Microbes Infect, 2003 Apr, 5(5), 449 - 56 Medical and economic impact of extraintestinal infections due to Escherichia coli: focus on an increasingly important endemic problem; Russo TA et al.; Escherichia coli is probably the best-known bacterial species and one of the most frequently isolated organisms from clinical specimens . Despite this, underappreciation and misunderstandings exist among medical professionals and the lay public alike regarding E . coli as an extraintestinal pathogen . Underappreciated features include (i) the wide variety of extraintestinal infections E . coli can cause, (ii) the high incidence and associated morbidity, mortality, and costs of these diverse clinical syndromes, (iii) the pathogenic potential of different groups of E . coli strains for causing intestinal versus extraintestinal disease, and (iv) increasing antimicrobial resistance . In this era in which health news often sensationalizes uncommon infection syndromes or pathogens, the strains of E . coli that cause extraintestinal infection are an increasingly important endemic problem and underappreciated "killers" . Billions of health care dollars, millions of work days, and hundreds of thousands of lives are lost each year to extraintestinal infections due to E . coli . New treatments and prevention measures will be needed for improved outcomes and a diminished disease burden. Nat Prod Res, 2003 Jun, 17(3), 207 - 14 Synthesis and antimicrobial activities of some isocoumarin and dihydroisocoumarin derivatives; Hussain M et al.; A new series of the isocoumarin derivatives of ibuprofen, fluribiprofen, naproxen, valproic acid and 1-naphthoic acid have been synthesized via condensation of homophthalic acid with their respective acid chlorides . The conversion of the latter two isocoumarins into (dl)-3,4-dihydroisocoumarins has also been achieved . Most of the synthesized compounds showed antifungal and antibacterial activities. Cell Mol Life Sci, 2003 Mar, 60(3), 536 - 49 Phylogeny, processing and expression of the rat cathelicidin rCRAMP: a model for innate antimicrobial peptides; Termen S et al.; A database search identified a rat cDNA clone which phylogenetic analysis revealed to encode a cathelicidin most similar to mouse cathelicidin CRAMP . The analysis also showed that the evolutionary pattern of the cathelicidin family is lineage specific . The rat cathelicidin is called rCRAMP . Its peptide was isolated from granulocytes, and determined to be 43 amino acids long by mass spectrometry and N-terminal sequencing . Synthetic rCRAMP had antimicrobial activity . The expression of rCRAMP was investigated by reverse-transcriptase polymerase chain reaction followed by Southern hybridization and by Western blot analysis . rCRAMP was identified in granulocytes, thymus, testis, lung, mouth mucosa, tongue, oesophagus, colon, caecum and small intestine, a distribution similar to cathelicidins of mouse and human . The rat is a small laboratory animal with additional disease models available compared to the mouse . Our results open up the possibility to use the rat as a model system to study responses connected to cathelicidin expression in health and disease. Int J Prosthodont, 2003 Mar-Apr, 16(2), 141 - 4 Cytotoxicity of antimicrobial tissue conditioners containing silver-zeolite; Abe Y et al.; PURPOSE: The purpose of this study was to determine the effects of incorporating antimicrobial silver-zeolite on the in vitro cytotoxicities of five tissue conditioners against the living dermal model, which consisted of normal human dermal fibroblasts in a collagen lattice . MATERIALS AND METHODS: Samples of each tissue conditioner containing silver-zeolite (2 wt% and 5 wt%) were prepared; samples without silver-zeolite were used as a control . Cytotoxicity of each sample against the living dermal model was evaluated by cell viability with MTT assay . RESULTS: Cell viabilities for Visco-Gel, GC Soft-Liner, FITT, and SR-Ivoseal decreased with increasing silver-zeolite content . Cell viability for Shofu Tissue Conditioner showed no significant difference with degree of silver-zeolite incorporation and had higher values than other tissue conditioners . CONCLUSION: Individual compounds eluted from tissue conditioners and silver ions released from silver-zeolite might cause such cytotoxicity . The results suggest that Shofu Tissue Conditioner is a possible candidate for a novel antimicrobial tissue conditioner containing silver-zeolite because it showed the highest cell viability and the smallest influence of incorporating silver-zeolite. Med Microbiol Immunol (Berl), 2003 May, 192(2), 71 - 8 Epub 2002 Jun 20. Antifungal activity of macrophages engineered to produce IFNgamma: inducibility by picolinic acid; Mucci A et al.; Macrophages are important antimicrobial effectors, whose efficacy is greatly enhanced by interferon-gamma (IFNgamma) . We recently engineered a mouse macrophage cell line to express the IFNgamma gene in a inducible manner . Such macrophages, Mphi10, include a construct containing the IFNgamma gene under the control of the synthetic promoter HRE3x-Tk . Picolinic acid (PA) is a catabolite of tryptophan, known to exert costimulatory activities on macrophages and expected to act on transcriptional elements within HRE3x-Tk promoter . Since evidence exists on the efficacy of engineered macrophages as carriers of therapeutic genes against tumors, we tested Mphi10, under basal conditions and following exposure to PA, as IFNgamma-producing cells in in vitro models of fungal infection . We found that Mphi10 constitutively exhibited anticryptococcal and anticandidal activity, low but detectable levels of IFNgamma mRNA and undetectable levels of nitric oxide synthase (iNOS) transcripts . Treatment with PA caused time-dependent enhancement of antifungal activity . The phenomenon was associated with the induction of both IFNgamma and iNOS gene expression and was followed by IFNgamma and NO production . The effect of the Mphi10-produced IFNgamma on other cells was also investigated by a transwell co-culture system . A major enhancement of phagocytosis and antifungal activity was observed in BV2 microglial cells that had been co-cultured with Mphi10 . Such an increase was only evident when Mphi10 had been pretreated with PA and was abrogated by concomitant addition of anti-IFNgamma antibodies . In conclusion, we show that Mphi10 respond to PA with the production of IFNgamma, which retains the ability to induce antifungal activity in the producing macrophages as well as in other macrophage populations . The potential use of Mphi10 as vectors for therapeutic genes in infectious diseases is discussed. Chem Pharm Bull (Tokyo), 2003 May, 51(5), 612 - 3 Bahiensol, a new glycerolipid from a cultured myxomycete Didymium bahiense var . bahiense; Misono Y et al.; Bahiensol (1), a new glycerolipid with antimicrobial activity has been isolated from a cultured plasmodium of myxomycete Didymium bahiense var . bahiense and its planar structure was elucidated by spectral data. Proc Natl Acad Sci U S A, 2003 May 27, 100(11), 6724 - 9 Epub 2003 May 07. Adoptive transfer of apoptotic splenocytes worsens survival, whereas adoptive transfer of necrotic splenocytes improves survival in sepsis; Hotchkiss RS et al.; In sepsis, both necrotic and apoptotic cell death can occur . Apoptotic cells induce anergy that could impair the host response, whereas necrotic cells cause immune activation that might result in enhanced antimicrobial defenses . We determined whether adoptive transfer of apoptotic or necrotic cells impacted survival in a clinically relevant sepsis model . We also evaluated the effects of adoptive transfer of apoptotic or necrotic cells on the prototypical TH1 and TH2 cytokines IFN-gamma and IL-4, respectively . C57BL6/J mice had adoptive transfer of apoptotic (irradiated) or necrotic (freeze thaw) splenocytes . Controls received saline . Apoptotic cells greatly increased mortality, whereas necrotic splenocytes markedly improved survival, P < or = 0.05 . The contrasting effects that apoptotic or necrotic cells exerted on survival were mirrored by opposite effects on splenocyte IFN-gamma production with greatly decreased and increased production, respectively . Importantly, either administration of anti-IFN-gamma antibodies or use of IFN-gamma knockout mice prevented the survival benefit occurring with necrotic cells . This study demonstrates that the type of cell death impacts survival in a clinically relevant model and identifies a mechanism for the immune suppression that is a hallmark of sepsis . Necrotic cells (and likely apoptotic cells) exert their effects via modulation of IFN-gamma Urology, 2003 May, 61(5), 1037 - 41 Preventive effect of pentoxifylline on renal scarring in rat model of pyelonephritis; Yagmurlu A et al.; OBJECTIVES: To evaluate the efficiency of pentoxifylline (PTX), a methyl xanthine derivative, in preventing renal scar formation after the induction of pyelonephritis in an experimental rat model with delayed antimicrobial therapy . METHODS: An inoculum of 1 x 10(9) colony-forming units/0.1 mL of the K-12 strain of Escherichia coli, which has both type 1 and P pili, was injected directly into both renal parenchyma of Wistar rats (n = 40) . Group 1 (control) received isotonic saline instead of bacterial solution (n = 10) . Four equal groups were then formed: group 2 was not treated and group 3 was treated only with ciprofloxacin for 5 days, starting 3 days after bacterial inoculation; in group 4, 50 mg/kg of PTX, and in group 5, PTX (50 mg/kg) and ciprofloxacin (15 mg/kg) together were administered intraperitoneally for 5 days, starting 3 days after bacterial inoculation . Six weeks after bacterial inoculation, all the rats were killed, and both kidneys were examined histopathologically for renal scarring . RESULTS: Delayed treatment with antibiotics had no effect on scarring compared with the untreated controls . However, the addition of PTX to the delayed antibiotic therapy significantly inhibited renal scarring compared with the untreated or antibiotic-only groups (P <0.05) . CONCLUSIONS: These results suggest that PTX is effective in preventing renal scar formation in pyelonephritis when the initiation of antimicrobial treatment is delayed in this rat model of pyelonephritis. Nat Prod Rep, 2003 Apr, 20(2), 252 - 73 Recent advances in antimicrobial nucleoside antibiotics targeting cell wall biosynthesis; Kimura K et al.; The biosynthesis of peptidoglycan is essential for all bacteria and has no counterpart in eukaryotic cells . It is one of the prime targets for antibiotic chemotherapy, especially phospho-MurNAc-pentapeptide translocase (translocase I) is a fascinating target in which there is no commercial antibiotic . In this review we will describe three nucleoside translocase I inhibitors, mureidomycin, tunicamycin and liposidomycin. Biol Proced Online, 2001 Aug 4, 3, 8 - 18 Cytochemical techniques and energy-filtering transmission electron microscopy applied to the study of parasitic protozoa; Vannier-Santos MA et al.; The study of parasitic protozoa plays a major role in cell biology, biochemistry and molecular biology . Numerous cytochemical techniques have been developed in order to unequivocally identify the nature of subcellular compartments . Enzyme and immuno-cytochemistry allow the detection of, respectively, enzymatic activity products and antigens in particular sites within the cell . Energy-filtering transmission electron microscopy permits the detection of specific elements within such compartments . These approaches are particularly useful for studies employing antimicrobial agents where cellular compartments may be destroyed or remarkably altered and thus hardly identified by standard methods of observation . In this regard cytochemical and spectroscopic techniques provide valuable data allowing the determination of the mechanisms of action of such compounds. J Clin Microbiol, 2003 May, 41(5), 1951 - 6 Validation of the automated reading and incubation system with Sensititre plates for antimicrobial susceptibility testing; Chapin KC et al.; The present study compared the antimicrobial susceptibility testing (AST) results generated by the Automated Incubation and Reading System (ARIS) with custom Sensititre plates (TREK Diagnostic Systems, Cleveland, Ohio) and MicroScan PC10 GP and NUMIC10 GN plates interpreted with the WalkAway-96 system (Dade Behring, West Sacramento, Calif.) for gram-positive (GP) and gram-negative (GN) organisms as part of an in-house validation . A total of 326 isolates (3,689 antimicrobial agent-organism combinations) were evaluated . Sensititre plates were inoculated according to the instructions of the manufacturer with a suspension adjusted to a 0.5 McFarland standard, while the Prompt Inoculation System was used for the MicroScan plates . ARIS and the WalkAway system were used for automated reading of the Sensititre and MicroScan plates, respectively, at 18 to 24 h . The results were analyzed for essential (+/-1 twofold dilution) and categorical (sensitive, intermediate, or resistant) agreements . Plates that resulted in ARIS interpretations with major (falsely resistant) or very major (falsely susceptible) errors compared to the results obtained with the WalkAway system were read manually to corroborate instrument readings . Isolates for which very major or major errors were obtained and for which the results were not resolved by manual reading were retested in parallel . Isolates for which very major or major errors were obtained and for which the results were not resolved upon repeat testing were tested by the National Committee for Clinical Laboratory Standards M7-A5 frozen reference microdilution method . Essential agreement was 95.8% for 246 GN isolates . The following categorical error rates were obtained for the GN isolates: 1.3% minor errors, 0% major errors, and 0.4% very major errors . For 95 GP isolates, there was 93.5% essential agreement . Categorical error rates for GP isolates were 0.9% minor errors, 0.6% major errors, and 0.4% very major errors . ARIS-Sensititre is a diagnostic system feasible for use for automated AST in a clinical laboratory. Genome Biol . 