measured, and in the flow cytometric assay for respiratory burst, oxidation of a dye by reactive oxygen species was recorded . In the classical assays, bactericidal effect on opsonized, live bacteria was quantified by the conversion of an indicator substance, superoxide anion production was assayed by the reduction of cytochrome c, whereas myeloperoxidase activity was determined with a radioactive iodination assay . The results showed that the Phagotest, Bursttest, cytochrome c and iodination assays gave repeatable results when samples were run in the same setup on the same day . Although day-to-day variability was significant in all assays, the described methods comprise a panel of useful tests for the evaluation of phagocytosis and respiratory burst activity in bovine PMNs . The flow cytometric methods represent a convenient alternative to the classical methods for measurement of phagocytosis and respiratory burst in bovine blood PMNs.

Dev Comp Immunol, 2004 Apr, 28(4), 307 - 23
Cloning and analyses of a BPI/LBP cDNA of the Atlantic cod (Gadus morhua L.); Stenvik J et al.; Using the differential screening technique, a cDNA related to the mammalian family of lipid transfer/lipopolysaccharide-binding proteins was cloned from the Atlantic cod (Gadus morhua L.) . The gene is an ortholog of a recently identified gene of rainbow trout (Oncorhynchus mykiss) . Phylogenetic analyses suggest that teleost BPI/LBP are modern descendants of the ancestor of mammalian bactericidal/permeability-increasing protein (BPI) and lipopolysaccharide-binding protein (LBP), and a gene of the urochordate Ciona intestinalis is related to this gene family . Molecular modeling suggests that the structure of cod BPI/LBP is similar to mammalian BPI and LBP, while its highly basic character is similar to BPI . Cod BPI/LBP is constitutively expressed in head-kidney (HK) leukocytes . After intraperitoneal injection of bacterin high levels of cod BPI/LBP mRNA were detected also in peripheral blood cells and spleen, while moderate to low levels of transcript were found in heart, liver, gills, skin, brain, and intestine . We conclude that the patterns of charge and expression of cod BPI/LBP are more similar to mammalian BPI than to mammalian LBP.

Antimicrob Agents Chemother, 2004 Jan, 48(1), 69 - 74
Anti-Trypanosoma cruzi activity of green tea (Camellia sinensis) catechins; Paveto C et al.; The trypanocidal action of green tea catechins against two different developmental stages of Trypanosoma cruzi is reported for the first time . This activity was assayed with the nonproliferative bloodstream trypomastigote and with the intracellular replicative amastigote parasite forms . An ethyl acetate fraction from Camellia sinensis green tea leaves, which contains most of the polyphenolic compounds and the maximal trypanocidal activity, was obtained by fractionation of the aqueous extract with organic solvents . The active compounds present in this extract were further purified by LH-20 column chromatography and were identified by high-performance liquid chromatography analysis with a photo diode array detector and gas chromatography coupled to mass spectroscopy . The following flavan-3-ols derivatives, known as catechins, were identified: catechin, epicatechin, gallocatechin, epigallocatechin, catechin gallate, epicatechin gallate, gallocatechin gallate, and epigallocatechin gallate . The purified compounds lysed more than 50% of the parasites present in the blood of infected BALB/c mice at concentrations as low as 0.12 to 85 pM . The most active compounds were gallocatechin gallate and epigallocatechin gallate, with minimal bactericidal concentrations that inhibited 50% of isolates tested of 0.12 and 0.53 pM, respectively . The number of amastigotes in infected Vero cells decreased by 50% in the presence of each of these compounds at 100 nM . The effects of the catechins on the recombinant T . cruzi arginine kinase, a key enzyme in the energy metabolism of the parasite, were assayed . The activity of this enzyme was inhibited by about 50% by nanomolar concentrations of catechin gallate or gallocatechin gallate, whereas the other members of the group were less effective . On the basis of these results, we suggest that these compounds could be used to sterilize blood and, eventually, as therapeutic agents for Chagas' disease.

Vet Res Commun, 2003 Dec, 27(8), 625 - 31
In vitro effects of acellular milk on the bactericidal components of caprine polymorphonuclear neutrophils; Kumar S et al.; The effects of acellular milk on the activity of the microbicidal cationic enzymes of the polymorphonuclear cells of goats were studied in an attempt to explain the phenomenon by which PMN functions fail in mastitis . Assays were undertaken on the myeloperoxidase, lysozyme and elastase activities in a polymorphonuclear cell (PMN) lysate, both in the presence and absence of acellular milk from homologous species . There was a significant decrease (p < 0.05) in the activity of lysozyme, myeloperoxidase and elastase in the presence of acellular milk . Superoxide and H2O2 production following activation of caprine PMNs by lipopolysaccharide (LPS) was significantly reduced (p < 0.05) in the presence of acellular milk . Thus, the microbicidal function of PMNs is significantly impaired in the presence of acellular milk and this may contribute to the development of mastitis in dairy animals.

Tuberculosis (Edinb), 2004, 84(1-2), 29 - 44
M . tuberculosis persistence, latency, and drug tolerance; Gomez JE et al.; The success of Mycobacterium tuberculosis as a pathogen is largely attributable to its ability to persist in host tissues, where drugs that are rapidly bactericidal in vitro require prolonged administration to achieve comparable effects . Latency is a frequent outcome of untreated or incompletely treated M . tuberculosis infection, creating a long-standing reservoir of future disease and contagion . Although the interactions between the bacterium and its host that result in chronic or latent infection are still largely undefined, recent years have seen a resurgence of interest and research activity in this area . Here we review some of the classic studies that have led to our current understanding of M . tuberculosis persistence, and discuss the varied approaches that are now being brought to bear on this important problem.

Psychiatry Res, 2003 Dec 1, 121(2), 123 - 32
Overproduction of neutrophil radical oxygen species correlates with negative symptoms in schizophrenic patients: parallel studies on neutrophil chemotaxis, superoxide production and bactericidal activity; Sirota P et al.; Defective neutrophil function in schizophrenic patients has recently been reported . There are several lines of evidence to support the contribution of oxygen free radicals in schizophrenia, including increased lipid peroxidation, fatty acids and alterations in blood levels of anti-oxidant enzymes . Eighteen schizophrenic patients (DSM-IV) and 15 healthy controls were studied . Neutrophil chemotaxis, superoxide production and bactericidal activity were investigated . A statistically significant increase of superoxide anion release was found in schizophrenic patients compared with controls (mean+/-S.E.M., patients: 6.89+/-0.30 nmol O2-/10(6) cells/min, controls: 5.13+/-0.55 nmol O2-/10(6) cells/min) . Moreover, a significant positive correlation between superoxide production and negative symptoms as assessed by the Positive and Negative Syndrome Scale was demonstrated . No differences were detected in chemotaxis and phagocytosis between schizophrenic patients and healthy controls . The present findings of a positive correlation between superoxide generation and negative symptoms in schizophrenic patients support the hypothesis that superoxide anion may participate in the pathogenesis of schizophrenia, as an excess of free radicals could contribute to the deterioration phase of the disease . Further studies are required to establish the role of oxidative stress in the ethiopathogenesis of schizophrenia.

J Immunol, 2003 Dec 1, 171(11), 6198 - 205
Aberrant inflammation and lethality to septic peritonitis in mice lacking STAT3 in macrophages and neutrophils; Matsukawa A et al.; Stat3 is a transcription factor mediating anti-inflammatory properties of IL-10 . In the present study, we demonstrate a pivotal role of Stat3 expressed in innate immune cells during septic peritonitis induced by cecal ligation and puncture (CLP) . Mice with targeted disruption of Stat3 in macrophages and neutrophils were succumbed to septic peritonitis induced by CLP . The mice displayed an excessive local and systemic inflammation relative to the control mice, an event that was accompanied by substantial increases in the level of multiple cytokines . Hepatic and renal injury was significantly exacerbated in mice with Stat3 deficiency . Despite enhanced inflammatory responses, the mice failed to facilitate bacterial clearance as compared with the control mice . In addition, the mice exhibited an increased lethality after i.p . inoculation of live bacteria recovered from CLP-mice . In vitro, resident peritoneal macrophages from mice with Stat3 deficiency impaired bactericidal activity relative to the control whereas productions of inflammatory cytokines were significantly augmented when cells were stimulated with a synthetic lipopeptide, macrophage-activating lipopeptide-2 and LPS . Elicited macrophages and neutrophils with Stat3 deficiency also impaired bactericidal activity as compared with those with Stat3 . Lysosomal enzyme release, an effector molecule for bacterial clearance, was significantly decreased in elicited leukocytes with Stat3 deficiency while increasing the production of inflammatory cytokines . Altogether, these results suggest that macrophage/neutrophil-specific STAT3 is crucial in not only modulating multiple organ failure associated with systemic inflammation but also intensifying the bactericidal activity, which highlight the significance of cell-specific Stat3 in the protective immunity during sepsis.

Eksp Klin Gastroenterol, 2003, (5), 25 - 30, 192
{Therapeutic effect of gastrozolum in stomach ulcers}; Sukhareva GV et al.; Gastrozolum is the proprietary name of a drug made in Saint Petersburg . Its international nonproprietary name is Omeprazole . The absorption rate is not related to food . Its pharmacotherapeutic action becomes apparent as an inhibitor of the proton pump leading to the inhibition of H+/K(+)-ATPase of the secretory membrane of parietal cells of the stomach mucous membrane and blocking of the concluding stage of hydrochloric acid secretion . The entire action leads to the decrease of the level of basal and induced secretion regardless of the nature of stimulus . As a result of this, symptoms of stomach ulcer decrease, and gastroduodenal ulcers heal faster . Penetrating into the stomach mucous membrane cells, the drug also has a cytoprotective action . The maximum blood concentration (0.6-1.5 mg/l) is found 2-3 hours after a single intake of 40 mg of the drug . It was determined that after the intake of 20 mg of Gastrozolum its action lasts for 24 hours and provides for the inhibition of both night and day secretion . The ricochet syndrome does not take place when the treatment is over . It was proved that Gastrozolum has a bactericidal action on Helicobacter pylori due to the sharp increase of stomach pH, which contributes to the realization of the effect of used components of the anti-helicobacter therapy . The experiment failed to establish any teratogenic or poisonous action on the embryos . The dosage form is a capsule containing 20 mg of Omeprazole in the form of pellets.

J Antimicrob Chemother, 2003 Dec, 52(6), 1025 - 8 Epub 2003 Nov 12.
In vitro activity of C-8-methoxy fluoroquinolones against mycobacteria when combined with anti-tuberculosis agents; Lu T et al.; OBJECTIVES: To examine the effect of first-line and second-line anti-tuberculosis agents on the ability of fluoroquinolones to kill mycobacteria . METHODS: A clinical isolate of Mycobacterium tuberculosis and a laboratory strain of Mycobacterium smegmatis were grown in liquid medium and treated with a fluoroquinolone in the presence or absence of anti-tuberculosis agents . Bacterial survival was determined by viable colony counts on agar medium . RESULTS: When moxifloxacin activity was examined in two-drug combinations containing traditional anti-tuberculosis agents, activity was greater than either compound alone with isoniazid, capreomycin and low, but not high, concentrations of rifampicin . Cycloserine contributed no additional activity, and ethambutol interfered with the lethal action of moxifloxacin and gatifloxacin . Experiments with M . smegmatis confirmed that both rifampicin and ethambutol reduce fluoroquinolone lethality . Moreover, ethambutol increased the recovery of fluoroquinolone-resistant mutants newly created by ethyl methanesulphonate treatment . CONCLUSIONS: The intrinsic bactericidal activity of C-8-methoxy fluoroquinolones can be adversely affected by some agents currently used for treatment of tuberculosis.

Klin Khir, 2003 Aug, (8), 41 - 3
{Surgical laser application in thoracic surgery . Potential and critical evaluation}; Grubnik VV et al.; A 25-year experience of pulmonary and pleural surgery in 348 patients, reconstructive esophageal operations in 74 patients, bronchoscopic interventions using different laser techniques--in 925 patients, and thoracoscopic operations of traumatic and spontaneous pneumothorax--in 150 patients has been summarized . The methods of laser pulmonary resection, processing of bronchial stump, fibrous capsule, pleurodesis, stitching of esophagogastric, esophagointestinal and interintestinal anastomoses and of bronchoscopic photo destruction of tracheal and bronchial tumors have been described . Haemostatic, bactericidal and ablastic properties of laser have been described . Lasers were used in miniinvasive thoracic surgery for thoracoscopic and videothoracoscopic interventions.

Zhongguo Wei Zhong Bing Ji Jiu Yi Xue, 2003 Nov, 15(11), 646 - 50
{Changes in Toll-like receptor 2 and 4 gene expression in vital organs in septic rats and their regulation mechanisms}; Yao YM et al.; OBJECTIVE: To investigate the changes in Toll-like receptor (TLR) 2 and 4 gene expression in vital organs in septic rats and their potential regulation mechanism . METHODS: One hundred Wistar rats were randomly divided into normal controls (n=10), sham-operated group (n=10), septic group (n=60), and recombinant bactericidal/permeability increasing protein (BPI)-treated group (n=20) . Severe sepsis was replicated by cecal ligation and puncture (CLP) . Animals were sacrificed at different time points after CLP, tissue TLR2/4 mRNA expression in the liver, lungs, kidneys as well as intestine were measured by reverse transcription-polymerase chain reaction (RT-PCR) . Plasma levels of tumor necrosis factor-alpha(TNF-alpha) and interleukin-10 (IL-10) were also determined by enzyme linked immunoadsorbent assay (ELISA) . RESULTS: TLR2/4 mRNA could be detected in various tissues with low values both in normal controls and sham-operated group, but they were markedly up-regulated at 2 hours after CLP, peaking at 6-12 hours . Tissue TLR4 mRNA was gradually down-regulated 24 hours later, while TLR2 mRNA levels maintained high values up to 72 hours . In comparison with the CLP group, treatment with BPI significantly decreased TLR2 mRNA in various tissues at 12 and 24 hours (P<0.05 or P<0.01), also tissue TLR4 mRNA at 12 hours (P<0.05 or P<0.01), without marked influence on TLR4 mRNA expression at 24 hours in liver, lungs and small intestine (P>0.05) . In addition, treatment with BPI could significantly lower plasma TNF-alpha levels at 12 hours post-CLP, on the other hand markedly elevate plasma IL -10 levels 24 hours later (P<0.01) . CONCLUSION: These data suggest that severe peritoneal infection could result in up-regulation of TLR2/4 mRNA expression in vital organs, which might play important roles in mediating proinflammatory cytokine synthesis and release . Endotoxemia appears to be involved in the activation of tissue TLR2/4 expression associated with CLP-induced sepsis.

J Surg Oncol, 2003 Nov, 84(3), 137 - 42
Effects of major liver resection, with or without recombinant bactericidal/permeability-increasing protein (rBPI21), on the angiogenic profile of patients with metastatic colorectal carcinoma; Wu FP et al.; BACKGROUND: Surgery induces a process of wound healing, which has immunological and angiogenic aspects . Bactericidal/permeability-increasing protein (BPI) is found in azurophilic granules of human neutrophils, which is bactericidal and neutralizes lipo-polysaccharide (LPS) . This may reduce postoperative infectious complications . In addition, BPI has been shown to be an inhibitor of angiogenesis . METHODS: A total of 18 patients with metastasized colorectal carcinoma to the liver were double blind randomized . The levels of the pro-angiogenic factors interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) and the angiogenesis inhibitor endostatin were investigated after liver surgery with perioperative administration of either rBPI(21) or placebo . RESULTS: The highest IL-6 levels were found during the first 24 hr and reached peak levels already at 2 hr postoperatively in both groups . In both groups VEGF levels decreased sharply in the postoperative hours, returning to baseline levels in the days afterwards . In both groups, an immediate decrease in endostatin levels was observed which remained significantly low . RBPI(21) transiently influenced IL-6 and VEGF . CONCLUSIONS: RBPI(21) only marginally affected IL-6 and VEGF levels . Surgery per se induced an immediate immune response (IL-6) and an immediate angiogenic response, reflected in an initial VEGF decrease and a longer lasting decrease of endostatin . These findings demonstrate the dynamics of tissue responses in the first phase of wound healing .

Environ Sci Technol, 2003 Oct 15, 37(20), 4785 - 9
Bactericidal activity of copper-deposited TiO2 thin film under weak UV light illumination; Sunada K et al.; The bactericidal activity of copper-deposited titanium dioxide thin film (Cu/TiO2) was investigated under very weak ultraviolet (UV) light illumination . To elucidate the roles of the film photocatalyst and the deposited copper in the bactericidal activity, cells from a copper-resistant Escherichia coli (E . coli) strain were utilized . A decrease in survival rate was not observed with the copper-resistant cells under dark conditions, but when illuminated with a very weak UV intensity of 1 microW/cm2, the survival rate decreased, suggesting photocatalytic bactericidal activity . The decay curve of survival on the Cu/TiO2 film under very weak UV light illumination consisted of two steps, similar to the survival change of normal E . coli on TiO2 films under rather strong UV illumination . The first step is due to the partial decomposition of the outer membrane in the cell envelope by a photocatalytic process, followed by permeation of the copper ions into the cytoplasmic membrane . The second step is due to a disorder of the cytoplasmic membrane caused by the copper ions, which results in a loss of the cell's integrity . These processes explain why the Cu/TiO2 film system shows an effective bactericidal activity even under very weak UV light illumination.

Biotechnol Lett, 2003 Oct, 25(19), 1661 - 5
Making thin polymeric materials, including fabrics, microbicidal and also water-repellent; Lin J et al.; A procedure is developed and validated for making a non-functionalized polyolefin fabric/film highly bactericidal and fungicidal which involves a free-radical grafting of maleic anhydride, followed by an attachment of polyethylenimine (PEI) and its subsequent N-alkylation . Separately, cotton fabric coated with a micron layer of a hydrophobic polymer using hot-filament chemical vapor deposition is rendered markedly hydrophobic; if this coating is preceded by immobilization of N-alkyl-PEI, the fabric becomes both water-repellent and bactericidal.

Mol Immunol, 2003 Nov, 40(7), 451 - 6
Host anti-microbial response to Helicobacter pylori infection; George JT et al.; beta-Defensin peptides are known to be potent anti-bacterials with a wide spectrum of activity . They, therefore, represent an important aspect of innate immunity . In the present study, we have extended our understanding of the regulation of the beta-defensins in response to Helicobacter pylori (H . pylori) infection . We found elevated levels of hBD2 and hBD3 transcripts within gastric cells following infection . This was reflected by increased secretion of the corresponding peptide . The relative bactericidal potency of the beta-defensins was also assessed . Our findings show that hBD3 was the most potent peptide tested followed by hBD2 and hBD1 . Relatively modest synergy between hBD1 and hBD2 was also noted . More importantly, we observed endogenous production of putative anti-microbial factors by infected gastric epithelial cells . Our study highlights the active participation of the epithelium in protection against potential pathogens.

Jpn J Thorac Cardiovasc Surg, 2003 Sep, 51(9), 413 - 9
A new sterilization technique with balloon-tube thoracostomy for thoracic empyema; Kadoyama C et al.; OBJECTIVE: Failure or prolongation of treatment for refractory thoracic empyema by the current chest-tube drainage technique is often due to sterilization difficulties . Insufficient sterilization prolongs hospitalization, and is often associated with life-threatening complications and/or additional invasive surgical procedures . A new chest-tube sterilization technique aimed at making it less invasive and shortening the therapy is proposed . METHODS: Following pretreatment for complications including loculation, bronchopleural fistula, or corticated lung, a double-lumen trocar catheter was introduced at the bottom of the empyemic cavity through the lateral chest wall . Then, a Foley balloon urethra-catheter was inserted and attached just inside the anterior chest wall at the top of the cavity for the evacuation of intrathoracic air . After irrigation of the cavity with distilled water once or twice, the cavity was completely filled with a bactericidal solution which was left in place for 30-60 minutes, followed by an antibiotic solution for more than 20 hours . RESULTS: Among the five treated post-lobectomy or pneumonectomy cases, sterilization was obtained after only one treatment in four cases and after two courses in the other . Catheterization duration from the initial treatment was 2-13 days . Neither recurrence nor treatment-related major complications were observed . CONCLUSIONS: This balloon-tube thoracostomy technique is simple, minimally invasive and cost-effective, due to shortening of the treatment time with minimal manpower and equipment requirements . It is thus a promising therapeutic approach to thoracic empyema and has the potential for application to other intrathoracic disorders.

