Peptides, 2004 Apr, 25(4), 675 - 83 The interaction of an antimicrobial decapeptide with phospholipid vesicles; Choi MJ et al.; Previously, by using combinatorial peptide libraries, we have identified activity-optimized decapeptide (KSL, KKVVFKVKFK-NH(2)), which exhibited a broad spectrum of the activity against bacteria and fungi without hemolytic activity . In order to examine lipid requirements and to understand the mode of KSL action, we investigated interactions of the peptide with vesicles consisting of various lipid compositions . KSL increased the permeability of negatively charged but not zwitterionic phospholipid membranes, and the leakage was independent on the size of encapsulated molecules (calcein, 1-aminonaphthalene-3,6,8-trisulfonic acid (ANTS)/N,N'-p-xylene bis(pyridinium) bromide (DPX), and fluorescein isothiocyanate (FITC)-dextran with different molecular weight), indicating that the peptide did not form pores or channels in this leakage process . KSL ability to permeabilize vesicles with negatively charged surface was dramatically reduced upon the addition of zwitterionic phospholipid rather than cholesterol, which revealed that the surface charge of lipid membranes played a major role in the activity and selectivity of KSL . Moreover, KSL diastereomer did not increase the permeability of negatively charged vesicles, indicating that the secondary structure of KSL was also required for membrane perturbation activity . Interestingly, KSL had an ability to cause aggregation and subsequent fusion of the acidic vesicles, which seemed to be related to the biological action . Structural studies performed by circular dichroism (CD) spectroscopy indicated that in the presence of acidic vesicles, the beta sheet structure of KSL must be required for the ability to (1) induce a leakage of dye from the acidic vesicles (2) to fuse the acidic vesicles . Mol Microbiol, 2004 Jun, 52(5), 1291 - 302 Role of KatG catalase-peroxidase in mycobacterial pathogenesis: countering the phagocyte oxidative burst; Ng VH et al.; Reactive nitrogen species (RNS) play an essential role in host defence against Mycobacterium tuberculosis (MTB) in the mouse model of tuberculosis (TB), as evidenced by the increased susceptibility of mice deficient in the inducible isoform of nitric oxide synthase (NOS2) . In contrast, the role of reactive oxygen species (ROS) in protection against MTB is less clear, and mice defective in the ROS-generating phagocyte NADPH oxidase (Phox) are relatively resistant . This suggests that MTB might possess efficient mechanisms to evade or counter the phagocyte oxidative burst, effectively masking the impact of this host defence mechanism . In order to assess the role of ROS detoxification pathways in MTB virulence, we generated a katG null mutant of MTB, deficient in the KatG catalase-peroxidase-peroxynitritase, and evaluated the mutant's ability to replicate and persist in macrophages and mice . Although markedly attenuated in wild-type C57Bl/6 mice and NOS2(-/-) mice, the DeltakatG MTB strain was indistinguishable from wild-type MTB in its ability to replicate and persist in gp91(Phox-/-) mice lacking the gp91 subunit of NADPH oxidase . Similar observations were made with murine bone marrow macrophages infected ex vivo: growth of the DeltakatG MTB strain was impaired in macrophages from C57Bl/6 and NOS2(-/-) mice, but indistinguishable from wild-type MTB in gp91(Phox-/-) macrophages . These results indicate that the major role of KatG in MTB pathogenesis is to catabolize the peroxides generated by the phagocyte NADPH oxidase; in the absence of this host antimicrobial mechanism, KatG is apparently dispensable. J Nat Prod, 2004 May, 67(5), 811 - 6 Tolaasins A--E, five new lipodepsipeptides produced by Pseudomonas tolaasii; Bassarello C et al.; Pseudomonas tolaasii, the causal organism of brown blotch disease of Agaricus bisporus and of the yellowing of Pleurotus ostreatus, was shown to produce in culture tolaasin I (1), tolaasin II (2), and five other minor metabolites, tolaasins A, B, C, D, and E (3-7) . These compounds were demonstrated to be important in the development of the disease symptoms . This paper reports on the structural elucidation, based essentially on NMR studies and MS spectra, and biological activity of the above lipodepsipeptides (3-7) . All the above analogues showed differences in the peptide moiety, as observed in other lipodepsipeptides of bacterial origin, and maintained the beta-hydroxyoctanoyl phi chain at the N-terminus, except tolaasin A, in which the acyl moiety was a gamma-carboxybutanoyl phi moiety . Among the target microorganisms used (fungi, yeast, and bacteria) the Gram-positive bacteria were the most sensitive, although the antimicrobial activity appeared to be correlated to the structural modification in the different analogues . The structure-activity relationships of these toxins are discussed. Ageing Res Rev, 2004 Jan, 3(1), 31 - 54 HIV infection and advanced age emerging epidemiological, clinical, and management issues; Manfredi R; While the mean age of HIV/AIDS patients at first diagnosis is progressively rising, no updated epidemiological estimates, controlled clinical data, and randomized therapeutic trials, are available regarding clinical and laboratory response to antiretroviral therapy, safety of anti-HIV compounds and their associations, potential drug-drug interactions, short- and long-term toxicity, consequences on underlying disorders, or interactions with concomitant pharmacological regimens, in the elderly . The life expectancy of HIV-infected persons treated with highly active