Peptides, 2003 Apr, 24(4), 523 - 30 Antimicrobial peptides in the first line defence of human colon mucosa; Tollin M et al.; Antimicrobial peptides and proteins are effector molecules in the protection of epithelial surfaces . We have evaluated the presence of antimicrobial peptides/proteins that can participate in human colonic defence against microbes . A peptide/protein extract of normal human colon mucosa was found to be active against Gram-positive bacteria, Gram-negative bacteria, and fungi . Four polypeptides with antimicrobial activity were isolated from this material and they were identified by N-terminal amino acid sequence analysis as ubiquicidin, histone H2B, eosinophil cationic protein, and phospholipase A(2) (PLA(2)) . Using immunodetection and mass spectrometry, LL-37, HNP1-3, and HBD-1 were also identified . Combined, these results indicate that the colon mucosa is protected by a complex mixture of polypeptides, able to kill invading microbes and working in synergy as a barrier against bacterial invasion. Int Immunopharmacol, 2003 Aug, 3(8), 1159 - 67 Combinatorial immunotherapies for infectious diseases; Hengel H et al.; Combating pathogenic organisms by combinatorial approaches involving appropriate immune response molecules and antimicrobial drugs represents a progessively more apparent and successful therapeutic paradigm for the treatment of acute and chronic persistent infectious diseases . This review explores areas of current innovation and provides an update of the present state of knowledge concerning combination of chemotherapy with several immune-based interventions in infections . In the future, a better understanding of microbial immune modulation and evasion may continue to open new avenues of inquiry and carefully targeted application of adjunctive immunotherapies. J Infect, 2003 Aug, 47(2), 110 - 6 Q fever: epidemiology, clinical features and prognosis . A study from 1983 to 1999 in the South of Spain; Alarcon A et al.; OBJECTIVES: Clinical polymorphism is a main feature of Q fever and, depending upon the geographic location, differences in its clinical picture have been described . The objective of this study was to determine the epidemiology, clinical features and prognosis of acute Q fever in our area . METHODS: From 1985 to 1999, consecutive cases of Q fever, presented as febrile syndrome and attended in a tertiary teaching hospital in Sevilla, Spain, were included and followed prospectively . RESULTS: Two hundred and thirty-one cases of acute Q fever were included . A non-focalized febrile syndrome lasting from 7 to 28 days (fever of intermediate duration) was the most frequent presentation (n=208, 90%) . One hundred and forty-eight patients had hepatitis . Overall, 53% of the cases were urban and contact with animals was referred in 39% of the patients . No relationship between clinical presentation and possible route of infection was observed . Prognosis was excellent (100% cured), although in 18 patients fever was prolonged more than 28 days and three patients developed life-threatening organ affection . Antimicrobial treatment was more effective if it was administered in the first two weeks (median defervescence of fever: 3 days versus 5.5 days, p<0.01) . CONCLUSIONS: Acute Q fever is a common cause of fever of intermediate duration, even in urban areas . Elevation of hepatic enzymes was the most frequent laboratory finding . Severe organ affection is uncommon and the overall prognosis of the disease is excellent . Early treatment seems to shorten the duration of the disease. Vet Microbiol, 2003 Aug 29, 95(1-2), 75 - 89 Interleukin 6, serum amyloid A and haptoglobin as markers of treatment efficacy in pigs experimentally infected with Actinobacillus pleuropneumoniae; Hulten C et al.; The possibility to use acute phase proteins to monitor the elimination of a bacterial infection in pigs would facilitate an objective assessment of treatment with various antimicrobial substances . To examine this possibility, the acute phase response (IL-6, serum amyloid A (SAA), and haptoglobin) elicited by Actinobacillus pleuropneumoniae and its reduction on treatment with various antibiotics was studied in serum from specific pathogen free (SPF) pigs . Pigs were infected intranasally with A . pleuropneumoniae serotype 2, and either left as non-treated control pigs or treated with different antibiotics intramuscularly at onset of respiratory disease (20h post-infection) . Pigs responded to the infection with prominent increases in activity and concentrations of IL-6, SAA, and haptoglobin . These responses were to a certain extent overlapping and covered the time span from a few hours after infection until development of detectable levels of specific antibodies (7-10 days post-infection in untreated pigs) . The haptoglobin response lasted until the end of the study on day 17 and thereby partly coincided with the antibody response . Treatment with antimicrobials that effectively reduced establishment of the infection with A . pleuropneumoniae also reduced the duration of all three acute phase responses, and reduced the concentration of serum haptoglobin . In contrast, less efficacious treatments did not reduce these acute phase responses . Thus, acute phase reactants can be applied to monitor therapeutic effects of antimicrobial drugs in the pig and measurements of IL-6, SAA and haptoglobin could add valuable information about the stage of infection during a disease outbreak. Biochem Biophys Res Commun, 2003 Jul 25, 307(2), 369 - 74 Yeast sphingolipid bypass mutants as indicators of antifungal agents selectively targeting sphingolipid synthesis; Nagiec MM et al.; Standard methods for evaluating the target specificity of antimicrobial agents often involve the use of microorganisms with altered expression of selected targets and thus either more resistant or more susceptible to target specific inhibitors . In this study we present an alternative approach that utilizes physiological bypass mutants . The Saccharomyces cerevisiae sphingolipid bypass mutant strain AGD is able to grow without making sphingolipids and importantly, tolerates loss-of-function mutations in the otherwise essential genes for both serine palmitoyltransferase (SPT) and inositol phosphorylceramide (IPC) synthase . We found that strain AGD was >1000-fold more resistant than the wild-type strain to selective inhibitors of SPT and IPC synthase . In contrast, strain AGD, which due to abnormal composition of the plasma membrane is sensitive to a variety of environmental stresses, was more susceptible than the wild-type to amphotericin B, voriconazole, and to cycloheximide . We show that in a simple growth assay the AGD strain is an appropriate and useful indicator for inhibitors of IPC synthase, a selective antifungal target. Biochemistry, 2003 Jul 22, 42(28), 8434 - 44 Recognition and resistance in TEM beta-lactamase; Wang X et al.; Developing antimicrobials that are less likely to engender resistance has become an important design criterion as more and more drugs fall victim to resistance mutations . One hypothesis is that the more closely an inhibitor resembles a substrate, the more difficult it will be to develop resistant mutations that can at once disfavor the inhibitor and still recognize the substrate . To investigate this hypothesis, 10 transition-state analogues, of greater or lesser similarity to substrates, were tested for inhibition of TEM-1 beta-lactamase, the most widespread resistance enzyme to penicillin antibiotics . The inhibitors were also tested against four characteristic mutant enzymes: TEM-30, TEM-32, TEM-52, and TEM-64 . The inhibitor most similar to the substrate, compound 10, was the most potent inhibitor of the WT enzyme, with a K(i) value of 64 nM . Conversely, compound 10 was the most susceptible to the TEM-30 (R244S) mutant, for which inhibition dropped by over 100-fold . The other inhibitors were relatively impervious to the TEM-30 mutant enzyme . To understand recognition and resistance to these transition-state analogues, the structures of four of these inhibitors in complex with TEM-1 were determined by X-ray crystallography . These structures suggest a structural basis for distinguishing inhibitors that mimic the acylation transition state and those that mimic the deacylation transition state; they also suggest how TEM-30 reduces the affinity of compound 10 . In cell culture, this inhibitor reversed the resistance of bacteria to ampicillin, reducing minimum inhibitory concentrations of this penicillin by between 4- and 64-fold, depending on the strain of bacteria . Notwithstanding this activity, the resistance of TEM-30, which is already extant in the clinic, suggests that there can be resistance liabilities with substrate-based design. J Cosmet Sci, 2003 May-Jun, 54(3), 251 - 61 N,N',N"-tris(dihydroxyphosphorylmethyl)-1,4,7-triazacyclononane (Deofix) - a high-affinity, high-specificity chelator for first transition series metal cations with significant deodorant, antimicrobial, and antioxidant activity; Laden K et al.; Deofix, N,N',N"-tris(dihydroxyphosphorylmethyl)-1,4,7-triazacyclononane, is a high-affinity, high-specificity chelator for first transition series cations such as iron, zinc, manganese, and copper . A 1% solution in 50% ethanol was found to be significantly better at reducing underarm malodor than a solution of 0.3% Triclosan in 50% ethanol . Compared to a 50% alcohol control, Deofix was found to produce a significant reduction in malodor for at least 48 hours . Deofix appears to work by reducing the concentration of first transition series metal ions below the levels needed for microbial cell reproduction and by inhibiting oxidative processes by interfering with catalytic formation of free radicals . Deofix has very low levels of toxicity when measured via a number of screening techniques. J Biol Chem, 2003 Sep 19, 278(38), 36250 - 6 Epub 2003 Jul 11. Macrophage-independent fungicidal action of the pulmonary collectins; McCormack FX et al.; Histoplasma capsulatum (Hc) is a facultative intracellular fungal pathogen that causes acute and chronic pneumonia . In this study, we investigated the role of the pulmonary collectins, surfactant proteins (SP) A and D, in the clearance of Hc yeast from the lung . Exposure of yeast to either collectin induced a dose-dependent decrease in {3H}leucine incorporation by several strains of Hc . This decrement was attributed to killing of the collectin-exposed yeast since it failed to grow on agar medium . Exposure to SP-A or -D resulted in increased yeast permeability based on a leak of protein from the organism and enhanced access of an impermeant substrate to intracellular alkaline phosphatase . Inbred and outbred SP-A null (-/-) mice were modestly more susceptible to pulmonary infection with Hc than strain and age-matched SP-A (+/+) control mice . The increase in susceptibility was associated with a decrement in the number of CD8+ cells in the lungs of SP-A-/- mice . Neither SP-A nor SP-D inhibited the growth of macrophage-internalized Hc . We conclude that the SP-A and SP-D are antimicrobial proteins that directly inhibit the growth of Hc by increasing permeability of the organism and that Hc gains asylum from collectin-mediated killing by rapid entry into pulmonary macrophages. J Biomol Screen, 2003 Jun, 8(3), 340 - 6 Amplified detection of transcriptional and translational inhibitors in bioluminescent Escherichia coli K-12; Galluzzi L et al.; The excessive prescription of antimicrobial agents and their use as animal growth promoters lead to the spread of resistance among pathogenic bacteria . Consequently, unnecessary use should be minimized, and new chemicals with novel mechanisms of action are needed . The authors have developed a fast method to measure the activity of antibiotics by means of a genetically engineered strain of Escherichia coli K-12 . The system is based on the full-length bacterial luciferase operon coupled to the tetracycline-inducible tetA promoter in the reporter plasmid pTetLux1 . Sublethal doses of tetracycline are used to start the luciferase synthesis in cultures that were previously incubated with the antibiotic under investigation . After a variable time frame-from 1 to 4 h, depending on the antimicrobial mode of action-the level of light emission from treated cultures is compared to the level obtained in control cultures . The gap in bioluminescence outlines the antibiotic interference in bacterial metabolism . Throughout this study, freeze-dried sensor cells were used to avoid repeated cultures from day to day . The authors show the results of 10 model antibiotics, representing different molecular structures and mechanisms of action . The results show that no actively dividing cells are needed for sensitive responses, especially when transcriptional and translational inhibitors, directly interfering with the luciferase production, are tested . The assay can be easily automated for high-throughput screening purposes of pharmaceutical industry. Pharmazie, 2003 Jun, 58(6), 372 - 7 Synthesis and antimicrobial testing of some new S-substituted-thiopyridines, thienopyridines, pyridothienopyrimidines and pyridothienotriazines; Abdel-Rahman AE et al.; The reaction of 5-acetyl-4-aryl-3-cyano-6-methylpyridine-2(1H)-thiones (1a, b) with 4-methylphenacyl bromide, chloro-N-arylacetamides or chloroacetonitrile gave the corresponding S-substituted thiopyridines 2a-c, 4a-f and 6a-c, respectively . The latter compounds underwent intramolecular Thorpe-Ziegler cyclization to give 2-substituted 5-acetyl-3-amino-4-aryl-6-methylthieno{2,3-b}pyridines 3a-c, 5a-f and 7a-c . Compounds 5a-f and 7b, c are key intermediates in the synthesis of the target compounds . Some compounds showed remarkable antimicrobial activity. Plant Mol Biol, 2003 May, 52(2), 433 - 46 A maize defense-inducible