FEMS Immunol Med Microbiol, 2003 Oct 15, 38(3), 181 - 91
Anti-adhesion therapy of bacterial diseases: prospects and problems; Ofek I et al.; The alarming increase in drug-resistant bacteria makes a search for novel means of fighting bacterial infections imperative . An attractive approach is the use of agents that interfere with the ability of the bacteria to adhere to tissues of the host, since such adhesion is one of the initial stages of the infectious process . The validity of this approach has been unequivocally demonstrated in experiments performed in a wide variety of animals, from mice to monkeys, and recently also in humans . Here we review various approaches to anti-adhesion therapy, including the use of receptor and adhesin analogs, dietary constituents, sublethal concentrations of antibiotics and adhesin-based vaccines . Because anti-adhesive agents are not bactericidal, the propagation and spread of resistant strains is much less likely to occur than as a result of exposure to bactericidal agents, such as antibiotics . Anti-adhesive drugs, once developed, may, therefore, serve as a new means to fight infectious diseases.

Cardiovasc Res, 2003 Oct 15, 60(1), 170 - 4
Nitric oxide synthase plays a role in Chlamydia pneumoniae-induced atherosclerosis; Chesebro BB et al.; OBJECTIVE: Chlamydia pneumoniae infection has been associated with atherosclerosis, although the mechanisms by which C . pneumoniae contribute to atherogenesis remain unclear . Altered production of nitric oxide, a known bactericidal and anti-inflammatory agent, represents one possible mechanistic link . To examine this issue, a diet-induced, hyperlipidemic mouse model of early atherosclerosis was used . METHODS: A series of intranasal inoculations of C . pneumoniae strain AR-39 were administered to mice lacking endothelial or inducible nitric oxide synthase and to normal controls . After 18 weeks on an atherogenic diet, atherosclerotic lesion area in the aortic sinus was measured using computer-assisted morphometry . RESULTS: In the absence of C . pneumoniae infection, diet-fed eNOS(-/-) mice developed enlarged fatty streak lesions of borderline significance in comparison to uninfected, wild-type mice, while the lesion area in uninfected, diet-fed iNOS(-/-) mice did not differ significantly from lesion area in wild-type animals . In contrast, lesion area in infected eNOS(-/-) mice increased slightly, but not significantly in comparison to uninfected eNOS(-/-) mice . Lesion area in the infected iNOS(-/-) mice was significantly enlarged when compared to both uninfected iNOS(-/-) mice as well as to infected wild-type mice . CONCLUSIONS: These data suggest that production of nitric oxide by eNOS protects against development of fatty streak lesions in uninfected hyperlipidemic mice, but does not offer additional protection in infected hyperlipidemic mice, while iNOS may play a protective role, thus limiting chlamydial exacerbation of fatty streak lesions.

Blood, 2004 Feb 1, 103(3), 1099 - 104 Epub 2003 Sep 25.
Granzyme B and perforin: constitutive expression in human polymorphonuclear neutrophils; Wagner C et al.; Polymorphonuclear neutrophils (PMNs) produce an abundance of bactericidal and cytotoxic molecules consistent with their role as first-line defense against bacterial infection . PMNs, however, also cause efficient cellular cytotoxicity when targeted through Fc receptors to appropriate antibody-coated target cells . Although this so-called antibody-dependent cellular cytotoxicity (ADCC) was described many years ago, the mechanism of killing is still elusive . We now have found that PMNs contain perforin and granzyme B, the 2 molecules known as the cytotoxic entity of natural killer cells and of cytotoxic T lymphocytes as well . Lysates of PMNs were lytic for chicken erythrocytes in a time-, temperature-, and Ca(2+)-dependent manner . Moreover, apoptosis of Jurkat cells was induced, consistent with the observation that the PMN lysates contain enzymatically active granzyme B . Taken together, our data provide evidence for the presence of perforin and granzyme B within the cytotoxic arsenal of PMNs.

Antimicrob Agents Chemother, 2003 Oct, 47(10), 3240 - 6
Essential oils as components of a diet-based approach to management of Helicobacter infection; Bergonzelli GE et al.; An increased density of Helicobacter pylori in the gastric mucosa can be associated with more severe gastritis and an increased incidence of peptic ulcers . Therefore, people with asymptomatic gastritis would certainly benefit from a nutritional approach to help them manage the infection and therefore decrease the risk of development of associated pathologies . We analyzed the activities of 60 essential oils against H . pylori P1 and identified 30 oils that affected growth, with in vitro inhibition zones ranging between 0.7 and 6.3 cm in diameter . We further analyzed the effects of 16 oils with different activities on H . pylori P1 viability . Fifteen showed strong bactericidal activities, with minimal bactericidal concentrations after 24 h ranging from 0.02 to 0.1 g/liter at pH 7.4 . Even though slight variations in activities were observed, the essential oils that displayed the strongest bactericidal potentials against H . pylori P1 were also active against other Helicobacter strains tested . Among the pure constituents of different essential oils tested, carvacrol, isoeugenol, nerol, citral, and sabinene exhibited the strongest anti-H . pylori activities . Although oral treatment of H . pylori SS1-infected mice with carrot seed oil did not result in significant decreases in the bacterial loads in the treated animals compared to those in the control animals, in all experiments performed, the infection was cleared in 20 to 30% of carrot seed oil-treated animals . Our results indicate that essential oils are unlikely to be efficient anti-Helicobacter agents in vivo . However, their effects may not be irrelevant if one plans to use them as food additives to complement present therapies.

J Hosp Infect, 2003 Sep, 55(1), 68 - 72
Hospital use of decontaminating mats; Marchetti MG et al.; Decontaminating mats made of several layers of adhesive sheets (water-based acrylic 6 g/m2) supplemented with a bactericidal agent (3-1 benzoisothiazolin) at a concentration of 25% were placed in the passages providing access to the operating rooms of an orthopaedic service . Contact plates containing tryptone soy agar were used to assess bacterial concentration at specific points in front of and beyond the mats . For trolley passageways two areas were defined: central and lateral paths, corresponding to the areas walked upon by the personnel pushing the trolleys and to the paths covered by the trolley wheels, respectively . In order to exclude a simple mechanical effect, a comparison of bacterial loads at defined sites beyond the mats was carried out in the presence and in the absence of decontaminating mats . Bacterial colony counts in the presence of decontaminating mats were substantially and statistically significantly reduced compared with the absence of mats . The lower mean number of colony-forming units detected at points located beyond the mats parallels this finding; this difference is also statistically significant . We thus conclude that decontaminating mats are potentially useful in decreasing micro-organism carry-over due to personnel or the passage of trolleys into areas at high risk of infection such as operating rooms.

J Microbiol Methods, 2003 Oct, 55(1), 173 - 86
Fluoro-luminometric real-time measurement of bacterial viability and killing; Lehtinen J et al.; The viability and killing of Escherichia coli was measured on a real-time basis using a fluoro-luminometric device, which allows successive measurements of fluorescence and bioluminescence without user intervention . Bacteria were made fluorescent and bioluminescent by expression of gfp and insect luciferase (lucFF) genes . The green fluorescent protein (GFP) is a highly fluorescent, extremely stable protein, which accumulates in cells during growth, and therefore the measured fluorescence signal was proportional to the total number of cells . The luciferase reaction is dependent of ATP produced by living cells, so that the bioluminescence level was a direct measure of the viable cells . In contrast to the bacterial luciferase, the insect luciferase uses a water-soluble and nonvolatile substrate, which makes automated multi-well microplate assay possible . For the validation of the assay, the proportion of living and dead cell populations was experimentally modified by incubating E . coli cells in the presence of various ethanol concentrations . Bacterial viability and killing measured by a fluoro-luminometric assay correlated fairly well with the reference methods: conventional plate counting, optical density measurement and various flow cytometric analyses . The real-time assay described here allows following the changes in bacterial cultures and assessing the bactericidal and other effects of various chemical, immunological and physical agents simultaneously in large numbers of samples.

Lett Appl Microbiol, 2003, 37(4), 318 - 23
Genetically modified filamentous phage as bactericidal agents: a pilot study; Hagens S et al.; AIMS: To evaluate the ability of a filamentous phage encoding lethal proteins to kill bacteria without host-cell lysis . METHODS AND RESULTS: Bacterial survival was determined after infection of a growing Escherichia coli culture with phage M13 encoding either the restriction endonuclease BglII gene or modified phage lambda S holin genes . The genetically engineered phage exerted a high killing efficiency while leaving the cells structurally intact . When compared with a lytic phage, the release of endotoxin was minimized after infection with the genetically modified phages . CONCLUSIONS: Genetically engineered phage can be used for efficient killing, concomitantly minimizing endotoxin release . SIGNIFICANCE AND IMPACT OF THE STUDY: This feasibility study provides a possible strategy for the use of genetically engineered phage as bactericidal agents by optimizing the advantages and minimizing potential risks such as release of pyrogenic cell wall components.

Clin Diagn Lab Immunol, 2003 Sep, 10(5), 797 - 801
Antibody responses to Escherichia coli O157 and other lipopolysaccharides in healthy children and adults; Navarro A et al.; In Mexico, diarrheal disease due to different serotypes of Escherichia coli is highly prevalent, with only sporadic isolation of O157 non-H7 strains . This could be due to exposure to the O157 or related E . coli lipopolysaccharide (LPS), such as O7 or O116, at an early age . By using enzyme-linked immunosorbent assay (ELISA) and Western blotting, the present study analyzed 605 serum samples from Mexican adults and infants without clinical symptoms of disease for the presence of antibodies to these three E . coli LPSs . The bactericidal activities of homologous and heterologous rabbit and human serum samples against O7, O116, and O157 E . coli LPSs were also determined . By using a cutoff point of 0.7, it was found by the ELISAs that 28 of 562 (5%) of the serum samples from adolescents and adults and 2 of 43 (5%) of the serum samples from infants less than 1 year of age reacted with the O157 LPS . By using cutoff points between 0.4 and 0.699, the proportion of serum samples from both age groups that reacted with the O157 LPS increased to 20% . Western blotting analysis of selected serum samples that showed an intermediate response against the O157 LPS by the ELISAs showed that 61 of 88 (69%) reacted with the same LPS . A similar result was observed for maternal milk samples . The bactericidal activities of rabbit serum samples against the O7, O116, and O157 LPSs showed that they were positive for both homologous and heterologous antigens . Similar results were observed with the human serum samples . O157 non-H7 strains were identified in only 10% of the E . coli strains isolated from 263 Mexican children with and without diarrhea over the past 15 years . This absence of O157:H7 strains in Mexico may be associated with the presence of antibodies against O157 or related E . coli LPSs.

Am J Physiol Cell Physiol, 2003 Oct, 285(4), C723 - 34
Rac regulates cardiovascular superoxide through diverse molecular interactions: more than a binary GTP switch; Gregg D et al.; The small G protein Rac has been implicated in multiple cardiovascular processes . Rac has two major functions: 1) it regulates the organization of the actin cytoskeleton, and 2) it controls the activity of the key enzyme complex NADPH oxidase to control superoxide production in both phagocytes and nonphagocytic cells . In phagocytes, superoxide derived from NADPH has a bactericidal function, whereas Rac-derived superoxide in the cardiovascular system has a diverse array of functions that have recently been a subject of intense interest . Rac is differentially activated by cellular receptors coupled to distinct Rac-activating adapter molecules, with each leading to pathway-specific arrays of downstream effects . Thus it may be important to investigate not just whether Rac is activated but also where, how, and for what effector . An understanding of the biochemical functions of Rac and its effectors lays the groundwork for a dissection of the exact array of effects produced by Rac in common cardiovascular processes, including cardiac and vascular hypertrophy, hypertension, leukocyte migration, platelet biology, and atherosclerosis . In addition, investigation of the spatiotemporal regulation of both Rac activation and consequent superoxide generation may produce new insights into the development of targeted antioxidant therapies for cardiovascular disease and enhance our understanding of important cardiovascular drugs, including angiotensin II antagonists and statins, that may depend on Rac modulation for their effect.

Front Biosci, 2003 Sep 01, 8, s1266 - 79
Diversity of voltage gated proton channels; Morgan D et al.; Voltage gated proton channels were first discovered in snail neurons and recently have been found in many mammalian cells . As their name suggests, H+ channels are sensitive to voltage, with an open probability that increases with membrane depolarization . Many properties that are shared by voltage-gated proton channels make them unique among ion channels . They show high selectivity for protons, strongly pH dependent gating, and a tiny single channel conductance . Although they are inhibited by divalent cations, including zinc and cadmium, no effective blockers exist . There is sufficient evidence to suggest that they are not water filled pores, unlike many other membrane bound ion channels . Instead, protons probably are conducted by a "hydrogen bonded chain" mechanism that resembles the Grotthuss mechanism in water . Differences in activation and deactivation kinetics of H+ currents in different cells suggest that there may be at least 4 isoforms of voltage gated proton channels . Gating kinetics may reflect specific functions . Voltage gated proton channels are well suited to extrude acid from cells and also may function in the extrusion of metabolic acid in the form of CO2 from the lungs . The best established function of H+ channels is in mammalian phagocytes, where they extrude protons to compensate for the charge separation created by the movement of electrons across the membrane by the bactericidal enzyme NADPH oxidase.

Mol Immunol, 2003 Sep, 40(5), 269 - 78
Molecular cloning of a novel bactericidal permeability-increasing protein/lipopolysaccharide-binding protein (BPI/LBP) from common carp Cyprinus carpio L . and its expression; Kono T et al.; A novel bactericidal permeability-increasing protein/lipopolysaccharide (LPS)-binding protein (BPI/LBP) was isolated from common carp Cyprinus carpio L . by EST analysis . This gene showed structural similarity with BPI/LBP gene in mammals . The isolated gene was composed of 1638 bp, which translated to a protein of 473 amino acid residues . The predicted signal peptide was 18 amino acid residues and the LPS-binding domain is conserved in this sequence consisted of residues 57-121 amino acid residues . This LPS-binding domain had high identity (76.9%) to that of rainbow trout LBP/BPI-2 . Phylogenetic analysis showed that carp BPI/LBP was similar to BPI or LBP of lipid-interactive protein family in mammals . Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that carp BPI/LBP gene was expressed in normal liver, head kidney, spleen, intestine, gill, heart and brain . Liver and head kidney stimulated with LPS (10 microg/ml) for 3, 6, 12, 24 and 48 h expressed carp BPI/LBP gene at all stimulation time periods . Expression level of liver was higher than that of head kidney and high expressions in each tissues was recorded at 3h post-LPS stimulation . After 3h, BPI/LBP gene expression level was gradually become less in the stimulated times.

Leg Med (Tokyo), 2003 Jun, 5(2), 73 - 86
Immunohistochemical detection of sepsis-induced lung injury in human autopsy material; Tsokos M; This review addresses our present-day knowledge on the role of different cellular adhesion molecules, cytokines and glycoproteins for the detection of sepsis-induced injury in the microvasculature of the human lung using immunohistochemistry . Through the induction and modulation of endothelial cell adhesion molecules, such as E-selectin (CD 62E), the vascular endothelium controls leukocyte extravasation into tissue . E-Selectin, not expressed by unstimulated endothelium, is activated by cytokines and initiates neutrophil recruitment in sepsis-induced lung injury . Since E-selectin is strongly expressed in the pulmonary microvasculature in sepsis-associated fatalities, the immunohistochemical detection of an intense expression of E-selectin in lung tissue is a valuable diagnostic tool in the forensic postmortem elucidation of death due to sepsis . VLA-4 (CD49d/CD29) is strongly expressed on intravascular, interstitial and intra-alveolar leukocytes in sepsis-associated fatalities, whereas in non-septic fatalities an irregular weak immunoreactivity can be observed on interstitial leukocytes and no positive immunohistochemical expression can be observed on intravascular or intra-alveolar leukocytes . ICAM-1 (CD54) is strongly expressed on endothelial cells of the pulmonary microvasculature and on pulmonary macrophages and lymphocytes in sepsis-associated fatalities . In contrast, an infrequent weak immunohistochemical reaction for ICAM-1 is found on pulmonary endothelium and on perivascular leukocytes in non-septic fatalities . The up-regulation of both cellular adhesion molecules can be considered as an useful immunohistochemical postmortem marker of sepsis . Lactoferrin (LF) is an iron-binding glycoprotein located in specific (secondary) granules of leukocytes and plays a central role in the host response to infectious stimuli in providing both bacteriostatic and bactericidal protection . There is a statistically significant association between an enhanced expression of LF on pulmonary leukocytes in sepsis-related fatalities in contrast to non-septic controls . The immunohistochemical detection of an enhanced expression of LF can contribute to the postmortem discrimination between sepsis and non-septic fatalities . Application of carbohydrate-specific lectins (ConA, UEA, GSA I, GSA II, MPA, PNA, Jac, WGA, MAA, LPA, SNA) on deparaffinated lung tissue sections from sepsis-associated fatalities and control cases results to some extent in different staining patterns of alveolar epithelial cells and subepithelial seromucous glands of the bronchi . Apart from differences in binding sites for alpha-mannose, N-acetyl-neuraminic acid and alpha-(2-6)-galactose (as detected by different expression for ConA, MAA and SNA), the main finding is that no binding sites for alpha-N-acetyl-galactosamine (as investigated by MPA immunoreactivity) can be detected on alveolar epithelial cells and mucous parts of subepithelial seromucous glands in sepsis cases in contrast to the presence of such binding sites in controls . Since most intracellular pathogens persist in macrophages and epithelial cells during infection, it is likely that these pathogens contribute to a continual deprivation or consumption, respectively, of glycoproteins physiologically secreted by alveolar epithelial and glandular cells at different time points and stages of infection and may, among other mechanisms, by reducing pathogen clearance amplify the inflammatory response . Vascular endothelial growth factor (VEGF), an angiogenic and chemotactic peptide, is abundantly expressed in normal lung tissue, especially in alveolar and bronchial epithelium, glandular cells of the bronchi, and activated alveolar macrophages . Pulmonary VEGF immunostaining differs in sepsis when compared to healthy individuals . In the latter a preponderant strong VEGF immunoreaction can be found on alveolar epithelium (predominately type II pneumocytes), bronchial epithelium and glandular cells of the bronchi and bronchioli, and activated alveolar macrophages . In contrast, in sepsis no VEGF immunopositivity can beivity can be observed on bronchial epithelium or glandular cells of the bronchi and bronchioli, and no or relatively sparse VEGF immunoreactivity is found on alveolar epithelial cells . The precise mechanisms of the decreased pulmonary VEGF expression in septic patients under conditions of intensive care medicine are not clear at present . During the complex cascade of excessive pro-inflammatory and anti-inflammatory mediator release involved in the host's systemic inflammatory response in the development of sepsis-induced lung injury, VEGF expression may be suppressed in sepsis by a hitherto not identified agent or the interaction of different mediators of cellular inflammation . For the detection of sepsis-induced lung injury the aforementioned markers can be used sufficiently, e.g . to give immunohistochemical evidence of a previously undiagnosed sepsis and to confirm or rule out a presumed diagnosis of a sepsis-associated fatality . The employment of the presented immunohistochemical methods will be particularly helpful when macroscopical and routine histological autopsy findings in cases of suspected fatal sepsis are unspecific or unconvincing, respectively, and clinical data on the patient's previous history are not available . Referring to the forensic argumentation regarding causality on the subject of possibly fatal septic complications, e.g . in the sequel of diagnostic or therapeutic iatrogenic injection procedures or being relevant to pressure sore-associated fatalities, aetiopathogenetic conclusions can be optimized on the basis of the described micromorphological investigations.

J Infect Dis, 2003 Sep 1, 188(5), 738 - 42 Epub 2003 Aug 05.
Crucial role of antibodies to pertactin in Bordetella pertussis immunity; Hellwig SM et al.; Pertussis, a serious infectious disease of the respiratory tract caused by Bordetella pertussis, is reemerging in vaccinated populations . Efforts to curtail this disease are hampered by limited insight into the basis of protective immunity . Opsonophagocytosis was recently found to play a central role in cellular bactericidal activity against B . pertussis . In the present study, we studied the specificity of opsonic antibodies . Anti-pertactin antibodies, but not anti-pertussis toxin, anti-fimbriae, or anti-filamentous hemagglutinin antibodies, were found to be crucial for B . pertussis phagocytosis . These data are consistent with field studies showing that levels of antibodies to pertactin correlate with protection.