2003;4(5):R31 . Epub 2003 Apr 17. Duplication and selection in the evolution of primate beta-defensin genes; Semple CA et al.; BACKGROUND: Innate immunity is the first line of defense against microorganisms in vertebrates and acts by providing an initial barrier to microorganisms and triggering adaptive immune responses . Peptides such as beta-defensins are an important component of this defense, providing a broad spectrum of antimicrobial activity against bacteria, fungi, mycobacteria and several enveloped viruses . Beta-defensins are small cationic peptides that vary in their expression patterns and spectrum of pathogen specificity . Disruptions in beta-defensin function have been implicated in human diseases, including cystic fibrosis, and a fuller understanding of the variety, function and evolution of human beta-defensins might form the basis for novel therapies . Here we use a combination of laboratory and computational techniques to characterize the main human beta-defensin locus on chromosome 8p22-p23 . RESULTS: In addition to known genes in the region we report the genomic structures and expression patterns of four novel human beta-defensin genes and a related pseudogene . These genes show an unusual pattern of evolution, with rapid divergence between second exon sequences that encode the mature beta-defensin peptides matched by relative stasis in first exons that encode signal peptides . CONCLUSIONS: We conclude that the 8p22-p23 locus has evolved by successive rounds of duplication followed by substantial divergence involving positive selection, to produce a diverse cluster of paralogous genes established before the human-baboon divergence more than 23 million years ago . Positive selection, disproportionately favoring alterations in the charge of amino-acid residues, is implicated as driving second exon divergence in these genes. Pharmacoepidemiol Drug Saf, 2003 Apr-May, 12(3), 243 - 7 Trends in antimicrobial utilization at a Spanish general hospital during a 5-year period; Hermosilla Najera L et al.; PURPOSE: Antimicrobials are a major part of hospital pharmacy budgets and must be considered in resource planning and spending projections . This study describes the profile of antibiotic use at a medium-sized hospital (by examining the ICU separately) and analyses its evolution over the period 1996-2000 . METHODS: Descriptive and retrospective study . Pharmacy records were reviewed to identify oral and parenteral antimicrobial agents administered to inpatients . Results were expressed in Daily Defined Doses (DDD) per 100 stays and day . RESULTS: During the 5-year study period 176.162 DDD/100 s-d of antibiotics were consumed in the ICU, whereas in the rest of the hospital usage was much lower (54.438 DDD/100 s-d) . Aminoglycosides, cephalosporins, penicillins, glycopeptides and carbapenems were the most commonly used groups of antimicrobials in the ICU, and penicillins, cephalosporins, trimethoprim/sulfonamide combinations, aminoglycosides and quinolones in the rest of the hospital . CONCLUSIONS: ICUs have some special features which make them different to the other inpatient areas . Because of that fact we consider it important to study this specific patient-care area separately. Pharmacoepidemiol Drug Saf, 2003 Apr-May, 12(3), 221 - 5 A study of drug use during pregnancy in a teaching hospital in western Nepal; Das B et al.; Information on the use of drugs during pregnancy is scarce and rather anecdotal . Careful consideration of the benefit to the mother and the risk to the fetus is required when prescribing drugs during pregnancy . The aim of this study was to gain knowledge on this issue in western Nepal . 2156 prescriptions of pregnant women were collected at random from the antenatal care (ANC) in obstetrics out-patient department (OPD) at Manipal Teaching Hospital (MTH), Nepal and analyzed for this study . The mean maternal age and hemoglobin concentration were 25 years and 12.21 g/dl, respectively . Twenty-three percent women attended obstetric OPD due to maternal disorders other than routine ANC (77%) . Problem oriented drug use was due to nausea/vomiting (4.7%), dyspepsia (3.1%), and per vaginal spotting/bleeding (3.4%), mainly . Most of the women got 2-3 drugs and commonly included nutritional supplementation and tetanus toxoid . The average number of drugs/prescription was 2.00, 15.37% and 64.8% drugs were prescribed by generic name and as fixed dose combinations, respectively . The most commonly prescribed drugs were nutritional supplements like iron, folate, calcium, vitamins (72.8%), followed by tetanus toxoid (12.4%), gastrointestinals (5%), antimicrobials (4.6%), etc . Though, the selection of drugs was rational in most of the cases, some anomalies were observed and discussed with the clinicians . Our data reflect the general extent and prescribing pattern for those Nepalese pregnant women attending hospital in western Nepal. Dent Today, 2003 Apr, 22(4), 107 - 13 Periodontal disease and diabetes mellitus . Bidirectional relationship; Mealey BL et al.; Periodontitis is a common problem in patients with diabetes . The relationship between these 2 maladies appears bidirectional--insofar that the presence of one condition tends to promote the other, and that the meticulous management of either may assist treatment of the other . Both diabetes and periodontitis can stimulate the chronic release of proinflammatory cytokines that have a deleterious effect on periodontal tissues . The chronic systemic elevation of proinflammatory cytokines caused by periodontitis may even predispose individuals to the development of type 2 diabetes . Mechanical treatment of periodontitis (scaling and root planing), when combined with short-term administration of therapeutic levels of tetracycline-type antimicrobials, can temporarily improve glycemic control in diabetic patients, especially in those with advanced forms of periodontitis and poor glycemic control before treatment . The biochemical mechanisms suggested by these studies imply that other periodontal procedures designed to rid patients of periodontal pathogens may also improve the management of diabetes . Therefore, the authors suggest that periodontal patients with diabetes be treated in consultation with a periodontist (Figures 3a through 4b). Infez Med, 1998, 6(1), 48 - 53 Type I necrotising fasciitis of heel in a diabetic patient; Gokalan I et al.; Necrotising fasciitis is a potentially fatal clinical disease caused by infection with various bacteria . Diabetes mellitus, one of the predisposing factors for necrotising fasciitis is also an important factor in the progression of the illness by increasing its morbidity and mortality rate . In this case report, a 54 year old diabetic woman with type-I necrotising fasciitis of the heel was treated with extensive debridments, broad spectrum antimicrobial therapy and intensive regulation of blood glucose levels . The resulting new positive data pertaining to the morbidity and mortality of this severe disease suggest that the old management of this disease should be re-examined. Infez Med, 1998, 6(1), 8 - 17 {Infectious diarrhoeas}; Sagnelli E et al.; The infective diarrhoea show a high morbility in infants of all countries and a high mortality in infants and children of developing countries . In developed countries there is a high frequency of "traveler's diarrhoea" . The different pathogenesis of infectious diarrhoeas (intestinal invasion, intestinal adherence, citonecrotic or citotonic toxins) influences the clinical presentation (watery diarrhoea or diarrhoea with blood and mucos, presence or assence of fever) . The key factor in the management of acute watery diarrhoea is the restoration of water and electrolyte balance; but frequently an antimicrobial treatment is needed. J Exp Med, 2003 May 5, 197(9), 1213 - 9 Deficiencies of GM-CSF and interferon gamma link inflammation and cancer; Enzler T et al.; Chronic inflammation contributes to carcinogenesis, but the underlying mechanisms are poorly understood . We report that aged granulocyte-macrophage colony stimulating factor (GM-CSF)-deficient mice develop a systemic lupus erythematosis (SLE)-like disorder associated with the impaired phagocytosis of apoptotic cells . Concurrent deficiency of interferon (IFN)-gamma attenuates the SLE, but promotes the formation of diverse hematologic and solid neoplasms within a background of persistent infection and inflammation . Whereas activated B cells show a resistance to fas-induced apoptosis, antimicrobial therapy prevents lymphomagenesis and solid tumor development . These findings demonstrate that the interplay of infectious agents with cytokine-mediated regulation of immune homeostasis is a critical determinant of cancer susceptibility. J Biol Chem, 2003 Oct 3, 278(40), 38557 - 65 Epub 2003 May 05. Molecular cloning and characterization of CYP719, a methylenedioxy bridge-forming enzyme that belongs to a novel P450 family, from cultured Coptis japonica cells; Ikezawa N et al.; Two cytochrome P450 (P450) cDNAs involved in the biosynthesis of berberine, an antimicrobial benzylisoquinoline alkaloid, were isolated from cultured Coptis japonica cells and characterized . A sequence analysis showed that one C . japonica P450 (designated CYP719) belonged to a novel P450 family . Further, heterologous expression in yeast confirmed that it had the same activity as a methylenedioxy bridge-forming enzyme (canadine synthase), which catalyzes the conversion of (S)-tetrahydrocolumbamine ((S)-THC) to (S)-tetrahydroberberine ((S)-THB, (S)-canadine) . The other P450 (designated CYP80B2) showed high homology to California poppy (S)-N-methylcoclaurine-3'-hydroxylase (CYP80B1), which converts (S)-N-methylcoclaurine to (S)-3'-hydroxy-N-methylcoclaurine . Recombinant CYP719 showed typical P450 properties as well as high substrate affinity and specificity for (S)-THC . (S)Scoulerine was not a substrate of CYP719, indicating that some other P450, e.g . (S)-cheilanthifoline synthase, is needed in (S)-stylopine biosynthesis . All of the berberine biosynthetic genes, including CYP719 and CYP80B2, were highly expressed in selected cultured C . japonica cells and moderately expressed in root, which suggests coordinated regulation of the expression of biosynthetic genes. Appl Environ Microbiol, 2003 May, 69(5), 2563 - 7 Enhanced killing of Acanthamoeba cysts with a plant peroxidase-hydrogen peroxide-halide antimicrobial system; Hughes R et al.; The activity of H(2)O(2) against the resistant cyst stage of the pathogenic free-living amoeba Acanthamoeba was enhanced by the addition of KI and either horseradish peroxidase or soybean peroxidase or, to a lesser degree, lactoperoxidase . This resulted in an increase in the cysticidal activity of 3% (wt/vol) H(2)O(2), and there was >3-log killing in 2 h, compared with the 6 h required for comparable results with the peroxide solution alone (P < 0.05) . With 2% H(2)O(2), enhancement was observed at all time points (P < 0.05), and total killing of the cyst inoculum occurred at 4 h, compared with 6 h for the peroxide alone . The activity of sublethal 1% H(2)O(2) was enhanced to give 3-log killing after 8 h of exposure (P < 0.05) . No enhancement was obtained when KCl or catalase was used as a substitute in the reaction mixtures . The H(2)O(2) was not neutralized in the enhanced system during the experiments . However, in the presence of a platinum disk used to neutralize H(2)O(2) in contact lens care systems, the enhanced 2% H(2)O(2) system gave 2.8-log killing after 6 h or total cyst killing by 8 h, and total neutralization of the H(2)O(2) occurred by 4 h . In contrast, 2% H(2)O(2) alone resulted in <0.8-log killing of cysts in the presence of the platinum disk due to rapid (<1 h) neutralization of the peroxide . Our observations could result in significant improvement in the efficacy of H(2)O(2) contact lens disinfection systems against Acanthamoeba cysts and prevention of acanthamoeba keratitis. J Microbiol Methods, 2003 Jul, 54(1), 111 - 5 Potassium and tetraphenylphosphonium ion-selective electrodes for monitoring changes in the permeability of bacterial outer and cytoplasmic membranes; Yasuda K et al.; A tetraphenylphosphonium ion (TPP(+))-selective electrode, originally developed as a membrane potential indicator, is useful for measuring increases in the permeability of bacterial outer membranes induced by antimicrobial agents . The combination of this electrode with a potassium ion-selective electrode enabled us to determine changes in the permeability of bacterial outer and cytoplasmic membranes simultaneously . Outer membrane permeabilization induced by antimicrobial agents, chlorhexidine and polyhexamethylene biguanide (PHMB), as monitored with the TPP(+) electrode, correlated closely with the ability of the agents to release lipopolysaccharide (LPS) from the outer membrane. J Nutr, 2003 May, 133(5 Suppl 2), 1656S - 1660S Infection-related morbidities in the mother, fetus and neonate; Bergstrom S; Only partially understood host defense mechanisms operate against infections affecting maternal and fetal morbidity . Subclinical ascending infections through the lower female genital tract are predominant worldwide . Important micronutrient deficiencies may prevail in low-income countries where these infections