antiretroviral therapy (HAART) now approximates that of general population matched for age, while also AIDS definition itself has lost most of its epidemiological and clinical significance, thanks to the immunoreconstitution resulting from the large-scale use of potent HAART regimens . The increased survival of HIV-infected patients, the late recognition of other subjects with missed or delayed diagnosis are responsible for a further expected rise of mean age of HIV-infected individuals, so that the patient population aged 60-70 years or more is expected to increase in coming years . Unfortunately, the majority of therapeutic trials involving antiretroviral therapy, as well as antimicrobial chemoprophylaxis for AIDS-related opportunistic complications, have advanced age and/or concurrent end-organ disorders among main exclusion criteria, or the design of these studies does not allow to extrapolate data regarding older patients, compared with younger ones . The very limited data presently available seem to demonstrate that HAART has a virological efficacy in the elderly comparable with that of younger adults, but immunological recovery is often slower and blunted, although several studies clearly demonstrated that thymic function is preserved until late adult age . When facing an HIV-infected patient with advanced age, health care givers have to pay careful attention to eventual end-organ disorders, all possible pharmacological interactions, overlapping toxicity due to concurrent drug administration . All these issues may significantly interfere with HAART activity, patient's adherence to prescribed medications, and frequency and severity of untoward effects . The guidelines of antiretroviral therapy and those of treatment and prophylaxis of AIDS-related diseases deserve appropriate updates, paralleling the increasing mean age of HIV-infected population . Moreover, epidemiological figures need an increased focus on older age, while clinical trials specifically targeting on the elderly population are mandatory to have reliable data on all aspects of HAART administration in advanced age. Int J Food Microbiol, 2004 Jun 15, 93(3), 267 - 79 Characterization of an E2-type colicin and its application to treat alfalfa seeds to reduce Escherichia coli O157:H7; Nandiwada LS et al.; Several outbreaks of Escherichia coli O157:H7 infections have been associated with contaminated alfalfa seeds . A recently isolated E . coli strain Hu194 was capable of inhibiting 22 strains of E . coli O157:H7 and this inhibition was mediated by the production of a colicin named Hu194 . The objectives of this study were to test the efficacy of treating alfalfa seeds with colicin Hu194 against E . coli O157:H7 strains, and to characterize this antimicrobial protein . Significant reductions (approximately 5 log CFU ml-1) in the viable cell counts of strains 43890 and 43895 were observed after 1-day incubation with semi-crude colicin, and after 2 days for strain 3081 . Strain 43890 was successfully eliminated (5 log CFU g-1) from inoculated alfalfa seeds after soaking in a colicin suspension at a concentration of 10,000 AU/g . Treatment of alfalfa seeds inoculated with strains 43895 and 3081 required 20-fold higher concentrations of colicin Hu194 to achieve as much as 3 log CFU g-1 reductions . The genes encoding the colicin Hu194 operon were located on a 6 kb plasmid, and the sequence analysis revealed that this colicin was an E-type DNAse . From the sequence data, the estimated molecular masses of colicin Hu194, its immunity protein and lysis protein were 61.3, 10.0 and 4.8 kDa, respectively . Based on DNA and protein sequence comparisons with other E-type colicin, colicin Hu194 belonged to the type E2-colicin cluster . However, cross-immunity tests between E-group colicins suggested that Hu194 colicin was divergent from the previously characterized E2 colicins. Phytother Res, 2004 Apr, 18(4), 339 - 41 The antimicrobial activity of essential oils of Hypericum scabrum, Hypericum scabroides and Hypericum triquetrifolium; Kizil G et al.; The essential oils of Hypericum scabrum, Hypericum scabroides and Hypericum triquetrifolium were studied for the first time for their antimicrobial activity against nine organisms . All the essential oils exhibited some broad spectrum antibacterial activity, at a concentration of 80 microg/mL . The essential oils of Hypericum species showed antibacterial activity against the tested organisms and a yeast . J Agric Food Chem, 2004 Jun 2, 52(11), 3530 - 5 chemical composition, plant genetic differences, antimicrobial and antifungal activity investigation of the essential oil of Rosmarinus officinalis L; Angioni A et al.; The chemical composition of the essential oil of the Sardinian Rosmarinus officinalis L . obtained by hydro distillation and steam\hydro distillation was studied using GC-FID and MS . Samples were collected at different latitude and longitude of Sardinia (Italy) . The yields ranged between 1.75 and 0.48% (v/w, volume/dry-weight) . A total of 30 components were identified . The major compounds in the essential oil were alpha-pinene, borneol, (-) camphene, camphor, verbenone, and bornyl-acetate . Multivariate analysis carried out on chemical molecular markers, with the appraisal of chemical, pedological, and random amplified polymorphic DNA data, allows four different clusters to be distinguished . The antimicrobial and antifungal tests showed a weak activity of Sardinian rosemary . On the other hand, an inductive effect on fungal growth, especially toward Fusarium graminearum was observed. J Agric Food Chem, 2004 Jun 2, 52(11), 3309 - 12 In vitro antioxidant, antimicrobial, and antiviral activities of the essential oil and various extracts from herbal parts and callus cultures of Origanum acutidens; Sokmen M et al.; The essential oil and various extracts obtained from Origanum acutidens and methanol extracts (MeOH) from callus cultures have been evaluated for their antioxidative, antimicrobial, and antiviral properties . The essential oil exhibited strong antimicrobial activity with a significant inhibitory effect against 27 (77%) of the 35 bacteria, 12 (67%) of the 18 fungi, and a yeast tested and moderate antioxidative capacity in DPPH and beta-carotene/linoleic acid assays . GC and GC-MS analyses of the oil resulted in the identification of 38 constituents, carvacrol being the main component . The MeOH extracts obtained from herbal parts showed better antioxidative effect than that of butylated hydroxytoluene (BHT), whereas callus cultures also exhibited interesting antioxidative patterns . Concerning antiviral activity, none of the extracts inhibited the reproduction of influenza A/Aichi virus in MDCK cells . The MeOH extracts from herbal parts inhibited the reproduction of HSV-1, and also callus cultures exerted slight antiherpetic effect. J Am Acad Orthop Surg, 2004 May-Jun, 12(3), 155 - 63 Spinal epidural abscess in adults; Bluman EM et al.; Spinal epidural abscess is a potentially life-threatening disease that can cause paralysis by the accumulation of purulent material in the epidural space . Although modern diagnostic and management methods have improved the prognosis, morbidity and mortality remain significant . Outcome usually is determined by the rapidity of the diagnosis and initiation of appropriate treatment . A high index of suspicion is warranted when a patient presents with spinal pain or a neurologic deficit in conjunction with fever or an elevated erythrocyte sedimentation rate . Gadolinium-enhanced magnetic resonance imaging should be done in suspected cases to localize and define the abscess . For spinal epidural abscess associated with neurologic compromise, the treatment of choice is emergent surgical decompression and debridement (with or without spinal stabilization), followed by long-term antimicrobial therapy . In the absence of a neurologic deficit, medical management is an alternative to surgery when the risk of neurologic complications is low based on the location and morphology of the abscess, immune status of the patient, and virulence of the organism. J Pept Sci, 2004 May, 10(5), 304 - 11 Antinematodal effect of antimicrobial peptide, PMAP-23, isolated from porcine myeloid against Caenorhabditis elegans; Park Y et al.; The antinematodal activity and mechanism of a 23-mer antimicrobial peptide, PMAP-23, derived from pig myeloid was investigated . PMAP-23 displayed a strong antinematodal activity against the eggs and worms of Caenorhabditis elegans . To investigate the antinematodal mechanism of PMAP-23, fluorescence activated flow cytometry and confocal laser scanning microscopy were performed . C . elegans treated with PMAP-23 showed higher fluorescence intensity by propidium iodide (PI) staining than normal cells . Confocal microscopy showed that the peptide was localized in the egg's shell and cell membrane . The action of the peptide against C . elegans membranes was examined by testing the membrane disrupting activity using liposome (PC/PS; 3:1, w/w) . The result suggests that PMAP-23 may exert its antinematodal activity by disrupting the structure of the cell membrane via pore formation or via direct interaction with the lipid bilayers. J Pept Sci, 2004 May, 10(5), 298 - 303 Structure-antiviral activity relationships of cecropin A-magainin 2 hybrid peptide and its analogues; Lee DG et al.; In order to elucidate the structure-antiviral activity relationship of cecropin A (1-8)-magainin 2 (1-12) (termed CA-MA) hybrid peptide, several analogues with amino acid substitutions were synthesized . In a previous study, it was shown that serine at position 16 in CA-MA hybrid peptide was very important for antimicrobial activity . Analogues were designed to increase the hydrophobic property by substituting a hydrophobic amino acid residue (S --> A, V, F or W, position 16) in the CA-MA hybrid peptide . In this study, the structure-antiviral activity relationships of CA-MA and its analogues were investigated . In particular, substitution of Ser with a hydrophobic amino acid, Val, Phe or Trp at position 16 caused a dramatic increase in the virus-cell fusion inhibitory activity . These results suggested that the hydrophobicity at position 16 in the hydrophobic region of CA-MA is important for potent antiviral activity. Acta Crystallogr D Biol Crystallogr, 2004 Jun, 60(Pt 6), 1121 - 4 Epub 2004 May 21. Purification, characterization and preliminary crystallographic studies of a novel plant defensin from Pachyrrhizus erosus seeds; Song X et al.; Defensins are small cysteine-rich antimicrobial peptides that are widely distributed in plants, insects and mammals . As potent defenders in protecting plants from pathogenic fungal attack, plant defensins are presumed to play an important role in the innate immunity of plants and are expected to find applications in the production of transgenic crops . A novel plant defensin protein SPE10 from Pachyrrhizus erosus seeds was purified and partially sequenced . Crystallization screening using the hanging-drop vapour-diffusion method resulted in two crystal forms, from one of which a diffraction data set was collected to 0.98 A resolution . The crystal belongs to space group P2(1), with unit-cell parameters a = 