Biochem Biophys Res Commun, 2003 Sep 5, 308(4), 764 - 9
Selective role of PI3K delta in neutrophil inflammatory responses; Sadhu C et al.; Although members of the class I phosphoinositide 3-kinases (PI3Ks) have been implicated in neutrophil inflammatory responses, the contribution of the individual PI3K isoforms in neutrophil activation has not been tractable with the non-selective inhibitors, LY294002 and wortmannin . We have developed a novel series of PI3K inhibitors that is selective for PI3K delta, an isoform expressed predominantly in hematopoietic cells . In addition to being selective between members of class I PI3Ks, representatives of these inhibitors such as IC980033 and IC87114 did not inhibit any protein kinases tested . Utilizing these inhibitors we report here a novel role for PI3K delta in neutrophil activation . Inhibition of PI3K delta with IC980033 and IC87114 blocked both fMLP- and TNF1 alpha-induced neutrophil superoxide generation and elastase exocytosis . The PI3K delta inhibitor IC87114 also blocked TNF1 alpha-stimulated elastase exocytosis from neutrophils in a mouse model of inflammation . To our knowledge, this is the first in vivo efficacy demonstration of a PI3K delta inhibitor in an animal model . Inhibition of PI3K delta, however, had no effect on in vitro neutrophil bactericidal activity and Fc gamma R-stimulated superoxide generation . Thus, PI3K delta plays an essential role in certain signaling pathways of neutrophil activation and appears to be an attractive target for the development of an anti-inflammatory therapeutic.

J Exp Med, 2003 Aug 18, 198(4), 653 - 9
Tuberculosis toxin blocking phagosome maturation inhibits a novel Ca2+/calmodulin-PI3K hVPS34 cascade; Vergne I et al.; The capacity of Mycobacterium tuberculosis to infect latently over one billion people and cause two million fatalities annually rests with its ability to block phagosomal maturation into the phagolysosome in infected macrophages . Here we describe how M . tuberculosis toxin lipoarabinomannan (LAM) causes phagosome maturation arrest, interfering with a new pathway connecting intracellular signaling and membrane trafficking . LAM from virulent M . tuberculosis, but not from avirulent mycobacteria, blocked cytosolic Ca2+ increase . Ca2+ and calmodulin were required for a newly uncovered Ca2+/calmodulin phosphatidylinositol (PI)3 kinase hVPS34 cascade, essential for production of PI 3 phosphate (PI3P) on liposomes in vitro and on phagosomes in vivo . The interference of the trafficking toxin LAM with the calmodulin-dependent production of PI3P described here ensures long-term M . tuberculosis residence in vacuoles sequestered away from the bactericidal and antigen-processing organelles in infected macrophages.

BMC Microbiol . 2003 Aug 12;3(1):16.
Effects of disruption of heat shock genes on susceptibility of Escherichia coli to fluoroquinolones; Yamaguchi Y et al.; BACKGROUND: It is well known that expression of certain bacterial genes responds rapidly to such stimuli as exposure to toxic chemicals and physical agents . It is generally believed that the proteins encoded in these genes are important for successful survival of the organism under the hostile conditions . Analogously, the proteins induced in bacterial cells exposed to antibiotics are believed to affect the organisms' susceptibility to these agents . RESULTS: We demonstrated that Escherichia coli cells exposed to levofloxacin (LVFX), a fluoroquinolone (FQ), induce the syntheses of heat shock proteins and RecA . To examine whether the heat shock proteins affect the bactericidal action of FQs, we constructed E . coli strains with mutations in various heat shock genes and tested their susceptibility to FQs . Mutations in dnaK, groEL, and lon increased this susceptibility; the lon mutant exhibited the greatest effects . The increased susceptibility of the lon mutant was corroborated by experiments in which the gene encoding the cell division inhibitor, SulA, was subsequently disrupted . SulA is induced by the SOS response and degraded by the Lon protease . The findings suggest that the hypersusceptibility of the lon mutant to FQs could be due to abnormally high levels of SulA protein resulting from the depletion of Lon and the continuous induction of the SOS response in the presence of FQs . CONCLUSION: The present results show that the bactericidal action of FQs is moderately affected by the DnaK and GroEL chaperones and strongly affected by the Lon protease . FQs have contributed successfully to the treatment of various bacterial infections, but their widespread use and often misuse, coupled with emerging resistance, have gradually compromised their utility . Our results suggest that agents capable of inhibiting the Lon protease have potential for combination therapy with FQs.

Vet Immunol Immunopathol, 2003 Aug 15, 94(3-4), 113 - 21
Immune modulation following immunization with polyvalent vaccines in dogs; Strasser A et al.; A decline in T-cell-mediated immunity and transient state of immunosuppression after immunization has been reported in dogs . Nevertheless, dogs are still routinely vaccinated with polyvalent live vaccines and severe disease does not generally occur . In order to investigate these effects on the canine immune system and to elucidate possible mechanisms we determined the following immune parameters in the blood of 33 clinically sound German shepherd dogs before and after standard vaccination with a polyvalent vaccine against distemper, parvovirus, viral hepatitis, leptospirosis, kennel cough and rabies: white and differential blood cell count, the serum concentrations and/or activities of IL-1, IL-2, IFN-gamma, TNF-alpha, neopterin and IgG, natural killer (NK) cell activity, bactericidal activity and complement hemolytic activity, lymphocyte proliferation test (LPT) and nitroblue tetrazolium test (NBT).Our major findings were that significant postvaccinal decreases in T-cell mitogenic response to PHA and in neutrophil function and neopterin serum concentration were accompanied by simultaneous increase in plasma IgG and hemolytic complement activity . This suggests a transient shift in the balance between cell-mediated and humoral (T(H)1/T(H)2) immunity rather than immunosuppression.These results do not imply that dogs should not receive live vaccines, as the response to vaccines just seems to create a state of altered homeostasis when immunization elicits protection by humoral and cell-mediated immunity . However, these recognized compromises of immune function should be considered and vaccines still be applied only in healthy animals and strictly according to the rules and regulations given by the manufacturer.

Acta Physiol Hung, 2003, 90(2), 83 - 95
Semmelweis' discovery and its Finnish follow-up; Hanninen O et al.; Professor Ignac Semmelweis (1818-1865) is one of the great personalities of medical history . He insisted on washing hands with chlorine water before any obstetrical intervention, he was the first to demonstrate its importance in preventing puerperal fever . Thus, the principle of asepsis was introduced prior to the discovery of bacteria and bacterial diseases . Semmelweis carefully documented his findings and in this way pioneered the scientific analysis of clinical data Medical community of that time misinterpreted Semmelweis' great ideas, he died abandoned and forgotten . A Finnish doctor Josef Adam Joachim Pippingskold was one of the first obstetricians who had realized the importance of Semmelweis' work . In 1861, in his letter to Semmelweis he reported about his own findings and favorable results in prevention of puerperal fever in Helsinki . Two decades earlier, Dr . Ehrstrom in the University of Helsinki had submitted his thesis on pathophysiology of puerperal fever that was similar to the ideas of Semmelweis . Long before modern times in Finland, mothers traditionally had their babies delivered in smoke saunas, where heating and smoke of bactericidal phenols created a clean, rather aseptic environment . Hand washing was self-evident necessity . However, the situation was quite different in the Central European universities and departments of obstetrics, where the medical training and clinical practice took place side by side . Semmelweis' life and his contribution to medicine was appreciated even in the theatrical circles of Finland . The piece "Semmelweis" of Norwegian playwright Jens Bjorneboe got its World Premier in the Swedish Theatre in Turku, former capital of Finland, in September 1969.

Zhonghua Yi Xue Za Zhi, 2003 Jun 10, 83(11), 932 - 5
{Antineutrophil cytoplasmic antibody associated vasculitis induced by antithyroid agents}; Guo XH et al.; OBJECTIVE: To investigate the significance of antineutrophil cytoplasmic antibody (ANCA) in hyperthyroidism at different stages and treated with different drugs and to investigate the relationship between the ANCA specific target antigens and the clinical manifestations of vasculitis . METHODS: Two hundred and sixteen patients with hyperthyroidism were divided into four groups: untreated (n = 34), treated with propylthiouracil (PTU) (n = 62), treated with methimazole (MMI) (n = 77), and treated with both PTU and MMI (n = 43) . Sera were collected from the 216 patients . Indirect immunofluorescence (IIF) test was used to detect the ANCA and antinuclear antibody (ANA) . Antigen-specific ELISA was used to detect the existence of 7 specific target antigens of ANCA: myeloperoxidase (MPO), proteinase 3 (PR3), lactoferrin (LF), human leukocyte elastase (HLE), azurocidin (AZU), cathepsinG (CG), and bactericidal/permeability-increasing protein (BPI) in the sera positive in ANCA and ANA determined by IIF . RESULTS: The IIF positive rate was 5.9% (2/34) in the untreated group; 22.6% (14/62) in the patients treated with PTU, all IIF-ANCA positive; 6.5% (5/77) in the patients treated with MMI, all IIF-ANA positive; and 27.9% (12/43) in the patients treated with both PTU and MMI, 8 IIF-ANA positive and 3 IIF-ANCA positive . The IIF-ANCA positive rate in the patients receiving PTU was significantly higher than that in the untreated patients (P < 0.017) . The IIF-ANA and IIF-ANCA positive rates of the patients treated with PTU were both significantly higher than those of the patients treated with MMI (P < 0.017) . Six of the IIF-positive 31 patients administered with drugs (19.4%) had signs and symptoms associated with vasculitis . Of the six patients, 4 were MPO antibody positive (66.7%), 2 were PR3 antibody positive (33.3%), 4 were LF antibody positive, 3 were HLE antibody positive, 3 were AZU antibody positive, and 2 were CG antibody positive . None was anti-BPI antibody positive . Two of the six patients were both anti-MPO and anti-PR3 negative . All MPO-ANCA positive patients had clinical signs and symptoms of vasculitis . CONCLUSION: PTU is associated with the production of ANCA in patients with hyperthyroidism . PTU induced ANCA is caused by polyclonal activation of B cells . Anti-MPO antibody may be related to the occurrence of clinical vasculitis.

Pancreatology, 2003, 3(4), 329 - 35
Nitric oxide regulates bacterial translocation in experimental acute edematous pancreatitis; Cevikel MH et al.; BACKGROUND/AIMS: The role of nitric oxide (NO) in bacterial translocation (BT) associated with acute pancreatitis is controversial . We investigated the effects of the NO synthase substrate, L-arginine, and the NO synthase inhibitor, N-nitro-L-arginine methyl ester (L-NAME), on BT in caerulein-induced acute pancreatitis in rats . METHODS: Acute pancreatitis was induced by subcutaneous injections of caerulein (12 microg/kg) at 6-hour intervals for 2 days . Subcutaneous injections of L-arginine (100 mg/kg) or L-NAME (10 mg/kg) were administeredonce daily for 2 days . At 48 h, pancreatic injury and BT to the mesenteric lymph nodes (MLN), liver, and peritoneum were assessed . RESULTS: Compared with controls, rats that received caerulein injections alone had increased BT to the MLN and pancreatic inflammatory changes . L-Arginine significantly reduced the inflammation and BT caused by caerulein . L-NAME did not significantly alter pancreatic inflammation . Although caerulein + L-NAME-treated rats had increased BT to the peritoneum, MLN, and liver compared with controls, rates of BT did not significantly differ between caerulein alone- and caerulein + L-NAME-treated rats . CONCLUSION: In acute edematous pancreatitis, BT is increased and is regulated by NO . NO substrates limit BT and pancreatic inflammation associated with acute pancreatitis, probably by their bactericidal actions and ability to improve pancreatic blood flow .

J Biol Chem, 2003 Oct 17, 278(42), 40542 - 9 Epub 2003 Jul 29.
Myeloperoxidase-derived hypochlorous acid antagonizes the oxidative stress-mediated activation of iron regulatory protein 1; Mutze S et al.; Hypochlorous acid (HOCl) is a highly reactive product generated by the myeloperoxidase reaction during the oxidative burst of activated neutrophils, which is implicated in many bactericidal and cytotoxic responses . Recent evidence suggests that HOCl may also play a role in the modulation of redox sensitive signaling pathways . The short half-life of HOCl and the requirement for a continuous presence of H2O2 as a substrate for its myeloperoxidase-catalyzed generation make the study of HOCl-mediated responses very difficult . We describe here an enzymatic model consisting of glucose/glucose oxidase, catalase, and myeloperoxidase (GOX/CAT/MPO) that allows the controlled generation of both HOCl and H2O2 and thus, mimics the oxidative burst of activated neutrophils . By employing this model we show that HOCl prevents the H2O2-mediated activation of iron regulatory protein 1 (IRP1), a central post-transcriptional regulator of mammalian iron metabolism . Activated IRP1 binds to (R)iron-responsive elements" (IREs) within the mRNAs encoding proteins of iron metabolism and thereby controls their translation or stability . The inhibitory effect of HOCl is not a result of a direct modification of IRP1 by this oxidant . Kinetics experiments provide evidence that HOCl intervenes with the signaling cascade, which results in the activation of IRP1 . We further demonstrate that HOCl antagonizes the H2O2-mediated increase in the levels of transferrin receptor, which is a downstream target of IRP1 . Our findings suggest that HOCl can modulate signaling pathways in a concerted action with H2O2 . The GOX/CAT/MPO system provides a valuable tool for studying the regulatory function of HOCl.

Biochem Soc Trans, 2003 Aug, 31(Pt 4), 806 - 9
Comparative analysis of the PLUNC (palate, lung and nasal epithelium clone) protein families; Bingle CD et al.; PLUNC (palate, lung and nasal epithelium clone) is a small, secreted protein that is expressed in the oropharynx and upper airways of humans, mice and rats . We have described a family of at least 14 PLUNC genes localized on chromosome 20 (in humans), 2 (in mice) or 3 (in rats) . These rapidly evolving proteins are structurally related to lipopolysaccharide-binding protein (LBP) and bactericidal/permeability-increasing protein (BPI) . In the present analysis we comment on the comparative aspects of this protein family, which may function to protect epithelial surfaces from pathogenic micro-organisms.

Biochem Soc Trans, 2003 Aug, 31(Pt 4), 801 - 5
Four BPI (bactericidal/permeability-increasing protein)-like genes expressed in the mouse nasal, oral, airway and digestive epithelia; Leclair EE; A cluster of related genes whose products show structural identity with bactericidal permeability-increasing protein (BPI) has been identified in the genomes of both mice (on chromosome 2) and humans (on chromosome 20) . Genes in the cluster include those encoding parotid secretory protein (PSP), von Ebner minor salivary gland protein (VEMSGP) and sequences in the PLUNC (palate, lung and nasal epithelium clone) family, among others . This mini-review addresses the tissue-specific expression of these genes in the mouse.

Biochem Soc Trans, 2003 Aug, 31(Pt 4), 791 - 4
Structure of human BPI (bactericidal/permeability-increasing protein) and implications for related proteins; Beamer LJ; Human bactericidal/permeability-increasing protein (BPI) belongs to a family of mammalian lipopolysaccharide-binding and lipid transport proteins . Recent sequence database searches indicate that several other protein families, including the palate, lung and nasal epithelial clone (PLUNC), parotid secretory protein (PSP) and BPI-like proteins, are likely to share the BPI fold, which was determined through X-ray crystallographic studies . As the single representative of its fold family of known structure, the three-dimensional model of BPI suggests structural features that are likely to be conserved across this large and varied group of proteins.

Biochem Soc Trans, 2003 Aug, 31(Pt 4), 781 - 4
Bovine parotid secretory protein: structure, expression and relatedness to other BPI (bactericidal/permeability-increasing protein)-like proteins; Wheeler TT et al.; Members of the family of BPI (bactericidal/permeability-increasing protein)-like proteins are as yet incompletely characterized, particularly in cattle, where full-length sequence information is available for only three of the 13 family members known from other species . Structural bioinformatic analyses incorporating bovine homologues of several members of the BPI-like protein family, including two forms of bovine parotid secretory protein (PSP), showed that this family is also present in cattle . Expression analyses of several members of the BPI-like protein family in cattle, including PSP (Bsp30), von Ebner's minor salivary gland protein (VEMSGP) and lung-specific X protein (LUNX), showed a restricted pattern of expression, consistent with earlier hypotheses that these proteins function in the innate immune response to bacteria . The possible role of bovine PSP in susceptibility to pasture bloat in cattle is discussed.

Biochem Soc Trans, 2003 Aug, 31(Pt 4), 777 - 80
Psp and Smgb: a model for developmental and functional regulation in the rat major salivary glands; Ball WD et al.; This paper summarizes past work detailing the developmental expression, cell and organ localization and biochemical features of the proteins parotid secretory protein (PSP) and isoforms of submandibular gland protein B (SMGB), and describes the molecular characterization of the genes that encode them, Psp and Smgb . These genes appear to be related to the BPI (bactericidal/permeability-increasing protein)/LBP (lipopolysaccharide-binding protein)/PLUNC (palate, lung and nasal epithelial clone) gene family found in the oral and respiratory organs of humans, rodents and cattle . We have emphasized the diverse patterns of expression of these genes among the submandibular, sublingual and parotid salivary glands of the rat, and their potential usefulness in defining and identifying genomic regulatory mechanisms of salivary development . While Psp is expressed similarly in the mouse, the putative Smgb gene of the mouse seems not to be expressed, apparently due to the insertion, between exons 1 and 2, of a gene for a retroviral protein.

Biol Chem, 2003 Jun, 384(6), 911 - 20
Isolation and characterization of a novel and potent inhibitor of Arg-gingipain from Streptomyces sp . strain FA-70; Kadowaki T et al.; Arg-gingipain (Rgp) is a major cysteine proteinase produced by the oral bacterium Porphyromonas gingivalis, which is a major pathogen of advanced periodontal diseases . This enzyme is important for the bacterium both to exhibit its virulence and to survive in periodontal pockets . The development of Rgp inhibitors thus provides new therapeutic approaches to periodontal diseases . In this study, we first isolated and purified a novel and potent inhibitor of Rgp from the culture supernatant of Streptomyces species strain FA-70, now designated as FA-70C1 . This compound was found to be an antipain analog composed of phenylalanyl-ureido-citrullinyl-valinyl-cycloarginal (C27H43N9O7) . The Ki value was calculated to be 4.5x10(-9) M when benzyloxycarbonyl-phenylalanyl-arginine-4-methly-coumaryl-7-amide was used as a substrate . This compound also inhibited cathepsins B, L, and H, though their Ki values were much higher than that of Rgp . FA-70C1 had little or no inhibitory activity on Lys-gingipain, another cysteine proteinase of P . gingivalis . The Rgp-induced degradation of various human proteins was completely blocked by this inhibitor . Disruption of both the bactericidal activity of polymorphonuclear leukocytes and the viability of human fibroblasts and umbilical vein endothelial cells induced by the culture supernatant of P . gingivalis was suppressed by the inhibitor in a dose-dependent manner . The enhancement of vascular permeability induced by in vivo administration of the culture supernatant of P . gingivalis was strongly inhibited by the inhibitor . Furthermore, the growth of P . gingivalis was suppressed by FA-70C1 in a dose-dependent manner . These results strongly suggest that FA-70C1 is a useful tool to prevent the virulence of P . gingivalis.

Glycobiology, 2003 Nov, 13(11), 755 - 63 Epub 2003 Jul 24.
Galectin-8 modulates neutrophil function via interaction with integrin alphaM; Nishi N et al.; The members of the galectin family are associated with diverse cellular events, including immune response . We investigated the effects of galectin-8 on neutrophil function . Human galectin-8 induced firm and reversible adhesion of peripheral blood neutrophils but not eosinophils to a plastic surface in a lactose-sensitive manner . Other human galectins, galectins-1, -3, and -9, showed low or negligible effects on neutrophil adhesion . Confocal microscopy revealed actin bundle formation in the presence of galectin-8 . Cytochalasins inhibited both actin assembly and cell adhesion induced by galectin-8 . Affinity purification of galectin-interacting proteins from solubilized neutrophil membrane revealed that N-terminal carbohydrate recognition domain (CRD) of galectin-8 bound promatrix metalloproteinase-9 (proMMP-9), and C-terminal CRD bound integrin alphaM/CD11b and proMMP-9 . A mutant galectin-8 lacking the carbohydrate-binding activity of N-terminal CRD (galectin-8R69H) retained adhesion-inducing activity, but inactivation of C-terminal CRD (galectin-8R233H) abolished the activity . MMP-3-mediated processing of proMMP-9 was accelerated by galectin-8, and this effect was inhibited by lactose . Galectins-1 and -3 did not affect the processing . Superoxide production, an essential event in bactericidal function of neutrophils, was stimulated by galectin-8 to an extent comparable to that induced by fMLP . Galectin-8R69H but not galectin-8R233H could stimulate superoxide production . Taken together, these results suggest that galectin-8 is a novel factor that modulates the neutrophil function related to transendothelial migration and microbial killing.