are much more common than in high-income countries . Important morbidities related to poor perinatal outcome both for the mother and for the fetus and newborn comprise preterm birth, prelabor rupture of membranes, placental abruption (predelivery detachment of the placenta), postpartum sepsis and maternal anemia . In the fetus, sepsis and intrauterine growth retardation are suspected to be consequences of ascending maternal infections . In the newborn, septicemia and respiratory disorders as well as some neurological disorders seem to be consequences of such ascending genital infections in the pregnant woman . It is concluded that much more attention should be given to efforts to elucidate the host defense mechanisms and antimicrobial barriers from the vagina through the cervix, fetal membranes and amniotic fluid including the early fetal immunocompetence in the second and the third trimester of pregnancy. Appl Microbiol Biotechnol, 2003 Jul, 62(1), 21 - 6 Epub 2003 May 01. epsilon-Poly-L-lysine: microbial production, biodegradation and application potential; Yoshida T et al.; epsilon-Poly-L-lysine (epsilon-PL) is a homo-poly-amino acid characterized by the peptide bond between the carboxyl and epsilon-amino groups of L-lysine . epsilon-PL shows a wide range of antimicrobial activity and is stable at high temperatures and under both acidic and alkaline conditions . The mechanism of the inhibitory effect of epsilon-PL on microbial growth is the electrostatic adsorption to the cell surface of microorganisms on the basis of its poly-cationic property . Due to this antimicrobial activity, epsilon-PL is now industrially produced in Japan as a food additive by a fermentation process using Streptomyces albulus . In spite of the practical application of epsilon-PL, the biosynthetic mechanisms of epsilon-PL have not been clarified at all . epsilon-PL producers commonly possess membrane-bound epsilon-PL-degrading aminopeptidase, which might play a role in self-protection. Infez Med, 1999, 7(1), 39 - 42 Conservative treatment of diabetic foot infections; Kara Gokalan I et al.; Foot infections commonly occur in diabetic patients, constituting a serious problem . 15% of diabetic have foot infections during their life-time . In previous years these life- threatening infections in 50-70% of patients resulted in limb amputation . Today, using an aggressive surgical approach, broad spectrum antimicrobial chemotherapy and intensive regulation of blood glucose level, the amputation rate has decreased to 5% . During the last 2 years, we evaluated 13 diabetic foot infections . 6 of the presented cases were grade 4 and 5, 3 of them grade 3 and 4 of them grade 2 . All of the patients were managed by multidisciplinary approach consisting of plastic and reconstructive surgery, endocrinology and infectious diseases specialists . All of the patients were monitored with daily wound care, regulation of blood glucose levels and administration of broad spectrum antibiotics . Only one of them required amputation . For 12 of the cases after the infection was controlled, the defects were reconstructed by plastic surgical procedures . We retrospectively analysed them to review diabetic foot infections and factors affecting prognosis. Pediatrics, 2003 May, 111(5 Pt 1), e586 - 9 Paranasal sinus findings in children during respiratory infection evaluated with magnetic resonance imaging; Kristo A et al.; OBJECTIVE: The spreading of acute respiratory infection into the paranasal sinuses in children is poorly defined . The main objective of this study was to evaluate the frequency and spontaneous resolution of paranasal sinus abnormalities in children with acute respiratory infection using magnetic resonance imaging (MRI) . METHODS: We examined 60 children with MRI (mean age: 5.7 years) with symptoms of acute respiratory infection . Twenty-six children with major abnormalities in the first MRI scan had a follow-up MRI taken 2 weeks later . RESULTS: The children had had symptoms of uncomplicated acute respiratory infection for an average of 6 days before the first examination (mean duration: 6.5; standard deviation: 3.0) . Approximately 60% of the children had major abnormalities in their maxillary and ethmoidal sinuses, 35% in the sphenoidal sinuses, and 18% in the frontal sinuses . The most common abnormal finding was mucosal swelling . The mean overall MRI scores correlated significantly with the symptom scores (r(s) = 0.3) . Of the individual symptoms, nasal obstruction, nasal discharge, and fever were significantly related to the MRI scores . Among the 26 children with major abnormalities in the first MRI, the findings subsequently improved significantly (mean {standard deviation} score: 12.7 {5.6} to 5.7 {5.2}), irrespective of the resolution of symptoms . CONCLUSIONS: These observations indicate that acute respiratory infection mostly spreads into the paranasal sinuses of children in the form of mucosal edema and that these abnormalities tend to resolve spontaneously without antimicrobial treatment. Pediatrics, 2003 May, 111(5 Pt 1), 1061 - 7 Antibiotic treatment of acute otorrhea through tympanostomy tube: randomized double-blind placebo-controlled study with daily follow-up; Ruohola A et al.; OBJECTIVE: The role of routine antimicrobial treatment of acute middle-ear infections is under debate, because the efficacy of antimicrobials in the resolution of middle-ear fluid has not been unambiguously proven . Acute tube otorrhea is regarded as evidence of acute otitis media, and for methodologic reasons it was chosen to provide objectivity for diagnostics and outcome assessment . The objective of this study was to assess whether amoxicillin-clavulanate accelerates the resolution of acute tube otorrhea . DESIGN AND SETTING: Randomized, double-blind, placebo-controlled study in outpatient setting . PATIENTS: Volunteer sample of basically healthy 6- to 72-month-old children with a tympanostomy tube . Eligibility required having acute tube otorrhea of <48 hours' of duration and no prior treatment within the last 2 weeks . The mean age of the participants was 25 months; they had a history of 3 episodes of acute otitis media (median), and 99% had manifestations of a concomitant respiratory infection . Of 79 randomized patients, 7 were withdrawn because of adverse events; 66 patients completed the study . INTERVENTIONS: Amoxicillin-clavulanate (N = 34; 45 mg/kg/d) or matching placebo (N = 32) for 7 days and daily suction of middle-ear fluid through tympanostomy tube . MAIN OUTCOME MEASURES: Duration of acute tube otorrhea and duration of bacterial growth in middle-ear fluid . RESULTS: The median duration of tube otorrhea was significantly shorter in amoxicillin-clavulanate than in the placebo group (3 vs 8 days) . At the end of the 7-day medication period, tube otorrhea was resolved in 28 of 34 children receiving amoxicillin-clavulanate compared with 13 of 32 children on placebo (treatment-control difference 41%; 95% confidence interval, 20%-63%; number needed to treat, 2.4) . The median duration of bacterial growth in middle-ear fluid was shorter in amoxicillin-clavulanate than in the placebo group (1 vs 8 days) . CONCLUSIONS: Oral antibiotic treatment significantly accelerates the resolution of acute tube otorrhea by reducing bacterial growth in middle-ear fluid. Fitoterapia, 2003 Apr, 74(3), 308 - 11 Antimicrobial and cytotoxic constituents of Loranthus globosus; Sadik G et al.; (+)-Catechin, 3,4-dimethoxycinnamyl alcohol and 3,4,5-trimethoxycinnamyl alcohol were isolated from the barks of Loranthus globosus . All compounds showed significant antimicrobial and cytotoxic activities. Fitoterapia, 2003 Apr, 74(3), 298 - 301 Molluscicidal and antimicrobial activity of Solanum aculeastrum; Wanyonyi AW et al.; The methanolic extract of the fresh root bark and berries of Solanum aculeastrum showed significant activity against host snails of schistosomiasis . The berries extract was more potent with 100% snail kill at 50 ppm . Fractionation increased activity with 100% mortality at 20 ppm . The aqueous and methanolic extracts of the berries showed moderate antimicrobial activity, increasing by fractionation. Fitoterapia, 2003 Apr, 74(3), 291 - 3 Antimicrobial activity of Ixora coccinea leaves; Annapurna J et al.; Ether and methanol extracts of Ixora coccinea dry leaves were tested for their antimicrobial activity . Ether fraction was found to be more active than the methanol fraction. Fitoterapia, 2003 Apr, 74(3), 284 - 7 Antimicrobial study of essential oils of Ocimum gratissimum leaves and Zanthoxylum xanthoxyloides fruits from Cameroon; Ngassoum MB et al.; An investigation of antimicrobial activities of essential oils of fresh leaves of Ocimum gratissimum and the essential oil of the dried fruits of Zanthoxylum xanthoxyloides was carried out . The essential oils showed extensive inhibition zones and are, therefore, effective antimicrobial systems. Int J Antimicrob Agents, 2003 May, 21(5), 488 - 91 In vitro susceptibility of Arcobacter butzleri and Arcobacter cryaerophilus to different antimicrobial agents; Fera MT et al.; Seventeen strains of Arcobacter butzleri and thirteen of Arcobacter cryaerophilus, were tested for their antimicrobial susceptibility to 26 antimicrobial agents . Among beta-lactams agents in this study, imipenem was the most active agent against both A . butzleri and A . cryaerophilus isolates with MIC(90) values of 2 and 4 mg/l, respectively . The most active cephalosporin tested was cefepime, although it was more active against A . butzleri (MIC(90) 8 mg/l) than A . cryaerophilus (MIC(90) 64 mg/l) . Levofloxacin, marbofloxacin, enrofloxacin and ciprofloxacin were the best-performing fluoroquinolones against these species . Of the aminoglycosides, amikacin was the most active agent against both A . butzleri and A . cryaerophilus strains with MIC(90) values of 64 and 16 mg/l, respectively . All isolates showed high levels of resistance to penicillins, macrolides, chloramphenicol, trimethoprim and vancomycin. Int J Antimicrob Agents, 2003 May, 21(5), 434 - 40 Susceptibilities of different Actinobacillus actinomycetemcomitans strains to lactoperoxidase-iodide-hydrogen peroxide combination and different antibiotics; Ihalin R et al.; Actinobacillus actinomycetemcomitans has an important aetiological role in localized juvenile periodontitis and in progressive periodontitis in adults . A . actinomycetemcomitans is found mainly in periodontal pockets but also in whole saliva, a potential transmission medium . It is sensitive to peroxidase-halide systems, but the differences between periodontitis associated clinical isolates and type strains are unclear . The sensitivities of these 2 strain groups to lactoperoxidase (LP)-iodide (I(-))-hydrogen peroxide (H(2)O(2)) combinations were investigated, and the sensitivities were compared with the susceptibilities to four antibiotics . There was great variation between the sensitivities of different strains, but the 2 strain groups responded similarly . The LP (75 microg)-I(-) (100 nmol)-H(2)O(2) (1000 nmol) combination produced a similar degree of inhibition as 2 microg ampicillin . The LP-I(-) system might be a potential antimicrobial agent against A . actinomycetemcomitans transmission via saliva. J Am Vet Med Assoc, 2003 May 1, 222(9), 1241 - 7 West Nile virus encephalomyelitis in horses: 46 cases (2001); Porter MB et al.; OBJECTIVE: To determine signalment, clinical findings, results of diagnostic testing, outcome, and postmortem findings in horses with West Nile virus (WNV) encephalomyelitis . DESIGN: Retrospective study . ANIMALS: 46 horses with WNV encephalomyelitis . PROCEDURE: Clinical data were extracted from medical records of affected horses . RESULTS: On the basis of clinical signs and results of serologic testing, WNV encephalomyelitis was diagnosed in 46 of 56 horses with CNS signs . Significantly more males than females were affected . Increased rectal temperature, weakness or ataxia, and muscle fasciculations were the most common clinical signs . Paresis was more common than ataxia, although both could be asymmetrical and multifocal . Supportive treatment included anti-inflammatory medications, fluids, antimicrobials, and slinging of recumbent horses . Results of the IgM capture ELISA and the plaque reduction neutralization test provided a diagnosis in 43 horses, and only results of the plaque reduction neutralization test were positive in 3 horses . Mortality rate was 30%, and 71% of recumbent horses were euthanatized . One horse that had received 2 vaccinations for WNV developed the disease and was euthanatized . Follow-up communications with 19 owners revealed that most horses had residual deficits at 1 month after release from the hospital; abnormalities were resolved in all but 2 horses by 12 months after release . CONCLUSIONS AND CLINICAL RELEVANCE: Our findings were similar to those of previous WNV outbreaks in horses but provided additional clinical details from monitored hospitalized horses . Diagnostic testing is essential to diagnosis, treatment is supportive, and recovery rate of discharged ambulatory horses is < 100%. Microbiol Immunol, 2003, 47(3), 191 - 8 Coenzyme Q1-catalyzed luminol chemiluminescent assay for rapid antimicrobial susceptibility testing of Mycobacterium bovis; Yamashoji S; Coenzyme Q1 is herein proposed as the best catalyst among coenzymes Q and vitamins K for quinone-catalyzed luminol chemiluminescent assays applied to rapid determination of viability or rapid antimicrobial susceptibility tests of Mycobacterium bovis . Luminol chemiluminescence intensity (LCI) was determined 10 min after the incubation of M . bovis with coenzyme Q1, and was proportional to CFU (colony-forming unit)/ml in the range of 9,000 