gene is a major facilitator superfamily member related to bacterial multidrug resistance efflux antiporters; Simmons CR et al.; A defense-inducible maize gene was discovered through global mRNA profiling analysis . Its mRNA expression is induced by pathogens and defense-related conditions in various tissues involving both resistant and susceptible interactions . These include Cochliobolus heterostrophus and Cochliobolus carbonum infection, ultraviolet light treatment, the Les9 disease lesion mimic background, and plant tissues engineered to express flavonoids or the avirulence gene avrRxv . The gene was named Zm-mfs1 after it was found to encode a protein related to the major facilitator superfamily (MFS) of intregral membrane permeases . It is most closely related to the bacterial multidrug efflux protein family, typified by the Escherichia coli TetA, which are proton motive force antiporters that export antimicrobial drugs and other compounds, but which can be also involved in potassium export/proton import or potassium re-uptake . Other related plant gene sequences in maize, rice, and Arabidopsis were identified, three of which are introduced here . Among this new plant MFS subfamily, the characteristic MFS motif in cytoplasmic TM2-TM3 loop, and the antiporter family motif in transmembrane domain TM5 are both conserved, however the TM7 and the cytoplasmic TM8-TM9 loop are divergent from those of the bacterial multidrug transporters . We hypothesize that Zm-Mfs1 is a prototype of a new class of plant defense-related proteins that could be involved in either of three nonexclusive roles: (1) export of antimicrobial compounds produced by plant pathogens; (2) export of plant-generated antimicrobial compounds; and (3) potassium export and/or re-uptake, as can occur in plant defense reactions. Microbiology, 2003 Jul, 149(Pt 7), 1893 - 900 EST analysis of genes expressed by the zygomycete pathogen Conidiobolus coronatus during growth on insect cuticle; Freimoser FM et al.; Conidiobolus coronatus (Zygomycota) is a facultative saprobe that is a pathogen of many insect species . Almost 2000 expressed sequence tag (EST) cDNA clones were sequenced to analyse gene expression during growth on insect cuticle . Sixty percent of the ESTs that could be clustered into functional groups (E<or=10(-5)) had their best BLAST hits among fungal sequences . These included chitinases and multiple subtilisins, trypsin, metalloprotease and aspartyl protease activities with the potential to degrade host tissues and disable anti-microbial peptides . Otherwise, compared to the ascomycete entomopathogen Metarhizium anisopliae, Con . coronatus produced many fewer types of hydrolases (e.g . no phospholipases), antimicrobial agents, toxic secondary metabolites and no ESTs with putative roles in the generation of antibiotics . Instead, Con . coronatus produced a much higher proportion of ESTs encoding ribosomal proteins and enzymes of intermediate metabolism that facilitate its rapid growth . These results are consistent with Con . coronatus having adapted a modification of the saprophytic ruderal-selected strategy, using rapid growth to overwhelm the host and exploit the cadaver before competitors overrun it . This strategy does not preclude specialization to pathogenicity, as Con . coronatus produces the greatest complexity of proteases on insect cuticle, indicating an ability to respond to conditions in the cuticle. Microbiology, 2003 Jul, 149(Pt 7), 1719 - 27 Rapid identification of Gram-positive anaerobic coccal species originally classified in the genus Peptostreptococcus by multiplex PCR assays using genus- and species-specific primers; Song Y et al.; Here, a rapid and reliable two-step multiplex PCR assay for identifying 14 Gram-positive anaerobic cocci (GPAC) species originally classified in the genus Peptostreptococcus (Anaerococcus hydrogenalis, Anaerococcus lactolyticus, Anaerococcus octavius, Anaerococcus prevotii, Anaerococcus tetradius, Anaerococcus vaginalis, Finegoldia magna, Micromonas micros, Peptostreptococcus anaerobius, Peptoniphilus asaccharolyticus, Peptoniphilus harei, Peptoniphilus indolicus, Peptoniphilus ivorii and Peptoniphilus lacrimalis) is reported . Fourteen type strains representing 14 GPAC species were first identified to the genus level by multiplex PCR (multiplex PCR-G) . Since three of these genera (Finegoldia, Micromonas and Peptostreptococcus) contain only a single species, F . magna, M . micros and P . anaerobius, respectively, these organisms were identified to the species level directly by using the multiplex PCR-G . Then six species of the genus Anaerococcus (A . hydrogenalis, A . lactolyticus, A . octavius, A . prevotii, A . vaginalis and A . tetradius) were further identified to the species level using multiplex PCR assays (multiplex PCR-Ia and multiplex PCR-Ib) . Similarly, five species of the genus Peptoniphilus (Pn . asaccharolyticus, Pn . harei, Pn . indolicus, Pn . ivorii and Pn . lacrimalis) were identified to the species level using multiplex PCR-IIa and multiplex PCR-IIb . The established two-step multiplex PCR identification scheme was applied to the identification of 190 clinical isolates of GPAC species that had been identified previously to the species level by 16S rRNA sequencing and phenotypic tests . The identification obtained from multiplex PCR assays showed 100 % agreement with 16S rDNA sequencing identification, but only 65 % (123/190) agreement with the identification obtained by phenotypic tests . The multiplex PCR scheme established in this study is a simple, rapid and reliable method for the identification of GPAC species . It will permit a more accurate assessment of the role of various GPAC species in infection and of the degree of antimicrobial resistance in each of the group members. Blood, 2003 Nov 1, 102(9), 3396 - 403 Epub 2003 Jul 10. Eosinophil-derived neurotoxin (EDN), an antimicrobial protein with chemotactic activities for dendritic cells; Yang D et al.; Recent publications have highlighted the chemotactic activities of antimicrobial proteins derived from the granules of neutrophils and basophils . Eosinophil granules also contain antimicrobial proteins . One of them is eosinophil-derived neurotoxin (EDN), a protein belonging to the ribonuclease A (RNase A) superfamily, which has recently been found to have antiviral activity in vitro . We found that EDN was selectively chemotactic for dendritic cells (DCs) . The DC chemotactic activity of EDN was inhibited by either pretreatment of DCs with pertussis toxin or by simultaneous addition of placental RNase inhibitor to inhibit the activity of EDN . EDN was not chemotactic for leukocytes other than DCs . Mouse eosinophil-associated RNase 2 (mEAR2), one of a cluster of divergent orthologs of human EDN, was also chemotactic for human as well as mouse DCs . Sequence and mutational analysis demonstrated the importance of the N-terminal region of mEAR2 in mediating its chemotactic effect on DCs . EDN also induced the activation of p42/44 mitogen-activated protein kinase (MAPK) in DCs . Furthermore, injection of mEAR2 into the air pouches of mice resulted in the recruitment of DCs into the air pouches . Thus, EDN and its mouse ortholog, mEAR2, are eosinophil granule-derived antimicrobial RNases that function as chemoattractants for DCs in vitro and in vivo. Ann Transplant, 2002, 7(4), 42 - 5 Interaction of E . coli with dendritic cells--a model for studies of graft infections; Stanislawska J et al.; The rejection process of skin allografts is mediated by dendritic cells (DC) and lymphocytes . The recipient DCs are engaged not only into an allogenic but also antibacterial reaction to the penetrating bacteria . The capacity of these cells to sample sites of pathogen entry, respond to microbial signals and activate naive T cells suggests a critical role for DC in initiating antimicrobial immunity . In our study, we investigated the ability of the green fluorescent protein (Gfp) labelled E . coli to infect DC . We studied kinetics of in vitro and in vivo adherence and incorporation of E . coli by rat spleen and bone marrow (BM) DC . Bacterial adherence to the cell surface was observed after 2 h incubation of DC with bacteria . A 24-hour culture of DC from both sources was followed by bacterial adherence to all cells and engulfment by at least 50% of cells . There was an increased expression of the phenotypic markers on the DC cultured with E.coli . The Gfp-labelled E.coli should be useful for studies of the activation of dendritic cells . The method will allow to study the process of simultaneous activation of DC by allo- and bacterial antigens. Biochim Biophys Acta, 2003 Jul 14, 1638(2), 157 - 63 Limited proteolysis of surfactant protein D causes a loss of its calcium-dependent lectin functions; Griese M et al.; Surfactant protein D (SP-D) is a multimeric collagenous lectin that mediates the clearance of pathogens and modulates immune cell functions via its C-terminal carbohydrate recognition domain (CRD) . We hypothesized that extracellular proteolysis of SP-D may result in a loss of its functional properties . Multimeric SP-D was partially digested by human leukocyte elastase (HLE) dose- and time-dependently . Physiologic concentrations of calcium slowed, but did not protect from degradation . In solution, both native and degraded SP-D had an apparent molecular weight of 650 to >1000 kDa . Under reducing conditions, the degraded SP-D monomers run at 10 kDa less than native SP-D . Amino acid sequencing located all major cleavage sites into the CRD . Functional studies showed that degraded SP-D had lost its calcium-dependent lectin properties, i.e . neither bound to mannose nor agglutinated bacteria . These studies demonstrate that elastase results in the limited proteolysis of SP-D with loss of its CRD-dependent activities and suggest that proteases at concentrations observed in various lung diseases may impair the antimicrobial and immunomodulatory roles of SP-D. Hematology, 2002 Oct, 7(5), 281 - 9 Oral mucositis: time for more studies; Alvarado Y et al.; Oral Mucositis (OM) is a frequent cause of severe morbidity in patients receiving chemotherapy and/or radiotherapy . The pathophysiology of OM involves direct cytotoxic effects, local inflammatory responses, and alterations in oral microflora . There are currently no approved agents for the prevention or treatment of OM . In this review we briefly describe current knowledge of the pathophysiology, clinical presentation, and management of OM . We then discuss investigational agents being studied in OM with a particular focus on local antimicrobial agents, hemopoietic growth factors, and cytokines . Measures to reduce the incidence of OM and/or alleviate its clinical sequelae should be incorporated into all chemotherapy or radiotherapy studies. Zhonghua Liu Xing Bing Xue Za Zhi, 2003 Jun, 24(6), 447 - 8 {In-vitro activity of rabeprazole, lansoprazole, and esomeprazole against Helicobacter pylori}; He LH et al.; OBJECTIVE: To investigate the antimicrobial activity of Pariet, Tekpron, Nexium, respectively, against Helicobacter pylori (H . pylori) in vitro . METHODS: Antimicrobial effects of these medicines were evaluated through detection of MICs for 3 H . pylori strains isolated from different countries . RESULTS: The MIC(99) contents were 2.25 mg/L, 42.5 mg/L and 360 mg/L, respectively, for the three medicines . The strains under testing exhibited the same susceptibility to each medicine . Nexium did not inhibit the bacteria under the concentration of 3.6 - 36 mg/L with more and bigger H . pylori colonies seen when compared with controls . CONCLUSIONS: The growth inhibitory activity appeared to be different among the three PPI medicines under investigation, with Rabeprazole the most potential agent of the three . Data suggested that the action of growth inhibition in vitro was resting on the characteristic of the given PPI as well as the supplements of the medicine. Clin Microbiol Infect, 2003 May, 9(5), 380 - 7 A series of infections due to Capnocytophaga spp in immunosuppressed and immunocompetent patients; Bonatti H et al.; OBJECTIVE: To investigate the epidemiology, microbiology and outcome of infections caused by Capnocytophaga spp . at a single center . METHODS: We report on ten documented infectious episodes caused by Capnocytophaga observed between 1994 and 1999 at the Innsbruck University Hospital . RESULTS: In seven of ten patients, Capnocytophaga septicemia was diagnosed during periods of neutropenia . In contrast, the remaining three patients had normal white blood cell counts when acquiring Capnocytophaga septicemia (one) and pleural empyema (two) . Blood cultures containing long, slender, Gram-negative rods, which grew slowly under anaerobic conditions and lacked susceptibility to metronidazole, were subcultivated in a CO2-enriched atmosphere (5%) . Subcultivation yielded Capnocytophaga in all ten cases within 2-12 days . The patients were then placed on appropriate antibiotic therapy, with or without additional surgical intervention, and the organism was eradicated . CONCLUSION: Identification of Capnocytophaga facilitates appropriate, and in most cases effective, antimicrobial therapy. Clin Microbiol Infect, 2003 Jun, 9(6), 531 - 7 Changing epidemiology and predictors of mortality in patients with spontaneous bacterial peritonitis at a liver transplant unit; Singh N et al.; OBJECTIVES: To determine whether antimicrobial resistance in pathogens and outcome in patients with spontaneous bacterial peritonitis (SBP) has evolved over time . METHODS: Sixty-one consecutive episodes of SBP were studied in patients with end-stage liver disease undergoing evaluation for liver transplantation between 1991 and 2001 . Patients were dichotomized into a cohort between 1991 and 1995 (the earlier cohort) and 1996-2001 (the later