32.71, b = 28.11, c = 54.85 angstroms, beta = 103.78 degrees . Preliminary crystallographic studies revealed two subunits in the asymmetric unit . Fitoterapia, 2004 Jun, 75(3-4), 409 - 11 Antimicrobial activity of Memecylon malabaricum leaves; Hullatti KK et al.; The petroleum ether, chloroform and methanol extracts of Memecylon malabaricum leaves were tested for antimicrobial activity . Only methanol extract has shown activity against bacteria both Gram (+) and Gram (-), and fungi . Fitoterapia, 2004 Jun, 75(3-4), 395 - 7 Antimicrobial activity of the macrofungus Pholiota adiposa; Dulger B; The 60% methanolic extract of Pholiota adiposa exhibited antimicrobial activity . Fitoterapia, 2004 Jun, 75(3-4), 368 - 70 Antimicrobial and cytotoxic activities of the fruit essential oil of Xylopia aethiopica from Nigeria; Asekun OT et al.; The fruits essential oil of Xylopia aethiopica showed activity against four microorganisms and cytotoxicity to carcinoma cells (Hep-2 cell line) at 5 mg/ml concentration . Bioorg Med Chem, 2004 Jun 15, 12(12), 3135 - 9 Synthesis and in vitro antimicrobial activities of 2-hydroxy-6-methyl-7-(arylamino)-1,7-dihydropurin-8-ones; Sharma P et al.; A number of 2-hydroxy-6-methyl-7-(arylamino)-1,7-dihydropurin-8-ones have been synthesized . 3-Oxo-2-(arylhydrazono)butyric acid ethyl ester were acetylated and treated with triethyl amine and formamide in presence of 1,4-dioxane to yield N-(5-acetyl-4-ethoxy-2-oxo-2,5-dihydro-imidazol-1-yl)-N-arylacetamide, which on refluxation with urea and freshly prepared sodium ethoxide yielded the title compound . All the newly synthesized compounds have been characterized by spectroscopic and elemental analysis data . The synthesized compounds were screened against a representative panel of susceptible and resistant Gram-positive and Gram-negative bacteria using a standard antibiotic drug purinthol as control . Quantitative structure-activity relationship has also been interpreted in terms of correlation of biological activity with molecular refractive index parameters (M(R)) and Hammett substituent constant (sigma). Biochim Biophys Acta, 2004 Jun 1, 1699(1-2), 221 - 7 Identification and structural analysis of the antimicrobial domain in hipposin, a 51-mer antimicrobial peptide isolated from Atlantic halibut; Birkemo GA et al.; Hipposin is a potent 51-mer antimicrobial peptide (AMP) from Atlantic halibut with sequence similarity to parasin (19-mer catfish AMP), buforin I (39-mer toad AMP), and buforin II (an active 21-mer fragment of buforin I), suggesting that the antimicrobial activity of these peptides might all be due to a common antimicrobial sequence motif . In order to identify the putative sequence motif, the antimicrobial activity of hipposin fragments against 20 different bacteria was compared to the activity of hipposin, parasin and buforin II . Neither parasin nor the 19-mer parasin-like fragment HIP(1-19) (differs from parasin in only three residues) that is derived from the N-terminal part (residues 1-19) of hipposin had marked antimicrobial activity . In contrast, the fragment HIP(16-36) (identical to buforin II) that is derived from the middle part of hipposin (residues 16-36) had such activity, indicating that this part of hipposin contained an antimicrobial sequence motif . The activity was enhanced when the parasin-like N-terminal sequence was also present, as the fragment HIP(1-36) which consists of residues 1-36 in hipposin was more potent than HIP(16-36) . Extending HIP(1-36) with three C-terminal residues-thereby constructing the buforin I-like peptide HIP(1-39) (differs from buforin I in only three residues)-increased the activity further . Also, the presence of the C-terminal part of hipposin (residues 40-51) increased the activity, as hipposin was clearly the most potent of all the peptides that were tested . Circular dichroism structural analysis of the peptides revealed that they were all non-structured in aqueous solution . However, trifluoroethanol and the membrane-mimicking entities dodecylphosphocholine micelles and negatively charged liposomes induced (amphiphilic) alpha-helical structuring in hipposin . Judging from the structuring of the individual fragments, the tendency for alpha-helical structuring appeared to be greater in the C-terminal and the buforin II-like middle region of hipposin than in the parasin-like N-terminal region. Lancet, 2004 May 22, 363(9422), 1683 - 8 Zinc for severe pneumonia in very young children: double-blind placebo-controlled trial; Brooks WA et al.; BACKGROUND: Pneumonia is a leading cause of morbidity and mortality in young children . Early reversal of severity signs--chest indrawing, hypoxia, and tachypnoea--improves outcome . We postulated that zinc, an acute phase reactant, would shorten duration of severe pneumonia and time in hospital . METHODS: In a double-blind placebo-controlled clinical trial in Matlab Hospital, Bangladesh, 270 children aged 2-23 months were randomised to receive elemental zinc (20 mg per day) or placebo, plus the hospital's standard antimicrobial management, until discharge . The outcomes were time to cessation of severe pneumonia (no chest indrawing, respiratory rate 50 per min or less, oxygen saturation at least 95% on room air) and discharge from hospital . Discharge was allowed when respiratory rate was 40 per minute or less for 24 consecutive hours while patients were maintained only on oral antibiotics . FINDINGS: The group receiving zinc had reduced duration of severe pneumonia (relative hazard {RH}=0.70, 95% CI 0.51-0.98), including duration of chest indrawing (0.80, 0.61-1.05), respiratory