to 2,250,000 . LCI depended on the the production of the superoxide anion (O2-) rather than H2O2 during a 10-min incubation of M . bovis with coenzyme Q1, as superoxide dismutase reduced LCI more effectively than catalase . The minimal inhibitory concentrations (MICs) of 10 kinds of antituberculous agents estimated on the basis of decrease in LCI after one or two days' cultivation were in good agreement with MICs determined by turbidity analysis, which requires upwards of 1 week to complete. J Pharm Pharmacol, 2003 Mar, 55(3), 291 - 300 Antimicrobial activity of beta-lactam antibiotics against clinical pathogens after molecular inclusion in several cyclodextrins . A novel approach to bacterial resistance; Athanassiou G et al.; Recognition and uptake by specific cellular receptors and transport systems for cyclodextrins have been demonstrated . Based on this concept, natural and synthetically modified cyclodextrins were used as drug carriers . Several beta-lactam antibiotics were selected and their inclusion complexes with different cyclodextrins were prepared (molar ratio ranging from 1:1 to 1:3) . The complex formation, in aqueous solution, was monitored and optimum complexation conditions were selected . The inclusion of the active molecules in the cyclodextrin cavity was confirmed by (1)H NMR spectroscopy . Specific HPLC methods for the quantitation of antibiotics in the presence of cyclodextrins were developed and their chemical stability under complexation conditions was confirmed . Antimicrobial activity of drug-cyclodextrin complexes, in terms of minimum inhibitory concentration (MIC), were compared with the corresponding values of uncomplexed free molecules . A wide range of clinical pathogens and known beta-lactamase-producing strains were tested . The activity of the cyclodextrin-included antibiotics was increased, particularly against Gram-negative clinical strains . The nature and degree of substitution on cyclodextrin macromolecules may be the predominant factor in the observed improvement in antimicrobial activity . We believe that the proposed methodology is a novel approach to the microbial resistance problem and will trigger research towards the development of new cyclodextrin derivatives bearing the ability to increase the uptake of included antimicrobial molecules through intensification of the corresponding molecular recognition phenomena. J Med Chem, 2003 May 8, 46(10), 1824 - 30 Novel pyrazolo{3,4-d}pyrimidine-based inhibitors of Staphlococcus aureus DNA polymerase III: design, synthesis, and biological evaluation; Ali A et al.; 6-Anilinopyrazolo{3,4-d}pyrimidin-4-ones are novel dGTP analogues that inhibit the replication-specific enzyme DNA polymerase III (DNA pol III) of Staphlococcus aureus and other Gram-positive (Gr+) bacteria . To enhance the potential of these inhibitors as antimicrobial agents, a structure-activity relationship was developed involving substitutions at the 2, 4, and pyrazolo NH positions . All of the new inhibitors were tested for their ability to inhibit S . aureus DNA pol III and the growth of several other Gr+ bacteria in culture . 2-Anilino groups with small hydrophobic groups in the meta or para position enhanced both antipolymerase and antimicrobial activity . 2-Benzyl-substituted inhibitors were substantially less active . Substitution in the 4-position by oxygen gave the optimal activity, whereas substitution at the pyrazolo NH was not tolerated . These pyrazolo{3,4-d}pyrimidine derivatives represent a novel class of antimicrobials with promising activities against Gr+ bacteria. Phytother Res, 2003 Apr, 17(4), 299 - 305 Pharmacological and toxicological properties of Nigella sativa; Ali BH et al.; The seeds of Nigella sativa Linn . (Ranunculaceae), commonly known as black seed or black cumin, are used in folk (herbal) medicine all over the world for the treatment and prevention of a number of diseases and conditions that include asthma, diarrhoea and dyslipidaemia . This article reviews the main reports of the pharmacological and toxicological properties of N . sativa and its constituents . The seeds contain both fixed and essential oils, proteins, alkaloids and saponin . Much of the biological activity of the seeds has been shown to be due to thymoquinone, the major component of the essential oil, but which is also present in the fi ed oil . The pharmacological actions of the crude extracts of the seeds (and some of its active constituents, e.g . volatile oil and thymoquinone) that have been reported include protection against nephrotoxicity and hepatotoxicity induced by either disease or chemicals . The seeds/oil have antiinflammatory, analgesic, antipyretic, antimicrobial and antineoplastic activity . The oil decreases blood pressure and increases respiration . Treatment of rats with the seed extract for up to 12 weeks has been reported to induce changes in the haemogram that include an increase in both the packed cell volume (PCV) and haemoglobin (Hb), and a decrease in plasma concentrations of cholesterol, triglycerides and glucose . The seeds are characterized by a very low degree of toxicity . Two cases of contact dermatitis in two individuals have been reported following topical use . Administration of either the seed extract or its oil has been shown not to induce significant adverse effects on liver or kidney functions . It would appear that the beneficial effects of the use of the seeds and thymoquinone might be related to their cytoprotective and antioxidant actions, and to their effect on some mediators of inflammation . Biophys J, 2003 May, 84(5), 3052 - 60 MSI-78, an analogue of the magainin antimicrobial peptides, disrupts lipid bilayer structure via positive curvature strain; Hallock KJ et al.; In this work, we present the first characterization of the cell lysing mechanism of MSI-78, an antimicrobial peptide . MSI-78 is an amphipathic alpha-helical peptide designed by Genaera Corporation as a synthetic analog to peptides from the magainin family . (31)P-NMR of mechanically aligned samples and differential scanning calorimetry (DSC) were used to study peptide-containing lipid bilayers . DSC showed that MSI-78 increased the fluid lamellar to inverted hexagonal phase transition temperature of 1,2-dipalmitoleoyl-phosphatidylethanolamine indicating the peptide induces positive curvature strain in lipid bilayers . (31)P-NMR of lipid bilayers composed of MSI-78 and 1-palmitoyl-2-oleoyl-phosphatidylethanolamine demonstrated that the peptide inhibited the fluid lamellar to inverted hexagonal phase transition of 1-palmitoyl-2-oleoyl-phosphatidylethanolamine, supporting the DSC results, and the peptide did not induce the formation of nonlamellar phases, even at very high peptide concentrations (15 mol %) . (31)P-NMR of samples containing 1-palmitoyl-2-oleoyl-phosphatidylcholine and MSI-78 revealed that MSI-78 induces significant changes in the bilayer structure, particularly at high peptide concentrations . At lower concentrations (1-5%), the peptide altered the morphology of the bilayer in a way consistent with the formation of a toroidal pore . Higher concentrations of peptide (10-15%) led to the formation of a mixture of normal hexagonal phase and lamellar phase lipids . This work shows that MSI-78 induces significant changes in lipid bilayers via positive curvature strain and presents a model consistent with both the observed spectral changes and previously published work. Arch Intern Med, 2003 Apr 28, 163(8), 972 - 8 Unnecessary use of antimicrobials in hospitalized patients: current patterns of misuse with an emphasis on the antianaerobic spectrum of activity; Hecker MT et al.; BACKGROUND: Unnecessary use of antimicrobials contributes to the emergence and dissemination of antimicrobial-resistant nosocomial pathogens in part through elimination of normal anaerobic bacterial flora that inhibit overgrowth of pathogenic microorganisms . METHODS: A prospective observational study was conducted in a 650-bed, university-affiliated hospital . All adult nonintensive care inpatients for whom new antimicrobials were prescribed during a 2-week period were monitored throughout their hospitalization . We examined how often antimicrobials, in particular those with antianaerobic activity, were used unnecessarily . The reasons for unnecessary therapy were assessed and common patterns of unnecessary use were identified . RESULTS: A total of 1941 antimicrobial days of therapy were prescribed for 129 patients . A total of 576 (30%) of the 1941 days of therapy were deemed unnecessary . The most common reasons for unnecessary therapy included administration of antimicrobials for longer than recommended durations (192 days of therapy), administration of antimicrobials for noninfectious or nonbacterial syndromes (187 days of therapy), and treatment of colonizing or contaminating microorganisms (94 days of therapy) . Antianaerobic agents accounted for 203 (35%) of the 576 unnecessary antimicrobial days of therapy, and these agents were also frequently prescribed (98 days of therapy) when equally efficacious alternative regimens with minimal antianaerobic activity were available . CONCLUSIONS: In our institution, hospitalized patients frequently received unnecessary antimicrobial therapy, and antianaerobic agents were often prescribed when this spectrum of activity was not indicated. Int J Biol Macromol, 2003 Mar, 32(1-2), 1 - 9 Conformational changes of pediocin in an aqueous medium monitored by fourier transform infrared spectroscopy: a biological implication; Gaussier H et al.; Fourier transform infrared (FTIR) spectroscopy was used to investigate the secondary structure of pediocin PA-1 in different aqueous media in relation to its antimicrobial activity . The experiments were performed at pD (pH meter corrected for deuterium isotope effect) 6, 7, and 8 and during a heating-cooling cycle of 20-80 degrees C . At pD 6, (i.e . pediocin's most active form), the FTIR results show that pediocin adopts an unordered structure with a small contribution of beta-turn . After a heating-cooling cycle, thermally-induced changes in pediocin are reversed and its activity is maintained . Increasing the pD to 7 and 8 leads to a more ordered secondary structure . For these two pD values, an increase in temperature induces an irreversible aggregation of protein as revealed by the amide I' band . The analysis of the Tyr region provides more insight into the aggregation process . In fact, it appears to be a two-step process, involving first the C (carboxy)-terminus of pediocin and then the N (amino)-terminus . This study reveals two major points: (1) the preservation of pediocin flexibility is essential for maintaining its activity; and (2) the aggregation of its C-terminus is sufficient to induce a loss of activity, suggesting that this region plays an important role in the activity of pediocin. Int J Infect Dis, 2002 Sep, 6(3), 182 - 6 Brucellosis in children: clinical observations in 115 cases; Shaalan MA et al.; OBJECTIVE: Brucellosis is endemic in Saudi Arabia . This report summarizes the epidemiology of brucellosis in children . METHOD: A retrospective review was made of medical records of all patients admitted to King Fahad National Guard Hospital with brucellosis during the period from 1984 to 1995 . RESULTS: Children < or =12 years constituted 115/545 (21%) of the total brucellosis admissions . The mean age was 5.8 years and 64% of the patients were males . Consumption of unpasteurized milk (often from camel) was the main source of infection . In 70% the clinical picture was dominated by arthritis, 20% of patients presented with a non-specific febrile illness without localizing signs, and 10% had a febrile illness with uncommon presentations . Brucella serology was most helpful in making an early diagnosis . Initial titers of >1:640 were found in 90% of the cases . Bacteremia was observed in 45% and of the isolates speciated, 96% were Brucella melitensis . No increase in resistance to commonly used antimicrobials was noted during the 12-year study period . A combination of rifampin plus co-trimoxazole with or without streptomycin was used in two thirds of the patients . The overall rate of relapse was 9% and one patient died from neurobrucellosis . CONCLUSION: Brucellosis presents in various ways and should be included in the differential diagnosis of arthritis in endemic countries . Prevention should rely on education including on boiling raw milk. World J Gastroenterol, 2003 May, 9(5), 1126 - 8 Type 2 diabetes mellitus affects eradication rate of Helicobacter pylori; Sargyn M et al.; AIM: To study the eradication rate of Helicobacter pylori (Hp) in a group of type 2 diabetes and compared it with an age and sex matched non-diabetic group . METHODS: 40 diabetic patients (21 females, 19 males; 56+/-7 years) and 40 non-diabetic dyspeptic patients (20 females, 20 males; 54+/-9 years) were evaluated . Diabetic patients with dyspeptic complaints were referred for upper gastrointestinal endoscopies; 2 corpus and 2 antral gastric biopsy specimens were performed on each patient . Patients with positive Hp results on histopathological examination comprised the study group . Non-diabetic dyspeptic patients seen at the Gastroenterology Outpatient Clinic and with the same biopsy and treatment protocol formed the control group . A triple therapy with amoxycillin (1 g b.i.d), clarithromycin (500 mg b.i.d) and omeprazole (20 mg b.i.d.) was given to both groups for 10 days . Cure was defined as the absence of Hp infection assessed by corpus and antrum biopsies in control upper gastrointestinal endoscopies performed 6 weeks after completing the antimicrobial therapy . RESULTS: The eradication rate was 50 % in the diabetic group versus 85 % in the