cohort) . RESULTS: Overall, 19% of all bacteria were multiply-antibiotic resistant . The frequency of multiple-antibiotic resistance in bacteria increased from 8.3% to 38.5% in the earlier as compared to the later cohort (P = 0.07) . Overall, mortality at 30 days in the study patients was 26% and had remained unchanged between the two cohorts . The mortality rate was significantly higher in patients with multiply-antibiotic-resistant bacteria than in those with other bacteria (P = 0.045) . However, the Child-Pugh score (P = 0.003), and renal failure (P = 0.04) were the only independently significant predictors of mortality in patients with SBP . CONCLUSIONS: Mortality in patients with end-stage liver disease who developed SBP has remained unchanged over the last decade . Although multiple-antibiotic resistance in bacteria causing SBP has increased over time, the severity of hepatic and renal dysfunction were the predominant determinants of outcome in these patients. Clin Microbiol Infect, 2003 Jun, 9(6), 467 - 74 Multicenter evaluation of the reproducibility of the proposed antifungal susceptibility testing method for fermentative yeasts of the Antifungal Susceptibility Testing Subcommittee of the European Committee on Antimicrobial Susceptibility Testing (AFST-EUCAST); Cuenca-Estrella M et al.; OBJECTIVE: To evaluate the intra- and inter-laboratory reproducibility of a new standard for susceptibility testing of fermentative yeasts . This standard is based on the M27-A procedure of the National Committee for Clinical Laboratory Standards (NCCLS), but incorporates several modifications, including spectrophotometric growth-dependent endpoint reading . METHODS: Nine laboratories participated in the study . Common material lots were used to test six Candida species (one each of C . albicans, C . tropicalis, C . parapsilosis, C . glabrata, C . krusei, and C . lusitaniae), and two quality control strains (C . krusei ATCC6258 and C . parapsilosis ATCC22019) . Triplicate testing on three separate days was performed in microtiter format with RPMI-2% glucose, pH 7.0 . Flucytosine, fluconazole and itraconazole were tested . In total, 3888 MIC values were included in the analyses . Reproducibility was calculated by means of agreement (percentage of MICs within one two-fold dilution of the mode) and intraclass correlation coefficient (ICC, maximum value of 1) . RESULTS: The average intra-laboratory agreements were 99% and 96% after 24 h and 48 h of incubation, respectively, with ICCs of 0.98 and 0.97 (P < 0.05) . Two strains exhibiting a trailing effect showed intra-laboratory agreement of 92% and ICCs of < 0.91 at 48 h . The inter-laboratory agreement was 94% and 88% after 24 h and 48 h, respectively, with ICCs of 0.93 and 0.91 (P < 0.05) . Lower values of agreement and ICCs were obtained for strains exhibiting trailing after 48 h of incubation . Itraconazole yielded the lowest values of reproducibility . CONCLUSION: The new procedure of EUCAST for antifungal susceptibility testing is a reproducible method within and between laboratories and offers several advantages over the NCCLS approved method. J Agric Food Chem, 2003 Jul 16, 51(15), 4225 - 32 Rapid confirmatory assay for determining 12 sulfonamide antimicrobials in milk and eggs by matrix solid-phase dispersion and liquid chromatography-mass spectrometry; Bogialli S et al.; A rapid confirmatory method for determining 12 sulfonamide (SAs) antibacterials in whole milk and eggs is presented . This method is based on the matrix solid-phase dispersion technique with hot water as extractant followed by liquid chromatography (LC)-mass spectrometry (MS) . The LC-MS instrument was equipped with an electrospray ion source and a single quadrupole . After 4 mL of a milk sample containing the analytes had been deposited on sand (crystobalite), this material was packed into an extraction cell . SAs were extracted by flowing 4 mL of water through the cell heated at 75 degrees C . With some modifications, this procedure was applied also to eggs . After pH adjustment and filtration, 0.5 mL of the final extracts was then injected into the LC column . MS data acquisition was performed in the positive-ion mode and by monitoring at least three ions for each target compound . The in-source collision-induced dissociation process produced confirmatory ions . At the 50 ng/g level, recovery of the analytes in milk and eggs was 77-92% with relative standard deviations ranging between 1 and 11% . Estimated limits of quantification (S/N = 10) were 1-3 ng/g of SAs in milk and 2-6 ng/g in eggs . With both matrices, attempts to reduce the analysis time by using a short chromatographic run time caused severe ion signal suppression for the early-eluted SAs . This effect was traced to competition effects by polar endogenous coextractives, maybe proteinaceous species, which are eluted in the first part of the chromatographic run . This unwelcome effect was almost completely removed by simply adopting more selective chromatographic conditions. Acta Pol Pharm, 2003 Jan-Feb, 60(1), 97 - 100 Chemoprevention of cancerogenesis--the role of sulforaphane; Solowiej E et al.; Isothiocyanates are a group of naturally occurring compounds with interesting medical properties, such as antimicrobial, antioxidant and antitumor activities . In our work we were trying to present the tumoricidal activity of new synthesized derivatives of one isothiocyanate: 1-isothio-cyanato-(4R)-(methylsulfinyl) butane {sulforaphane} . Many chemical substances derived from plants, undoubtedly have protective properties . The effectiveness of sulforaphane is based on induction of hepatic detoxifying enzymes. Acta Pol Pharm, 2003 Jan-Feb, 60(1), 31 - 9 Technology of eye drops containing aloe (Aloe arborescens Mill.--Liliaceae) and eye drops containing both aloe and neomycin sulphate; Kodym A et al.; Eye drops made of aloe are a sterile, aqueous extract of fresh leaves of Aloe arborescens Mill., containing necessary additives and neomycin sulphate . The aim of the studies was to establish the technology of eye drops containing biologically active aloe substances and those containing both chemical constituents of aloe and neomycin sulphate . Within the studies, the formulary content and the way of preparing eye drops were determined, criteria were defined and methods of qualitative assessment of drops were proposed . On the basis of the proposed analytical methods, the physicochemical and microbiological stability of the eye drops stored at a temperature of 20-25 degrees C was studied . As the criteria of qualitative assessment of the eye drops, the following analyses were considered: sterility, appearance of the eye drops (clarity), pH, osmotic pressure, density, viscosity, TLC analysis, content of aloenin and aloin, studies of anti-microbial activity of neomycin in the drops, and preservative efficiency of thiomersal in the eye drops . The studies showed that the additives such as: sodium chloride, benzalkonium chloride, chlorhexidine diacetate and digluconate, phenylmercuric borate and Nipagins M and P could not be used to prepare the eye drops because they were involved in pharmaceutical interactions with chemical constituents of aloe in the eye drops . The eye drops containing: aqueous extract of fresh leaves of aloe, boric acid, thiomersal, sodium pyrosulphite, disodium EDTA, beta-phenylethyl alcohol and neomycin sulphate, both freshly prepared and after two years of storage, met the requirements of the Polish Pharmacopoeia (PPh V) mentioned in the monograph Guttae ophthalmicae . They were sterile, clear, their osmotic pressure approximated the osmotic pressure of lacrimal fluid and they were characterized by appropriate pH . Aloenin in the drops was much more stable than aloin . Neomycin after two years of storage retained almost 98% of its starting antimicrobial activity which allows the conclusion that the biologically active aloe substances did not decrease the stability of neomycin in the drops . The preservation assay showed that thiomersal, both in the freshly prepared drops and after two years of storage, maintained antimicrobial activity, which was in accordance with PPh V. Orv Hetil, 2003 Jun 1, 144(22), 1077 - 83 {Eradication of Helicobacter pylori infection in Hungary (1993-2002): a meta-analysis}; Buzas GM; INTRODUCTION: Meta-analysis is a useful tool in obtaining sound data in evidence-based medicine . The guidelines of eradication have been proposed in the I . Maastricht consensus (1996), which was modified in 2000 . AIM OF STUDY: The scope of present paper is the meta-analysis of eradication studies performed in Hungary between 1993-2002 . METHODS: Articles/abstracts dealing with eradication, published in peer-reviewed Hungarian and international journals, meetings of the Hungarian Society of Gastroenterology and its Endoscopic Section and international congresses were identified from major databases and by manual search . The data were classified into groups based on eradication schedules, type of antibiotic used, and provenience of the study . The pooled eradication rates were calculated and compared with the international data . RESULTS: The pooled eradication rate of proton pump inhibitor-based triple therapies was 82.9% (confidence interval: 72.1-93.7%); omeprazole, pantoprazole and lansoprazole-based therapies given b.i.d . were equally efficient . H2-blockers based triple combinations lasting 10-14 days obtained a pooled eradication rate of 86.3% (confidence interval: 72.2-99.8%), insignificantly higher (p = 0.07) than the rate obtained with proton pump inhibitors . Classical triple therapy's (bismuth + 2 antimicrobials) pooled eradication rate is significantly lower as that of proton pump inhibitor (p = 0.01) or H2 blocker-based combinations (p = 0.008) . Clarithromycin, nitroimidazoles, amoxicillin and doxycyclin-based therapies obtained pooled eradication rates of 79.4%, 74.7%, 79.3% and 73.0%, respectively . The results obtained in randomized controlled trials and university centers were better than those achieved in county and urban hospitals . CONCLUSIONS: The results are in part contrasting with the recommendations of European and national consensus . Proton pump inhibitor-based triple regimens are proposed as first-line treatment, which is in agreement with our results, but H2-blockers based regimens seem to be of same efficacy in open studies, although these represent a lower level of evidence . Classical triple therapies, also recommended, did not reach acceptable rates of eradication . Antibiotic-based analysis did not reflect the local resistance profile . A multicenter, randomized Hungarian study is warranted to assess the real efficacy of the eradication schedules. J Immunol, 2003 Jul 15, 171(2), 964 - 70 Early self-regulatory mechanisms control the magnitude of CD8+ T cell responses against liver stages of murine malaria; Hafalla JC et al.; Following immunization with Plasmodium yoelii sporozoites, the CD8(+) T cell population specific for the SYVPSAEQI epitope expressed in sporozoite and liver stages of this malaria parasite revealed the existence of a short term Ag presentation process that translated into a single clonal burst . Further expansion of this CD8(+) T cell population in conditions of sustained Ag exposure and additional supply of naive cells was inhibited by regulatory mechanisms that were developed as early as 24-48 h after priming . Studies using mouse models for Plasmodium or influenza virus infections revealed that this mechanism is Ag specific and is mediated by activated CD8(+) T cells that inhibit the priming of naive cells . This interference of the priming of naive cells appeared to result from limited access to Ag-presenting dendritic cells, which become disabled or are eliminated after contact with activated cells . Thus, concomitantly with the development of their effector antimicrobial capacity, CD8(+) T cells also acquire a self-regulatory role that is likely to represent one of the earliest mechanisms induced in the course of an immune response and that limits the magnitude of the early expansion of CD8(+) T lymphocytes reactive to microorganisms. J Immunol, 2003 Jul 15, 171(2), 931 - 7 Inactivation of human beta-defensins 2 and 3 by elastolytic cathepsins; Taggart CC et al.; beta-Defensins are antimicrobial peptides that contribute to the innate immune responses of eukaryotes . At least three defensins, human beta-defensins 1, 2, and 3 (HBD-1, -2, and -3), are produced by epithelial cells lining the respiratory tract and are active toward Gram-positive (HBD-3) and Gram-negative (HBD-1, -2, and -3) bacteria . It has been postulated that the antimicrobial activity of defensins is compromised by changes in airway surface liquid composition in lungs of patients with cystic fibrosis (CF), therefore contributing to the bacterial colonization of the lung by Pseudomonas and other bacteria in CF . In this report we demonstrate that HBD-2 and HBD-3 are susceptible to degradation and inactivation by the cysteine proteases cathepsins B, L, and S . In addition, we show that all three cathepsins are present and active in CF bronchoalveolar lavage . Incubation of HBD-2 and -3 with CF bronchoalveolar lavage leads to their degradation, which can be completely (HBD-2) or partially (HBD-3) inhibited by a cathepsin inhibitor . These results suggest that beta-defensins are susceptible to degradation and inactivation by host proteases, which may be important in the regulation of beta-defensin activity . In chronic lung diseases associated with infection, overexpression of cathepsins may lead to increased degradation of HBD-2 and -3, thereby favoring bacterial infection and colonization. Can J Gastroenterol, 2003 Jun, 17 Suppl B, 41B - 45B Eradication therapy should be different for dyspeptic patients than for ulcer patients; de Boer WA; Physicians should