rate more than 50 per min (0.74, 0.57-0.98), and hypoxia (0.79, 0.61-1.04), and overall hospital duration (0.75, 0.57-0.99) . The mean reduction is equivalent to 1 hospital day for both severe pneumonia and time in hospital . All effects were greater when children with wheezing were omitted from the analysis . INTERPRETATION: Adjuvant treatment with 20 mg zinc per day accelerates recovery from severe pneumonia in children, and could help reduce antimicrobial resistance by decreasing multiple antibiotic exposures, and lessen complications and deaths where second line drugs are unavailable. J Colloid Interface Sci, 2004 Jul 1, 275(1), 177 - 82 Silver nanoparticles as antimicrobial agent: a case study on E . coli as a model for Gram-negative bacteria; Sondi I et al.; The antimicrobial activity of silver nanoparticles against E . coli was investigated as a model for Gram-negative bacteria . Bacteriological tests were performed in Luria-Bertani (LB) medium on solid agar plates and in liquid systems supplemented with different concentrations of nanosized silver particles . These particles were shown to be an effective bactericide . Scanning and transmission electron microscopy (SEM and TEM) were used to study the biocidal action of this nanoscale material . The results confirmed that the treated E . coli cells were damaged, showing formation of "pits" in the cell wall of the bacteria, while the silver nanoparticles were found to accumulate in the bacterial membrane . A membrane with such a morphology exhibits a significant increase in permeability, resulting in death of the cell . These nontoxic nanomaterials, which can be prepared in a simple and cost-effective manner, may be suitable for the formulation of new types of bactericidal materials. Microbes Infect, 2004 May, 6(6), 536 - 41 Induced expression of the antimicrobial peptide melittin inhibits experimental infection by Mycoplasma gallisepticum in chickens; Lazarev VN et al.; The in vivo action of the antimicrobial peptide melittin, expressed from a recombinant plasmid vector, on chickens experimentally infected with Mycoplasma gallisepticum was studied . The plasmid vector pBI/mel2/rtTA includes the melittin gene under the control of an inducible tetracycline-dependent human cytomegalovirus promoter and the gene coding for the trans-activation protein rtTA . Aerosol administration of the vector, followed by infecting the chickens with M . gallisepticum 1226, is shown to inhibit development of infection . The inhibitory action was confirmed by a complex of clinical, pathomorphological, histological and serological studies, and also by comparing the M . gallisepticum reisolation frequency from the respiratory tract and internal organs . The data suggest that plasmid vectors expressing genes of antimicrobial peptides can be considered as potential agents for the prevention and treatment of mycoplasma infections in poultry farming. Trends Biotechnol, 2004 Jun, 22(6), 286 - 94 The contribution of farm animals to human health; Kues WA et al.; Farm animals and their products have a longstanding and successful history of providing significant contributions to human nutrition, clothing, facilitation of labour, research, development and medicine and have thus been essential in improving life expectancy and human health . With the advent of transgenic technologies the potential of farm animals for improving human health is growing and many areas remain to be explored . Recent breakthroughs in reproductive technologies, such as somatic cloning and in vitro embryo production, and their merger with molecular genetic tools, will further advance progress in this field . Here, we have summarized the contribution of farm animals to human health, covering the production of antimicrobial peptides, dietary supplements or functional foods, animals used as disease models and the contribution of animals to solving urgent environmental problems and challenges in medicine such as the shortage of human cells, tissues and organs and therapeutic proteins . Some of these areas have already reached the level of preclinical testing or commercial application, others will be further advanced only when the genomes of the animals concerned have been sequenced and annotated . Provided the necessary precautions are being taken, the transmission of pathogens from animals to humans can be avoided to provide adequate security . Overall, the promising perspectives of farm animals and their products warrant further research and development in this field. Pediatr Clin North Am, 2004 Jun, 51(3), 819 - 27, xii Anomalies, abnormalities, and care of the umbilicus; Pomeranz A; This article discusses various anomalies and abnormalities of the umbilical cord and issues related to cord care . The issue of screening newborns with an isolated single umbilical artery for renal abnormalities is addressed . The clinical presentation of infants with omphalomesenteric and urachal duct remants along with the more common umbilical granuloma is reviewed . All three of these abnormalities can present with a wet or draining cord . The need for umbilical cord treatment with antimicrobial/antiseptic agents versus dry cord care is discussed, as are serious infections that involve the cord. Insect Mol Biol, 2004 Jun, 13(3), 283 - 91 Transcriptional regulation in cowpea bruchid guts during adaptation to a plant defence protease inhibitor; Moon J et al.; Cowpea bruchid, when fed on a diet containing the soybean cysteine protease inhibitor soyacystatin N (scN), activates an array of counter-defence genes to adapt to the negative effects of the inhibitor and