non-diabetic control group (P<0.001) . CONCLUSION: Type 2 diabetic patients showed a significantly lower eradication rate than controls which may be due to changes in microvasculature of the stomach and to frequent antibiotic usage because of recurrent bacterial infections with the development of resistant strains. J Pediatr Gastroenterol Nutr, 2003 May, 36(5), 616 - 22 Culturing Helicobacter pylori from clinical specimens: review of microbiologic methods; Ndip RN et al.; Helicobacter pylori is probably the most common chronic bacterial infection of humankind, and is usually acquired first in childhood . Microbiologic culture of H . pylori is the "gold standard" for diagnosis in a patient with suspected infection . Although not currently recommended for routine use, culture allows testing for susceptibility to antimicrobials, especially in populations with a high prevalence of drug resistance . Gastric biopsies are the specimens most commonly used to culture H . pylori, but stool, vomitus, saliva, and dental plaque offer opportunities . This review examines the current methods used to culture H . pylori from biologic specimens and suggests useful hints to enhance its recovery rate. Mol Biol Evol, 2003 Jun, 20(6), 914 - 23 Epub 2003 Apr 25. Molecular population genetics of inducible antibacterial peptide genes in Drosophila melanogaster; Lazzaro BP et al.; Insects respond to septic infection in part by producing a suite of antimicrobial peptides that may be subject to host-pathogen coevolutionary dynamics . In order to infer population genetic forces acting on Drosophila antibacterial peptide genes, we examine global properties of polymorphism and divergence in the Drosophila melanogaster defensin, drosocin, metchnikowin, attacin C, diptericin A, and cecropin A, B, and C genes . As a functional class, antibacterial peptides exhibit low levels of interspecific amino acid divergence . There are multiple amino acid polymorphisms segregating within D . melanogaster, however, a high proportion of which change the charge or polarity of the variable residue . These polymorphisms are particularly prevalent in processed signal and propeptide domains . We find that models of coevolutionary "arms races" and selectively maintained hypervariability do not adequately describe the population dynamics of mature antibacterial peptides in D . melanogaster, but that a highly significant excess of high-frequency derived polymorphisms coupled with substantial intralocus linkage disequilibrium suggests that positive selection may act on antibacterial peptide genes . Some attributes of the data may be consistent with a simple demographic model of population founding followed by expansion, but departures from the equilibrium null tend to be more pronounced in the peptide genes than at other loci around the genome. Am J Clin Nutr, 2003 May, 77(5), 1234 - 40 Potential host-defense role of a human milk vitamin B-12-binding protein, haptocorrin, in the gastrointestinal tract of breastfed infants, as assessed with porcine haptocorrin in vitro; Adkins Y et al.; BACKGROUND: Limited information exists on the biological role of a vitamin B-12-binding protein, haptocorrin, in human milk . The expression of haptocorrin by human mammary epithelial cells and its presence in human milk suggest a potential physiologic function in breastfed infants . OBJECTIVE: We investigated the extent to which haptocorrin could withstand proteolytic degradation and exert antimicrobial activity under in vitro conditions designed to simulate the gastrointestinal tract of breastfed infants . DESIGN: An in vitro model that simulates infant gastric and intestinal digestion was developed . The structural stability of porcine haptocorrin after exposure to digestive enzymes (pepsin and pancreatin) was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot analysis, column chromatography, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) . The antimicrobial activity of haptocorrin was determined by incubating haptocorrin with enteropathogenic Escherichia coli O127 strain 2348/69 and monitoring bacterial growth . RESULTS: The structural analysis of haptocorrin exposed to enzymes did not show a decrease in molecular weight, which indicated that haptocorrin can survive proteolytic degradation . Both haptocorrin exposed to digestive enzymes and undigested haptocorrin inhibited the growth of enteropathogenic E . coli and did so to a similar extent . Thus, haptocorrin in vitro not only retains its structure after exposure to proteases but also exhibits antimicrobial activity . CONCLUSION: These results suggest that haptocorrin may exert a host-defense function against pathogens in the gastrointestinal tracts of breastfed infants. J Environ Sci Health B, 2003 May, 38(3), 391 - 404 Analysis of microbiological screen test data for antimicrobial residues in food animals; Dey BP et al.; This study analyzes the National Residue Program (NRP) of the Food Safety and Inspection Service (FSIS), United States Department of Agriculture (USDA), data for the years 1983-1998 to determine the effectiveness of all three microbiological screen tests that were developed and used by the FSIS to control antimicrobial residues in food animals . The Swab Test On Premises (STOP) was the first screen test introduced in slaughterhouses, followed by the Calf Antibiotic Sulfonamide Test (CAST) and the Fast Antimicrobial Screen Test (FAST) . The data for STOP indicates that during 1983-1998, the rate of food animal carcasses with violative levels of antimicrobial residues reduced from 2.33% to 0.45% under the monitoring plan and under the surveillance plan, the rate reduced from 55.1% to 0.56% . Similarly, the data for CAST indicates that the rate of calf carcasses with violative levels of antimicrobial residue also declined significantly during those years . Because of its higher sensitivity and shorter analytical time, the use of FAST started in 1995 . By 1999, it had practically replaced the use of STOP and CAST in bovine species . The use of only one test such as FAST instead of different tests has removed confusion for testing different species of food animals and thereby has enhanced the efficiency of the NRP. Mycopathologia, 2002, 156(1), 1 - 7 Susceptibility and features of the ultrastructure of Prototheca zopfii following exposure to copper sulphate, silver nitrate and chlorexidine; Melville PA et al.; One of the most important forms of the occurrence of protothecosis is bovine mastitis . Studies on the "in vivo" and "in vitro" susceptibility to antimicrobials have shown that the microorganism is resistant to most of them . Looking for alternative treatments this study aimed to study the susceptibility to copper sulphate (which has an important algicide effect) and silver nitrate (used in dairy cattle breeding for the cauterization of mammary glands) and also to chlorexidine (an important post-dipping anti-septic used in dairy practice), and the effect of these antimicrobials in the ultrastructure of Prototheca zopfii before and after the exposure to these drugs . The "in vitro" susceptibility tests to chlorexidine, silver nitrate and copper sulphate of the strains of Prototheca zopfii for the determination of their minimal microbicidal concentrations (MMC), were performed using the tube dilution method in Sabouraud dextrose broth and evaluation of colony growth after plating in Sabouraud dextrose agar . The MMCs of chlorexidine, copper sulphate and silver nitrate of the 50 strains tested were 0.01%, 0.1% and 0.3%, respectively . The tubes containing the material used in the antimicrobial susceptibility tests were prepared for the examination in an electron microscope . The untreated controls of P . zopfii showed a similar ultrastructural appearance with the typical characteristics of the microorganism . Cells exposed to silver nitrate showed changes suggesting thickness of the cell wall . Cells exposed to chlorexidine showed changes suggesting degradation of intra-cellular organelles present in the cytoplasm . P . zopfii treated with copper sulphate showed changes suggesting fibrilation of inner layer of cell wall. J Antibiot (Tokyo), 2003 Feb, 56(2), 190 - 6 Standards of the in vitro mutation frequency study and the antimicrobial activity study in gut; Fukuyasu T et al.; In view of the recent rapid increase in incidence of infection with antimicrobial resistant bacteria in human medicine, there is international controversy as to the medical risk that is created by transfer of antimicrobial resistant bacteria and antimicrobial resistant genes, which may be produced through the processes of administration of antimicrobials to food-producing animals, via the food chain . Accordingly, International Cooperation on Harmonization of Technical Requirements for Registration of Veterinary Medicinal Products (VICH) provides the comprehensive guidelines for characterizing selection of antimicrobial resistance by bacteria which may adversely affect human health in order to establish the system of registry of antimicrobial drugs to be administered to food- producing animals . Currently, Japanese Society of Antimicrobials for Animals recognizes the lack of technical test standards in compliance with VICH guidelines in the world and has established the test standards for "in vitro mutation frequency studies" to evaluate the appearance frequency and resistant level of resistant bacteria and those for studies on the "antimicrobial activity in gut" to estimate the effects on intestinal flora from professional aspects. Chemotherapy, 2003 May, 49(1-2), 76 - 84 Procalcitonin and C-reactive protein in infective endocarditis: correlation with etiology and prognosis; Kocazeybek B et al.; BACKGROUND: The aim of this study was to investigate the diagnostic values of serum procalcitonin (PCT) and C-reactive protein (CRP) levels in infective endocarditis (IE) and to correlate them with the etiology of the disease and the prognosis of the patients . METHODS: Fifty patients who were diagnosed as having IE based on Duke criteria (major and/or minor) were included in the study at the Istanbul University Cardiology Institute and Florence Nightingale Hospital . Forty patients with bacteremia (non-IE) and 50 healthy blood donors were also included in the study as the control group . During the 45 days of medical follow-up, in those patients who had a response to medical therapy based on the results of left ventricular function tests, transesophageal echocardiography (TEE) and culture, among other factors, PCT and CRP levels were measured in 5-cm(3) blood samples obtained without anticoagulant when they were first admitted (day 0), as well as 24 h and 15, 30 and 45 days after admission . In the patients who had valve replacement, 5-cm(3) blood samples without anticoagulant were similarly obtained on the day of admission, after 24 h and/or on the 15th day, and 1 day before and on the 2nd and 5th days after the operation . RESULTS: In this study, a significant difference (p < 0.001) was found between the IE group and the healthy control group with respect to their serum PCT and CRP levels at the time of admission . No significant difference was found between IE and non-IE groups (p > 0.05) . The sensitivity of PCT in comparison to CRP was found to be lower (84 vs . 100%); however, its specificity was determined to be higher (88 vs . 72%) . The median values of serum PCT in the nonoperated and operated cases at the time of admission, after 24 h and on the 15th day were 3.71, 5.35 and 0.44, and 2.45, 4.28 and 4.22 ng/ml, respectively, and those of CRP were 9.30, 10.95 and 10.65, and 9.5, 10.9 and 10.2 mg/dl, respectively . The median values of serum PCT were found to be higher in cases with IE and non-IE related to gram-negative bacteria than those related to gram-positive bacteria (p < 0.02) . This was found to be insignificant for CRP (p > 0.05) . CONCLUSIONS: As a result, this study suggests that in the diagnosis of IE, it would be beneficial to use PCT, besides TEE, culture and other clinical criteria, for its high specificity and positive predictive value in comparison to CRP . This study also suggests that in determining the response to medical treatment in the follow-up period, PCT could be a more valuable parameter than CRP, as PCT has a high prognostic value and is a good indicator for valve replacement in addition to the major criteria . Furthermore, serum PCT levels may help the physician to decide on the antimicrobial therapy combination before obtaining the culture results, or in situations in which the agent could not be isolated yet . Invest Ophthalmol Vis Sci, 2003 May, 44(5), 1807 - 13 Characterization of mucins in human lacrimal sac and nasolacrimal duct; Paulsen FP et al.; PURPOSE: Mucins are polymers that may reduce drag and enhance tear outflow . Mucin expression and distribution in human efferent tear ducts were tested in the physiological state, and potential differences in the expression pattern were investigated in the presence of primary acquired dacryostenosis (PANDO) . METHODS: Expression of mucins in human lacrimal sac and nasolacrimal ducts was monitored by reverse transcription-polymerase chain reaction analysis . The presence and distribution of MUC1, -2, -4, -5AC, -5B, -6, and -7 in epithelia of the efferent tear duct passage are assessed with antisera to mucin peptide cores . Twenty normal tissues from cadavers and surgical specimens from 20 patients with PANDO were tested . RESULTS: mRNAs for all mucins investigated were detected in healthy human lacrimal sacs and nasolacrimal ducts . MUC6 mRNA was detected in only about half of the investigated samples . A reduced level of MUC2, -5AC, and -5B mRNAs was observed in PANDO . Immunohistochemistry revealed MUC2 