try to achieve an optimal cure rate with their initial Helicobacter pylori eradication therapy . Most physicians use the same treatment in all their patients . H pylori infection in patients with peptic ulcer disease (PUD) is more likely to be cured than that in patients with functional dyspepsia (FD) . Differences in cure rates of 5% to 15% are usually reported, which is considered to be clinically relevant . A plausible biological explanation for this finding suggests that different strains (virulent {cagA+, vacA type s1} compared with nonvirulent strains {cagA-, vacA type s2}) in PUD and FD induce different changes in the gastric mucosa, and this facilitates or impairs antimicrobial efficacy . Physicians should be aware that most published treatment studies have included only PUD patients . This means that in clinical practice cure rates obtained in patients with FD or perhaps uninvestigated dyspepsia are usually lower than those reported in the literature . This has implications for the choice of treatment . Physicians should consider prolonging the duration of initial Helicobacter eradication therapy from seven to 10 to 14 days in patients without ulcers. Can J Gastroenterol, 2003 Jun, 17 Suppl B, 30B - 32B Are there geographical and regional differences in Helicobacter pylori eradication? Vakil N. An important area of controversy in Helicobacter pylori eradication is the apparent difference in eradication rates seen in different countries and populations . A recent meta-analysis showed that several factors may affect the outcome of therapy . Individuals residing in northeast Asia had higher eradication rates than those residing in Europe or other areas of Asia . Triple and quadruple drug therapies had significantly higher eradication rates than did dual drug therapies . Treatment regimens that lasted longer than 14 days were better than those that lasted less than seven days, but there was no significant advantage for 10 day therapy over seven day therapy . A number of factors may play a role in determining the regional and geographical differences in H pylori eradication therapy . Included in these factors are genetic differences in the metabolism of the proton pump inhibitor, which can alter the availability of antimicrobials in the stomach . Regional differences in antimicrobial resistance also affect the outcome of therapy . Some studies suggest that the degree of gastritis and the nature of the underlying disease may affect the outcome of therapy, but the data are controversial . Understanding the regional and geographical differences in H pylori eradication can help physicians select the optimal treatment regimen in different regions. Drugs Today (Barc), 2000 Dec, 36(12), 829 - 34 Slowly resolving and nonresolving pneumonias; Cunha BA; Nonresolving or slowly resolving pulmonary infiltrates are a clinical diagnostic challenge for physicians . It is important to differentiate slowly resolving from nonresolving pneumonias because the causes of each of these two clinical entities are different . In general, slowly resolving pneumonias are due to antimicrobial or host defense factors . Nonresolving pneumonias are usually noninfectious and usually require invasive diagnostic techniques to confirm the diagnosis . The most common clinical error made in approaching patients with nonresolving or slowly resolving pulmonary infiltrates is to treat the patient with different antibiotics over an extended period of time . Non-resolving or slowly resolving pneumonia should prompt the clinician to intensify diagnostic efforts to arrive at an etiologic diagnosis . The response should not be extended to polypharmacy since antibiotic-related causes of failure in treating problems resulting in slowly resolving pneumonia are one of the least common reasons for this clinical presentation. Drugs Today (Barc), 2000 Nov, 36(11), 785 - 91 Pneumonia in the elderly; Cunha BA; Pneumonia is a frequent cause of mortality and morbidity in the elderly . This article discusses pneumonia in the elderly based on causative organisms and sites where the pneumonia was acquired . Community-acquired pneumonia, nursing home-acquired pneumonia and nosocomial pneumonia in the elderly are reviewed from a diagnostic and therapeutic perspective . Optimal antimicrobial therapy is based on knowledge of the pulmonary pathogens associated with community-acquired, nursing home-acquired and nosocomial pneumonias . Antibiotic dosing should take into account the functional action of the liver and kidneys in individuals of advanced age. Nature, 2003 Jul 17, 424(6946), 277 - 83 Epub 2003 Jun 29. Genes that act downstream of DAF-16 to influence the lifespan of Caenorhabditis elegans; Murphy CT et al.; Ageing is a fundamental, unsolved mystery in biology . DAF-16, a FOXO-family transcription factor, influences the rate of ageing of Caenorhabditis elegans in response to insulin/insulin-like growth factor 1 (IGF-I) signalling . Using DNA microarray analysis, we have found that DAF-16 affects expression of a set of genes during early adulthood, the time at which this pathway is known to control ageing . Here we find that many of these genes influence the ageing process . The insulin/IGF-I pathway functions cell non-autonomously to regulate lifespan, and our findings suggest that it signals other cells, at least in part, by feedback regulation of an insulin/IGF-I homologue . Furthermore, our findings suggest that the insulin/IGF-I pathway ultimately exerts its effect on lifespan by upregulating a wide variety of genes, including cellular stress-response, antimicrobial and metabolic genes, and by downregulating specific life-shortening genes. Urol Int, 2003, 71(1), 124 - 6 Amoxicillin-induced nephritis and tubulitis in a child; Ferrara P et al.; The occurrence of interstitial nephritis in patients receiving antimicrobial therapy has frequently been reported in adults while it has rarely been described in children . We report the case of a patient treated with amoxicillin who presented hallucinations and serosanguineous blisters during treatment and developed renal failure a few days after discontinuation of the drug . On renal biopsy an interstitial nephritis with tubulitis was identified . Rev Chir Orthop Reparatrice Appar Mot, 2003 Jun, 89(4), 287 - 96 {Use of combined gallium-technetium scintigraphy to determine the interval before second-stage prosthetic reimplantation in hip arthroplasty infection: a consecutive series of 30 cases}; Piriou P et al.; PURPOSE OF THE STUDY: We report a series of 30 consecutive patients with chronically infected total hip replacement in a prospective treatment protocol that included two-stage revision surgery and scintigraphic monitoring . The serial bone scans were used to evaluate the course of infection, but not for diagnosis . Negative scintigraphic results were required before the second-stage prosthesis reinsertion after laboratory, clinical, and radiographic normalization were achieved . MATERIAL AND METHODS: Between 1987 and 1997, we prospectively followed thirty patients, who had a chronically infected hip arthroplasty treated by the conventional two-stage revision procedure using scintigraphic verification . For the present series, negative bone scan results were achieved in the resected hip before reinsertion of the prosthesis in all patients except one . The labels used were in every case gallium-67 and technetium-99m MDP with early and late (after 30 hours) scans . A scintigraphic result was considered positive if more gallium than technetium was fixed at a site . Our conventional medical and surgical protocol consisted of an initial complete excision of all foreign bodies with systematic parenteral administration of two antibiotics after having searched for the causative organism . A spacer was never used . Tibial pin traction was always applied during the duration of drainage of the wound . The antimicrobial regimen was administered to all of these patients for 3 months . The prosthesis was reinserted when C-reactive protein (CRP) levels returned to normal and negative scintigraphic results were obtained after a period with no antibiotic therapy . Reimplantation of the prosthesis was always performed with preventive antibiotic therapy selected according to the susceptibility of the initial organisms and begun after collecting new intraoperative bacteriological culture specimens . This antibiotic therapy was pursued only for the duration of the postoperative drainage . RESULTS: This follow-up based on combined technetium-gallium bone scans demonstrated two major advantages . First, no recurrence of infection was observed except in the single patient for whom the protocol was not observed . The second advantage was to permit nonarbitrary determination of the moment of reimplantation of the prosthesis, as there is no clear consensus regarding the interval before reinsertion in the literature . The patients underwent the second-stage of hip reconstruction after a mean interval of 9 months . The mean delay before negative scintigraphic results was 7 months . DISCUSSION: This method, which determines the optimum delay before reimplantation reducing the risk of reinfection to a minimum, gave promising results in this prospective study of 30 patients. J Clin Microbiol, 2003 Jul, 41(7), 3175 - 80 Environmental isolation of Balamuthia mandrillaris associated with a case of amebic encephalitis; Schuster FL et al.; This report describes the first isolation of the ameba Balamuthia mandrillaris from an environmental soil sample associated with a fatal case of amebic encephalitis in a northern California child . Isolation of the ameba into culture from autopsied brain tissue confirmed the presence of Balamuthia: In trying to locate a possible source of infection, soil and water samples from the child's home and play areas were examined for the presence of Balamuthia: The environmental samples (plated onto nonnutrient agar with Escherichia coli as a food source) contained, in addition to the ameba, a variety of soil organisms, including other amebas, ciliates, fungi, and nematodes, as contaminants . Presumptive Balamuthia amebas were recognized only after cultures had been kept for several weeks, after they had burrowed into the agar . These were transferred through a succession of nonnutrient agar plates to eliminate fungal and other contaminants . In subsequent transfers, axenic Naegleria amebas and, later, tissue cultures (monkey kidney cells) served as the food source . Finally, the amebas were transferred to cell-free axenic medium . In vitro, the Balamuthia isolate is a slow-growing organism with a generation time of approximately 30 h and produces populations of approximately 2 x 10(5) amebas per ml . It was confirmed as Balamuthia by indirect immunofluorescence staining with rabbit anti-Balamuthia serum and human anti-Balamuthia antibody-containing serum from the amebic encephalitis patient . The environmental isolate is similar in its antimicrobial sensitivities and identical in its 16S ribosomal DNA sequences to the Balamuthia isolate from the deceased patient. J Clin Microbiol, 2003 Jul, 41(7), 3142 - 6 Evaluation of the NCCLS extended-spectrum beta-lactamase confirmation methods for Escherichia coli with isolates collected during Project ICARE; Tenover FC et al.; To determine whether confirmatory tests for extended-spectrum beta-lactamase (ESBL) production in Escherichia coli are necessary, we selected 131 E . coli isolates that met the National Committee for Clinical Laboratory Standards (NCCLS) screening criteria for potential ESBL production from the Project ICARE (Intensive Care Antimicrobial Resistance Epidemiology) strain collection . For all 131 isolates, the broth microdilution (BMD) MIC of at least one extended-spectrum cephalosporin was >/=2 micro g/ml . For 21 of 131 (16%) isolates, the ESBL confirmatory test was positive; i.e., the BMD MICs of ceftazidime or cefotaxime decreased by >/=3 doubling dilutions in the presence of clavulanic acid (CA) or the disk diffusion zone diameters increased by >/=5 mm around ceftazidime or cefotaxime disks in the presence of CA . All 21 isolates were shown by PCR to contain at least one of the genes bla(TEM), bla(SHV), and bla(OXA), and in isoelectric focusing (IEF) tests, all isolates demonstrated at least one beta-lactamase band consistent with a TEM, SHV, or OXA enzyme . Of the 21 isolates, 3 showed a CA effect for cefotaxime by BMD but not by disk diffusion testing . A total of 59 (45%) of the 131 isolates demonstrated decreased susceptibility to cefpodoxime alone (MIC = 2 to 4 micro g/ml), and none had a positive ESBL confirmatory test result . These were classified as false positives according to ESBL screen test results . For the remaining 51 (39%) isolates, the cefpodoxime MICs ranged from 16 to >128 micro g/ml and the MICs for the other extended-spectrum cephalosporins were highly variable . All 51 isolates gave negative ESBL confirmatory test results . Most showed IEF profiles consistent with production of both a TEM and an AmpC beta-lactamase, and representative isolates of several phenotypic groups showed changes in porin profiles; these 51 isolates were considered true negatives . In all, only 16% of 131 E . coli isolates identified as potential ESBL producers by the current NCCLS screening criteria were confirmed as ESBL producers . Thus, changing the interpretation of extended-spectrum cephalosporins and aztreonam results from the susceptible to the resistant category without confirming the presence of an ESBL phenotype would lead to a large percentage of false resistance results and is not recommended . However, by increasing the cefpodoxime MIC screening breakpoint to >/=8 micro g/ml, 45% of the false-positive results could be eliminated . NCCLS has incorporated this change in the cefpodoxime screening breakpoint