Antimicrob Agents Chemother, 2003 Aug, 47(8), 2666 - 8
Alpha-defensin 1 (human neutrophil protein 1) as an antichemotactic agent for human polymorphonuclear leukocytes; Grutkoski PS et al.; Medium conditioned by tumor necrosis factor alpha (TNF-alpha)-stimulated polymorphonuclear leukocytes (PMN) (CM-TNF) suppresses PMN migration . Therefore, we wished to identify the agent(s) in CM-TNF that mediated antichemotactic activity . CM-TNF was fractionated by high-performance liquid chromatography, and one fraction with antichemotactic activity contained the bactericidal protein human neutrophil protein 1 (HNP-1) . We showed that HNP-1 suppresses PMN migration to formyl-methionyl-leucyl-phenylalanine but not to interleukin 8.

Antimicrob Agents Chemother, 2003 Aug, 47(8), 2646 - 8
In vitro susceptibilities of seven Leptospira species to traditional and newer antibiotics; Hospenthal DR et al.; Human leptospirosis is generally treated with penicillin or doxycycline . We studied the susceptibilities of 11 serovars (seven species) of Leptospira to 14 antibiotics . With the exception of chloramphenicol, all tested agents were at least as potent as penicillin and doxycycline, with the macrolide and ketolide drugs producing the lowest MICs (and minimal bactericidal concentrations).

Aquat Toxicol, 2003 Sep 10, 64(4), 375 - 91
Stimulation of defense factors for oysters deployed to contaminated sites in Pensacola Bay, Florida; Fisher WS et al.; A positive association between chemical contaminants and defense factors has been established for eastern oysters (Crassostrea virginica) from Florida, but it is unknown whether such factors can be stimulated through short-term exposure to contaminants in the field . Hatchery oysters were deployed at two contaminated sites and one reference site near Pensacola, Florida, during spring and summer in 1998 . Putative defense measurements, notably hemocyte count and bactericidal activity, were significantly elevated after 12-week deployment during summer at the most contaminated site . This site exhibited a dramatic increase in chemical concentrations in oyster tissue relative to both the initial concentrations in hatchery oysters and to oysters deployed at the reference site . Hemocyte activity was not stimulated after 16-week deployment of hatchery oysters in spring, despite similar increases in tissue chemical concentrations, so defense activation by short-term exposure may covary with other unmeasured environmental or physiological parameters . Using the converse approach, Pensacola Bay oysters were collected from two contaminated sites and deployed at the reference site for 16 weeks during spring . Results from this converse deployment were ambiguous; serum lysozyme concentrations were reduced for oysters transplanted from both sites, but hemocyte activities were not significantly changed . The principal outcome from this study was the demonstration of enhanced defense activities for oysters upon short-term summer deployment at a contaminated site.

Aquat Toxicol, 2003 Sep 10, 64(4), 363 - 73
Greater hemocyte bactericidal activity in oysters (Crassostrea virginica) from a relatively contaminated site in Pensacola Bay, Florida; Oliver LM et al.; Bivalve mollusks such as Crassostrea virginica inhabiting polluted estuaries and coastal areas may bioaccumulate high concentrations of contaminants without apparent ill effects . However, changes in putative internal defense activities have been associated with contaminant accumulation in both experimental and long-term field exposures . In an effort to elucidate these relationships, 40 oysters were collected from Bayou Chico (BC) and East Bay (EB) in Pensacola Bay, FL, two estuaries known to differ in the type and magnitude of chemical contaminants present . Oyster tissue concentrations of metals, tri- and dibutyltin (TBT, DBT), polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) were measured in individual oysters, as were hemocyte counts (HCs), hemocyte bacterial killing indices (KI), serum lysozyme (LYS) and serum protein (PRO) levels . Average HC, KI, LYS and PRO were significantly higher in BC oysters, which also had significantly higher tissue concentrations of total trace metals, butyltins (BTs), PAHs, PCBs, pesticides, and Mn, Cu, Zn and Sn . EB oysters had low organic contaminant levels and no detectable BTs, but significantly higher concentrations of Al, Cr, Fe, Ag, Cd, and Hg . Simple correlation analysis between specific defense measurements and specific chemical analytes showed specific positive relationships that corroborated previous findings in other FL estuaries . Canonical correlation analysis was used to examine relationships between defense measurements and tissue metals using linearly combined sets of variables . Results were also consistent with previous findings-the highest possible canonical correlation was positive: r=0.864, P<0.0019 among canonical variables composed of HC, KI and LYS for defense, and Fe, Cu, Ag, Cd, Sb, Sn, Ni, Pb and Hg for metals.

Infez Med, 1996, 4(4), 228 - 33
{In vitro and in vivo effects of cefodizime and ceftriaxone on oxygen-dependent killing of neutrophils of elderly patients with exacerbation of chronic bronchitis}; Papa L et al.; The immunomodulating activity of some chemotherapeutic agents, is particularly interesting . Our study has evaluated the in vitro and in vivo effects of two third generation cephalosporins: cefodizime and ceftriaxone, on the chemiluminescence of polymorphonuclears of 20 elderly patients affected by acute exacerbation of bronchitis . Twenty healthy patients have been also evaluated in vitro . Antibiotics have been used in vitro at the concentration of 50 g/ml while in vivo a group of 10 patients have been treated with cefodizime (2g/daily in 2 divided doses), another group of 10 patients with ceftriaxone (2g/daily as a single dose); in both groups the antibiotic treatment was given for 7-10 days . A significant potentiation of chemiluminescence has been shown in both groups of patients treated in vivo; whereas the preincubation in vitro of the polymorphonuclear suspension, both in healthy patients and in elderly bronchopathic ones, with antibiotics, has not changed the activity of oxygen dependent killing . Finally, we believe that bactericidal properties of the antibiotic still remain now the most important criteria of choice in order to assure effective control of infections.

Clin Diagn Lab Immunol, 2003 Jul, 10(4), 702 - 9
Changes in avidity and level of immunoglobulin G antibodies to Mycobacterium tuberculosis in sera of patients undergoing treatment for pulmonary tuberculosis; Arias-Bouda LM et al.; Much is known about specific antibodies and their titers in patients with tuberculosis . However, little is known about the avidity of these antibodies or whether changes in avidity occur during the progression of the disease or during treatment . The aims of this study were to determine the avidity of antibodies to Mycobacterium tuberculosis in patients with pulmonary tuberculosis, to explore the value of avidity determination for the diagnosis of tuberculosis, and to study changes in levels of antibodies and their avidity during treatment . Antibody avidity was measured by an enzyme-linked immunosorbent assay with thiocyanate elution . Avidity indices and serum levels of immunoglobulin G to M . tuberculosis were determined for 22 patients with pulmonary tuberculosis before and during treatment and for 24 patients with other pulmonary diseases . Antibody levels and avidity were both significantly higher in untreated tuberculosis patients than in the controls . Avidity determination had more diagnostic potential than determination of the antibody levels . Tuberculosis patients with a long duration of symptoms had higher antibody avidity than those with a recent onset of symptoms, indicating affinity maturation of specific antibodies during active disease . In the early phase of treatment, a decrease in antibody avidity was observed for 73% of all tuberculosis patients, accompanied by an initial increase in antibody levels in 36% of these patients . These phenomena could be explained by an intense stimulation of the humoral response by antigens released from killed bacteria, reflecting early bactericidal activity of antituberculous drugs leading to the production of low-affinity antibodies against these released antigens.

Biochim Biophys Acta, 2003 Jul 14, 1638(2), 164 - 72
Preparation of recombinant rat eosinophil-associated ribonuclease-1 and -2 and analysis of their biological activities; Ishihara K et al.; Rat eosinophils contain eosinophil-associated ribonucleases (Ears) in their granules . Ears are thought to be synthesized as pre-forms and stored in the granules as mature forms . However, the N-terminal amino acid of mature Ear-1 and Ear-2 is still controversial . Therefore, we prepared two recombinant mature forms of Ear-1 and Ear-2 in which the N-terminal amino acids are Ser24 (S) {Ear-1 (S) and Ear-2 (S)} and Gln26 (Q) {Ear-1 (Q) and Ear-2 (Q)}, and analyzed their biological activities by comparing them with those of pre-form Ear-1 and pre-form Ear-2 . The four mature Ears showed RNase A activity as well as bovine pancreatic RNase A activity, but pre-Ear-1 and pre-Ear-2 showed no RNase A activity . Mature Ear-1 (Q) and mature Ear-2 (Q) showed more potent RNase A activity than mature Ear-1 (S) and mature Ear-2 (S), respectively . The RNase A activities of mature Ear-1 (Q) and mature Ear-2 (Q) were reduced by treatment at 96 degrees C for 20 min or with RNase inhibitor . The growth of Escherichia coli was inhibited by both pre-Ears and mature Ears in a concentration-dependent manner, and was almost completely suppressed at 1.0 microM . The bactericidal activities of mature Ear-1 (Q) and mature Ear-2 (Q) were not inhibited by RNase inhibitor, but was increased by treatment at 96 degrees C for 20 min.

Kidney Int, 2003 Aug, 64(2), 728 - 36
High-dose parenteral iron sucrose depresses neutrophil intracellular killing capacity; Deicher R et al.; BACKGROUND: Iron is essential for the formation of hemoglobin . During long-term treatment with human recombinant erythropoietin (rhEPO), the majority of end-stage renal disease (ESRD) patients will not respond adequately to rhEPO unless substituted with intravenous iron . However, concern exists about possible detrimental effects of parenteral iron on cellular host defense and iron-mediated increments of oxidative stress . METHODS: We analyzed phagocytic functions of polymorphonuclear leukocytes (PMN) isolated from 20 ESRD patients on peritoneal dialysis in response to 300 mg of iron sucrose or placebo administered intravenously over two hours in a randomized, double-blind manner . We evaluated Fc gamma R-dependent phagocytosis and killing (primary outcome variable) of opsonized Escherichia coli, Fc gamma R-dependent oxidative burst capacity, and complement receptor 3 (CR3, Mac1, CD11b/CD18)/tumor necrosis factor alpha (TNFalpha)-mediated release of bactericidal lactoferrin before, during, one hour, and two days after administration . RESULTS: The absolute count and the percentage of E . coli killed by PMN of iron sucrose-treated peritoneal dialysis patients decreased significantly over time in comparison to placebo-treated patients (F = 3.48, df = 4, P = 0.008; F = 3.99, df = 4, P = 0.006, respectively) . All secondary outcome variables were not different between both groups over time . CONCLUSIONS: Killing capacity of PMN isolated from ESRD patients decreases in response to high-dose parenteral iron sucrose, possibly in part explaining reported higher hospitalization rates and lower survival rates of dialysis patients receiving frequent and high-dose parenteral iron.

Appl Environ Microbiol, 2003 Jul, 69(7), 4278 - 81
Mode of bactericidal action of silver zeolite and its comparison with that of silver nitrate; Matsumura Y et al.; The properties of the bactericidal action of silver zeolite as affected by inorganic salts and ion chelators were similar to those of silver nitrate . The results suggest that the contact of the bacterial cell with silver zeolite, the consequent transfer of silver ion to the cell, and the generation of reactive oxygen species in the cell are involved in the bactericidal activity of silver zeolite.

Zhonghua Shao Shang Za Zhi, 2003 Apr, 19(2), 86 - 8
{The expression and analysis of its activity of anti-bacterial peptide gloverin in COS-7 cells}; Zhou H et al.; OBJECTIVE: To investigate the expression and analysis of its activity of anti-bacterial peptide gloverin in COS-7 cells . METHODS: The appearance frequency of all genetic codes in the cDNA sequence from the same species of protein Attacin A was analyzed, and its cDNA sequence was synthesized by PCR overlapping extension method in conjunction with the designation of the known protein sequence of gloverin . The genes were inserted into pCDSI, an eukaryotic vector, after being identified correctly . As a result, the vector pBZHG was constructed . Thereafter, the liposome FuGENE( trade mark ) 6 was employed as the vector, and the COS-7 cells were transfected with liposome pBZHG and blank vector pCDSI . The normal cells were taken as the control . The supernatant was collected for the detection of its bactericidal activity after 72 PBHs . RESULTS: The gloverin cDNA sequence designed artificially was expressed in COS-7 cells . The supernatant of the cells transfected by pBZHG exhibited bactericidal activity to E . coli J5 when compared with that from normal cells and in cells transfected with blank vectors . CONCLUSION: The designed cDNA sequence of gloverin was proved to be genuine, and it provided the basis for future study of its antibiotic and anti-endotoxin activities.

Vector Borne Zoonotic Dis, 2002 Winter, 2(4), 249 - 54
Diversity of Ixodes-borne Borrelia species--clinical, pathogenetic, and diagnostic implications and impact on vaccine development; van Dam AP; Among Borrelia spirochetes carried by hard ticks belonging to the various Ixodes species, at least 10 species can be distinguished . Of these, Borrelia burgdorferi sensu stricto is involved in human Lyme borreliosis in North America and Europe, and Borrelia garinii and Borrelia afzelii in human disease in Europe and Asia . The pathogenetic significance of the other species is uncertain . Although some of the Borrelia species are restricted to certain tick species, Ixodes ricinus, the vector of Lyme borreliosis in Europe, can be infested by at least five different species, including all three pathogenic species . There is evidence that different Borrelia species are preferentially found in different hosts: In Europe, B . afzelii is frequently found in small mammals, whereas B . garinii and Borrelia valaisiana are often found in birds . This could very well be related to differential sensitivity of these species to complement-mediated bactericidal activity of different hosts . Borrelial complement regulator acquiring proteins, among them OspE or Erp proteins, bind to host factor H and related proteins, and this binding protects against activation of complement by the spirochetal surface . The binding is different for proteins originating from different species and is also depending on the host origin of factor H . In Europe, B . garinii is mainly found in neuroborreliosis, whereas in skin disease B . afzelii is more frequently found . The reason is unclear . The majority of human sera cross-react between proteins of different Borrelia species, but some sera react only with proteins from one of the species . This holds especially for reactivity with OspC . A vaccine against B . burgdorferi sensu stricto has been licensed, but was recently redrawn from the market because of commercial reasons . A vaccine protecting against all three pathogenic species is not yet available.

Int J Tuberc Lung Dis, 2003 Jun, 7(6), 575 - 9
Impact of iron loading on the activity of isoniazid or ethambutol in the treatment of murine tuberculosis; Lounis N et al.; OBJECTIVE: To assess the impact of iron loading on the activity of isoniazid and ethambutol in the treatment of murine tuberculosis . DESIGN: Iron-loaded and iron-normal female Balb/C mice infected with 1.5 x 10(7) colony forming units of Mycobacterium tuberculosis were treated with either isoniazid or ethambutol for 28 days . RESULTS: For both treatments, the outcome was impaired by the iron loading: bactericidal activity of isoniazid was partially but significantly reduced and ethambutol bactericidal activity was totally inhibited . CONCLUSION: The treatment of tuberculosis in patients with iron loading should be longer than for normal patients or should contain an additional drug.

J Infect Dis, 2003 Jun 15, 187(12), 1977 - 80 Epub 2003 Jun 04.
Mefloquine, moxifloxacin, and ethambutol are a triple-drug alternative to macrolide-containing regimens for treatment of Mycobacterium avium disease; Bermudez LE et al.; Macrolides are the core of effective drug regimens for the treatment of Mycobacterium avium complex (MAC) disease . Mefloquine (MFQ), moxifloxacin (MXF), and ethambutol (EMB), in combination, were evaluated against both clarithromycin-resistant (CLR-R) and CLR-susceptible (CLR-S) MAC; MFQ (40 mg/kg), MXF (100 mg/kg), or EMB (100 mg/kg/day) was given to mice for 4 weeks . MFQ was bactericidal, whereas MXF and EMB were bacteriostatic against both MAC 101 CLR-S and CLR-R . The combination of MFQ and EMB reduced (P<.05, for comparison with controls), and the combination of MFQ and MXF significantly reduced, the load of CLR-R in both the liver and the spleen . Treatment with all 3 drugs was associated with approximately 1-log reduction of CLR-R after 1 week, 2.1-log reduction of CLR-R after 4 weeks, and 2.17-log reduction in MAC/mL blood . Treatment of MAC 101 CLR-S strain had comparable results.

Probl Tuberk, 2003, (2), 27 - 31
{Modalities of complex therapy in different categories of patients with pulmonary tuberculosis}; Krasnov VA et al.; The paper proposes the standards of combined treatment involving intermittent intravenous bactericidal chemotherapy for different groups of patients . It describes treatment regimens at the inpatient stage, defines the scope and time of control examination and correction of a treatment protocol by taking into account the pattern, phase, and dissemination of a process and the duration of the disease.

J Rheumatol, 2003 Jun, 30(6), 1248 - 52
Bactericidal/permeability-increasing protein and cathepsin G are the major antigenic targets of antineutrophil cytoplasmic autoantibodies in systemic sclerosis; Khanna D et al.; OBJECTIVE: To study the prevalence and antigenic specificity of antineutrophil cytoplasmic autoantibodies (ANCA) in patients with systemic sclerosis (SSc) . METHODS: Sera from 68 patients with SSc were screened for ANCA by indirect immunofluorescence (IIF) assay and for antibodies to myeloperoxidase (MPO) by ELISA . All sera positive for ANCA on IIF were analyzed for reactivity against antigenic targets other than MPO {bactericidal/permeability-increasing protein (BPI), cathepsin G, lysozyme, elastase, PR3, and lactoferrin} . Twenty-three sera negative for ANCA were also tested for antibodies to BPI and cathepsin G using ELISA . RESULTS: The study included 33 patients with diffuse and 35 with limited SSc . ANCA was detected in 24 of the 68 sera (35.3%) . In these 24 sera the antigenic targets were BPI in 14, cathepsin G in 13, and MPO in 8 . Sera of 11 patients had reactivity against both BPI and cathepsin G . In sera, that were negative for ANCA, antibodies to BPI (4/23), cathepsin G (3/23), and MPO (1/44) were found in a small proportion of patients . Patients with antibodies to BPI had lower skin score, whereas no patient with antibodies to MPO had renal disease . CONCLUSION: BPI and cathepsin G are the major antigenic targets of ANCA seen in patients with SSc . Patients with antibodies to BPI had lower skin scores.

Eur J Histochem, 2003, 47(2), 173 - 6
Demonstration of glucose-6-phosphate dehydrogenase in rat Kupffer cells by a newly-developed ultrastructural enzyme-cytochemistry; Matsubara S et al.; Although various tissue macrophages possess high glucose-6-phosphate dehydrogenase (G6PD) activity, which is reported to be closely associated with their phagocytotic/bactericidal function, the fine subcellular localization of this enzyme in liver resident macrophages (Kupffer cells) has not been determined . We have investigated the subcellular localization of G6PD in Kupffer cells in rat liver, using a newly developed enzyme-cytochemical (copper-ferrocyanide) method . Electron-dense precipitates indicating G6PD activity were clearly visible in the cytoplasm and on the cytosolic side of the endoplasmic reticulum of Kupffer cells . Cytochemical controls ensured specific detection of the enzymatic activity . Rat Kupffer cells abundantly possessed enzyme-cytochemically detectable G6PD activity . Kupffer cell G6PD may play a role in liver defense by delivering NADPH to NADPH-dependent enzymes . G6PD enzyme-cytochemistry may be a useful tool for the study of Kupffer cell functions.

Zhonghua Jie He He Hu Xi Za Zhi, 2003 Jan, 26(1), 30 - 3
{Rearrangement and altered expression of actin in macrophages induced by Mycobacterium tuberculosis}; Xu YZ et al.; OBJECTIVE: To investigate the effect of M . tuberculosis infection on actin in host-cells . METHODS: The form and distribution of fibrous actin and changes of actin expression were observed by confocal microscopy and Western blot analysis in macrophages infected with either M . tuberculosis H(37)R(a) or H(37)R(v) at 6 h, 12 h and 24 h after infection . Non-infected macrophages or macrophages treated with dead M . tuberculosis H(37)R(v) served as controls . RESULTS: F-actin aggregated and the actin expression was suppressed in macrophages infected with H(37)R(a) or H(37)R(v), and the effect appeared earlier in cells infected by the virulent H(37)R(v) strain than in cells infected by the avirulent H(37)R(a) strain . In macrophages treated with the dead H(37)R(v), actin was not affected . CONCLUSION: The results suggest that in the process of infection, M . tuberculosis evades the bactericidal mechanisms possibly by secretion of certain proteins or factors which affect the host-cell actin.