in goblet cells and single epithelial cells . Both MUC5AC and -5B were detected in goblet cells forming intraepithelial mucous glands . MUC7 was present only in columnar epithelial cells of the efferent tear duct system . No immunoreactivity was observed with antibodies against MUC1, -4, and -6 peptide cores . CONCLUSIONS: Human efferent tear ducts express and produce a broad spectrum of mucins that is partly comparable with that in the conjunctiva and the salivary glands . The mucin diversity of the efferent tear ducts could enhance tear transport and antimicrobial defense . Reduced levels of mucin mRNA in a nonfunctioning though patent segment of the lacrimal passage, which is associated with epiphora, suggests that mucins ease tear flow through the efferent tear ducts. Am J Respir Cell Mol Biol, 2003 Sep, 29(3 Pt 1), 344 - 51 Epub 2003 Apr 24. Induction of regulated upon activation, normal T cells expressed and secreted (RANTES) and transforming growth factor-beta 1 in airway epithelial cells by Mycoplasma pneumoniae; Dakhama A et al.; Mycoplasma pneumoniae infection exacerbates asthma in children and may play a role in the pathogenesis of chronic asthma . Because the airway epithelium is a preferential site for M . pneumoniae infection and a major source of the chemokine regulated on activation, normal T cells expressed and secreted (RANTES) and transforming growth factor (TGF)-beta1, we postulated that this microorganism may contribute to the disease by inducing these mediators through direct interaction with airway epithelial cells . We investigated the effects of M . pneumoniae on RANTES and TGF-beta1 production in primary cultures of normal human bronchial epithelial (NHBE) cells and small airway epithelial (SAEC) cells . Both cell types were permissive to M . pneumoniae infection in vitro, but their responses were different . TGF-beta1 was induced at higher levels in NHBE than in SAEC cultures, whereas RANTES was induced in SAEC cultures but not in NHBE cultures . These effects were attenuated by erythromycin and dexamethasone . In vitro adherence assays further indicated that the effects of erythromycin were mediated through its antimicrobial action, resulting in diminished adherence of the pathogen, whereas the effects of dexamethasone did not appear to be by inhibition of adherence . These results suggest that M . pneumoniae infection may contribute to the pathogenesis of chronic asthma at different levels of the airways, by inducing TGF-beta1 in large airways and the chemokine RANTES in small airways. J Clin Pharm Ther, 2003 Apr, 28(2), 123 - 9 Duration of antimicrobial therapy for nosocomial pneumonia: possible strategies for minimizing antimicrobial use in intensive care units; Dugan HA et al.; OBJECTIVE: To review published data evaluating shorter courses of antibiotic therapy for nosocomial pneumonia and provide recommendations for minimizing antimicrobial use in intensive care units . DATA SOURCE: Literature was identified through MEDLINE (1966 through 6/2002) and a manual search of critical care, infectious disease, and pharmacy journals was conducted to identify relevant abstracts . DATA SYNTHESIS: Antibiotic use may be decreased by discontinuing therapy after 3 days in patients with low likelihood of nosocomial pneumonia . In addition, clinical guidelines or invasive diagnostic procedures may be effectively instituted to reduce duration of antibiotic therapy . CONCLUSION: Shorter-course antibiotic therapy may be beneficial in decreasing lengths of hospital and intensive care units stays, antimicrobial resistance, and total hospital costs . Further research is needed to determine the optimal duration of therapy in patients with nosocomial pneumonia. Biochem J, 2003 Jul 15, 373(Pt 2), 621 - 8 Oncorhyncin III: a potent antimicrobial peptide derived from the non-histone chromosomal protein H6 of rainbow trout, Oncorhynchus mykiss; Fernandes JM et al.; The partial N-terminal amino acid sequence of the antimicrobial peptide reported in the present paper has been submitted to the TrEMBL database under the accession number P83338 . A 6.7 kDa antimicrobial peptide was isolated from trout skin secretions using acid extraction followed by cation-exchange chromatography, (t)C(18) solid-phase extraction, and C(18) reversed-phase HPLC . The molecular mass of this peptide, which is tentatively named oncorhyncin III, is 6671 Da, as determined by matrix-assisted laser-desorption ionization MS . N-terminal amino acid sequencing revealed that the first 13 residues of oncorhyncin III are identical with those of the non-histone chromosomal protein H6 from rainbow trout . Hence these data combined with the MS results indicate that oncorhyncin III is likely to be a cleavage product of the non-histone chromosomal protein H6 (residues 1-66) and that it probably contains two methylated residues or one double methylation . The purified peptide exhibits potent antibacterial activity against both Gram-positive and Gram-negative bacteria, with minimal inhibitory concentrations in the submicromolar range . The peptide is sensitive to NaCl, and displays no haemolytic activity towards trout erythrocytes at concentrations below 1 microM . Scanning electron microscopy revealed that oncorhyncin III does not cause direct disruption of bacterial cells . Reconstitution of the peptide in planar lipid bilayers strongly disturbs the membranes, but does not induce the formation of stable ion channels . Taken together, these results support the hypothesis that oncorhyncin III plays a role in mucosal innate host defence. Zh Mikrobiol Epidemiol Immunobiol, 2000 Jul-Aug, (4 Suppl), 95 - 9 {Persistence properties of microflora of open water reservoirs and drinking water}; Misetov IA et al.; As revealed in this study, the characteristics of the bacterial persistence of water microflora are suitable for the sanitary and hygienic characterization of natural water reservoirs and drinking water, as well as for evaluating the effectiveness of the antimicrobial action of disinfectants. Biopolymers, 2003, 71(1), 77 - 84 Transmembrane segment peptides with double D-amino acid replacements: helicity, hydrophobicity, and antimicrobial activity; Maeda M et al.; The adoption of a helical conformation in a membrane environment effectively increases the "apparent hydrophobicity" of a peptide segment by satisfying the backbone H-bonding potential, thus stabilizing it in this environment . Here we sought to explore whether destabilizing the helical conformation would have a measurable effect on the apparent hydrophobicity of such segments in both aqueous and membrane-mimetic environments . In order to uncouple peptide hydrophobicity from helicity, we used the prototypic KKAAAAAAAAAAAAWAAAAAAKKKKNH(2) peptide as a template, and performed pairwise DD-scanning mutagenesis over the length of the sequence . Studies on this library of 13 peptides show that the DD replacements at positions near the center of peptide sequence had the most significant effects on the peptides' retention time in high performance liquid chromatography experiments . Decreased retention times correlate well with decreased helicity as measured by CD spectroscopy in the aqueous environment . Trp fluorescence measurements indicated that the peptides displayed a significant red shift in LPC (but not LPG) with peptides having DD replacements near the middle of the peptide sequence, emphasizing the importance of the anionic membrane in promoting peptide insertion . When tested against a laboratory strain of Escherichia coli, antimicrobial activity of the DD-peptides correlated with the apparent hydrophobicity but not with the overall micelle-based helical content of the peptides per se . Further analysis of the DD-positional dependence of the antimicrobial activity suggests that the presence of a local, uninterrupted stretch of helical structure (10-12 residues) may be a prerequisite for peptide biological activity . The overall findings support the notion that one should distinguish between the hydrophobicity of individual residues and the apparent hydrophobicity of the peptide as a whole, as the latter will ultimately have a greater influence on the properties of the full-length species . Infez Med, 2000, 8(3), 139 - 155 {A proposal for guidelines of antimicrobial prophylaxis in surgery and surveillance of their application}; Camporese A et al.; Not available Acta Trop, 2003 Apr, 86(1), 19 - 24 Genital mycoplasmas in women attending a family planning clinic in Guiné-Bissau and their susceptibility to antimicrobial agents; Domingues D et al.; A study on the prevalence of genital mycoplasmas and their susceptibility to the most common antimicrobial agents used for treating the infection was conducted on 94 women attending a family planning clinic in Guine-Bissau . Fifty-four women (57.4%) were positive for Mycoplasma hominis and/or Ureaplasma urealyticum . M . hominis and U . urealyticum separately isolated from infected women yielded frequencies of 31.5 and 27.8%, respectively, the remainder were infected with both species . No strain was found to be resistant to all three commonly employed antibiotics for the management of these infections (erythromycin, tetracycline and ofloxacin), although multiple resistance to two antibiotics was frequent, especially when both genital mycoplasmas were present . Some 90.7 and 24.1% of all isolates were resistant to erythromycin and tetracycline, respectively . No resistance was observed to ofloxacin, although 50% of the strains had intermediate resistance . The high prevalence of genital mycoplasmas in women attending a family planning clinic in Guine-Bissau, as demonstrated in this study, appears to be associated with trichomonosis and bacterial vaginosis . These infections were also found to be highly resistant to erythromycin and tetracycline and to have intermediate resistance to ofloxacin . However, further studies are necessary to establish the burden of infection due to antibiotic resistant genital mycoplasmas. Exp Mol Pathol, 2003 Apr, 74(2), 190 - 5 Activity of Novispirin G-10, a novel antimicrobial peptide against Chlamydia trachomatis and vaginosis-associated bacteria; Yasin B et al.; We tested the activity of Novispirin G-10, a novel antimicrobial alpha-helical octadecapeptide structurally related to cathelicidins and other innate immunity peptides, against Chlamydia trachomatis serovars L2, D, and E and three organisms associated with bacterial vaginosis (BV) . The peptide's activity against C . trachomatis was measured in 48-h shell vial assays with McCoy cell targets . Exposure to 100 micro g/ml of Novispirin G-10 reduced the infectivity of serovars D and E by 99.4-100% and serovar L2 by 91.7-99.1% . At the same concentration of 100 micro g/ml, Novispirin G-10 rapidly killed >99% of Mobiluncus curtisii, Gardnerella vaginalis, and Prevotella bivia, in standard colony-forming unit (CFU) assays . Given its simple structure and relative lack of cytotoxic and hemolytic activity, Novispirin G-10 may be a useful component of microbicide preparations designed to prevent chlamydial infection and/or remediate the abnormal vaginal flora associated with BV. Exp Mol Pathol, 2003 Apr, 74(2), 180 - 2 In human epidermis, beta-defensin 2 is packaged in lamellar bodies; Oren A et al.; The skin presents a mechanical, as well as an immunological barrier to infection, and displays considerable innate immune capacity . Recently, cultured human keratinocytes were described to produce and export a microbicidal peptide human beta-defensin 2 (HBD-2) . Immunogold was used to label ultrathin cryosections of stimulated, cultured human epidermis . HBD-2 was found to be stored in the lamellar bodies (LBs) of the stimulated keratinocytes of the spinous layer of the epidermis . HBD-2 was also found in the intercellular space . These findings suggest that HBD-2 is released with the contents of the LBs . Along with other investigations, our findings indicate that the lipid "permeability" barrier of the skin contains antimicrobial substances. J Matern Fetal Neonatal Med, 2003 Jan, 13(1), 2 - 21 Antimicrobial peptides in amniotic fluid: defensins, calprotectin and bacterial/permeability-increasing protein in patients with microbial invasion of the amniotic cavity, intra-amniotic inflammation, preterm labor and premature rupture of membranes; Espinoza J et al.; OBJECTIVE: Neutrophil defensins (HNP 1-3), bactericidal/permeability-increasing protein (BPI) and calprotectin (MRP8/14) are antimicrobial peptides stored in leukocytes that act as effector molecules of the innate immune response . The purpose of this study was to determine whether parturition, premature rupture of the membranes (PROM) and microbial invasion of the amniotic cavity (MIAC) are associated with changes in amniotic fluid concentrations of these antimicrobial peptides . STUDY DESIGN: Amniotic fluid was retrieved by amniocentesis from 333 patients in the following groups: group 1, mid-trimester with a subsequent normal pregnancy outcome (n = 84); group 2, preterm labor and intact membranes without MIAC who delivered at term (n = 36), or prematurely (n = 52) and preterm labor with MIAC (n = 26); group 3, preterm PROM with (n = 26) and without (n = 26) MIAC; and group 4, term with intact membranes in the absence of MIAC, in labor (n = 52) and not in labor (n = 31) . The concentrations of HNP 1-3, BPI and calprotectin in amniotic fluid were determined by specific and sensitive immunoassays . Placentae of patients in both preterm labor with intact membranes and preterm PROM groups who delivered within 72 h of amniocentesis were examined . Non-parametric statistics, receiver-operating characteristic (ROC) curves and Cox regression models were used for analysis . A p