in its recent documents. J Biol Chem, 2003 Sep 19, 278(38), 36859 - 67 Epub 2003 Jul 03. Solution structure of the recombinant penaeidin-3, a shrimp antimicrobial peptide; Yang Y et al.; Penaeidins are a family of antimicrobial peptides of 47-63 residues isolated from several species of shrimp . These peptides display a proline-rich domain (N-terminal part) and a cysteine-rich domain (C-terminal part) stabilized by three conserved disulfide bonds whose arrangement has not yet been characterized . The recombinant penaeidin-3a of Litopenaeus vannamei (63 residues) and its {T8A}-Pen-3a analogue were produced in Saccharomyces cerevisiae and showed similar antimicrobial activity . The solution structure of the {T8A}-Pen-3a analogue was determined by using two-dimensional 1H NMR and simulated annealing calculations . The proline-rich domain, spanning residues 1-28 was found to be unconstrained . In contrast, the cysteine-rich domain, spanning residues 29-58, displays a well defined structure, which consists of an amphipathic helix (41-50) linked to the upstream and the downstream coils by two disulfide bonds (Cys32-Cys47 and Cys48-Cys55) . These two coils are in turn linked together by the third disulfide bond (Cys36-Cys54) . Such a disulfide bond packing, which is in agreement with the analysis of trypsin digests by ESI-MS, contributes to the highly hydrophobic core . Side chains of Arg45 and Arg50, which belong to the helix, and side chains of Arg37 and Arg53, which belong to the upstream and the downstream coils, are located in two opposite parts of this globular and compact structure . The environment of these positively charged residues, either by hydrophobic clusters at the surface of the cysteine-rich domain or by sequential hydrophobic residues in the unconstrained proline-rich domain, gives rise to the amphipathic character required for antimicrobial peptides . We hypothesize that the antimicrobial activity of penaeidins can be explained by a cooperative effect between the proline-rich and cysteine-rich features simultaneously present in their sequences. Int J Antimicrob Agents, 2003 Jul, 22(1), 77 - 80 Production and resolution of cantharidin-induced inflammatory blisters; Maglio D et al.; While inflammatory blisters have long been utilized as a means of evaluating antimicrobial disposition to aid in the development of new treatments for skin and skin structure infections, sparse data are available regarding the healing of the blisters once the experiment has been completed . We report the blister induction technique and resolution time in ten volunteers enrolled in a pharmacokinetic study using the cantharidin-induced inflammatory blister technique. Int J Antimicrob Agents, 2003 Jul, 22(1), 1 - 6 Empirical treatment of acute cystitis in women; Nicolle LE; Empirical antimicrobial treatment for acute cystitis in women requires continuing reassessment as the antimicrobial susceptibility of community isolates of Escherichia coli evolves . Current recommendations for 3 days trimethoprim or trimethoprim/sulphamethoxazole are compromised by increasing resistance of community E . coli to these agents . Fluoroquinolones are an alternate 3-day therapy, but increasing resistance is being reported from some countries, and widespread community use may promote resistance, limiting effectiveness of these agents for more serious infections . Alternate regimens supported by recent clinical trials suggest pivmecillinam given twice daily for 7 days is as effective as 3 days of quinolone therapy, while microbiological cure is 80% with 3 days therapy twice daily, and 90% with 3 days therapy thrice daily . Nitrofurantoin given for 7 days has a cure rate of 80-85% . Fosfomycin trometamol as a single dose has cure rates of 75-85% . All these agents are effective, but a compromise in efficacy or duration of therapy compared with current 3-day regimens may have to be considered. Aging Clin Exp Res, 2003 Feb, 15(1), 12 - 8 Risks for frequent antimicrobial-treated infections in postmenopausal women; Boudreau DM et al.; BACKGROUND AND AIMS: Infections are a major cause of morbidity and mortality in older adults . Little is known about factors that alter the susceptibility to infection in the older population . This study in postmenopausal women examines health-related conditions and behavioral factors that may increase the risk of frequent infections, defined as having, on average, one or more infections per year . METHODS: A prospective cohort study with 5 years of follow-up was conducted in 1320 women aged 55 to 80 years . The subjects were Group Health Cooperative of Puget Sound (GHC) enrollees screened for a large fracture prevention trial who also participated in a survey of dietary and supplemental vitamin use . The main outcome, total number of infection events per subject, was derived from a new method of identifying outpatient infections based on the antimicrobial prescription fills recorded in GHC automated pharmacy records . RESULTS: Prevalent lung disease (OR = 6.1, 95% CI 2.8-13.4), receiving a prescription for vitamin C (OR = 2.1, 95% CI 1.4-3.4), and the second and third tertiles of the Chronic Disease Score (OR = 1.7, 95% CI 1.1-2.7 and OR = 2.4, 95% CI 1.5-3.9, respectively) were associated with 5 or more antimicrobial-treated infections during follow-up . A body mass index (BMI) of less than 22 kg/m2 (OR = 0.6, 95% CI 0.3-1.0) was suggestive of an association . CONCLUSIONS: The study provides new information on risk factors for outpatient infections and raises new questions regarding the susceptibility to frequent infections in older women . In addition, the automated pharmacy record method used in this study offers a low-cost alternative for use in future epidemiologic research. Semin Respir Infect, 2003 Jun, 18(2), 122 - 8 When can empiric therapy for intensive care unit-acquired pneumonia be withheld or withdrawn? Koeman M, Bonten MJ. Diagnosing ventilator-associated pneumonia (VAP) is difficult, creating important clinical dilemmas for intensive care physicians . Adequate empiric antimicrobial therapy is crucial because VAP is associated with increased morbidity and mortality, especially when initial treatment is inappropriate . Because VAP is the most frequent occurring nosocomial infection, it is, to a large extent, responsible for the high antibiotic consumption in ICUs, which is an important cause for selection and induction of antibiotic resistance . In addition, antibiotics may have adverse effects and their costs should be considered . Therefore, a balance should be found between the obvious necessary therapeutic benefits and the negative effects (selection of resistant pathogens, costs, and adverse effects) of antibiotics in the treatment of VAP . Although guidelines for initial antimicrobial therapy have been established, no such recommendations exist for withholding or withdrawing antimicrobial treatment, and little is known about the optimal duration of therapy. Semin Respir Infect, 2003 Jun, 18(2), 112 - 21 Effectiveness of programs to decrease antimicrobial resistance in the intensive care unit; Hall CS et al.; Resistance of microbes to antibiotics is an increasing problem in intensive care units (ICUs) with a prevalence of 86% in some isolates . Resistance results in increased morbidity, mortality, and increased costs . Risk factors associated with the development of resistance and strategies to combat resistance are discussed . Risk factors include increased antibiotic use, host factors including severity of illness and length of stay, and lack of adherence to infection control practices . Multiple strategies to decrease resistance have been studied . Changing antimicrobial practices via guideline development, antibiotic restriction, use of information systems technology, crop rotation, narrowing spectrum of empiric antibiotics, multidisciplinary approaches, and selective decontamination have had variable results . Prevention of horizontal transmission via handwashing, glove and gown use, alternatives to soap, and improving the workload and facilities for health care workers is discussed . Primary prevention via decreased length of stay, selective digestive decontamination, vaccine development, and decreased use of invasive devices also plays a role. Proc Natl Acad Sci U S A, 2003 Jul 22, 100(15), 8880 - 5 Epub 2003 Jul 02. Engineering disulfide bridges to dissect antimicrobial and chemotactic activities of human beta-defensin 3; Wu Z et al.; Human defensins form a family of small, cationic, and Cys-rich antimicrobial proteins that play important roles in innate immunity against invading microbes . They also function as effective immune modulators in adaptive immunity by selectively chemoattracting T lymphocytes and immature dendritic cells . On the basis of sequence homology and the connectivity of six conserved Cys residues, human defensins are classified into alpha and beta families . Structures of several beta-defensins have recently been characterized, confirming the disulfide connectivity conserved within the family, i.e., Cys1-Cys5, Cys2-Cys4, and Cys3-Cys6 . We found that human beta-defensin 3 (hBD3), a recently described member of the growing beta family, did not fold preferentially into a native conformation in vitro under various oxidative conditions . Using the orthogonal protection of Cys1-Cys5 and of Cys1-Cys6, we chemically synthesized six topological analogs of hBD3 with predefined disulfide connectivities, including the (presumably) native beta pairing . Unexpectedly, all differently folded hBD3 species exhibited similar antimicrobial activity against Escherichia coli, whereas a wide range of chemotactic activities was observed with these analogs for monocytes and cells transfected by the chemokine receptor CCR6 . Furthermore, whereas substitution of all Cys residues by alpha-aminobutyric acid completely abolished the chemotactic activity of hBD3, the bactericidal activity remained unaffected in the absence of any disulfide bridge . Our findings demonstrate that disulfide bonding in hBD3, although required for binding and activation of receptors for chemotaxis, is fully dispensable for its antimicrobial function, thus shedding light on the mechanisms of action for human beta-defensins and the design of novel peptide antibiotics. Appl Environ Microbiol, 2003 Jul, 69(7), 4190 - 1 A selective medium for quantitative reisolation of Trichoderma harzianum from Agaricus bisporus compost; Williams J et al.; We adapted a selective medium, previously developed for reisolation of Trichoderma spp . from soil, for quantitative determination of growth of T . harzianum from commercial Agaricus bisporus composts . This medium enables comparisons of aggressive (sensu inhibition of A . bisporus yield) with nonaggressive T . harzianum groups . The resulting medium contains the antimicrobials chloramphenicol, streptomycin, quintozene, and propamocarb and was highly selective, allowing the recovery of T . harzianum, as viable conidia and hyphal fragments, in compact colonies with the absence of visible microbial contaminants. Int J Infect Dis, 2003 Mar, 7 Suppl 1, S5 - 12 Why do we need to eradicate pathogens in respiratory tract infections? Garau J. Evidence from studies in otitis media, acute bacterial sinusitis and acute exacerbations of chronic bronchitis indicate that clinical efficacy is dependent on bacterial eradication . Failure to eradicate bacterial pathogens increases the potential for clinical failure, incurring further costs, and may also select and maintain bacteria that are resistant to a wide range of antimicrobials . Bacteriologically confirmed clinical failures have been reported in pneumococcal pneumonia with both macrolides and older fluoroquinolones (ciprofloxacin, ofloxacin, and levofloxacin) . These failures were due to the involvement of resistant pathogens (macrolides) or suboptimal pharmacokinetics/pharmacodynamics (PK/PD) (quinolones) . However, persistent positive blood cultures have not been reported during therapy with adequate doses of benzylpenicillins or aminopenicillins . Treatment failure, driven by the failure to eradicate pathogens, leads to both economic and environmental costs, hospitalization being the major cost driver . Failure to achieve bacterial eradication may also lead to the development and spread of resistance . Different types of antimicrobials appear to be driving resistance to different extents, and this may be due to suboptimal PK/PD . In conclusion, factors to consider when prescribing include an accurate diagnosis, knowledge of local epidemiology, the role of PK/PD principles in antimicrobial choice, clinical outcomes in relation to bacteriologic efficacy, and resistance and its bacteriologic and clinical impact . The vicious cycle of infection, inappropriate therapy, bacteriologic failure, selection/spread of resistance and further infection needs to be broken by the use of appropriate treatments to achieve bacterial eradication. Int J Infect Dis, 2003 Mar, 7 Suppl 1, S1 - 4 Introduction: the goals of antimicrobial therapy; Song JH; Antimicrobial agents are generally evaluated in preclinical studies assessing in vitro activity, animal models demonstrating in vivo bacteriologic efficacy, and clinical trials primarily investigating safety and clinical efficacy . However, large sample sizes are required to detect any differences in outcomes between antimicrobials in clinical trials, and, generally, studies are powered to show only clinical equivalence . In addition, diagnosis is often based on clinical symptoms, rather than microbiological evidence of bacterial infection, and the patients most likely to have resistant pathogens are often excluded . Clinical efficacy can be achieved in some bacterial infections in which antimicrobials are suboptimal or even not prescribed . However, bacterial eradication