regain its normal rate of feeding and development . A collection of 1920 cDNAs was obtained by differential subtraction with cDNAs prepared from guts of the 4th instar larvae of scN-adapted (reared on scN-containing diet) and scN-unadapted (reared on regular scN-free diet) cowpea bruchids . Subsequent expression profiling using DNA microarray and Northern blot analyses identified ninety-four transcript species from this collection that are responsive to dietary scN . scN-adapted insects induced genes encoding protein and carbohydrate digestive enzymes, probably to help meet their carbon and nitrogen requirements . Up-regulation of antimicrobial and detoxification protein genes may represent a generalized defence response . Genes down-regulated by scN reflected physiological adjustments of the cowpea bruchids to scN challenge . A large portion of the responsive genes, presumably involved in carrying out the counter-defence response, were of unknown function . The full-length cDNA of an scN-inducible cathepsin B-like cysteine protease was obtained . Its transcriptional response to scN during larval development contrasts with the pattern of the cathepsin L family, the major digestive enzymes . These results suggest cathepsin B-like cysteine proteases may play a crucial role in cowpea bruchid adaptation to dietary scN. Biochemistry, 2004 Jun 1, 43(21), 6393 - 403 Effect of drastic sequence alteration and D-amino acid incorporation on the membrane binding behavior of lytic peptides; Papo N et al.; The amphipathic alpha-helix is a common motif found in many cell lytic peptides including antimicrobial peptides . We have recently shown that significantly altering the amphipathic structure of a lytic peptide by reshuffling its sequence and/or replacing a few l-amino acids with their D-enantiomers did not significantly affect the antimicrobial activity of the peptides nor their ability to bind and permeate negatively charged (PE/PG) membranes . However, a pronounced effect was observed regarding their hemolytic activity and their ability to bind and permeate zwitterionic (PC/Cho) membranes . To shed light on these findings, here we used surface plasmon resonance (SPR) with mono- and bilayer membranes . We found that the L-amino acid (aa) peptides bound 10-25-fold stronger to PC/Cho bilayers compared with monolayers, whereas the diastereomers bound similarly to both membranes . A two-state reaction model analysis of the data indicated that this difference is due to the insertion of the L-aa peptides into the PC/Cho bilayers, whereas the diastereomers are surface-localized . In contrast, only an approximately 2-fold difference was found with negatively charged membranes . Changes in the amphipathicity markedly affected only the insertion of the L-aa peptides into PC/Cho bilayers . Furthermore, whereas the all-L-aa peptides bound similarly to the PC/Cho and PE/PG membranes, the diastereomers bound approximately 100-fold better to PE/PG compared with PC/Cho membranes, and selectivity was determined only in the first binding step . The effect of the peptides on the lipid order determined by using ATR-FTIR studies supported these findings . Besides shedding light on the mode of action of these peptides, the present study demonstrates SPR as a powerful tool to differentiate between non-cell-selective compared with bacteria-selective peptides, based on differences in their membrane binding behavior. Clin Infect Dis, 2004 Jun 1, 38(11), 1570 - 8 Epub 2004 May 13. Clinical outcomes of pneumococcal pneumonia caused by antibiotic-resistant strains in asian countries: a study by the Asian Network for Surveillance of Resistant Pathogens; Song JH et al.; To evaluate the clinical outcomes of pneumococcal pneumonia caused by antibiotic-resistant strains in Asian countries, we performed a prospective observational study of 233 cases of adult pneumococcal pneumonia in 9 Asian countries from January 2000 to June 2001 . Among 233 isolates, 128 (55%) were not susceptible to penicillin (25.3% were intermediately susceptible, and 29.6% were resistant) . Clinical severity of pneumococcal pneumonia was not significantly different between antibiotic-resistant and antibiotic-susceptible groups . Mortality rates among patients with pneumococcal pneumonia caused by penicillin-, cephalosporin-, or macrolide-resistant strains were not higher than those with antibiotic-susceptible pneumococcal pneumonia . Bacteremia and mechanical ventilation were significant risk factors for death, but any kind of antibiotic resistance was not associated with increased mortality due to pneumococcal pneumonia . Outcome of pneumococcal pneumonia was not significantly affected by drug resistance, and current antimicrobial regimens are mostly effective in the treatment of pneumococcal pneumonia, despite the widespread emergence of in vitro resistance. Orthopade, 2004 Jul, 33(7), 822 - 8 {Guidelines on antimicrobial therapy in situations of periprosthetic THR infection}; Frommelt L; Periprosthetic infection is nowadays a rare complication in artificial joint replacement . The infection of joint prostheses is a foreign body associated infection at the site of bone tissue which is difficult to treat with antimicrobial agents . Apart from cases with early, non-established infections, the surgical removal of the foreign material and radical debridement of bone and soft tissue is necessary . The surgical revision is performed preferentially using a one or two stage exchange of the prosthesis . In some cases, only the removal of the prosthesis is necessary . In these cases, a pseudoarthroplasty or an arthrodesis