value of < 0.05 was considered statistically significant . RESULTS: Intra-amniotic infection was associated with a significant increase in amniotic fluid concentrations of immunoreactive HNP 1-3, BPI and calprotectin in both women with preterm labor and intact membranes, and women with preterm PROM . Preterm PROM was associated with a significant increase in amniotic fluid concentrations of immunoreactive HNP 1-3, BPI and calprotectin . Preterm parturition was associated with a significant increase in amniotic fluid concentrations of immunoreactive HNP 1-3, BPI and calprotectin, while parturition at term was associated with a significant increase in amniotic fluid concentrations of immunoreactive HNP 1-3 . Among patients with preterm labor and intact membranes, elevation of amniotic fluid HNP 1-3, BPI and calprotectin concentrations was associated with intra-amniotic inflammation, histological chorioamnionitis and a shorter interval to delivery . CONCLUSION: MIAC, preterm parturition and preterm PROM are associated with increased amniotic fluid concentrations of immunoreactive HNP 1-3, BPI and calprotectin . Moreover, elevated amniotic fluid concentrations of BPI, immunoreactive HNP 1-3 and calprotectin are associated with intra-amniotic inflammation, histological chorioamnionitis and shorter amniocentesis-to-delivery interval in patients presenting with preterm labor with intact membranes. Z Naturforsch {C}, 2003 Mar-Apr, 58(3-4), 195 - 200 The bioactive essential oil of Heracleum sphondylium L . subsp . ternatum (Velen.) Brummitt; Iscan G et al.; The essential oil of Heracleum sphondylium L subsp . ternatun (Velen.) Brummit (Umbelliferae) was isolated from crushed seeds by means of hydrodistillation and analyzed by GC and GC/MS . Major components were identified as 1-octanol (50.3%), octyl butyrate (24.6%), and octyl acetate (7.3%) . Furthermore, antimicrobial activity of the oil was evaluated using microdilution broth and agar diffusion methods . The bioactive constituent of the essential oil was determined as 1-octanol by using a bioautography assay. Poult Sci, 2003 Apr, 82(4), 632 - 9 Perspectives on the use of organic acids and short chain fatty acids as antimicrobials; Ricke SC; Organic acids have a long history of being utilized as food additives and preservatives for preventing food deterioration and extending the shelf life of perishable food ingredients . Specific organic acids have also been used to control microbial contamination and dissemination of foodborne pathogens in preharvest and postharvest food production and processing . The antibacterial mechanism(s) for organic acids are not fully understood, and activity may vary depending on physiological status of the organism and the physicochemical characteristics of the external environment . An emerging potential problem is that organic acids have been observed to enhance survivability of acid sensitive pathogens exposed to low pH by induction of an acid tolerance response and that acid tolerance may be linked to increased virulence . Although this situation has implications regarding the use of organic acids, it may only apply to circumstances in which reduced acid levels have induced resistance and virulence mechanisms in exposed organisms . Evaluating effectiveness of organic acids for specific applications requires more understanding general and specific stress response capabilities of foodborne pathogens . Development and application of molecular tools to study pathogen behavior in preharvest and postharvest food production environments will enable dissection of specific bacterial genetic regulation involved in response to organic acids . This could lead to the development of more targeted strategies to control foodborne pathogens with organic acids. Poult Sci, 2003 Apr, 82(4), 618 - 21 Antibiotic residues in poultry tissues and eggs: human health concerns? Donoghue DJ. Antibiotics are used by the poultry industry to enhance the health and productivity of flocks . The use of antimicrobials is strictly regulated by the Food and Drug Administration (FDA) and the USDA to warrant their safety and efficacy . Prior to regulatory approval, the pharmacokinetics and tissue tolerances of an antimicrobial are determined to set the proper dosage . To ensure proper use, both the FDA and USDA have research, surveillance, and compliance programs to develop detection methods and monitor poultry tissues for antimicrobials . Unfortunately, there is the perception among many consumers that our food supply contains high concentrations of drug or hormone residues causing significant health concerns or problems . In fact, foods produced in this country (including poultry) are very safe and meet the highest standards to exclude chemical contaminants . An overview will be presented on the federal oversight and monitoring of antimicrobial residues in poultry tissues. Poult Sci, 2003 Apr, 82(4), 613 - 7 Observations on the history of the development of antimicrobials and their use in poultry feeds; Jones FT et al.; Antimicrobials are powerful tools, but controversy and conflict often follow power . The development of antimicrobials was marked by personal attacks, political intrigue, internal conflicts, and lawsuits . Such controversy and conflict has continued . The early history of supplementing animal feeds with antimicrobials parallels the isolation and identification of vitamin B12 . Vitamin B12 was isolated and characterized in 1948, but further research showed that several feed ingredients, including dried mycelia of certain fungi, were more potent as growth promoters in the diet of chicks than was vitamin B12 alone . The growth-promoting component in fungal mycelia was shown to have antimicrobial activity . A total of 32 antimicrobial compounds are approved for use in broiler feeds in the U.S . without a veterinary prescription . Fifteen compounds are listed for treatment of coccidiosis, 11 are listed as growth promotants, and six are listed for other purposes . Seven compounds are also used in human medicine . These compounds include bacitracin, chlotetracycline, erythromycin, lincomycin, novobiocin, oxytetracycline, and penicillin . No published estimates of antimicrobial use in animals exist at present, and estimates of that use differ markedly . The Union of Concerned Scientists (UCS) estimates usage at 30.6 million pounds, nearly 50% (49.85%) higher than the Animal Health Institute (AHI) estimate of 20.42 million pounds . AHI surveyed their members (the manufacturers of antimicrobials) to obtain their estimates, whereas USC calculated their estimates using published data and the following general formula: antimicrobial use = number of animals treated x average days treated x average dose. Planta Med, 2003 Apr, 69(4), 350 - 5 New antimicrobial alkaloids from the roots of Polyalthia longifolia var . pendula; Faizi S et al.; Bioassay-guided isolation studies on the root extract of Polyalthia longifolia var . pendula possessing significant antibacterial activity led to the isolation of three new alkaloids pendulamine A (1), pendulamine B (2) and penduline (3) along with stigmasterol 3-O-beta-D-glucoside, allantoin, the known diterpenoid kolavenic acid, and the azafluorene alkaloid isoursuline . The structures of these compounds have been elucidated with the help of spectroscopic studies including 2D NMR experiments . Compounds 1 and 2, which are the only protoberberine alkaloids having a monosubstituted A ring with a hydroxy group at C-3, were found to be the active antibacterial principles of the roots . Their MIC values ranged between 0.02-20 microg against the tested bacteria. Eur J Clin Microbiol Infect Dis, 2003 Apr, 22(4), 261 - 7 Epub 2003 Mar 28. Comparative in vitro surveillance of amoxicillin-clavulanic acid and four oral comparators against 21232 clinical isolates from europe; Hoban DJ et al.; The present study was conducted to determine the in vitro activity of amoxicillin-clavulanic acid compared to that of four newer antimicrobial agents (ampicillin, azithromycin, cefuroxime and trimethoprim-sulfamethoxazole) . All of the agents were tested against 21232 recent clinical isolates encompassing 37 species submitted from 16 European countries between 1997 and 1999 . After 20 years of clinical use, amoxicillin-clavulanic acid continues to retain much of its initial activity against targeted gram-positive organisms, selected gram-negative organisms and major respiratory pathogens. Pediatr Radiol, 2003 Apr, 33(4), 281 - 3 Epub 2003 Feb 12. Lemierre's syndrome: post-anginal septicemia; de Lima JE Jr et al.; Lemierre's syndrome is an obscure entity with very significant related morbidity . We describe a typical case and a never before reported complication . Increased awareness of this condition is necessary, since it is given little consideration even when patients present with the very characteristic clinical picture . US will depict internal jugular vein (IJV) thrombosis in most cases and should be the modality of choice for the initial evaluation of suspected thrombosis of the IJVs . Associated demonstration of cavitary pulmonary nodules establishes the diagnosis of Lemierre's syndrome . Early recognition and institution of adequate antimicrobial therapy are key points in the successful management of this syndrome. Antimicrob Agents Chemother, 2003 May, 47(5), 1732 - 5 Pharmacokinetics of intramuscularly administered ertapenem; Musson DG et al.; Ertapenem (INVANZ) is a new once-a-day parental beta-lactam antimicrobial agent that has been shown to be highly effective as a single agent for treatment of various community-acquired and mixed infections . The plasma pharmacokinetics of a 1-g intramuscular (i.m.) dose was compared with those of a 1-g intravenous (i.v.) dose infused over 30 min, the recommended rate of i.v . infusion for comparison, and over 120 min, which more closely mimicked the time course for absorption of the i.m . form . In a three-period crossover study (Part A), 26 healthy subjects received single doses of ertapenem administered i.m., i.v . infused over 30 min, and i.v . infused over 120 min . Blood for ertapenem analysis was collected over 24 h postdose for each treatment . In Part B, these fasted subjects received a 1-g i.m . dose of ertapenem once daily for 7 days . Following a 1-g i.m . dose and a 1-g i.v . dose infused over 120 min, the geometric mean area under the concentration curve from hour 0 to infinity (AUC(0- infinity )) was 541.8 micro g . hr/ml following i.m . administration and 591.4 micro g . hr/ml following a 120-min infusion; the geometric mean ratio was 0.92 with a 90% confidence interval of 0.88 to 0.95 . The geometric mean AUC(0- infinity ) was nearly identical when 1-g doses were infused over 30 or 120 min . Although the maximum concentration of drug in serum was somewhat lower following i.m . administration than following i.v . administration, the shape of the plasma concentration profiles was roughly comparable at later time points . Ertapenem did not accumulate after multiple 1-g i.m . daily doses over 7 days . The geometric mean ratio for AUC(0-24) (day 7/day 1) was 0.98 with a 90% confidence interval of 0.94 to 1.02 . Thus, the relative bioavailability of the 1-g i.m . dose was 92% . Ertapenem does not accumulate following multiple daily 1-g i.m . doses over 7 days. Antimicrob Agents Chemother, 2003 May, 47(5), 1614 - 20 Antimicrobial and immunologic activities of clarithromycin in a murine model of Mycoplasma pneumoniae-induced pneumonia; Hardy RD et al.; Because macrolide antibiotics are hypothesized to possess immunomodulatory activity independent of their antimicrobial activity, we evaluated the immunomodulatory effect of clarithromycin in a murine model of lung inflammation induced by either live or UV-killed Mycoplasma pneumoniae . BALB/c mice were intranasally inoculated once with live or UV-killed M . pneumoniae . Clarithromycin (25 mg/kg of body weight) or placebo was subcutaneously administered once daily in both groups of mice . In mice infected with live M . pneumoniae, clarithromycin treatment significantly reduced quantitative M . pneumoniae bronchoalveolar lavage (BAL) culture, pulmonary histopathologic scores (HPS), and airway resistance-obstruction (as measured by plethysmography) compared with placebo . Concentrations of tumor necrosis factor alpha, gamma interferon, interleukin-6 (IL-6), mouse KC (functional IL-8), JE/MCP-1, and MIP-1alpha in BAL fluid were also significantly decreased in mice infected with live M . pneumoniae given clarithromycin . In contrast, mice inoculated with UV-killed M . pneumoniae had no significant reduction in HPS, airway resistance-obstruction, or BAL cytokine or chemokine concentrations in response to clarithromycin administration . Clarithromycin therapy demonstrated beneficial effects (microbiologic, histologic, respiratory, and immunologic) on pneumonia in the mice infected with live M . pneumoniae; this was not observed in the mice inoculated with UV-killed M . pneumoniae. Eur J Biochem, 2003 May, 270(9), 2068 - 81 Antimicrobial peptides from hylid and ranin frogs originated from a 150-million-year-old ancestral precursor with a conserved signal peptide but a hypermutable antimicrobial domain; Vanhoye D et al.; The dermal glands of frogs produce antimicrobial peptides that protect the skin against noxious microorganisms and assist in wound repair . The sequences of these peptides are very dissimilar, both within and between species, so that the 5000 living anuran frogs may produce approximately 100 000 different antimicrobial peptides . The antimicrobial peptides of South American hylid frogs are derived from precursors, the preprodermaseptins, whose signal peptides and intervening sequences are remarkably conserved, but their C-terminal domains are markedly diverse, resulting in mature peptides with different lengths, sequences and antimicrobial spectra . We have used the extreme conservation in the preproregion of preprodermaseptin transcripts to identify new members of this family in Australian and South American hylids . All these peptides are cationic, amphipathic and alpha-helical . They killed a broad spectrum of microorganisms and acted in synergy . 