maximizes clinical efficacy and may also reduce the development and spread of resistant organisms . The goal of antimicrobial therapy is, therefore, to eradicate bacteria at the site of infection . Bacterial eradication is not usually assessed as a primary endpoint within the limits of currently recommended clinical trial design . However, pharmacokinetic (PK) (serum concentration profiles, penetration to site of infection) and pharmacodynamic (PD) (susceptibility, concentration- versus time-dependent killing, post-antimicrobial effects) criteria can be used to predict bacteriologic efficacy . PK/PD predictions should be confirmed during all phases of antimicrobial development and throughout clinical use in response to changing patterns of resistance . A clear rationale for dose recommendations can be determined preclinically based on PK/PD parameters, and correlated with efficacy, safety and resistance endpoints in clinical trials . The duration of treatment and dose should be the shortest that will reliably eradicate the pathogen(s), and that is safe and well tolerated . Currently available agents vary significantly in their ability to achieve PK/PD parameters necessary for bacteriologic eradication . Recommendations for appropriate antimicrobial therapy should be based on PK/PD parameters, with the aim of achieving the maximum potential for eradication of both existing and emerging resistant pathogens. Pediatrics, 2003 Jul, 112(1 Pt 2), 253 - 8 Antibiotic resistance: what is the impact of agricultural uses of antibiotics on children's health? Shea KM. Antimicrobial resistance has reached crisis stage in human medicine . The rapid acceleration of multidrug-resistant bacteria in the past 2 decades has overtaken new drug development, and patients and clinicians are faced with the prospect of untreatable infections . Although much of the problem stems from overuse and misuse of antimicrobial agents in human medicine, large-scale use of antimicrobials in agriculture also contributes to the crisis . Agricultural uses of antibiotics produce environmental exposures in a variety of reservoirs, which select for resistant microbes and microbial genes . This article presents the major lines of evidence documenting the risks to human health of some of the agricultural uses of antimicrobials . A brief review of the microbiologic antecedents of resistance is followed by a discussion of agricultural uses of antimicrobials and a targeted review of the literature, which provides the background knowledge and evidence necessary for pediatricians and other clinicians to be informed and to advocate for judicious use of antimicrobials in all sectors. Pediatrics, 2003 Jul, 112(1 Pt 1), 143 - 9 Diagnosis and treatment of acute otitis media: an assessment; Garbutt J et al.; OBJECTIVE: To assess compliance with the Centers for Disease Control and Prevention (CDC) evidence-based guidelines for the judicious use of antimicrobials in children with acute otitis media (AOM) . METHODS: Compliance with CDC's recommended diagnostic criteria and antimicrobial treatments for management of AOM was assessed by chart review and self-report for 29 community pediatricians in St . Louis, Missouri . For each physician, a simple random sample of AOM visits was selected and reviewed by trained reviewers . In addition, each physician completed a questionnaire . RESULTS: Compliance with recommended diagnostic criteria was 38% (95% confidence interval: 34%-42%; n = 573) by chart audit and 41% (95% confidence interval: 24%-61%; n = 29) by self-report . Antimicrobial selection assessed by chart audit was consistent with CDC guidelines in 68% (95% confidence interval: 64%-72%) of visits for a new infection, 63% (95% confidence interval: 47%-78%) of visits for treatment failure, and 50% (95% confidence interval: 33%-67%) for recurrent disease . Self-reported compliance with treatment guidelines for new infection was 100% (95% confidence interval: 88%-100%) and 83% (95% confidence interval: 64%-94%) for treatment failure . Noncompliance was most frequently attributable to overuse of broad-spectrum antimicrobials . Most patients treated with amoxicillin received a 10-day course (98%) . Subtherapeutic dosing occurred in 26% of patients treated with amoxicillin . CONCLUSIONS: Overdiagnosis of AOM is common . Efforts to improve the judicious use of antimicrobials for AOM should focus on improving diagnostic accuracy, limiting the use of broad-spectrum antimicrobials to cases where they offer clinical benefit, and ensuring that amoxicillin dosing regimens are optimal. J Antimicrob Chemother, 2003 Aug, 52(2), 224 - 8 Epub 2003 Jul 01. Antibiotic susceptibilities of Gram-positive anaerobic cocci: results of a sentinel study in England and Wales; Brazier JS et al.; OBJECTIVE: A sentinel study was carried out to determine the antimicrobial susceptibilities of Gram-positive anaerobic cocci (GPAC) freshly isolated from clinical material in diagnostic laboratories in England and Wales . METHODS: A total of 113 GPAC isolates consisting predominantly of current or former members of the genus Peptostreptococcus was obtained from 17 sentinel laboratories in England and one in Wales . Minimum inhibitory concentrations (MICs) of 10 antimicrobial agents were determined by the Etest method . The agents tested were: penicillin, tetracycline, erythromycin, cefoxitin, clindamycin, chloramphenicol, imipenem, co-amoxiclav, piperacillin/tazobactam and metronidazole . MIC50 and MIC90 values for each drug-species combination were calculated whenever suitable numbers of each species were obtained . RESULTS: Excellent spectra of activity (0% resistance) against GPAC were seen for metronidazole, piperacillin/tazobactam, cefoxitin, imipenem and chloramphenicol . Low degrees of resistance to co-amoxiclav (3.5%), clindamycin (7.1%), penicillin (7.1%) and significant degrees of resistance to tetracycline (41.6%) and erythromycin (27.4%) were detected . Some examples of putative macrolide-lincosamide linked resistance were noted in seven (6.2%) isolates of GPAC . CONCLUSION: This study is one of the largest susceptibility studies specifically on GPAC carried out to date and the resulting data may be of value to those involved in the empirical treatment of infections involving Gram-positive anaerobic cocci. Fitoterapia, 2003 Jul, 74(5), 476 - 8 Antimicrobial activity of Amomum cannicarpum; Mathew J et al.; The petroleum ether and methanol extracts of rhizomes of Amomum cannicarpum exhibited moderate inhibiting activity against both Gram-positive and Gram-negative bacteria . None of the extractives was active against the tested moulds. Lancet Infect Dis, 2003 Jul, 3(7), 405 - 12 Antimicrobial therapy to prevent or treat oral mucositis; Donnelly JP et al.; Oral mucositis represents a significant source of morbidity after chemotherapy and radiation therapy . Since infection may have an important role in the pathophysiology of oral mucositis, several antimicrobial agents have been investigated for their efficacy in preventing and treating this disease . We sought to establish the weight of evidence for antimicrobial treatment and identified 31 prospectively designed clinical trials of which 13 reported some benefit and 15 did not . No clear pattern was identified regarding patient type, cancer treatment, or type of antimicrobial agent used, and inconsistent assessment of oral mucositis made comparison of outcomes difficult . Newer drugs, such as the topical antimicrobial peptide iseganan HCl initially showed promise in reducing mucositis and the related oral pain but the results of a phase 3 trial were disappointing and the line of enquiry was abandoned altogether . Hence, there is a need to better understand the role of the microflora in the cause of oral mucositis if an antimicrobial agent for prevention and treatment of this disease is to be developed. Genomics, 2003 Aug, 82(2), 172 - 84 Expansion of the BPI family by duplication on human chromosome 20: characterization of the RY gene cluster in 20q11.21 encoding olfactory transporters/antimicrobial-like peptides; Andrault JB et al.; Antimicrobial peptides provide a defense system against microorganisms . One class of these molecules binds lipophilic substrates and is therefore directed against gram-negative bacteria . This family includes proteins related to bactericidal/permeability-increasing protein (BPI) . We characterized an approximately 100-kb cluster of three human genes named RYSR, RYA3, and RY2G5 that are related to the BPI family . The RY cluster maps to 20q11.21, >5 Mb upstream of the BPI cluster . The RY and BPI genes have similar exon structures, indicating that they were derived by duplication from a common ancestor . We identified mouse BPI-related and RY orthologues in syntenic regions, indicating that the gene family expanded before mouse and human diverged . Expression analyses show that RYs are strongly expressed in the olfactory epithelium, suggesting that they also could act as odorant transporters or detoxification agents in the olfactory system . Together, these data show how mammals diversified their antimicrobial defenses/olfactory pathways through a duplication-driven adaptive selection process. Med Microbiol Immunol (Berl), 2004 Feb, 193(1), 1 - 17 Epub 2003 Jun 27. The power of combinatorial immunology: IL-12 and IL-12-related dimeric cytokines in infectious diseases; Holscher C; Appropriate induction of a Th1 immune response is required for effective antimicrobial immunity . However, dysregulated Th1 immune responses after infection may also lead to immunopathology . Thus, cell-mediated immune responses have to be tightly regulated . Upon infection, the production of interleukin (IL)-12, a heterodimeric cytokine composed of a p35 and a p40 subunit, is the dominant factor in Th1 cell development . The recent discovery of novel dimeric cytokines closely related to IL-12 add now to our understanding of cellular immunity and the fine tuning of T cell responses . At the onset of infection, IL-27, a heterodimer composed of the IL-12p40-related protein EBI-3 (Epstein-Barr virus-induced gene 3) and the IL-12p35-related protein p28 induces the expression of a functional IL-12 receptor in naive CD4+ T cells, making these cells sensitive to IL-12-mediated Th cell development . Later during infection, IL-23, a heterodimer composed of the IL-12p40 subunit and the IL-12p35-related molecule p19, preferentially acts on Th1 effector/memory CD4+ T cells . The IL-12p40 subunit can also form a homodimer, IL-12p80, which act as an IL-12 and IL-23 antagonist by competing at their receptors . This review focuses on these IL-12-related cytokines contributing to fine tuning of T cell responses after infection with intracellular pathogens. Anal Sci, 2003 Jun, 19(6), 969 - 70 Crystal structure of 3,3-dichloro-N-p-methoxyphenyl-4-(2-phenylstryl)-2-azetidinone; Kabak M et al.; 3,3-Dichloro-N-p-methoxyphenyl-4-(2-phenylstryl)-2-azetidinone (C22H15Cl2NO2) was studied by X-Ray analysis, which indicated a monoclinic space group, P2(1)/c, with a = 9.619(5), b = 13.879(4), c = 14.161(5)A, beta = 100.16(3)degrees, V = 1860.8(13)A3, Z = 4, Dc = 1.414 g cm(-3), micro(Mo Kalpha) = 0.366 mm(-1) and F000 = 816 . The structure was solved by direct methods and refined to R = 0.041 for 4026 reflections {I > 2sigma(I) . The beta-lactam ring (2-azetidinone) has antimicrobial affects . The substituents of the methoxyphenyl and phenyl substituents do not change the activity property of the beta-lactam ring, and the activity properties depend on the planarity of the beta-lactam ring. Rev Argent Microbiol, 2003 Jan-Mar, 35(1), 29 - 40 {Consensus on antimicrobial sensitivity tests in gram-positive cocci . Subcommittee on Antimicrobials, SADEBAC (Argentinian Society of Clinical Bacteriology), Argentinian Association of Microbiology}; Famiglietti A et al.; Antimicrobial susceptibility testing is mainly performed in Argentina by disk diffusion method, following National Committee for Clinical Laboratory Standards (NCCLS) recommendations . We worked out new recommendations for the reporting and interpretation of this test when dealing with gram-positive cocci, in accordance to local trends and epidemiology . General considerations for performing the diffusion assay, quality control, and an update on susceptibility testing for gram-positive cocci are reported in this first document . The present update should be considered as a group of recommendations summarized by Argentinean experts and as the result of a consensus meeting coordinated by the Subcomision de Antimicrobianos of the Sociedad Argentina de Bacteriologia Clinica (Asociacion Argentina de Microbiologia) . Experts in antimicrobial agents were convened in order to prepare this final document . These recommendations take into account local needs, affordability and availability to be used in current practice, tending to contribute to the correct antimicrobial treatment election, according to the particular microorganism and the infection sites. J Formos Med Assoc, 2003 Apr, 102(4), 270 - 2 Peritonitis caused by Chryseobacterium meningosepticum in a patient undergoing continuous ambulatory peritoneal dialysis; Wu VC et al.; Chryseobacterium meningosepticum is rarely encountered as a pathogen causing peritonitis in adults . A 54-year-old woman who underwent continuous ambulatory peritoneal dialysis for 8 years developed peritonitis due to C . meningosepticum . Although she received intravenous antibiotics with good in vitro activity against the organism, the fever and signs of peritonitis persisted . The Tenckhoff catheter was finally removed on the 25th day of hospitalization and the fever subsided . Four isolates of C . meningosepticum recovered from 4 ascites samples drawn on the third, 13th, 18th, and 23rd hospitalization days had identical antibiograms and random