Zhongguo Zhong Yao Za Zhi, 2002 Jun, 27(6), 412 - 8
{Comparison among families of Mutong}; Ma HM et al.; OBJECTIVE: To distinguish families of Mutong correctly and direct effective and safe clinical administration . METHOD: Comparison among families of Mutong on Herbs, Taxology, Clinic, Pharmacology and Toxicology . RESULT: 1 . There are mainly three families of Mutong: Lardizabalaceae, Ranunculaceae, Aristolochiaceae, which were all included in China Pharmacopeia in 1963 . However only Mutong of Ranunculaceae and Aristolochiaceae family have been included in China Pharmacopeia since 1977, but Mutong of Lardizabalaceae family has not been included in China Pharmacopeia ever since . 2 . It was Mutong of Lardizabalaceae family that was used mainly through the ages without toxic records, and Mutong of Aristolochiaceae e.g . Caulis Aristolochia manshuriensis (CAM) was not put down in writing of past ages but is mainly used today with toxicity repeatedly . 3 . CAM contain aristolochic acid and aristololactam with high toxicity, which plays an uncertain role in diuresis with poor bactericidal power . Mutong of Lardizabalaceae family e.g . Akebia trifoliata (Thunb.) Koidz . var . australis (Diels) Rehd (ATKV) don't contain aristolochic acid and aristololactam, which has low toxicity and plays a certain role in diuresis with high bactericidal power . CONCLUSION: It may be quite safe to use ATKV instead of CAM in clinics . So we suggest that ATKV should be reused as first Mutong in China Pharmacopeia revised edition in order to ensure a correct understanding of the facts and reveal Mutong in its true colors, and CAM should be used as second Mutong strictly according to the rules in China Pharmacopeia revised edition.

Anaesthesist, 2003 May, 52(5), 442 - 52
{Opioids and immunosuppression . Clinical relevance?}; Welters I; First observations that opioids may have disadvantageous effects on the immune response have been made more than 100 years ago.Today the immunosuppressive effect of morphine is well established.Drug-induced immunomodulation is of growing importance in modern anesthetic concepts . The reduced stress response observed after morphine application contributes to this effect as well as direct impairment of immune effector cells such as bactericidal activity, intracellular killing,proliferative response or cytokine synthesis . Opioid-induced immunomodulation is mediated by opioid receptors found on immunocytes and in the central nervous system.A negative feedback mechanism via the hypothalamo-pituitary-adrenal axis may potentiate the direct inhibitory effect of morphine on the immune response.A final statement regarding the clinical relevance of opioid-induced immunosuppression cannot be made at this point, since the existing clinical data are preliminary and inconclusive.Therefore, further clinical studies are mandatory to elucidate the influence of opioid treatment on immune regulation in different clinical settings in anesthesia, critical care, pain therapy and emergency medicine.Further investigations may help to not only provide sufficient analgesia by application of opioids, but also to assess advantages and disadvantages on immune function.

Hepatogastroenterology, 2003 Mar-Apr, 50(50), 577 - 81
Inhibition of peptic ulcer relapse by ranitidine and ecabet independently of eradication of Helicobacter pylori: a prospective, controlled study versus ranitidine; Koizumi W et al.; BACKGROUND/AIMS: The recent increase in resistant strains of Helicobacter pylori has become a serious problem . Ecabet is a novel anti-ulcer agent that acts directly on the gastric mucosa, has bactericidal activity, and inhibits adhesion of Helicobacter pylori to the gastric mucosa . These actions result from inhibition of urease and ATPase in Helicobacter pylori, a mechanism distinct from that of antibiotics . METHODOLOGY: Sixty-three patients positive for Helicobacter pylori who had been cured of gastric ulcers and duodenal ulcers were randomly assigned to receive maintenance therapy with ranitidine alone or a combination of ranitidine and ecabet . Ulcer relapse was studied in these patients . RESULTS: The cumulative relapse rates in the ranitidine group and the ecabet plus ranitidine group were respectively, 29.6% and 4.4% after 1 year of treatment and 66.1% and 13.0%, after 2 years . These differences were significant (p = 0.006) . Multivariate analysis of factors potentially related to relapse showed that outcome was significantly related only to treatment (p = 0.020) and not to other characteristics, such as age, diagnosis, or sex . CONCLUSIONS: We conclude that maintenance therapy with a combination of ranitidine and ecabet prevents ulcer relapse in Helicobacter pylori-positive patients . Controlled studies comparing ulcer relapse rates between eradication treatment and maintenance therapy with ranitidine and ecabet are awaited.

Phytother Res, 2003 May, 17(5), 575 - 7
Two isoflavones and bioactivity spectrum of the crude extracts of Iris germanica rhizomes; Orhan I et al.; In vitro biological activities including bactericidal, fungicidal and insecticidal activities as well as phytotoxicity and brine shrimp toxicity of the petroleum ether, chloroform and ethyl acetate extracts of Iris germanica L . were determined . The bactericidal activity of the extracts was assayed by the agar well diffusion test . In the fungicidal test, the agar tube dilution method was used . The insecticidal activity was determined by the exposure method . The toxicity of the extracts was evaluated by the phytotoxicity test as well as the brine shrimp toxicity test . The chloroform and ethyl acetate extracts of I . germanica rhizomes exhibited bactericidal activity, while the petroleum ether extract did not exhibit any bactericidal, fungicidal and insecticidal activities . It was also inactive in the brine shrimp toxicity test, whereas it showed significant phytotoxicity against the plant Lemna aequinoctialis Welv . Two known isoflavones were isolated from the chloroform extract of the plant .

Eur J Appl Physiol, 2003 May, 89(3-4), 257 - 62 Epub 2003 Feb 28.
Effect of exercise to exhaustion on myeloperoxidase and lysozyme release from blood neutrophils; Morozov VI et al.; Exercise sessions (swimming in rats and treadmill running in humans) resulted in stimulation of neutrophil degranulation in the experiments with animals and in the human study . Myeloperoxidase (MPO) (+67%) and lysozyme (+51%) quantities in the plasma of rats increased significantly immediately after exercise . The blood plasma lysozyme concentration was increased by 41% at the 6th min of treadmill exercise in athletes . The blood concentrations of neutrophil proteins normalized both in humans and animals at rest . The neutrophil protein concentrations in blood increased in parallel with the decrease of their level in leukocytes . The neutrophil capacity for an oxidative burst was not changed by the exercise, but decreased for 3-6 h in the post-exercise period . Such dynamics of the oxidative burst activity suggest a lack of association between this parameter and the degranulation process . The neutrophil proteins that appear in blood during degranulation can be involved in enhancing the bactericidal potency of blood, the activation of granulopoiesis, neutrophil efflux from bone marrow, and the conditioning of blood endothelium for leukocyte extravasation.

J Mol Microbiol Biotechnol, 2003, 5(2), 105 - 22
Comparison of the changes in global gene expression of Escherichia coli induced by four bactericidal agents; Shaw KJ et al.; DNA microarrays provide a global view of the physiological state of the cell by parallel analysis of the expression levels of all the genes in an organism . The effects of four bactericidal agents on the expression pattern of Escherichia coli MG1655 were assessed . Compounds were chosen on the basis of their different mechanisms of action and included inhibitors of DNA replication and recombination, translation, transcription and cell wall biosynthesis . The addition of rifampin resulted in increased expression of the target, rpoB, as well as several genes involved in nucleotide salvage and purine biosynthesis . The addition of ampicillin resulted in overall changes in gene expression that showed some similarity to changes induced by rifampin . The addition of the antibiotics kanamycin or norfloxacin resulted in the induction of unique gene expression signatures: a heat shock response to kanamycin and an SOS response to norfloxacin . Several genes of unknown function showed expression profiles similar to the genes associated with the SOS or the heat shock response . Thus, these profiles define families of genes with similar expression phenotypes that can be tested for related function .

J Immunol, 2003 May 15, 170(10), 5276 - 80
Calcium-independent phospholipase A2 is required for lysozyme secretion in U937 promonocytes; Balboa MA et al.; As a part of their surveillance functions in the immune system, monocytes/macrophages secrete large amounts of the bactericidal enzyme lysozyme to the extracellular medium . We report here that lysozyme secretion in activated U937 promonocytes depends on a functional calcium-independent phospholipase A(2) (iPLA(2)) . Inhibition of the enzyme by bromoenol lactone or by treatment with a specific antisense oligonucleotide results in a diminished capacity of the cells to secrete lysozyme to the extracellular medium . Calcium-independent PLA(2) is largely responsible for the maintenance of the steady state of lysophosphatidylcholine (lysoPC) levels within the cells, as manifested by the marked decrease in the levels of this metabolite in cells deficient in iPLA(2) activity . Reconstitution experiments reveal that lysoPC efficiently restores lysozyme secretion in iPLA(2)-deficient cells, whereas other lysophospholipids, including lysophosphatidic acid, lysophosphatidylserine, and lysophosphatidylethanolamine, are without effect . Arachidonic acid mobilization in activated U937 cells is under control of cytosolic phospholipase A(2) (cPLA(2)) . Selective inhibition of cPLA(2) results in a complete abrogation of the arachidonate mobilization response, but has no effect on lysozyme secretion . These results identify iPLA(2)-mediated lysoPC production as a necessary component of the molecular machinery leading to lysozyme secretion in U937 cells and rule out a role for cPLA(2) in the response . Collectively, the results demonstrate distinct roles in inflammatory cell signaling for these two intracellular phospholipases.

Biosci Biotechnol Biochem, 2003 Feb, 67(2), 410 - 4
Antioxidative compounds from Crotalaria sessiliflora; Mun'im A et al.; Seven antioxidative compounds were isolated from the EtOAc extract of the aerial part of C . sessiliflora (Japanese name, tanukimame) by activity-guided fractionation with 2,2-diphenyl-1-picrylhydrazyl (DPPH) . Among the isolated compounds, hydroxyeucomic acid showed the strongest free radical-scavenging activity, which was almost identical to that of epigallocatechin gallate, against DPPH . Orientin and isoorientin showed strong anti-peroxidative activities toward linoleic acid and protective effects against the bactericidal action of the tert-butyl peroxyl radical . Their activities were nearly equal to that of epigallocatechin gallate.

Antibiot Khimioter, 2002, 47(12), 15 - 9
{Evaluation of levofloxacin antitubercular activity in vitro and in lung tissue culture}; Sokolova GB et al.; Levofloxacin in vitro demonstrated bactericidal effect against susceptible and multi-drug resistant Mycobacterium tuberculosis: range of MICs was 0.25-0.5 mcg/mL, MBC-0.5-1.0 mcg/mL . Postantibiotic effect after 24-hour exposition to bactericidal concentrations was 35-39 days . Levofloxacin possesed low toxicity when tested in mice lungs tissue culture--maximum safety concentration was 50 mcg/mL . Bactericidal effect of levofloxacin started three days after exposition and was maximal by 7 days of incubation: by this time mycobacterial microcolonies destruction started with detritus formation . It is emphasized that lung cells kept their viability completely . Combination of levofloxacin with isoniazide or pirazinamide resulted in strong synergistic effect obvious after 5 days of incubation, mycobacterial colonies destruction was registered by 7th day . Combination of levofloxacin with rifampicin resulted in antagonistic effect obvious by 7th day of the contact: the resulting effect was statistically significant and was manifested as microcolonies number and size enlargement when compared to data for single levofloxacin.

J Clin Endocrinol Metab, 2003 May, 88(5), 2141 - 6
Frequency of antineutrophil cytoplasmic antibody in Graves' disease patients treated with methimazole; Guma M et al.; Retrospective studies have shown antineutrophil cytoplasmic antibody (ANCA) positivity in patients treated for Graves' hyperthyroidism; ANCA has been attributed to either antithyroid drugs or to the disease itself . The aim of this study was to determine ANCA in Graves' disease patients at diagnosis and after treatment with methimazole and to evaluate the relationship between ANCA and hyperthyroidism evolution . Thirty patients recently diagnosed with Graves' hyperthyroidism were prospectively studied . ANCA were determined by indirect immunofluorescence . ANCA autoantibodies against specific antigens (proteinase 3, myeloperoxidase, bactericidal/permeability-increasing protein (BPI), cathepsin, lysozyme, elastase, and lactoferrin) were detected by ELISA . The median observation period was 22 months . Kaplan-Meier analysis was performed to identify ANCA as an outcome variable . Twenty patients (67%) were ANCA positive before the onset of treatment, and four (19%) remained positive after 1 yr of antithyroid drug treatment . No differences were observed in any clinical or analytical features between patients with or without positive ANCA . Before treatment, BPI-positive patients required radioiodine treatment or presented relapse more rapidly than BPI-negative patients (log-rank test P < 0.0002) . Patients with Graves' hyperthyroidism show positive ANCA before medical treatment, which points to a relationship with the autoimmune disease itself . Our results suggest that BPI-positive patients tend to relapse with antithyroid medication.

Am J Chin Med, 2003, 31(1), 141 - 8
Qi-training enhances respiratory burst function and adhesive capacity of neutrophils in young adults: a preliminary study; Lee MS et al.; The main objective of this study is to examine the effect of Qi-training on the immune system, especially neutrophil bactericidal function . Nine healthy male subjects were studied for the effects of one bout of ChunDoSunBup (CDSB) Qi-training on superoxide (O2- production and adhesion capacity of neutrophils at times immediately after (Post I) and 2 hours after the Qi-training (Post II) . The Qi-training enhanced O2- production, reaction velocity and neutrophil adhesion capacity and there were significant differences at Post I compared to before Qi-training (Pre) . In addition, the number of white blood cells (WBC), monocytes and lymphocytes were changed significantly through Qi-training.Therefore, it seems that CDSB Qi-training may increase the resistance of trained individuals against common infection and inflammation.

Nat Prod Res, 2003 Apr, 17(2), 115 - 25
Synthesis and biological screening of 7-hydroxy-4-methyl-2H-chromen-2-one, 7-hydroxy-4,5-dimethyl-2H-chromen-2-one and their some derivatives; Khan KM et al.; 7-Hydroxy-4-methyl-2H-chromen-2-one (2), 7-hydroxy-4,5-dimethyl-2H-chromen-2-one (15) and their some derivatives were synthesized for exploring selected biological screening . The compounds 9 and 13 had shown high degree of cytotoxic activity . Three compound 9, 10 and 13 showed high degree of bactericidal activity amongst the present series.

Biochemistry, 2003 Apr 22, 42(15), 4444 - 51
Structural and functional determinants of human plasma phospholipid transfer protein activity as revealed by site-directed mutagenesis of charged amino acids; Ponsin G et al.; Human plasma phospholipid transfer protein (PLTP) exchanges phospholipids between lipoproteins and remodels high-density lipoproteins (HDLs) . We determined phospholipid transfer activity and HDL binding ability in wild-type PLTP and in 16 PLTP variants created by replacing 12 charged amino acids by site-directed mutagenesis . The data were analyzed in relation to the structure of a member of the same gene family, bactericidal/permeability-increasing protein, which is a boomerang-shaped molecule containing two symmetrical, hydrophobic pockets that bind phospholipid molecules . When expressed in COS-7 cells, wild-type and all mutant PLTPs accumulated intracellularly to nearly the same extent . Relative to wild-type PLTP, substitution(s) for amino acids with a lateral position totally exposed to the solvent produced reductions in transfer activity proportional to the reductions in the level of HDL binding . Variants containing substitutions for charged amino acids on the concave surface of PLTP did not affect binding to HDL or specific transfer activity . A mutation in the C-terminal pocket (E270R) led to a decrease in both the specific transfer activity and the level of binding to HDLs, whereas mutations in the N-terminal pocket (R25E and D231R) resulted in a large decrease in specific transfer activity without affecting HDL binding . The data support a model of transfer in which N- and C-terminal pockets have different roles in HDL binding and transfer activity . The N-terminal pocket may be critical to PLTP transfer activity but may have no involvement in binding to lipoproteins, whereas amino acid substitutions in the C-terminal pocket might reduce PLTP activity by decreasing PLTP's affinity for HDLs.

J Clin Microbiol, 2003 Apr, 41(4), 1791 - 3
In vitro susceptibility testing of four antibiotics against Borrelia burgdorferi: a comparison of results for the three genospecies Borrelia afzelii, Borrelia garinii, and Borrelia burgdorferi sensu stricto; Sicklinger M et al.; MICs and minimal bactericidal concentrations (MBCs) were evaluated for the four antibiotics azithromycin, amoxicillin, ceftriaxone, and doxycycline against the three main genospecies of Borrelia burgdorferi sensu lato . In MBC testing, statistically significant differences between the genospecies could be found in 7 out of 12 comparative evaluations (P < 0.05).

Russ J Immunol, 2002 Apr, 7(1), 58 - 62
Local immunological markers of different rate of growth of uterine myoma; Malyshkina AI et al.; The aim of our work was to establish the peculiarities of phenotype profile and functional activity of peritoneal immunocompetent cells of women with different rate of myoma growth . Forty three women with uterine myoma (main group) and 16 healthy women (control group) were recruited for study . According to the growth rate and tumor's size the main group was divided into two clinical subgroups: (1) women with stable small size of uterine myoma (23 patients), (2) women with large size rapidly growing myoma (20 patients) . The expression of CD markers on the surface of peritoneal fluid (PF) lymphocytes and bactericidal and adhesive activity of macrophages were studied . It was found that in women with stable small myomas both lymphocytes and macrophages in PF were activated . In women with rapidly growing large myomas cellular reactions seemed to be inhibited and the impairment of maturation and differentiation of lymphocytes was observed . It can be suggested that local immune response is directly connected with the rate of myoma growth.

Russ J Immunol, 1999 Dec, 4(4), 333 - 336
Immunology of Early and Late Ontogenesis; Shabalin VN; Immunological interrelations are one of the most important factors, forming all the periods of ontogenetic body development . At early ontogenesis in the organism of mammals, including humans, the immunological tolerance is induced to own (self) antigens that prevents the development of autoimmune reactions in adult life . The system of immune recognition reaches its fine discriminative capacity in mammals and humans on the background of recognition of self HLA class I (CTL of CD8(+) phenotype) or class II (T helpers of CD4(+) phenotype) . The phenomenon of major histocompatibility complex (MHC) restriction reactivity of cytotoxic T lymphocytes (CTL) makes difficult the recognition in changes or mutations in HLA system . Natural killers (NK) control constantly the structure of self MHC class I molecules, and in contrast to CTL may recognize small antigenic changes of MHC molecules, induced by viruses, mutations and transformation . Immune system aging is displayed by both quantitative, and functional changes in lymphocyte subpopulations with prevalence of T helpers type 2 over type 1 cells, responsible for the development of DTH . According to immunological theory of aging, the role of immune system cells should be taken into consideration in relation to the synthesis of activated oxygen forms with bactericidal properties by monocytes and neutrophils . Overproduction of activated oxygen radicals may result in destruction of proteins, lipids, DNA, and in the body aging . Activated oxygen radicals are one of events of lymphocyte apoptosis in aerobic conditions.

Infect Control Hosp Epidemiol, 2003 Mar, 24(3), 191 - 4
Can Whipple's disease be transmitted by gastroscopes?
La Scola B, Rolain JM, Maurin M, Raoult D.
OBJECTIVE: To determine whether disinfection protocols currently used for gastroscopes are effective against cultures of Tropheryma whipplei . DESIGN: The bactericidal activity of 2% glutaraldehyde and two peracetic acids on the Twist-Marseille strain of T . whipplei grown in cell monolayers was determined . PATIENTS: Two patients who were diagnosed as having Whipple's disease 3 years after they had had intestinal biopsies . RESULTS: The disinfectants reduced bacteria by approximately 2 log to 3 log10 after 5 to 60 minutes of contact . CONCLUSION: The bactericidal activity of a disinfectant is usually considered significant if it causes a 5 log10 or greater reduction in viable bacterial titers . Disinfecting gastroscopes with 2% glutaraldehyde or peracetic acids for 20 minutes may be insufficient to prevent transmission of T . whipplei on the instruments or stop false-positive results on polymerase chain reaction.

J Chemother, 2003 Feb, 15(1), 47 - 52
Activities of sitafloxacin (DU-6859a), either singly or in combination with rifampin, against Mycobacterium ulcerans infection in mice; Dhople AM et al.; Efficacy of a new fluoroquinolone, sitafloxacin (DU-6859a), against Mycobacterium ulcerans was evaluated in vivo using the mouse footpad system . The growth of M . ulcerans in mouse footpads was completely inhibited when mice were fed with sitafloxacin at a dose of 25 mg/kg body weight per day; on the other hand similar effects were observed with ofloxacin at a dose of 100 mg/kg body weight per day . In the presence of rifampin, the above dose of sitafloxacin could be reduced by 75% to achieve total inhibition, while, under similar circumstances, the dose of ofloxacin could be reduced by only 50% . Either used singly or in combination with rifampin, the effects of sitafloxacin were bactericidal . The results suggest that sitafloxacin should be evaluated as a chemotherapeutic agent against M . ulcerans infection.