42 preprodermaseptin gene sequences from 10 species of hylid and ranin frogs were analyzed in the context of their phylogeny and biogeography and of geophysical models for the fragmentation of Gondwana to examine the strategy that these frogs have evolved to generate an enormous array of peptide antibiotics . The hyperdivergence of modern antimicrobial peptides and the number of peptides per species result from repeated duplications of a approximately 150-million-year-old ancestral gene and accelerated mutations of the mature peptide domain, probably involving a mutagenic, error-prone, DNA polymerase similar to Escherichia coli Pol V . The presence of antimicrobial peptides with such different structures and spectra of action represents the successful evolution of multidrug defense by providing frogs with maximum protection against infectious microbes and minimizing the chance of microorganisms developing resistance to individual peptides . The hypermutation of the antimicrobial domain by a targeted mutagenic polymerase that can generate many sequence changes in a few steps may have a selective survival value when frogs colonizing a new ecological niche encounter different microbial predators. Ann Pharmacother, 2003 May, 37(5), 646 - 51 An interventional program to improve antibiotic use; Feucht CL et al.; BACKGROUND: Large volume and often inappropriate use of specific antimicrobial agents increase selective pressure for emergence of resistant bacteria and place strain on the pharmacy budget . OBJECTIVE: To initiate a multidisciplinary program designed to align intravenous vancomycin and fluoroquinolone prescribing practices with guidelines for appropriate use of these agents . METHODS: A multidisciplinary, prospective interventional program was implemented to encourage early discontinuation of inappropriate vancomycin and fluoroquinolone therapy and decrease inappropriate duplicative gram-negative coverage using fluoroquinolones . A computerized review was performed for patients receiving intravenous vancomycin and fluoroquinolones for 1998 in a Veterans Affairs Medical Center . In June 1999, guidelines were disseminated and an interventional program was initiated, with a monthly conference for medical residents regarding antimicrobial resistance and local hospital practices . Concurrently, a prospective review of new orders was assessed by the clinical pharmacist and interventions performed when inappropriate use occurred . RESULTS: The interventional program was successful in reducing unnecessary duplicative gram-negative coverage with intravenous fluoroquinolones by 26% (p < 0.001) from 1998 to 2001 . Overall, a 43% reduction in the number of courses of intravenous fluoroquinolones was seen during these 4 years . Courses lasting >5 days were reduced by 22% (p < 0.001) . Vancomycin prescriptions deemed inappropriate that were administered >5 days were reduced by 16% (p < 0.001) during the same time period . The interventions performed by the clinical pharmacist were deemed successful, with a 76% acceptance rate by providers . CONCLUSIONS: Education of physicians through monthly conferences and personal interventions resulted in an increase in appropriate empiric antibiotic use, a decrease in the duration of inappropriate use, and a decrease in duplicate gram-negative coverage. Rinsho Byori, 2003 Mar, 51(3), 194 - 200 {Reliability of bioluminescent assay (ATP method) for testing antimicrobial susceptibility of Mycobacterium tuberculosis}; Yamazaki T et al.; To evaluate the reliability of our previously reported antimicrobial susceptibility test by ATP method, we have compared our ATP method to the reference test methods such as Mycobacteria Growth Indicator Tube(MGIT) method, minimum inhibitory concentration(MIC) method, NCCLS M24-T agar proportion method(M24-T method), and Vite spectrum method . The concentrations of drugs used for the assessment were isoniazid(INH) 0.1 microgram/ml, rifampicin(RFP) 2.0 micrograms/ml, ethambutol(EB) 2.5 micrograms/ml, streptomycin (SM) 2.0 micrograms/ml, and kanamycin (KM) 5.0 micrograms/ml . When six M . tuberculosis ATCC strains were subjected to 6 independent experiments by using ATP method, highly reproducible results were obtained on the fifth day of the incubation . We examined correlation among ATP method and reference test methods in drug susceptibility testing for 65 clinical isolates of M . tuberculosis . The correlation between ATP method and MGIT-, MIC-, M24-T method were more than 95% for all drugs . When ATP method and Vite spectrum method was compared, the correlation was 87.7% for INH, 98.5% for RFP, 90.8% for EB, 92.3% for SM, 96.9% for KM . The culture period for determining susceptibility between ATP method and MGIT method was compared by using ATCC reference strains and clinical isolates . Six M . tuberculosis ATCC strains were subjected to 6 independent experiments . By the MGIT method, 8 days were required to obtain the results, whereas 3 days were enough by the ATP method . For 65 clinical isolates, the MGIT method required 9 days for determining susceptibility of all isolates . The ATP method required only 5 days for the same strains . These data demonstrate that the improved ATP method that we reported, is simple, rapid, highly reproducible and nonradiometric, and could be used for the assessment of drug susceptibility for M . tuberculosis with high reliability. J Immunol, 2003 May 1, 170(9), 4708 - 16 Retrocyclin, an antiretroviral theta-defensin, is a lectin; Wang W et al.; Theta-defensins are circular octadecapeptides that contain an internal tridisulfide ladder . Because retrocyclin-1, an ancestral hominid theta-defensin, can protect human cells in vitro from infection by T- and M-tropic strains of HIV-1, we used surface plasmon resonance techniques to study its binding to glycoproteins and glycolipids implicated in HIV-1 entry . Retrocyclin-1 bound with high affinity to gp120 (K(d), 35.4 nM), CD4 (K(d), 31 nM), and galactosylceramide (K(d), 24.1 nM) . Neither a circular form of retrocyclin without its tridisulfide ladder nor its beta-hairpin precursor with these disulfides intact bound gp120 or CD4 effectively . Retrocyclin also bound fetuin, an extensively glycosylated protein, with high affinity, but it did not bind nonglycosylated gp120 or BSA . However, retrocyclin did bind to a neoglycoprotein, BSA, with covalently attached sugar residues . Experiments with glycosidase-treated fetuin, gp120, and CD4 revealed that both O-linked and N-linked sugars were used as binding sites . In a panel of retrocyclin variants, binding to immobilized gp120 and CD4 were highly correlated to each other and to the peptide's ability to protect human PBMCs from infection by HIV-1 . Although small, cysteine-rich antimicrobial peptides with lectin-like properties exist in plants, theta-defensins are the first such molecules to be identified in vertebrates . Retrocyclin's ability to recognize and bind carbohydrate-containing surface molecules is integrally related to its ability to protect cells from HIV-1 infection. Arch Dermatol, 2003 Apr, 139(4), 459 - 64 Effects of subantimicrobial-dose doxycycline in the treatment of moderate acne; Skidmore R et al.; OBJECTIVE: To determine if treatment with subantimicrobial-dose (SD) doxycycline hyclate (20-mg tablets taken twice daily) improved clinical outcome, had any detectable effect on skin flora, led to overgrowth or colonization of skin by opportunistic pathogens, or resulted in an increase in antibiotic resistance by the surface skin microflora in patients with moderate acne compared with placebo . DESIGN: Multicenter, double-blind, randomized, placebo-controlled, parallel-group trial . SETTING: Two university-based clinics . SUBJECTS: Adults (N = 51) with moderate facial acne . INTERVENTIONS: Patients were randomized to receive SD doxycycline (Periostat; CollaGenex Pharmaceuticals Inc, Newtown, Pa) or placebo twice daily for 6 months . MAIN EFFICACY OUTCOMES: Primary: changes from baseline in numbers of inflammatory, noninflammatory, and total lesions . Secondary: changes from baseline of individual counts of papules, pustules, and nodules and global assessments of clinical improvement by patient and physician . RESULTS: Forty patients completed 6 months of treatment . At 6 months, the SD doxycycline group had a significantly greater percent reduction in the number of comedones (P<.01), inflammatory and noninflammatory lesions combined (P<.01), and total inflammatory lesions (P<.05) than did the placebo group . They also had significantly greater improvement according to the clinician's global assessment (P =.03) . There were no significant differences in microbial counts between groups and no evidence of change in antibiotic susceptibility or colonization by potential pathogens . The treatment was well tolerated . CONCLUSIONS: Twice-daily SD doxycycline treatment significantly reduced the number of inflammatory and noninflammatory lesions in patients with moderate facial acne, was well tolerated, had no detectable antimicrobial effect on the skin flora, and did not result in any increase in the number or severity of resistant organisms. Trends Microbiol, 2003 Apr, 11(4), 179 - 84 Contamination of abiotic surfaces: what a colonizing bacterium sees and how to blur it; Lejeune P; Many microbes are able to interfere with solid surfaces and trigger highly sophisticated colonization responses that include expression of specific properties such as increased resistances to antimicrobial agents . An anticontamination strategy might be to prevent adhesion by interfering with the surface-sensing processes and repelling the pioneering cells, to maintain the cellular sensitivity to antimicrobial agents . Recent studies have shown that differences in the physiological state of free-floating and attached bacteria, which could explain the increased levels of resistance, are triggered very early during attachment . Two-component-mediated signalling mechanisms are involved in these surface-sensing processes . Drugs and surface treatments able to interfere with the stimulation factors of these sensing systems (water activity and accumulation of proteins within the periplasm) could "blind" the colonizing bacteria and delay surface contamination. Insect Biochem Mol Biol, 2003 May, 33(5), 541 - 59 Identification by subtractive suppression hybridization of bacteria-induced genes expressed in Manduca sexta fat body; Zhu Y et al.; Insect immune processes are mediated by programs of differential gene expression . To understand the molecular regulation of the immune response in the tobacco hornworm, Manduca sexta, the relevant subset of differentially expressed genes of interest must be identified, cloned and studied in detail . In this study, suppression subtractive hybridization, a PCR-based method for cDNA subtraction was performed to identify mRNAs from fat body of immunized larvae that are not present (or present at a low level) in control larvae . A subtracted cDNA library enriched in immune-inducible genes was constructed . Northern blot analysis of a sample of clones from our subtracted library indicated that >90% of the clones randomly selected from the subtracted library are immune inducible . Sequence analysis of 238 expressed sequence tags (ESTs) revealed that 120 ESTs, representing 54 distinct genes or gene families, had sequences identical or similar to previously characterized genes, some of which have been confirmed to be involved in innate immunity . These ESTs were categorized into seven groups, including pattern recognition proteins, serine proteinases and their inhibitors, and antimicrobial proteins . 112 ESTs, about 47.5% of the library, showed no significant similarity to any known genes . The sequences identified in this M . sexta library reflect our knowledge of insect immune strategies and may facilitate better understanding of insect immune responses. J Community Health, 2003 Apr, 28(2), 139 - 50 Short- and long-term effects of handwashing with antimicrobial or plain soap in the community; Larson E et al.; Little is known about effects of public use of antimicrobial handwashing soap . A double-blinded, randomized clinical trial of hands of primary caretakers in 238 inner city households was conducted in which effects of plain or antimicrobial (containing 0.2% triclosan) handwashing soap on bacterial counts of the hands were compared before and after a single wash and before and after handwashing following a year of product use . The randomly assigned product was provided without cost to each household during monthly home visits, and compliance with product use was monitored . Households were contacted by telephone weekly and with a home visit monthly for 11 months . Hand cultures were obtained before and after handwashing at baseline and after 11 months, using a modified glove juice technique . Overall, there were no significant differences in pre-to-post handwashing counts at baseline (p = 0.41), but by the end of one year, post-wash counts were significantly lower than pre-wash (p = 0.000) for those using either antimicrobial or plain soap . There were no significant differences in mean log counts either before or after handwashing between those using the antimicrobial or plain soap at baseline or after a year of u