amplified DNA polymorphism patterns generated by an arbitrarily primed polymerase chain reaction . Early removal of Tenckhoff catheter and appropriate antimicrobial therapy are crucial to the successful treatment of peritonitis due to C . meningosepticum. Acta Biochim Pol, 2003, 50(2), 461 - 9 Novel properties of antimicrobial peptides; Kamysz W et al.; Endogenous peptide antibiotics are known as evolutionarily old components of innate immunity . Due to interaction with cell membrane these peptides cause permeabilization of the membrane and lysis of invading microbes . However, some studies proved that antimicrobial peptides are universal multifunctional molecules and their functions extend far beyond simple antibiotics . In this review we present an overview of the general mechanism of action of antimicrobial peptides and discuss some of their additional properties, like antitumour activity, mitogenic activity, role in signal transduction pathways and adaptive immune response. Actas Urol Esp, 2003 Apr, 27(4), 305 - 7 {Meningitis caused by multiresistant E . coli after an echo-directed transrectal biopsy}; Rodriguez-Patron Rodriguez R et al.; Transrectal prostate biopsy is the most accurate method for prostate cancer diagnosis . Although an antimicrobial prophylaxis is employed in most cases, infectious complications are among the most severe . We present a case of E . coli multirresistant meningitis after transrectal prostate biopsy despite quinolone prophylaxis. Clin Infect Dis, 2003 Jul 1, 37(1), 65 - 72 Epub 2003 Jun 24. Do antimicrobial-impregnated central venous catheters prevent catheter-related bloodstream infection? McConnell SA, Gubbins PO, Anaissie EJ. Controversy surrounds the role of central venous catheters (CVCs) impregnated with antimicrobial agents in the prevention of catheter-related bloodstream infection (CRBSI) . We reviewed the current literature to evaluate the efficacy of antimicrobial-impregnated CVCs for preventing CRBSI . Eleven randomized studies published in article form were identified that included a control group that received nonimpregnated CVCs . We evaluated study methodologies, inclusion of key patient characteristics, use of clinically relevant end points, and molecular-relatedness studies . Review of these 11 trials revealed several methodological flaws, including inconsistent definitions of CRBSI, failure to account for confounding variables, suboptimal statistical and epidemiological methods, and rare use of clinically relevant end points . This review also failed to demonstrate any significant clinical benefit associated with the use of antimicrobial-impregnated CVCs for the purpose of reducing CRBSI or improving patient outcomes . More rigorous studies are required to support or refute the hypothesis that antimicrobial-impregnated CVCs reduce the rate of or prevent CRBSI. Clin Infect Dis, 2003 Jul 1, 37(1), 59 - 64 Epub 2003 Jun 23. Antibiotic combinations with redundant antimicrobial spectra: clinical epidemiology and pilot intervention of computer-assisted surveillance; Glowacki RC et al.; Redundant antibiotic combinations are a potentially remediable source of antibiotic overuse . At a public teaching hospital, we determined the incidence, cost, and indications for such combinations and measured the effects of a pharmacist-based intervention . Of 1189 inpatients receiving >or=2 antibiotics, computer-assisted screening identified 192 patients (16.1%) receiving potentially redundant combinations . Chart reviews showed that 137 episodes (71%) were inappropriate . Physician overprescribing errors were found in 77 episodes (56%); most involved redundant coverage for gram-positive or anaerobic organisms . In 76 episodes (55%), lapses in the medication ordering and distribution system led to the persistence in the pharmacy records of regimens no longer active according to the patient charts . The incidence of redundant antibiotic combinations was significantly higher in the intensive care unit and surgery services, compared with medical services . Interventions to discontinue redundant agents were successful in 134 (98%) of the 137 episodes . Potential drug cost savings and reduction in redundant antibiotic combination days were 10,800 dollars and 584 days, respectively; pharmacist time for patient review and intervention cost 2880 dollars . Use of redundant antibiotic combinations was common, and a pharmacist-based intervention was feasible, with a potential annualized cost savings of 48,000 dollars. Blood, 2003 Oct 1, 102(7), 2660 - 9 Epub 2003 Jun 26. Toll-like receptors stimulate human neutrophil function; Hayashi F et al.; The first immune cell to arrive at the site of infection is the neutrophil . Upon arrival, neutrophils quickly initiate microbicidal functions, including the production of antimicrobial products and proinflammatory cytokines that serve to contain infection . This allows the acquired immune system enough time to generate sterilizing immunity and memory . Neutrophils detect the presence of a pathogen through germ line-encoded receptors that recognize microbe-associated molecular patterns . In vertebrates, the best characterized of these receptors are Toll-like receptors (TLRs) . We have determined the expression and function of TLRs in freshly isolated human neutrophils . Neutrophils expressed TLR1, 2, 4, 5, 6, 7, 8, 9, and 10-all the TLRs except TLR3 . Granulocyte-macrophage colony-stimulating factor (GM-CSF) treatment increased TLR2 and TLR9 expression levels . The agonists of all TLRs expressed in neutrophils triggered or primed cytokine release, superoxide generation, and L-selectin shedding, while inhibiting chemotaxis to interleukin-8 (IL-8) and increasing phagocytosis of opsonized latex beads . The response to the TLR9 agonist nonmethylated CpG-motif-containing DNA (CpG DNA) required GM-CSF pretreatment, which also enhanced the response to the other TLR agonists . Finally, using quantitative polymerase chain reaction (QPCR), we demonstrate a chemokine expression profile that suggests that TLR-stimulated neutrophils recruit innate, but not acquired, immune cells to sites of infection. J Clin Virol, 2003 Jul, 27(2), 146 - 51 Varicella zoster and Borrelia burgdorferi are the main agents associated with facial paresis, especially in children; Jaamaa S et al.; BACKGROUND: The etiology of facial paresis (FP) often remains unresolved . Yet, a microbial association is frequently suspected . OBJECTIVE: To evaluate the infectious etiology of FP by using sensitive tests . STUDY DESIGN: We studied the serum and cerebrospinal fluid of 42 patients diagnosed with idiopathic peripheral facial paresis using sensitive serological methods and nucleic acid detection and for reference, 42 patients with other neurological disorders (OND) matched for age, sex, season and geographical area . RESULTS: Varicella zoster virus and Borrelia burgdorferi accounted for 56% of all associated agents in children with FP compared with 11% of OND (P=0.01) . In adults, the respective numbers were 29 and 13% . Other treatable etiological agents, Chlamydia pneumoniae and Mycoplasma pneumoniae, accounted for 11% in children and 8% in adults and with the same prevalence between patients with FP and OND . CONCLUSIONS: Microbes, with specific therapy available accounted for 52% of all associated agents in the patients with FP when compared with 26% in controls with OND (P=0.04) . Based on this, we conclude that the patients with FP may benefit from antimicrobial therapy. Exp Hematol, 2003 Jun, 31(6), 535 - 44 Unrelated umbilical cord blood transplants in adults: Early recovery of neutrophils by supportive co-transplantation of a low number of highly purified peripheral blood CD34+ cells from an HLA-haploidentical donor; Fernandez MN et al.; OBJECTIVE, METHODS, AND RESULTS: To reduce the period of posttransplant neutropenia and related early morbidity and mortality of cord blood (CB) transplants, we assessed the feasibility of co-infusion of a low number of highly purified peripheral blood CD34+ cells from a related haploidentical donor with a CB graft . Between March 1999 and May 2002, 11 patients with high-risk hematologic malignancies were transplanted using this strategy . The seven patients who received a haploidentical peripheral blood graft and a CB graft from a sibling (6) or the father (1) had prompt recovery (9-17 days, median 10) of the absolute neutrophil count (ANC) to greater than 0.5 x 10(9)/L . Analysis of DNA polymorphisms showed initial predominance of the haploidentical genotype both in granulocytes and in mononuclear cells, and subsequent progressive replacement by cells of CB genotype until final complete CB chimerism was achieved by patients who survived for sufficient periods of time . The four patients who received maternal haploidentical cells had no significant contribution of these to blood leukocytes, although complete CB chimerism was achieved by three of them and two reached engraftment of the CB on days +20 and +36 . Morbidity due to early bacterial or fungal infections was remarkably low in patients with prompt ANC recovery . CONCLUSION: Our data show that co-infusion of a CB unit and a low number of haploidentical CD34+ cells may result in a shortened period of posttransplant neutropenia . This is likely the result of prompt and transient engraftment of the haploidentical hematopoietic stem cells that may provide the patient antimicrobial protection until the later engraftment of the CB hematopoietic stem cells. Scand J Immunol, 2003 Jul, 58(1), 67 - 75 Interaction between Borrelia burgdorferi and immature human dendritic cells; Suhonen J et al.; Antigen uptake and the following maturation of dendritic cells (DCs) are pivotal to the initiation of specific antimicrobial immune responses . DCs also play an important role in the recruitment and activation of the cells of the innate immune system . We have examined the interactions of DCs with Borrelia burgdorferi to find explanations for the difficulties the human immune system has in dealing with the bacterium . Phagocytosis of B . burgdorferi by immature DCs and the effect of the bacterium on the maturation and interleukin-8 (IL-8) secretion of DCs were studied . Borreliae were phagocytized and processed into fragments by DCs; narrow tube-like pseudopods and broad pseudopods were used for the engulfment . The immature DC population gained a heterogeneous appearance within 2 h of incubation with the borreliae . A 24 h coculture with borreliae induced maturation and IL-8 secretion in the DCs in a manner comparable with the effect of lipopolysaccharides . All strains studied, including a mutant strain lacking outer surface proteins A and B, were capable of inducing these responses . Thus, our results did not show any clear inadequacy concerning the way DCs are dealing with B . burgdorferi . However, further studies on the subject are required. Am J Vet Res, 2003 Jun, 64(6), 694 - 9 Pharmacokinetics of imipenem in dogs; Barker CW et al.; OBJECTIVE: To determine the plasma pharmacokinetics of imipenem (5 mg/kg) after single-dose IV, IM, and SC administrations in dogs and assess the ability of plasma samples to inhibit the growth of Escherichia coli in vitro . ANIMALS: 6 adult dogs . PROCEDURE: A 3-way crossover design was used . Plasma concentrations of imipenem were measured after IV, IM, and SC administration by use of high-performance liquid chromatography . An agar well antimicrobial assay was performed with 3 E coli isolates that included a reference strain and 2 multidrug-resistant clinical isolates . RESULTS: Plasma concentrations of imipenem remained above the reported minimum inhibitory concentration for E coli (0.06 to 0.25 microg/mL) for a minimum of 4 hours after IV, IM, and SC injections . Harmonic mean and pseudo-standard deviation half-life of imipenem was 0.80 +/- 0.23, 0.92 +/- 0.33, and 1.54 +/- 1.02 hours after IV, IM, and SC administration, respectively . Maximum plasma concentrations (Cmax) of imipenem after IM and SC administration were 13.2 +/- 4.06 and 8.8 +/- 1.7 mg/L, respectively . Time elapsed from drug administration until Cmax was 0.50 +/- 0.16 hours after IM and 0.83 +/- 0.13 hours after SC injection . Growth of all 3 E coli isolates was inhibited in the agar well antimicrobial assay for 2 hours after imipenem administration by all routes . CONCLUSIONS AND CLINICAL RELEVANCE: Imipenem is rapidly and completely absorbed from intramuscular and subcutaneous tissues and effectively inhibits in vitro growth of certain multidrug-resistant clinical isolates of E coli. Eur J Clin Microbiol Infect Dis, 2003 Jul, 22(7), 427 - 30 Epub 2003 Jun 24. Mycobacterium elephantis: not an exceptional finding in clinical specimens; Tortoli E et al.; Following the recent report of new 16S rDNA sequences of Mycobacterium elephantis, three clinical strains suspected to belong to such species were investigated using biochemical and cultural tests, high performance liquid chromatography of cell wall mycolic acids and genetic sequencing . Antimicrobial susceptibility was also determined . The findings confirmed recent data concerning human isolates of this new mycobacterium and identified a new 16S rDNA sequevar for this species. Surg Neurol, 2003 Jun, 59(6), 509 - 11 Delirium in the elderly resulting from azithromycin therapy; Cone LA et al.; BACKGROUND: Azithromycin, a semi-synthetic azalide antibiotic, is a macrolide that thus far has not shared the neuropsychiatric side effects of other macrolides such as erythromycin and clarithromycin . METHODS: We now report significant delirium associated with conventional dosing of azithromycin in two geriatric patients who were being treated for lower respiratory tract infection . RESULTS: The onset of delirium was apparent within 72 hours of initiating azithromycin therapy and lasted 48 to 72 hours after discontinuing