J Int Acad Periodontol, 2003 Jan, 5(1), 23 - 8
Effects of Er:YAG laser on periodontal therapy; Ishikawa I et al.; The use of the laser in dentistry has been widening due to its increasing use in surgery and medicine . In the field of periodontology, the most commonly used lasers have been CO2 and Nd:YAG lasers . According to the last reviews reporting the use of lasers in periodontal therapy, these lasers were considered unsuitable for dental treatment, due to various shortcomings, such as the carbonization and severe thermal damages caused on the target and surrounding tissues, and therefore their use has been confined to soft tissue procedures . Technological advances and improvements have increased the choices of the available laser systems . Among them, Er:YAG lasers seem to be of promising use . The Er:YAG laser is a solid-state crystal laser operating in the infrared wavelength (2,940 nm) . Due to its high absorbability in water and hydroxyapatite, several studies have shown the effectiveness of this laser for both hard and soft tissue ablation, and its bactericidal effects with less or even no pain under clinical applications . The variety of the potential applications for this laser has been studied and the interest about its use in dental practice has increased among practitioners . This review includes studies regarding the use of Er:YAG laser on hard tissue procedures, such as calculus removal and osseous surgery as well as soft tissue management, like gingivectomy, gingival curettage and melanin removal.

Infect Immun, 2003 Apr, 71(4), 1843 - 8
Inactivation of the Moraxella catarrhalis 7169 ferric uptake regulator increases susceptibility to the bactericidal activity of normal human sera; Furano K et al.; Moraxella catarrhalis is a strict human pathogen and a significant cause of respiratory disease and otitis media . In direct response to these infections, research efforts have focused primarily on the identification of potential vaccine targets . The general biology of M . catarrhalis, however, including the mechanisms utilized to survive in the human host, remains poorly understood . Previous work has demonstrated that M . catarrhalis expresses iron-repressible proteins, suggesting the presence of iron acquisition systems under the control of a ferric uptake regulator (Fur) . In this study M . catarrhalis fur has been cloned and sequenced from strain 7169 . A deletion-insertion mutation of 7169 fur resulted in upregulation of iron-repressible outer membrane proteins in the absence and presence of iron . This mutant strain, 7169fur1, was significantly more sensitive to the bactericidal activity of normal human serum than the resistant wild-type strain . These data suggest that constitutive expression of iron-regulated proteins may provide multiple targets for human antibodies . In addition, the 7169 fur mutant provides an important tool for further investigation of the iron acquisition mechanisms utilized by M . catarrhalis.

Free Radic Res, 2003 Jan, 37(1), 85 - 90
Dietary nitrate inhibits stress-induced gastric mucosal injury in the rat; Miyoshi M et al.; Dietary nitrate is reduced to nitrite by some oral bacteria and the resulting nitrite is converted to nitric oxide (NO) in acidic gastric juice . The aim of this study is to elucidate the pathophysiological role of dietary nitrate in the stomach . Intragastric administration of nitrate rapidly increased nitrate and NO in plasma and the gastric headspace, respectively . Water-immersion-restraint stress (WIRS) increased myeloperoxidase (MPO) activity in gastric mucosa and induced hemorrhagic erosions by a nitrate-inhibitable mechanism . In animals that had received either cardiac ligation or oral treatment with povidone-iodine, a potent bactericidal agent, administration of nitrate failed to increase gastric levels of NO and to inhibit WIRS-induced mucosal injury . WIRS decreased gastric mucosal blood flow by a mechanism which was inhibited by administration of nitrate . These data suggested that the enterosalivary cycle of nitrate and related metabolites consisted of gastrointestinal absorption and salivary secretion of nitrate, its conversion to nitrite by oral bacteria and then to NO in the stomach might play important roles in the protection of gastric mucosa from hazardous stress.

Int Immunopharmacol, 2003 Mar, 3(3), 375 - 81
Stimulation of murine macrophages by native and heat-denatured lectin from Abrus precatorius; Tripathi S et al.; We have evaluated the immunostimulant activity of native agglutinin (NA) and heat-denatured agglutinin (HDA) obtained from Abrus precatorius seed kernels on murine macrophages . Activated macrophages play an important role in nonspecific immunostimulation in turn by activating the other immune cells in the cascade . Significant alterations are observed in the macrophage functions both by native and heat-denatured Abrus agglutinin . Increased production of nitric oxide and hydrogen peroxide and a high phagocytic and bactericidal activity is potentiated by both NA and HDA . It is also observed that activated macrophages also released interleukin-1 . These results suggest that both NA and HDA act as immunostimulants in vitro.

Eur Biophys J, 2003 Mar, 32(1), 22 - 32 Epub 2002 Nov 22.
Magainin 2 channel formation in planar lipid membranes: the role of lipid polar groups and ergosterol; Gallucci E et al.; Magainin 2, a polycationic peptide, displays bactericidal and tumoricidal activity, presumably interacting with negatively charged phospholipids in the membrane hosts . In this work, we investigate the role played by the lipid head-group in the interactions and self-association of magainin 2 during pore formation in lipid bilayers . Two methods are used: single-channel and macroscopic incorporation into planar lipid membranes . Single-channel incorporation showed that magainin 2 did not interact with zwitterionic membranes, while the addition of negatively charged dioleoylphosphatidylglycerol to the membrane leads to channel formation . On the other hand, magainin 2 did not form channels in membranes made up of dioleoylphosphatidylserine (DOPS), although the addition of ergosterol to DOPS membranes leads to channel formation . This finding could indicate that ergosterol may be a possible target of magainin 2 in fungal membranes . Further support for this hypothesis comes from experiments in which the addition of ergosterol to palmitoyloleoylphosphatidylcholine membranes induced channel formation . Besides the role of negatively charged membranes, this study has shown that magainin 2 also forms channels in membranes lacking heads, such as monoolein and oxidized cholesterol, indicating an interaction of magainin 2 with acyl chains and cholesterol, respectively . This finding provides further evidence that peptide binding and assembly in lipid membranes is a complex process driven by electrostatic and/or hydrophobic interactions, depending on the structure of the peptide and the membrane composition.

J Immunol, 2003 Mar 15, 170(6), 2811 - 5
Cutting edge: Mycobacterium tuberculosis blocks Ca2+ signaling and phagosome maturation in human macrophages via specific inhibition of sphingosine kinase; Malik ZA et al.; One-third of the world's population is infected with Mycobacterium tuberculosis (Mtb), and three million people die of tuberculosis each year . Following its ingestion by macrophages (MPs), Mtb inhibits the maturation of its phagosome, preventing progression to a bactericidal phagolysosome . Phagocytosis of Mtb is uncoupled from the elevation in MP cytosolic Ca(2+) that normally accompanies microbial ingestion, resulting in inhibition of phagosome-lysosome fusion and increased intracellular viability . This study demonstrates that the mechanism responsible for this failure of Ca(2+)-dependent phagosome maturation involves mycobacterial inhibition of MP sphingosine kinase . Thus, inhibition of sphingosine kinase directly contributes to survival of Mtb within human MPs and represents a novel molecular mechanism of pathogenesis.

Eksp Klin Gastroenterol, 2002, (5), 42 - 4, 127
{Clinico-immunologic effect of immunomodulin and bactim in duodenal ulcer under environmental pollution conditions}; Abdullaev RB; There was a study of the effect of immunomodulin and bactrim (biseptol-480) preparations in 53 patients with duodenal ulcer living in the ecologically polluted Southern Aral region . It was revealed that the application of immunomodulator immunomodulin and bactrim, a combined sulfanilamide preparation, in combination with the conventional anti-ulcer treatment promotes stronger bactericidal activity with respect to Helicobacter pylori, development of the immune status of the organism, acceleration of ulcer cicatrisation, reduction of the average period of staying in an in-patient hospital, which is an indication of its economic efficiency.

Eksp Klin Gastroenterol, 2002, (5), 7 - 13, 126
{Problems of functional study of the stomach in contemporary gastrology}; Gorshkov VA; In the assessment of the functional state of the stomach the interest to its secretory functioning has remained constant and in the center of attention of physiologists and clinicians during the past century . That is understandable . After all, the level of secretion of gastric juice enriched with the acid and pepsins defines numerous aspects in the functioning of this organ and its peptic, bactericidal and even evacuation functions . Moreover, under certain conditions active gastric juice can turn from a mediator in the normal peptic process into a solely pathogenetic factor and promote the development of a number of so-called acid-dependent diseases that have conventionally included stomach ulcer, reflux-esophagitis, postgastrectomy ulcers of the anastomosis and other more infrequent pathological states . A distinct positive reaction to the application of anti-acid preparations is common for all these diseases . At the same time, the role of HCI in their development remains comprehensible only in some aspects.

Eur J Immunol, 2003 Mar, 33(3), 708 - 19
Artificial-infection protocols allow immunodetection of novel Borrelia burgdorferi antigens suitable as vaccine candidates against Lyme disease; Wallich R et al.; Vaccination with recombinant outer surface protein A (OspA) from Borrelia burgdorferi provides excellent antibody-mediated protection against challenge with the pathogen in animal models and in humans . However, the bactericidal antibodies are ineffective in the reservoir host, since OspA is expressed by spirochetes only in the vector, but rarely, if at all, in mammals . Using an artificially generated immune serum (anti-10(8) spirochetes) with high protective potential for prophylactic and therapeutic treatment, we have now isolated from an expression library of B . burgdorferi (strain ZS7) three novel genes, zs7.a36, zs7.a66 and zs7.a68 . All three genes are located, together with ospA/B, on the linear plasmid lp54, and are expressed in vitro and in ticks . At least temporarily two of them, ZS7.A36 and ZS7.A66, are also expressed during infection . The respective natural antigens are poorly immunogenic ininfected normal mice but elicited antibodies in Lyme disease patients . We show that recombinant preparations of ZS7.A36, ZS7.A66 and ZS7.A68 induce functional antibodies in rabbits capable of protecting immunodeficient mice against subsequent experimental infection . These findings suggest that all three recombinant antigens represent potential candidates for a "second generation" vaccine to prevent and/or cure Lyme disease.

Probl Tuberk, 2002, (12), 56 - 8
{Action of ultraviolet laser radiation on extracellular and phagocytic Mycobacteria tuberculosis in vitro}; Faizullin DR et al.; Ultraviolet laser radiation (a wavelength of 248 nm) was examined in vitro for its bacteriostatic and bactericidal actions on M . tuberculosis H37Rv . Sterilization was achieved when a dose of 10 m/J/cm2 was applied . This indicates that this type of radiation suppresses both the formation of colonies upon exposure of free Mycobacteria in the suspension and the incorporation of 3H-uracil by the mycobacteria phagocytized with peritoneal macrophages.

Antimicrob Agents Chemother, 2003 Mar, 47(3), 1135 - 6
In vitro activities of rifamycin derivatives ABI-1648 (Rifalazil, KRM-1648), ABI-1657, and ABI-1131 against Chlamydia trachomatis and recent clinical isolates of Chlamydia pneumoniae; Roblin PM et al.; ABI-1648 (rifalazil) is a semisynthetic rifamycin with potent bactericidal activity against intracellular respiratory bacteria, including Mycobacterium tuberculosis, and a long half-life (approximately 60 h) and thus can be administered once weekly . We therefore tested the in vitro activities of ABI-1648, its derivatives ABI-1657 and ABI-1131, azithromycin, and levofloxacin against 10 strains of Chlamydia trachomatis and 10 recent clinical isolates of Chlamydia pneumoniae . The MICs at which 90% of the isolates were inhibited and the minimal bactericidal concentration at which 90% of the isolates were killed for ABI-1648, ABI-1657, and ABI-1131 were 0.0025 micro g/ml for C . trachomatis and 0.00125 to 0.0025 micro g/ml for C . pneumoniae . ABI-1648, ABI-1657, and ABI-1131 were 10- to 1,000-fold more active than azithromycin and levofloxacin.

Biochem Biophys Res Commun, 2003 Mar 7, 302(2), 193 - 200
Structural and functional analysis of BmjMIP, a phospholipase A2 myotoxin inhibitor protein from Bothrops moojeni snake plasma; Soares AM et al.; A protein, which neutralizes the enzymatic, toxic, and pharmacological activities of various basic and acidic phospholipases A(2) from the venoms of Bothrops moojeni, Bothrops pirajai, and Bothrops jararacussu, was isolated from B . moojeni snake plasma by affinity chromatography using immobilized myotoxins on Sepharose gel . Biochemical characterization of this myotoxin inhibitor protein (BmjMIP) showed it to be an oligomeric glycoprotein with a M(r) of 23,000-25,000 for the monomeric subunit . BmjMIP was stable in the pH range from 4.0 to 12.0, between 4 and 80 degrees C, even after deglycosylation . The role of the carbohydrate moiety was investigated and found not to affect the in vitro function of the inhibitor . The corresponding 500bp cDNA obtained by RT-PCR from the liver of the snake encodes a mature protein of 166 amino acid residues including a 19 amino acid signal peptide . The primary structure of BmjMIP showed a high similarity with other snake phospholipase A(2) inhibitors (PLIs) in which the carbohydrate recognition domain (CRD) and the glycosylation site (Asn103) are conserved . Circular dichroism spectroscopy indicated that no significant alterations in the secondary structure of either the BmjMIP or the target protein occur upon their interaction . BmjMIP has a wide range of inhibitory properties against basic and acidic PLA(2)s from Bothrops venoms (anti-enzymatic, anti-myotoxic, anti-edema inducing, anti-cytotoxic, anti-bactericidal, and anti-lethal) . However, the inhibitor showed a reduced ability to neutralize the biological activities of crotoxin B (CB), the PLA(2) homologue associated with crotapotin in Crotalus durissus terrificus snake venom . Finally, the purified PLA(2) inhibitor was shown to protect in vivo against the toxic and pharmacological effects of a homologous PLA(2) enzyme, suggesting that PLIs or a corresponding derived peptide may prove useful in the treatment of snakebite victims or, more importantly, in the treatment of the many human diseases in which these enzymes have been implicated.

Immunology, 2003 Mar, 108(3), 384 - 90
An abnormal but functionally active complement component C9 protein found in an Irish family with subtotal C9 deficiency; Orren A et al.; Two independently segregating C9 genetic defects have previously been reported in two siblings in an Irish family with subtotal C9 deficiency . One defect would lead to an abnormal C9 protein, with replacement of a cysteine by a glycine (C98G) . The second defect is a premature stop codon at amino acid 406 which would lead to a truncated C9 . However, at least one of two abnormal proteins was present in the circulation of the proband at 0.2% of normal C9 concentration . In this study, the abnormal protein was shown to have a molecular weight approximately equal to that of normal C9, and to carry the binding site for monoclonal antibody (mAb) Mc42 which is known to react with an epitope at amino acid positions 412-426, distal to 406 . Therefore, the subtotal C9 protein carries the C98G defect . The protein was incorporated into the terminal complement complex, and was active in haemolytic, bactericidal and lipopolysaccharide release assays . A quantitative haemolytic assay indicated even slightly greater haemolytic efficiency than normal C9 . Epitope mapping with six antihuman C9 mAbs showed the abnormal protein to react to these antibodies in the same way as normal C9 . However, none of these mAbs have epitopes within the lipoprotein receptor A module, where the C98G defect is located . The role of this region in C9 functionality is still unclear . In conclusion, we have shown that the lack of a cysteine led to the production of a protein present in the circulation at very much reduced levels, but which was fully functionally active.

Rev Prat, 2002 Dec 1, 52(19), 2111 - 4
{Mycobacterium tuberculosis and its host}; Vincent V et al.; Human beings are the only hosts of Mycobacterium tuberculosis . Owing to a peculiar architecture and composition of the cell wall, M . tuberculosis presents characteristic properties (acid fastness, high hydrophobicity) . Molecules, proteins and polysaccharides, present on the cell surface play a key role in the rapid bacterial phagocytosis by macrophages . M . tuberculosis is able to inhibit the intracellular bactericidal mechanisms . Potent thymodependent immune responses control bactericial growth through inflammatory reactions.

Dev Comp Immunol, 2003 Apr, 27(4), 313 - 21
CpG oligodeoxynucleotides activate grass carp (Ctenopharyngodon idellus) macrophages; Meng Z et al.; In mice and humans, B cells, antigen-presenting cells including monocytes, macrophages and dendritic cells and natural killer cells can be stimulated directly or indirectly by the bacterial DNA and oligodeoxynucleotides (ODN) containing the CpG motifs (CpG DNA) . Using head kidney macrophages of grass carp (Ctenopharyngodon idellus) as an in vitro model, we investigated the effects of several CpG-ODNs on fish immunocytes . The CpG-ODNs included the optimal motifs: the ODN-1826 (GACGTT) and -2006 (GTCGTT) for the mice and humans cells, the ODN-1670 (AACGTT) used in Atlantic salmon, the ODN-D containing two repeats motif of those in 1670 and the ODN-R with an inverted CpG . The results showed that CpG has an immunomodulatory role in grass carp, and all the ODNs except the ODN-R could activate macrophages, increasing the levels of superoxide anion (O(2)(-)), hydrogen peroxide (H(2)O(2)), acid phosphatase and bactericidal activity . New evidence was provided that CpG-ODN could induce the cell-mediated immunity in fish . Interestingly, no significant differences among the ODNs tested could be found and the ODN-D was not more efficient than 1670 . It suggests that the sequence which contains the unmethylated 'CG' dinucleotides could make contribute to this immunostimulatory effect . These findings indicate that CpG-ODNs could be useful tools for understanding the important anti-bacterial defense mechanism in fish . And it may have potential application in the minimizing the impact of fish diseases and enhancing the efficacy of antigen and DNA vaccines.

Vet Hum Toxicol, 2003 Feb, 45(1), 18 - 20
Structure-activity relationships among zearalenone and its derivatives based on bovine neutrophil chemiluminescence; Murata H et al.; We compared the effects of zearalenone (ZEN), an estrogenic mycotoxin produced by Fusarium fungi, and its derivatives--a- and b-zearalenols (Zel), zearalanone (ZAN), and a- and b-zearalanols (Zal)--on bovine neutrophils in vitro by using chemiluminescence, a bactericidal parameter . ZEN, a-Zel, and b-Zel suppressed luminol-dependent, phorbol 12-myristate 13-acetate-elicited chemiluminescence at a concentration of 10(-5) M, whereas ZAN, a-Zal and b-Zal did not . The suppressive zearalenols are derived from ZEN through reduction of the C6'-ketone into hydroxide, whereas the non-suppressive ZAN and Zal group possesses a hydrogenated C1'-2' bond in place of the double bond adopted in the macrolide ring or ZEN and the zearalenols . In consideration of these structure-activity relationships among ZEN and its derivatives, we conclude that possession of the C1'-2' double bond is essential for zearalenones to induce neutrophil suppression.

Proc Natl Acad Sci U S A, 2003 Feb 18, 100(4), 1490 - 3 Epub 2003 Feb 07.
Evidence for the production of trioxygen species during antibody-catalyzed chemical modification of antigens; Wentworth P Jr et al.; Recent work in our laboratory showed that products formed by the antibody-catalyzed water-oxidation pathway can kill bacteria . Dihydrogen peroxide, the end product of this pathway, was found to be necessary, but not sufficient, for the observed efficiency of bacterial killing . The search for further bactericidal agents that might be formed along the pathway led to the recognition of an oxidant that, in its interaction with chemical probes, showed the chemical signature of ozone . Here we report that the antibody-catalyzed water-oxidation process is capable of regioselectively converting antibody-bound benzoic acid into para-hydroxy benzoic acid as well as regioselectively hydroxylating the 4-position of the phenyl ring of a single tryptophan residue located in the antibody molecule . We view the occurrence of these highly selective chemical reactions as evidence for the formation of a short-lived hydroxylating radical species within the antibody molecule . In line with our previously presented hypothesis according to which the singlet-oxygen ((1)O*(2)) induced antibody-catalyzed water-oxidation pathways proceeds via the formation of dihydrogen trioxide (H(2)O(3)), we now consider the possibility that the hydroxylating species might be the hydrotrioxy radical HO(3)*, and we point to the remarkable potential of this either H(2)O(3)- or O(3)-derivable species to act as a masked hydroxyl radical HO* in a biological environment.

Hunan Yi Ke Da Xue Xue Bao, 2002 Feb 28, 27(1), 17 - 8
{Cloning of cDNA fragments of human defensins}; Liu SP et al.; In order to acquire the cDNA of a small molecular peptide with biological activity, the cDNA encoding human defensins was isolated from total RNAs of peripheral leukocyte in a patient with chronic myelogenous leukemia by the RT-PCR method, about 280 bp in length . The RT-PCR products were cloned into the pUCm-T vector, and may be used to construct a novel fusion protein, which contains human defensins and human bactericidal permeability increasing proteins.