without definitely implanted foreign material is performed . Amputation is carried out if all other forms of revision have failed . All of these surgical procedures are accomplished by antimicrobial therapy, whether systemically administered and/or topically by drug delivery systems . Controlled studies for the comparison of one and two stage exchange procedures are not currently available . For the therapy of periprosthetic infection, an infectious disease specialist who is experienced in this area is needed in addition to an experienced surgical team . Thus, the therapy of these infections should be performed exclusively in centres of competence. Methods Mol Biol, 2004, 268, 317 - 30 Flavonoids from Argentine Tagetes (Asteraceae) with antimicrobial activity; Tereschuk ML et al.; The flavonoids, constituting one of the most numerous and widespread groups of natural plant constituents, are important to humans not only because they contribute to plant colors but also because many members are physiologically active . These low-molecular-weight substances, found in all vascular plants, are phenylbenzopyrones . Over 4000 structures have been identified in plant sources, and they are categorized into several groups . Primarily recognized as pigments responsible for the autumnal burst of hues and the many shades of yellow, orange, and red in flowers and food, the flavonoids are found in fruits, vegetables, nuts, seeds, stems, flowers, and leaves as well as tea and wine and are important constituents of the human diet . They are prominent components of citrus fruits and other food sources . Flavonols (quercetin, myricetin, and kaempferol) and flavones (apigenin and luteolin) are the most common phenolics in plant-based foods . Quercetin is also a predominant component of onions, apples, and berries . Such flavanones as naringin are typically present in citrus fruit, and flavanols, particularly catechin, are present as catechin gallate in such beverages as green or black tea and wine . Some major sources of flavonoids are outlined in Table 1 . The daily intake of flavonoids in humans has been estimated to be approx 25 mg/d, a quantity that could provide pharmacologically significant concentrations in body fluids and tissues, assuming good absorption from the gastrointestinal tract.Biological activity of flavonoids was first suggested by Szent-Gyorgyi 1938, who reported that citrus peel flavonoids were effective in preventing the capillary bleeding and fragility associated with scurvy . The broad spectrum of biological activity within the group and the multiplicity of actions displayed by a certain individual members make the flavonoids one of the most promising classes of biologically active compounds. Methods Mol Biol, 2004, 268, 281 - 8 Testing disinfectants in the food factory: phenol coefficient method; Herrera AG; Contamination of foods by the environment has direct public health and keeping quality significance . The food factory environment (with raw materials and processing) governs the numbers and types of microorganisms in finished products . Use of the appropriate sampling procedures permits us to discover the magnitude and type of contamination . Microbiological sampling allows objective evaluation of the disinfectants and the sanitation practices and procedures used in the food factory.Disinfectants are antimicrobial pesticides that are primarily used on inanimate surfaces (such as floors, walls, and countertops) to kill infectious bacteria, fungi, and viruses . Antimicrobial pesticides are substances used to kill or suppress the growth of harmful microorganisms on inanimate objects and surfaces . Products intended for the control of microorganisms in or on people or animals are considered drugs, not pesticides, and are therefore regulated by the Food and Drug Administration (FDA) . Antimicrobial pesticides are divided into two broad use categories: 1 . Non-public health products include those used to control the growth of algae, odor-causing bacteria, and microorganisms causing spoilage, deterioration, and fouling of materials . Examples include antimicrobials used in cooling towers, paints, and paper products . 2 . Public health products are intended to control microorganisms infectious to people . Examples include sterilants, which are used to destroy or eliminate all forms of microbial life including fungi, viruses, and all forms of bacteria and their spores; disinfectants, which are used to destroy or irreversibly inactivate infectious fungi and bacteria, but not necessarily their spores; and sanitizers, which are used to reduce, but not necessarily eliminate microorganisms . Examples range from sterilants used to treat surgical instruments to disinfectants applied to hospital floors, walls, and bed linens and sanitizers used on carpets or in laundry additives. Methods Mol Biol, 2004, 268, 153 - 61 Viability of amoebae, fungal conidia, and yeasts: rapid assessment by flow cytometry; Noble-Wang JA et al.; Conventional methods for the evaluation of antimicrobials and disinfecting solutions with microorganisms involve culture-based techniques, which are time-consuming and underestimate the number of viable organisms . Rapid detection and viability measurements of microorganisms in homogenous and heterogenous microbial populations have been greatly enhanced by recent advances in the use of fluorescent stains in flow cytometry (FCM) . FCM has been applied to enumerate, differentiate, and identify microorganisms, determine protein and DNA content of cells, analyze the physiological state of individual cells, and analyze the interaction of