treatment with the drug . CONCLUSIONS: In contrast to the adverse central nervous system symptoms associated with clarithromycin, those induced by azithromycin seem to take longer to resolve, perhaps based upon the longer elimination half-life of the latter antimicrobial, particularly in geriatric women. Microvasc Res, 2003 Jul, 66(1), 38 - 48 CAP37, a neutrophil-derived inflammatory mediator, augments leukocyte adhesion to endothelial monolayers; Lee TD et al.; Cationic antimicrobial protein of molecular weight 37 kDa (CAP37) is a multifunctional inflammatory mediator that was originally isolated from human neutrophils and described to possess bactericidal and monocyte-activating functions . More recently its expression in endothelial and epithelial cells in response to inflammatory mediators and its ability to activate endothelial cells and alter permeability has been demonstrated . We hypothesize that CAP37 facilitates the process of transendothelial migration not only because of its potential to act as a chemoattractant but also through its ability to promote leukocyte adhesion to the endothelium by modulating adhesion molecule expression on the endothelium . Here we describe its ability to mediate neutrophil and monocyte adherence to endothelial monolayers in vitro . Using reverse transcriptase-polymerase chain reaction and flow cytometry, we demonstrate its ability to upregulate the adhesion molecules, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin in human umbilical vein and lung microvessel endothelial cells . The identity and kinetics of upregulation of the specific adhesion molecule was dependent on the endothelial cell type, suggesting that adhesion molecules on endothelial cells from different vascular beds are differentially regulated by CAP37 . The cell-specific kinetics of adhesion molecule upregulation by CAP37 may influence selective leukocyte migration in certain inflammatory situations. J Infect Dis, 2003 Jul 1, 188(1), 146 - 52 Epub 2003 Jun 16. Killing of African trypanosomes by antimicrobial peptides; McGwire BS et al.; Antimicrobial peptides are components of the innate immune systems of a wide variety of eukaryotic organisms and are being developed as antibiotics in the fight against bacterial and fungal infections . We explored the potential activities of antimicrobial peptides against the African trypanosome Trypanosoma brucei, a vector-borne protozoan parasite that is responsible for significant morbidity and mortality in both humans and animals . Three classes of mammalian antimicrobial peptides were tested: alpha-defensins, beta-defensins, and cathelicidins . Although members of all 3 classes of antimicrobial peptides showed activity, those derived from the cathelicidin class were most effective, killing both insect and bloodstream forms of the parasite . The mechanism of action of the cathelicidins against T . brucei involves disruption of surface membrane integrity . Administration of cathelicidin antimicrobial peptides to mice with late-stage T . brucei infection acutely decreased parasitemia and prolonged survival . These results highlight the potential use of antimicrobial peptides for the treatment of African trypanosomiasis. Med Electron Microsc, 2003 Jun, 36(2), 94 - 7 Localization of human beta-defensin 3 mRNA in normal oral epithelium, leukoplakia, and lichen planus: an in situ hybridization study; Nishimura M et al.; Human beta-defensin 3 (hBD-3), an antimicrobial peptide, is produced by various epithelial and some nonepithelial tissues . hBD-3 mRNA is widely expressed in oral tissues, including oral epithelium and the salivary glands . Although the localization of hBD-1 and hBD-2 has been well demonstrated in tissue sections, the localization pattern of hBD-3 has not yet been shown . In the present study, we investigated the expression pattern of hBD-3 mRNA by in situ hybridization using specific RNA probes; the signal for hBD-3 was detected in upper spinous and granular layers in normal oral epithelium . In cases of leukoplakia, a strong signal of hBD-3 mRNA was observed in the granular layer . In lichen planus, the signal was strongly detected in the spinous and suprabasal layers . The signals were stronger than those of either normal oral epithelium or leukoplakia . The results indicate that the localization pattern of hBD-3 is very similar to that of hBD-2 . hBD-2 and hBD-3 may function together or compensate each other for expressional loss. ORL J Otorhinolaryngol Relat Spec, 2003 Mar-Apr, 65(2), 117 - 20 Microbiology and management of deep facial infections and Lemierre syndrome; Brook I; This review describes the microbiology, diagnosis and management of deep facial infections and Lemierre syndrome . The origins of most of these infections are odontogenic infections that spread to fascial spaces of the lower head and upper neck . Other sources include pharyngotonsillar, nasal, otologic, salivary gland and dermatologic infections, hematogenic spread, cervical adenitis and trauma . These space infections can be divided into those around the face (masticatory, buccal, canine and parotid), the suprahyoid area (submandibular, sublingual and lateral pharyngeal) and the infrahyoid region or lateral neck (retropharyngeal and pretracheal spaces) . The organisms accounting for these infections are aerobic and anaerobic that arise from the oropharyngeal flora . Complications of these infections can be life threatening and can result from hematogenic or direct spread . Complications that arise following local extension include suppurative jugular thrombophlebitis, cavernous sinus thrombosis, carotid erosion, maxillary sinusitis and osteomyelitis of the jaws . Management includes surgical drainage and antimicrobial therapy . FASEB J, 2003 Aug, 17(11), 1502 - 4 Epub 2003 Jun 03. Dual oxidases represent novel hydrogen peroxide sources supporting mucosal surface host defense; Geiszt M et al.; Lactoperoxidase (LPO) is an enzyme with antimicrobial properties present in saliva, milk, tears, and airway secretions . Although the formation of microbicidal oxidants by LPO has been recognized for some time, the source of hydrogen peroxide (H2O2) for LPO-catalyzed reactions remains unknown . Reactive oxygen species produced by the phagocyte NADPH oxidase (phox) play a critical role in host defense against pathogens; however, analogous oxidant-generating systems in other tissues have not been associated with antimicrobial activity . Several homologues of gp91phox, the catalytic core of this enzyme, were described recently; dual oxidase (Duox)1/thyroid oxidase 1 and Duox2/thyroid oxidase 2 were identified in the thyroid gland and characterized as H2O2 donors for thyroxin biosynthesis . We examined Duox1 and Duox2 expression in secretory glands and on mucosal surfaces and give evidence for their presence and activity in salivary glands, rectum, trachea, and bronchium . Epithelial cells in salivary excretory ducts and rectal glands express Duox2, whereas tracheal and bronchial epithelial cells express Duox1 . Furthermore, we detected Duox1-dependent H2O2 release by cultured human bronchial epithelial cells . Our observations suggest that Duox1 and Duox2 are novel H2O2 sources that can support LPO-mediated antimicrobial defense mechanisms on mucosal surfaces. Int Endod J, 2003 Jul, 36(7), 453 - 63 Microbial causes of endodontic flare-ups; Siqueira JF Jr; LITERATURE REVIEW: Inter-appointment flare-up is characterized by the development of pain, swelling or both, following endodontic intervention . The causative factors of flare-ups encompass mechanical, chemical and/or microbial injury to the pulp or periradicular tissues . Of these factors, microorganisms are arguably the major causative agents of flare-ups . Even though the host is usually unable to eliminate the root canal infection, mobilization and further concentration of defence components at the periradicular tissues impede spreading of infection, and a balance between microbial aggression and host defences is commonly achieved . There are some situations during endodontic therapy in which such a balance may be disrupted in favour of microbial aggression, and an acute periradicular inflammation can ensue . Situations include apical extrusion of infected debris, changes in the root canal microbiota and/or in environmental conditions caused by incomplete chemo-mechanical preparation, secondary intraradicular infections and perhaps the increase in the oxidation-reduction potential within the root canal favouring the overgrowth of the facultative bacteria . Based on these situations, preventive measures against infective flare-ups are proposed, including selection of instrumentation techniques that extrude lesser amounts of debris apically; completion of the chemo-mechanical procedures in a single visit; use of an antimicrobial intracanal medicament between appointments in the treatment of infected cases; not leaving teeth open for drainage and maintenance of the aseptic chain throughout endodontic treatment . Knowledge about the microbial causes of flare-ups and adoption of appropriate preventive measures can significantly reduce the incidence of this highly distressing and undesirable clinical phenomenon. J Pept Res, 2003 Aug, 62(2), 53 - 62 From pro defensins to defensins: synthesis and characterization of human neutrophil pro alpha-defensin-1 and its mature domain; Wu Z et al.; Human neutrophil alpha-defensins (HNPs) are small, cationic, Cys-rich antimicrobial proteins that play important roles in innate immunity against infectious microbes such as bacteria, fungi and enveloped viruses . Synthesized as inactive precursors in vivo (pre-proHNPs), HNPs are activated through proteolytic removal of the inhibitory pro-peptide required for subcellular sorting and correct folding . We seek to understand the molecular basis for the recognition between the 45-residue pro-peptide and the C-terminal functional domain . Here we described, total chemical synthesis of the 75-residue human neutrophil pro alpha-defensin-1 (proHNP1) via native chemical ligation . After oxidative folding, proHNP1 is cleaved by cyanogen bromide at the Met45-Ala46 peptide bond to release the mature form . The native disulfide connectivity in HNP1, i.e . Cys1-Cys6, Cys2-Cys4 and Cys3-Cys5, is verified by mass mapping of peptide fragments generated by proteolytic digestion and Edman degradation . Fluorescence spectroscopy studies and antimicrobial activity assays further support that synthetic proHNP1 and HNP1 are correctly folded . While largely unstructured in aqueous solution, the pro-peptide binds to HNP1 intermolecularly with an apparent Kd value of 6.2 microM at pH 7.4, confirming the mode of intramolecular inactivation of human alpha-defensin precursors. Eur J Biochem, 2003 Jul, 270(13), 2805 - 13 Key role of the loop connecting the two beta strands of mussel defensin in its antimicrobial activity; Romestand B et al.; To elucidate the structural features of the mussel defensin MGD1 required for antimicrobial activity, we synthesized a series of peptides corresponding to the main known secondary structures of the molecule and evaluated their activity towards Gram-positive and Gram-negative bacteria, and filamentous fungi . We found that the nonapeptide corresponding to residues 25-33 of MGD1 (CGGWHRLRC) exhibited bacteriostatic activity once it was cyclized by a non-naturally occurring disulfide bridge . Longer peptides corresponding to the amino acid sequences of the alpha-helical part or to the beta-strands of MGD1 had no detectable activity . The bacteriostatic activity of the sequence 25-33 was strictly dependent on the bridging of Cys25 and Cys33 and was proportional to the theoretical isoelectric point of the peptide, as deduced from the variation of activity in a set of peptide analogues of the 25-33 sequence with different numbers of cationic charges . By using confocal fluorescence microscopy, we found that the cyclic peptides bound to Gram-positive bacteria without apparent lysis . However, by using a fluorescent dye, we observed that dead bacteria had been permeated by the cyclic peptide 25-33 . Sequence comparisons in the family of arthopod defensins indicate that MGD1 belongs to a subfamily of the insect defensins, characterized by the constant occurrence of both positively charged and hydrophobic amino acids in the loop . Modelling studies showed that in the MGD1 structure, positively charged and hydrophobic residues are organized in two layered clusters, which might have a functional significance in the docking of MGD1 to the bacterial membrane. Aliment Pharmacol Ther, 2003 Jun 15, 17(12), 1545 - 51 Is antimicrobial susceptibility testing necessary before second-line treatment for Helicobacter pylori infection? Miwa H, Nagahara A, Kurosawa A, Ohkusa T, Ohkura R, Hojo M, Enomoto N, Sato N. BACKGROUND: An antimicrobial susceptibility test for Helicobacter pylori before second-line treatment is often performed, although whether the test is truly necessary remains unknown . PATIENTS AND METHODS: Eighty-two patients with H . pylori infection for whom first-line treatment with a 1-week proton pump inhibitor/amoxicillin-clarithromycin (AC) regimen had failed were randomly assigned to two groups: those having or not having the susceptibility test before re-treatment . The cure rates for these two groups were compared . RESULTS: Five of the 82 patients were excluded from the analysis . For 38 patients in the susceptibility-test group, we used what we considered the best regimen based on susceptibility testing: 10 patients {no resistance to clarithromycin (CAM)} received the lansoprazole-amoxicillin-clarithromycin regimen, 22 patients {19 CAM resistant, metronidazole (MNZ) susceptible; three failure of culture} were given the lansoprazole-amoxicillin-metronidazole (LAM) regimen, and six patients (both MNZ and CA