J Biol Chem, 2003 Apr 18, 278(16), 13838 - 46 Epub 2003 Feb 06.
Interactions of mouse Paneth cell alpha-defensins and alpha-defensin precursors with membranes . Prosegment inhibition of peptide association with biomimetic membranes; Satchell DP et al.; The bactericidal activity of mouse alpha-defensins (cryptdins) requires proteolytic activation of inactive precursors by matrix metalloproteinase-7 (matrilysin, EC, MMP-7(a)) . To investigate mechanisms of cryptdin-4 (Crp4) peptide interactions with membrane bilayers and to determine whether MMP-7-mediated proteolysis activates the membrane disruptive activity of Crp4, associations of Crp4 and melittin with biomimetic lipid/polydiacetylene chromatic vesicles were characterized . The peptides differ in their sensitivity to vesicle lipid composition and their depth of bilayer penetration . Crp4 undergoes strong interfacial binding onto lipid bilayers with disruption of the bilayer head group region, unlike melittin, which inserts more deeply into the hydrophobic core of the bilayer . Colorimetric and tryptophan fluorescence studies showed that Crp4 insertion is favored by negatively charged phospholipids and that zwitterionic and Escherichia coli phospholipids promote stronger interfacial binding; melittin-membrane interactions were independent of either variable . In contrast to the membrane disruptive activity of Crp4, pro-Crp4 did not perturb vesicular membranes, consistent with the lack of bactericidal activity of the precursor, and incubation of Crp4 with prosegment in trans blocked Crp4 and G1W-Crp4 membrane interactions at concentrations that inhibit Crp4 bactericidal activity . CD measurements showed that Crp4 has an expected beta-sheet structure that is not evident in the pro-Crp4 CD trace or when Crp4 is incubated with prosegment, indicating that the beta-sheet signal is attenuated by proregion interactions or possibly disrupted by the prosegment . Collectively, the results suggest that the prosegment inhibits Crp4 bactericidal activity by blocking peptide-mediated perturbation of target cell membranes, a constraint that is relieved when MMP-7 cleaves the prosegment.

Biol Neonate, 2003, 83(1), 25 - 9
Evaluation of a netilmicin-loading dose in very low birthweight infants; Berger A et al.; BACKGROUND: The bactericidal efficacy of aminoglycosides is directly related to maximum serum concentrations, particularly the initial one . Therefore, several groups have recommended an aminoglycoside loading dose . Our goal was to develop a simplified dosage regimen for preterm infants which would result in therapeutic maximum serum concentrations early in the course of therapy . METHODS: Open, noncomparative study during November 2000 to April 2001 . The modified netilmicin-dosing protocol included a loading dose of 5 mg/kg in the first week of life, followed by a maintenance regimen of 3.5 mg/kg every 24 h . After the first week of life the corresponding doses were 6 (loading) and 5 mg/kg (maintenance) . A peak level was measured 30 min after the second dose, and a trough level immediately before the third dose . RESULTS: Thirty-five very low birthweight infants (mean birthweight 876 +/- 170, range 536-1,385 g; mean gestational age 26 +/- 1.8, range 23-30 weeks) who had 46 episodes of netilmicin treatment were included in the analysis . Mean netilmicin peak and trough values were 15.9 +/- 3.7 (range 8.9-28.9) and 3.4 +/- 1.3 (range 1.0-7.8) micromol/l, respectively . Ninety-one percent of all peak levels were within the targeted range of > or =10 micromol/l . Eleven trough values (24%) were > or =4 micromol/l: in 7 instances netilmicin was administered within the first week of life, 5 of these patients had concomitant indomethacin treatment . Only 1 of the 35 neonates had a rise in serum creatinine of > or =0.5 mg/dl during netilmicin therapy . Hearing evaluations were performed in 25 of the 29 surviving infants at discharge home, all of which gave normal results . CONCLUSIONS: The new netilmicin-dosing protocol yielded therapeutic maximum serum concentrations in 91% of cases after the second dose . However, a significant number of very low birthweight infants had elevated trough levels, particularly when netilmicin was administered in the first week of life with concomitant indomethacin treatment . We speculate that a longer interval between the loading dose and the first maintenance dose would result in fewer elevated trough levels with a similarly high number of therapeutic peak levels .

Shock, 2003 Jan, 19(1), 50 - 4
Glutamine improves impaired cellular exudation and polymorphonuclear neutrophil phagocytosis induced by total parenteral nutrition after glycogen-induced murine peritonitis; Ikeda S et al.; Clinical and laboratory evidence shows that enteral feeding significantly reduces pneumonia and intra-abdominal abscess formation after celiotomy for severe trauma . Supplementation of total parenteral nutrition (TPN) with glutamine (GLN) supports impaired immunity induced by TPN in several animal and human studies . This work investigates the peritoneal cellular response and polymorphonuclear neutrophil (PMN) bactericidal function after mouse chemical peritonitis after TPN with and without GLN . Thirty-three mice received chow, TPN, or 2% GLN-supplemented TPN (GLN-TPN) for 5 days . All mice then received 2 mL of a 1% glycogen solution intraperitoneally to induce cell exudation, and peritoneal exudative cells (PECs) were recovered 4 h later . Total and differential PEC numbers, as well as PMN phagocytosis, reactive oxygen intermediate production (ROI), CD11b (integrin aM chain) expression, and CD16/32 (Fcgamma II/III receptor) expression were measured . PMN, macrophage, and lymphocyte cell numbers were significantly lower with TPN than with chow or GLN-TPN groups, with no differences between chow and GLN-TPN . TPN significantly lowered peritoneal PMN phagocytosis compared with chow (P < 0.05) and approached significance with GLN-TPN (P = 0.06) . There were no significant differences in ROI production or CD11b and CD16/32 expression on peritoneal PMN . GLN supplementation improved the reduction in cell exudation and PMN phagocytosis induced by TPN after chemical peritonitis.

Wei Sheng Wu Xue Bao, 2001 Dec, 41(6), 669 - 73
{Gene cloning and expression of human bactericidal/permeability-increasing protein}; Xu J et al.; The gene of human bactericidal/permeability-increasing protein (BPI) was cloned from peripheral blood lymphocytes of a normal Chinese individual . The result of sequencing showed the gene is 1452 bp encoding a 27-residue signal peptide and a 456-residue matured protein, and it has six nucleotide variations compared with the sequence reported which results in 4 different amino acids . In order to get recombinant BPI, the gene was cloned into an expressing plasmid and expressed in CHO cells . The recombinant protein was purified using cation-exchange chromatography and its bioactivity was proved with bactericidal assays.

J Infect Dis, 2003 Jan 15, 187(2), 270 - 8 Epub 2003 Jan 06.
Whole blood bactericidal activity during treatment of pulmonary tuberculosis; Wallis RS et al.; The timely evaluation of new drugs that can be used to shorten tuberculosis (TB) treatment will require surrogate markers for relapse . This study examined bactericidal activity against intracellular Mycobacterium tuberculosis in whole blood culture (whole blood bactericidal activity; WBA) during TB treatment . In the absence of chemotherapy, immune mechanisms in patient blood resulted in bacteriostasis, whereas administration of oral chemotherapy resulted in bacillary killing . Total WBA per dose was greater during the intensive phase of treatment than during the continuation phase (mean, -2.32 vs . -1.67 log(10) cfu-days, respectively; P<.001) . Cumulative WBA throughout treatment was greater in subjects whose sputum cultures converted to negative by the eighth week of treatment than in those for whom conversion was delayed (mean, -365 vs . -250 log(10) cfu-days; P=.04) and correlated with the rate of decrease of sputum colony-forming unit counts during the first 4 weeks of treatment (P=.018), both of which are indicative of prognosis . These findings indicate that measurement of WBA may have a role in assessing the sterilizing activity of new anti-TB drugs.

Biochem Soc Trans, 2003 Feb, 31(Pt 1), 275 - 80
Phosphoinositide 3-kinase gamma: a key modulator in inflammation and allergy; Wymann MP et al.; Chronic inflammation and allergy involve the activation of tissue-resident cells and, later on, the invasion of effector cells . We have previously shown that the loss of phosphoinositide 3-kinase (PI3K) gamma impairs chemokine-dependent migration of neutrophils and macrophages both in vitro and in vivo . On the other hand, PI3K gamma is not required either during phagocytic processes or in the activation of bactericidal activities like granule secretion and particle-mediated respiratory burst in neutrophils . Tissue mast cells are key regulators in allergy and inflammation and release histamine upon clustering of their IgE receptors . We have demonstrated that murine mast cell responses are exacerbated in vitro and in vivo by autocrine signals, and require functional PI3K gamma . Adenosine, acting through the A(3) adenosine receptor, as well as other agonists of G(alpha i)-coupled receptors, transiently increased PtdIns(3,4,5) P (3) exclusively via PI3K gamma . PI3K gamma-derived PtdIns(3,4,5) P (3) was instrumental for initiation of a sustained influx of external Ca(2+) and degranulation . Mice that lacked PI3K gamma did not form oedema when challenged by passive systemic anaphylaxis . PI3K gamma thus relays inflammatory signals through various GPCRs, and is thus central to mast cell function . Taken together, this suggests that pharmaceutical targeting of PI3K gamma might alleviate inflammation at both early and late stages of the allergic response.

Br J Ophthalmol, 2003 Feb, 87(2), 163 - 7
Comparison of 5% povidone-iodine solution against 1% povidone-iodine solution in preoperative cataract surgery antisepsis: a prospective randomised double blind study; Ferguson AW et al.; BACKGROUND/AIM: Povidone-iodine (PI, Betadine) is routinely used as a preoperative topical antiseptic in cataract surgery as it has been shown to reduce the incidence of postoperative endophthalmitis . However, the concentration used clinically is variable . In vitro studies have shown that PI is paradoxically more effective at lower concentration . This study was undertaken to determine if this effect was reproducible in vivo . METHODS: A prospective randomised double blind study was carried out in the ophthalmic theatre in a district general hospital . 105 patients attending for routine cataract surgery were randomly allocated to have their conjunctival fornices irrigated preoperatively with either PI 1% (group A) or PI 5% (group B) . Conjunctival swabs were taken, in identical fashion, both before and 1 minute after irrigation . The number and species of bacterial colonies cultured from each swab was counted . The difference in the median number of bacterial colonies from pre-irrigation to post-irrigation cultures was then compared between the groups . RESULTS: Bacterial cultures were gained from 100 patients (33 male, 67 female, mean age 74 years, range 30-95 years) . Group B (5% PI) showed a decrease in median colony forming units (CFU) pre-irrigation from 100 to 40 CFU post-irrigation (a drop of 60%) . This was greater than in group A (1% PI) where the reduction was 120 CFU pre-irrigation to 100 CFU post-irrigation (a drop of 16.7%) (Mann-Whitney test, p<0.05) . At higher initial bacterial loads (CFU pre-irrigation >1000), the difference in median between the two groups became larger as the number of pre-irrigation bacteria increased . In group B pre-irrigation CFU reduced from 3340 to 110 post-irrigation (a drop of 96.7%) compared with group A: 5000 CFU pre-irrigation to 3000 post-irrigation (a drop of 40%) (Mann-Whitney test, p=0.0014) . CONCLUSION: Despite in vitro evidence of higher bactericidal efficacy of PI at more dilute concentrations, 5% PI is more effective than 1% PI in decreasing the human conjunctival bacterial flora in vivo, particularly in the presence of heavier initial bacterial load.

Antimicrob Agents Chemother, 2003 Feb, 47(2), 653 - 7
Sterilizing activities of fluoroquinolones against rifampin-tolerant populations of Mycobacterium tuberculosis; Hu Y et al.; The bactericidal activities of ciprofloxacin, ofloxacin, levofloxacin, moxifloxacin, and gatifloxacin were tested in three models of rifampin-tolerant Mycobacterium tuberculosis persisters . Model 1 was a 100-day-old, unshaken, anaerobically adapted culture in which serial dilutions of the quinolones were incubated for 5 days and CFU counts were then done In models 2 and 3, 100 mg of rifampin/liter was added to the 100-day culture for 5 or 7 days to produce tolerant organisms that did not grow on plates; the rifampin was then washed off, fresh medium was added to allow recovery of growth on plates, and the culture was incubated for 7 days before CFU counts . In model 2, the quinolones were added after rifampin had been washed off, whereas in model 3 the quinolones were added to the cultures containing rifampin . In models 1 and 2, ciprofloxacin had the least bactericidal activity, ofloxacin and levofloxacin had greater activities, and moxifloxacin and gatifloxacin had the greatest activities . In model 3, ofloxacin had no detectable activity whereas moxifloxacin killed about log(10) 0.279 CFU of the persisters per ml at concentrations attainable in lesions; isoniazid had virtually no activity . These findings predict that ofloxacin will not be found to have effective sterilizing activity in clinical studies now planned whereas moxifloxacin will be able to shorten treatment.

Antimicrob Agents Chemother, 2003 Feb, 47(2), 614 - 9
Culture and antibiotic susceptibility of Bartonella quintana in human erythrocytes; Rolain JM et al.; Bartonella quintana, the agent of trench fever, has recently been implicated in various diseases, in particular, bacteremia and endocarditis in homeless people . The host cell of Bartonella spp . is believed to be the erythrocyte, and in the present study we demonstrate that B . quintana can be cultured in vitro in human erythrocytes . The bacteria were found to be intraerythrocytic by laser confocal microscopy with Bartonella species-specific monoclonal antibodies . Infections with B . quintana decreased the life span of erythrocytes in culture from 8.6 to 4.8 days . In the culture system we found that most of the antibiotics that we tested (doxycycline, fluoroquinolone compounds, and beta-lactams) were not bactericidal . Gentamicin was bactericidal at 4 micro g/ml, as was rifampin, but to a lesser extent . At this concentration, gentamicin has been shown to enter erythrocytes slowly and to reach a peak level of 0.26 micro g/ml after 24 h . At 0.26 micro g/ml, however, we found that gentamicin was not able to kill extracellular B . quintana, even after 96 h of incubation . We hypothesize that erythrocytes may be a reservoir for B . quintana and that the bactericidal activity of gentamicin that we observed occurs mainly when the bacteria emerge from the erythrocytes and are found extracellularly . It would appear that gentamicin should be administered for at least 5 days to cure patients infected with B . quintana.

Scand J Immunol, 2003 Jan, 57(1), 2 - 10
Anti-inflammatory activities of human lactoferrin in acute dextran sulphate-induced colitis in mice; Haversen LA et al.; In this study, we investigated the anti-inflammatory effects of orally administered human lactoferrin (hLF) and two peptides, based on the bactericidal region of hLF (HLD1 and HLD2), on the course of experimental colitis . Acute colitis was induced in C57Bl/6 mice by giving 5% dextran sulphate (DX) in the drinking water . The mice were killed after 2 or 7 days of DX exposure . The animals were given hLF or the peptides orally twice a day (2 mg/dose/mouse) during the DX exposure . In the control animals, the hLF or the peptides were replaced by bovine serum albumin or water . The appearance of occult blood in the faeces and macroscopic rectal bleeding were significantly delayed and partly reduced in the hLF-treated animals compared with the control animals . The shortening of the colon, a pathological effect of DX exposure, was significantly less pronounced in the hLF-treated group compared with the control group . Also, the interleukin-1beta (IL-1beta) levels in the blood were significantly diminished in this group after 2 days of DX exposure . A significantly lower crypt score was observed in the distal part of the colon in the hLF-treated group compared with the control group . Also, significantly reduced numbers of CD4 cells, F4/80-positive macrophages and tumour necrosis factor-alpha-producing cells were detected by immunohistochemistry in the distal colon of the hLF-treated animals compared with the control animals after 7 days of DX exposure . A reduction was also observed concerning the IL-10-producing cells in the middle colonic submucosa . The HLD1 and HLD2 treatment, which was carried out for 2 days, only gave results almost identical to those of hLF, concerning clinical parameters after the 2 days of DX exposure . An even stronger effect was observed for HLD2, regarding decreased occult blood in the faeces and colon length . Our results show that perorally given hLF mediates anti-inflammatory effects on the DX-induced acute colitis, and further suggest that the bactericidal region of the hLF molecule may be involved in these activities.

Infect Immun, 2003 Feb, 71(2), 1016 - 9
Influence of extracellular bactericidal agents on bacteria within macrophages; Hamrick TS et al.; We employed gentamicin-sensitive and -resistant derivatives of Escherichia coli in a macrophage phagocytosis assay that compared lambda bacteriophage and gentamicin as extracellular bactericidal agents . Colony counts and direct microscopic examination of phagocytized E . coli supported the conclusion that gentamicin entered macrophages, even at low concentrations, and contributed to their bactericidal activity . Also, two E . coli strains differing in the ability to express the adhesin of type 1 pili (FimH) were distinguishably different in intracellular survival when lambda was used as the extracellular killing agent but were indistinguishable when gentamicin was employed.

Am J Respir Crit Care Med, 2003 May 15, 167(10), 1348 - 54 Epub 2003 Jan 06.
Bactericidal and sterilizing activities of antituberculosis drugs during the first 14 days; Jindani A et al.; Colony-forming units of Mycobacterium tuberculosis in sputum were counted at 2-day intervals in 100 patients treated with 22 regimens of isoniazid, rifampin, pyrazinamide, ethambutol, or streptomycin, given alone or in combinations . The exponential fall in colony-forming units was measured by linear regression coefficients of the log counts during the initial 2-day phase of rapid, drug-determined killing and during the subsequent 12 days of much slower sterilizing activity . The regression coefficients during the first 2 days varied significantly according to the drug; the greatest effects in multiple regression analyses were due to isoniazid (p < 0.001) and rifampin (p = 0.027) . The rapid kill obtained with isoniazid was unaffected by addition of other drugs, so that a change in activity after adding an unknown drug to isoniazid would not be measurable . In multiple regression analysis of the coefficients during Days 2-14, rifampin and streptomycin had significant effects (p = 0.007 and 0.006, respectively), indicating that both drugs had important sterilizing activity, streptomycin particularly early . Isoniazid and pyrazinamide had no significant effects . In analyses of combined drug regimens only, ethambutol had an effect (p = 0.01) in reverse direction to that of rifampin, suggesting it antagonized the sterilizing activity of other drugs.

Microbiol Immunol, 2002, 46(11), 761 - 6
Use of immunoglobulin enriched bovine colostrum against oral challenge with enterohaemorrhagic Escherichia coli O157:H7 in mice; Funatogawa K et al.; An immunoglobulin enriched bovine colostrum preparation, IMMULAC (New Zealand Dairy Group, Cambridge, New Zealand), contains antibodies against various bacterial antigens . In the present study, we investigated the protective effects of a commercial bovine colostrum preparation against infections with enterohaemorrhagic Escherichia coli (EHEC) O157:H7 in a murine model . Balb/c mice were given drinking water containing streptomycin for 3 days before and following oral challenge with streptomycin-resistant EHEC O157:H7 strain (O157-SM(R)) . In mice pretreated with streptomycin, EHEC O157:H7 maintained stable levels of bacterial colonization in the intestines for the 3-week experimental time period . Oral administration of colostrum resulted in rapid decrease in the bacteria numbers compared with administration of skim-milk . Colostrum showed no direct in vitro bactericidal properties against either EHEC O157:H7 . When sections prepared from cecum walls of streptomycin-pretreated mice were incubated in vitro with EHEC O157:H7, the colostrum significantly prevented the attachment of the organisms to the sections when compared with skim-milk . These results indicate that oral administration of bovine colostrum effectively protects mice against food-borne infections by inhibiting bacterial attachment to the intestinal mucous membrane, colonization and growth in the intestinal tract.

Zhonghua Shao Shang Za Zhi, 2002 Apr, 18(2), 95 - 8
{The study on the role of modeling peptides derived from bactericidal/permeability increasing protein on the endotoxin neutralization}; Zheng J et al.; OBJECTIVE: To observe the role of four modeling peptides (10342, 10343, 10344, 10345) derived from bactericidal/permeability increasing protein (BPI) in neutralizing endotoxin (LPS) in vitro and in vivo . METHODS: Quantitative limulus amoebocyte lysate assay was employed to evaluate the capacity of BPI peptides in neutralizing endotoxin in vitro . The protective capacity of the peptides was observed in mice challenged with endotoxin by intravenous administration via the tail vein . The influence of the peptides on serum TNFalpha and IL-6 levels in rats with endotoxemia were also observed . RESULTS: All of the four peptides possessed endotoxin-neutralizing capacity which was strengthened along with the increase in their concentration . Among the peptides, 10342 is the strongest one . All of the peptides had strong power to protect mice from endotoxin with 90% protective rate . In the rats with endotoxemia, the four peptides could reduce the levels of serum TNFalpha and IL-6 significantly at different time-points . CONCLUSION: Four BPI modeling peptides possessed not only endotoxin-neutralizing capacity in vitro, but also potential protective capacity in animals challenged with endotoxin.

Zhonghua Shao Shang Za Zhi, 2002 Apr, 18(2), 81 - 3
{An exploration of the preventive effects on lanthanum chloride on enteral bacterial translocation in scalded rats}; Liu Q et al.; OBJECTIVE: To explore the preventive effect of lanthanum chloride on enteral bacterial translocation in scalded rats . METHODS: Ninety Sprague-Dawley (SD) rats were employed in the study and randomly divided into three groups, i.e . normal control (A), burn control (B) and treatment (C) groups . Plasmid PUC19 labelled by JM109 was transfected to Escherichia coli (E . coli), so that restriction endonuclease finger - print image spectrum analysis could be applied to the tracing and quantification of the translocation of E . coli from intestine to mesenteric lymph nodes (MLNs) and blood . The intestinal tissue contents of endotoxin (ET), nitric oxide (NO), nitric oxide synthase (NOS), malondialdehyde (MDA) and superoxide dismutase (SOD) were determined . RESULTS: It was identified that the bacteria in MLNs and blood exhibited the same gene map with those from gastric gavage in B and C groups . But the bacterial quantity in MLNs in C group on 3 postburn day (PBD) was much lower than that in B group (P < 0.05) . The intestinal MDA content in C group on 1 and 3 PBDs was obviously higher than that in B group (P < 0.05) . CONCLUSION: Bacteria (E . coli) could be translocated from gut to MLNs and blood, which could be evidently alleviated by lanthanum chloride by means of its bactericidal property, inhibition of NOS activity, so that NO production decreased, and its ability to increase SOD activity leading to less production of MDA.

Clin Exp Rheumatol, 2002 Nov-Dec, 20(6), 783 - 9
Antineutrophil cytoplasmic antibody (ANCA)-associated systemic vasculitis after immunisation with bacterial proteins; Savige J et al.; OBJECTIVE: There is circumstantial evidence for a role for infections in the development of the small vessel vasculitides associated with antineutrophil cytoplasmic antibodies (ANCA) . The aim of this study was to determine whether the immunisation of rats with bacterial proteins could result in circulating ANCA, T cells with specificity for ANCA antigens, and a systemic vasculitis . METHODS: Adult male Wistar rats were immunised with pasteurised sonicated S . aureus (n = 7), E . coli (n = 8), purified protein derivative (PPD, n = 5), myeloperoxidase (MPO, n = 5) or phosphate-buffered saline (PBS, n = 5), in complete and in incomplete Freund's adjuvant . ANCA were assayed by indirect immunofluorescent (IIF) examination of normal rat neutrophils, and in ELISAs using human proteinase 3 (PR3), MPO and bactericidal/permeability-inreasing protein (BPI) . The T cell response to PR3, MPO and BPI was assessed by a whole blood T cell proliferative assay in vitro, and by a delayed type hypersensitivity (DTH) response in vivo . Kidney and bowel were examined histologically for evidence of vasculitis and colitis . RESULTS: One rat from each group immunised with S . aureus or E . coli developed pauciimmune segmental glomerular sclerosis . The rat immunised with E . coli had additionally an arteritis affecting renal interlobular and gut vessels . This rat had circulating C-ANCA, that produced granular cytoplasmic neutrophil fluorescence with central accentuation, but the target antigen could not be determined in ELISAs using human PR3, MPO or BPI . In animals immunised with S . aureus or E . coli, there was no significant T cell proliferative or DTH response specific for human PR3, MPO or BPI . CONCLUSION: The development of ANCA and vasculitis in a rat immunised with bacterial proteins indicates that the relationship between infections and ANCA should be investigated further.

J Immunol, 2003 Jan 1, 170(1), 14 - 8
Cutting edge: bacterial lipoprotein induces endotoxin-independent tolerance to septic shock; Wang JH et al.; Tolerance to bacterial cell wall components is an adaptive host response . Endotoxin/LPS tolerance is characterized by a survival advantage against subsequent lethal LPS challenge . However, it is uncertain whether LPS tolerance can afford protection against other septic challenges . In this study, we show that tolerance induced by bacterial lipoprotein (BLP) protects mice against not only BLP-induced lethality, but also LPS-, live bacteria-, and polymicrobial sepsis-induced lethality . In contrast, LPS tolerance offers no survival benefit against the latter two challenges . Furthermore, induction of BLP tolerance results in overexpression of complement receptor type 3 and FcgammaIII/IIR on neutrophils (polymorphonuclear neutrophils) and peritoneal macrophages, with increased bacterial recognition and bactericidal activity, whereas LPS-tolerized mice exhibit an impaired ability to ingest and to kill bacteria . These results indicate that BLP tolerance is a novel adaptive host response associated with a unique protective effect during septic shock.

Infect Immun, 2003 Jan, 71(1), 494 - 503
Role of avian pathogenic Escherichia coli virulence factors in bacterial interaction with chicken heterophils and macrophages; Mellata M et al.; Avian pathogenic Escherichia coli (APEC) cause extraintestinal disease in avian species via respiratory tract infection . Virulence factors associated with APEC include type 1 and P fimbriae, curli, aerobactin, lipopolysaccharide (LPS), K1 capsular antigen, temperature-sensitive hemagglutinin (Tsh), and an uncharacterized pathogen-specific chromosomal region (the 0-min region) . The role of these virulence factors in bacterial interaction with phagocytes was investigated by using mutants of three APEC strains, each belonging to one of the most predominant serogroups O1, O2, and O78 . Bacterial cell interaction with avian phagocytes was tested with primary cultures of chicken heterophils and macrophages . The presence of type 1 fimbriae and, in contrast, the absence of P fimbriae, K1 capsule, O78 antigen, and the 0-min region promoted bacterial association with chicken heterophils and macrophages . The presence of type 1 and P fimbriae, O78 antigen, and the 0-min region seemed to protect bacteria against the bactericidal effect of phagocytes, especially heterophils . The tested virulence factors seemed to have a limited role in intracellular survival for up to 48 h in macrophages . Generally, opsonized and nonopsonized bacteria were eliminated to the same extent, but in some cases, unopsonized bacteria were eliminated to a greater extent than opsonized bacteria . These results confirm the important role of type 1 fimbriae in promotion of initial phagocytosis, but nevertheless indicate a role for type 1 fimbriae in the protection of bacteria from subsequent killing, at least in heterophils . The results also indicate a role for K1 capsule, O78 antigen, P fimbriae, and the 0-min region in initial avoidance of phagocytosis, but demonstrate an additional role for O78 antigen, P fimbriae, and the 0-min region in subsequent protection against the bactericidal effects of phagocytes after bacterial association has occurred.

Zhonghua Nei Ke Za Zhi, 2002 Nov, 41(11), 766 - 8
{Antigens of antineutrophil cytoplasmic autoantibodies in recognizing a novel neutrophil granule antigen in lupus nephritis and its association with photosensitivity and oral ulceration}; Chen M et al.; OBJECTIVE: To investigate whether there are some unknown target antigens of antineutrophil cytoplasmic autoantibodies (ANCA) in patients with lupus nephritis (LN) and the prevalence and the clinical significance of these unknown target antigens . METHODS: Serum was collected from 72 renal-biopsy-proven lupus nephritis (LN) patients . Mixed neutrophil granules were separated from normal human peripheral neutrophils and soluble acid extracts in non-reducing conditions were used as antigens in Western-blot analysis to detect ANCA in the serum sample of LN patients . RESULTS: Serum from seme LN patients could blot a few bands . Interestingly, serum from 14/72 (19.4%) of the patients recognized a novel 69,000 protein band and that from 10/72 (13.9%) sera recognized a 55,000 protein band that might be the bactericidal/permeability-increasing protein (BPI) . The 69,000 target antigen was different from the known target ANCA antigens such as cathepsin G and lactoferrin . Further study revealed that the percentages of patients with photosensitivity and oral ulcer in anti-69,000 autoantibody positive group were significantly higher than those in the anti-69,000 autoantibody negative group (57.1% vs 12.1%, P < 0.005, and 50.0% vs 17.2%, P < 0.05, respectively) . CONCLUSIONS: A novel 69,000 protein in human neutrophil granules is identified as a target antigen of ANCA in LN patients . The anti-69,000 autoantibodies may be associated with photosensitivity and oral ulcer in LN patients.

J Biol Chem, 2003 Mar 7, 278(10), 7910 - 9 Epub 2002 Dec 13.
Structural determinants of procryptdin recognition and cleavage by matrix metalloproteinase-7; Shirafuji Y et al.; The bactericidal activity of mouse Paneth cell alphadefensins, or cryptdins, is dependent on processing of cryptdin precursors (pro-Crps) by matrix metalloproteinase-7 (MMP-7) (Wilson, C . L., Ouellette, A . J., Satchell, D . P., Ayabe, T., Lopez-Boado, Y . S., Stratman, J . L., Hultgren, S . J., Matrisian, L . M., and Parks, W . C . (1999) Science 286, 113-117) . To investigate the mechanisms of pro-Crp processing by this enzyme, recombinant pro-Crp4, a His-tagged chimeric pro-Crp (pro-CC), and site-directed mutant precursors of each were digested with MMP-7, and the cleavage products were analyzed by NH(2)-terminal peptide sequencing . Proteolysis of pro-Crp4 with MMP-7 activated in vitro bactericidal activity to the level of the mature Crp4 peptide by cleaving pro-Crp4 at Ser(43) downward arrow Ile(44) and Ala(53) downward arrow Leu(54) in the proregion and near the Crp4 peptide NH(2) terminus between Ser(58) downward arrow Leu(59) . Because the Crp4 NH(2) terminus occurs at Gly(61), not Leu(59), MMP-7 is necessary but insufficient to complete the processing of Crp4 . Crp activating proteolysis at S58 downward arrow L59 was unaffected by I44S/I44D or L54S/L54D loss-of-function mutations in pro-Crp4, and a (L59S)-pro-CC mutant was cleaved normally at Ser(43) downward arrow Val(44) and Ser(53) downward arrow Leu(54) sites but not at the peptide NH(2) terminus . C57BL/6 mice contain an abundant (L59S)-Crp4 mutant peptide with Leu(54) at its NH(2) terminus resulting from Ala(53) downward arrow Leu(54) cleavage and loss-of-function at the Ser(58) downward arrow Ser(59) cleavage site . Thus, alpha-defensins resulting from mutations at MMP-7 cleavage sites exist in mouse populations . A pro-CC substrate containing both L54S and L59S mutations resisted cleavage at Ser(43) downward arrow Val(44) completely, showing that cleavage at one or both downstream sites must precede proteolysis at Ser(43) downward arrow Val(44) . These findings show that MMP-7 activation of pro-Crps can occur without proteolysis of the proregion, and prosegment fragmentation depends, at least in part, on the release of the Crp peptide from the precursor.

Psychosom Med, 2002 Nov-Dec, 64(6), 963 - 70
Perceived stress and psychological well-being are associated with antibody status after meningitis C conjugate vaccination; Burns VE et al.; OBJECTIVE: Psychological stress has been associated with reduced immune response to a variety of vaccinations . This study is the first to examine antibody status after vaccination with a conjugate vaccine, in which a polysaccharide antigen is conjugated to a protein to elicit a thymus-dependent antibody response . METHODS: Sixty undergraduate students, who had received the meningitis C conjugate vaccine before recruitment, attended a single testing session . They provided a blood sample and completed a battery of questionnaires, including the Life Events Scale for Students, Perceived Stress Scale, and General Health Questionnaire (GHQ-28) . Both meningitis C-specific antibody titer and the serum bactericidal assay titer to whole meningitis C bacteria were assayed . RESULTS: High perceived stress, but not life events stress, was associated with low antibody titers . Poor antibody titers were also predicted by relatively low levels of psychological well-being as measured by the GHQ-28 . Of the GHQ-28 subscales, anxiety/insomnia and social dysfunction were associated with antibody status . No psychological variables emerged from bivariate analyses as predictive of the adequacy of bactericidal titer . CONCLUSIONS: This study provides the first evidence that antibody status after a conjugate vaccination may be susceptible to psychological influences.

Biochem Biophys Res Commun, 2002 Dec 13, 299(4), 613 - 20
Characterization of the specific cleavage of ceiE7-mRNA of the bactericidal ColE7 operon; Chang SJ et al.; Posttranscriptional control of the bactericidal ColE7 operon has been implicated by a feedback endonucleolytic cleavage of its own mRNA . The cleavage site has been located at the coding region of ceiE7, the second cistron of the ColE7 cea-cei-cel polycistronic transcript . Interestingly, Im7 protein, the translation product of ceiE7, is required for the specific cleavage . It was found that both sequence (GAUCUGAUU) flanking the cleavage site and the putative T1 stem-loop structure distal to the coding region of ceiE7 gene play a critical role for the specific cleavage of ceiE7-mRNA . Furthermore, we have verified that a di-nucleotide GG sequence located at the topmost position of the loop region of the putative stem-loop structure is essential for the specific cleavage of ceiE7-mRNA . Thus, our data reveal the existence of a novel mRNA degradative machinery for the regulation of the expression of ColE7 operon.

Nitric Oxide, 2002 Dec, 7(4), 283 - 8
Increased nitric oxide production by neutrophils from patients with chronic granulomatous disease on trimethoprim-sulfamethoxazole; Tsuji S et al.; Chronic granulomatous disease (CGD) is an inherited disease characterized by severe and recurrent bacterial and fungal infections . Phagocytic cells of CGD patients are unable to produce superoxide anion, and their efficiency in bacterial killing is significantly impaired . In these patients, the prophylactic and therapeutic validity of a long-term use of trimethoprim-sulfamethoxazole (TMP-SMX) has been well established . However a role of nitric oxide (NO) produced by phagocytic cells from CGD patients is unknown, and the mechanism of TMP-SMX in CGD is unclear . We have directly measured NO production in whole human blood by using 4,5-diaminofluorescein as a novel fluorescent indicator for intracellular NO . Intracellular NO production of gated neutrophils increased time dependently when stimulated by lipopolysaccharide (LPS) and calcium ionophore . Although all polymorphonuclear leukocyte (PMN) specimens from patients with CGD failed to generate hydrogen peroxide, NO production by CGD PMNs was significantly increased compared with that of control PMNs (p<0.05) . TMP-SMX with LPS significantly increased compared with LPS-stimulated samples at clinical (n=5, p<0.05) and 10-fold clinical concentrations (n=5, p<0.01) . TMP-SMX with LPS in CGD PMNs significantly increased the production of NO in comparison with the LPS stimulation at 10-fold clinical concentrations (n=5, p<0.05) . In conclusion, our data indicate the possibility that NO production by neutrophils from patients with CGD treated with TMP-SMX has a role of bactericidal activity instead of O(2)(-) in host defense mechanism.

J Immunol, 2002 Dec 1, 169(11), 6352 - 60
A novel interaction of outer membrane protein A with C4b binding protein mediates serum resistance of Escherichia coli K1; Prasadarao NV et al.; Escherichia coli is an important pathogen that causes meningitis in neonates . The development of bacteremia preceding the traversal across the blood-brain barrier is a prerequisite for this pathogen that obviously must survive the bactericidal activity of serum . Here we report that outer membrane protein A (OmpA) of Escherichia coli contributes to serum resistance by binding to C4b binding protein (C4bp), a complement fluid phase regulator . C4bp contains seven identical alpha-chains and one beta-chain linked together with disulfide bridges . We found that OmpA binds the alpha-chain of C4bp, which is composed of eight homologous complement control protein (CCP) modules . Binding studies using mutants of recombinant C4bp that lack one CCP at a time suggest that CCP3 is the major site of interaction with OmpA . Furthermore, we demonstrate that the N terminus of OmpA interacts with C4bp . Binding of C4bp to OmpA is not significantly inhibited in the presence of either C4b or heparin and is not salt sensitive, implying that it is hydrophobic in nature, suggesting a novel interaction between OmpA and C4bp . A compelling observation in this study is that synthetic peptides corresponding to CCP3 sequences block the binding of C4bp to OmpA and also significantly enhance serum bactericidal activity.

Biochim Biophys Acta, 2002 Dec 12, 1579(2-3), 92 - 100
The BSP30 salivary proteins from cattle, LUNX/PLUNC and von Ebner's minor salivary gland protein are members of the PSP/LBP superfamily of proteins; Wheeler TT et al.; Saliva influences rumen function in cattle, yet the biochemical role for most of the bovine salivary proteins (BSPs) has yet to be established . Two cDNAs (BSP30a and BSP30b) from bovine parotid salivary gland were cloned and sequenced, each coding for alternate forms of a prominent protein in bovine saliva . The BSP30 cDNAs share 96% sequence identity with each other at the DNA level and 83% at the amino acid level, and appear to arise from separate genes . The predicted BSP30a and BSP30b proteins share 26-36% amino acid identity with parotid secretory protein (PSP) from mouse, rat and human . BSP30 and PSP are in turn more distantly related to a wider group of proteins that includes lung-specific X protein, also known as palate, lung, and nasal epithelium clone (LUNX/PLUNC), von Ebner's minor salivary gland protein (VEMSGP), bactericidal permeability increasing protein (BPI), lipopolysaccharide binding protein (LBP), cholesteryl ester transfer protein (CETP), and the putative olfactory ligand-binding proteins RYA3 and RY2G5 . Bovine cDNAs encoding homologs of LUNX/PLUNC and VEMSGP were isolated and sequenced . Northern blot analysis showed that LUNX/PLUNC, BSP30 and VEMSGP are expressed in bovine salivary tissue and airways, and that they have non-identical patterns of expression in these tissues . The expression of both BSP30a and BSP30b is restricted to salivary tissue, but within this tissue they have distinct patterns of expression . The proximity of the human genes coding for the PSP/LBP superfamily on HSA20q11.2, their similar amino acid sequence, and common exon segmentation strongly suggest that these genes evolved from a common ancestral gene . Furthermore, they imply that the BSP30a and BSP30b proteins may have a function in common with other members of this gene family.

Biochimie, 2002 May-Jun, 84(5-6), 433 - 8
The modes of action of colicins E5 and D, and related cytotoxic tRNases; Masaki H et al.; Colicins E5 and D cleave the anticodon loops of distinct tRNAs of Escherichia coli both in vivo and in vitro, which accounts for their bactericidal actions through depletion of tRNAs and prevention of protein synthesis . The targets of colicin E5 are five tRNA species for four amino acids, tyrosine, histidine, asparagine and aspartic acid, and those of colicin D are four isoaccepting tRNAs for arginine . These two colicins represent a new class, the "tRNase-type", of the nuclease-type colicins, which previously comprised the DNase-type and ribotoxin-type (or rRNase-type) . On the other hand, a certain clinical E . coli strain produces a potentially suicidal "anticodon-nuclease", PrrC, in response to phage T4 infection, which specifically cleaves its own lysine tRNA . For these three tRNases, i.e . colicins E5 and D, and PrrC, the substrates and reaction products, as well as their physiological consequences, are very similar to each other, but so many molecular features are different that these three proteins are assumed to have acquired similar functions through evolutionary convergence from different origins.

Antibiot Khimioter, 2002, 47(6), 12 - 7
{Effect of levofloxacine on cell elements of the lung tissue and on the growth of drug resistant Mycobacteria tuberculosis}; Sokolova GB et al.; Levofloxacin effect on morphokinetic parameters of the murine lung tissue culture was investigated . This model allowed also to evaluate the levofloxacin activity against drug-resistant intracellular and extracellular Mycobacterium tuberculosis: MBC was 0.5-1.0 mg/l . Bactericidal effect started on the 3d day of incubation and was maximal on 7th day . Tissue cells vitality was not changed . Combination of levofloxacin with antimycobacterial drugs of first choice was investigated: isoniazid demonstrated synergistic effect, pyrazinamide also demonstrated synergistic effect . Combination of levofloxacin with rifampicin was antagonistic.

What Is Staphylococcus Aureus?, What Is Cell Biology?, What Is Biofilter?, What Is Amino Acid?, What Is Rhizobia?, r, Microbe, r, Microbes, n, Bacterium, r, Microorganism, r, Bacteria, r, Escherichia coli, i, Candida albicans, o, Pseudomonas aeruginosa, s, Bacteriophage, o, Anaerobe, c, Enterobacters, s, Bacillus subtilis, r, Bacillus subtilis, c, Bacillus, s, Bacteriophages, o, Streptomycin, e, Microbial, a, Pasteurella, e, Escherichia coli, s, Corynebacter, i, Haemophilus, o, Salmonella typhimurium, r, Bacillus, e, Bacteria, n, Antibiotics, s, Escherichia coli

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005