drugs, antibiotics, and antimicrobials with microbial cells . Four physiological states of cells can be distinguished by FCM: (1) reproductively viable, (2) metabolically active, (3) intact, and (4) permeabilized.FCM permits a rapid and quantitative measurement of the optical characteristics of cells as they pass through, in a single file, a focused beam of light . As cells are carried within a fast-flowing fluid stream and through the focus of exciting light, three parameters are measured: forward angle light scatter, side angle light scatter, and fluorescence emitted by dyes that have specific interaction with intracellular components of individual cells . FCM data that are presented in histogram and dot plots can be generated to give information on a variety of properties of interest among cells in the population as a whole.FCM offers major advantages in multiparameter data acquisition and multivariate data analysis, high-speed analysis, and cell-sorting capabilities . Disadvantages may be associated with the cost, which is usually over 100,000 (US Dollars) for a typical laser-based flow cytometer with just analyzing capabilities . Another disadvantage is that skilled personnel are usually required to operate these complex instruments so as to get optimum performance . A schematic overview of flow cytometry is presented in Fig . 1. Antimicrob Agents Chemother, 2004 Jun, 48(6), 2132 - 9 Interaction of antimycobacterial drugs with the anti-Mycobacterium avium complex effects of antimicrobial effectors, reactive oxygen intermediates, reactive nitrogen intermediates, and free fatty acids produced by macrophages; Sano K et al.; The profiles of the interaction of antimycobacterial drugs with macrophage (MPhi) antimicrobial mechanisms have yet to be elucidated in detail . We examined the effects of various antimycobacterial drugs on the anti-Mycobacterium avium complex (MAC) antimicrobial activity of reactive oxygen intermediates (ROIs), especially of an H(2)O(2)-halogen (H(2)O(2)-Fe(2+)-NaI)-mediated bactericidal system, reactive nitrogen intermediates (RNIs), and free fatty acids (FFAs), which are known as central antimicrobial effectors of host MPhis against mycobacterial pathogens . We have found that certain drugs, such as rifampin (RIF), rifabutin (RFB), isoniazid (INH), clofazimine (CLO), and some fluoroquinolones, strongly or moderately reduced the anti-MAC activity of the H(2)O(2)-Fe(2+)-NaI system, primarily by inhibiting the generation of hypohalite ions and in part by interfering with the halogenation reaction of bacterial cell components due to the H(2)O(2)-Fe(2+)-NaI system . This phenomenon is specific to the H(2)O(2)-Fe(2+)-NaI system, since these drugs did not reduce the anti-MAC activity of RNIs and FFAs . From the perspective of the chemotherapy of MAC infections, the present findings indicate an important possibility that certain antimycobacterial drugs, such as rifamycins (RIF and RFB), INH, CLO, and also some types of fluoroquinolones, may interfere with the ROI-mediated antimicrobial mechanisms of host MPhis against intracellular MAC organisms. Antimicrob Agents Chemother, 2004 Jun, 48(6), 1983 - 8 The action of bismuth against Helicobacter pylori mimics but is not caused by intracellular iron deprivation; Bland MV et al.; Helicobacter pylori is highly susceptible to bismuth, a heavy metal with antimicrobial activity linked to its effect on bacterial iron uptake . Three strains of H . pylori were analyzed for indicators of iron limitation following exposure to the MIC of colloidal bismuth subcitrate (MIC(CBS)) . Similar morphologic and outer membrane changes were observed following growth in iron-limiting medium and at the MIC(CBS) that inhibited the growth of all three strains . These changes, which were also observed for iron-limited bacteria, were alleviated by the addition of iron to the cultures . H . pylori ATP levels, reduced in iron-limiting medium, were below the limits of detection in two of the three strains following exposure to bismuth . The addition of iron partially restored bacterial ATP levels in these two strains, although not to normal concentrations . In contrast, exposure of the same strains to the MIC(CBS) failed to deplete intracellular levels of iron, which were significantly reduced by culturing in iron-limiting medium . Thus, the antimicrobial effect of bismuth and of iron limitation on H . pylori may be similar . However, the respective mechanisms of intracellular action would appear to be mediated by different pathways within the cell. Biotechnol Appl Biochem, 2004 Jun, 39(Pt 3), 339 - 45 Recombinant antimicrobial peptides efficiently produced using novel cloning and purification processes; Metlitskaia L et al.; Endogenous antimicrobial peptides are ubiquitous components of animal and plant host defences . These peptides, usually cationic and amphipathic, kill target cells rapidly and are efficacious against antibiotic-resistant and clinically relevant pathogens . A practical challenge in the development of cationic peptides as therapeutics is to meet the production requirements for large quantities of highly purified drug substance at competitive costs . While chemical peptide synthesis can be used to manufacture cationic peptides, we have developed cost-effective methods for recombinant production by expressing fusion proteins comprised of multiple copies of the peptides . The fusion proteins accumulate in Escherichia coli inclusion bodies and constitute over 50% of the total cellular proteins . Active antimicrobial peptides are released by chemical reagents and purified by chromatography, combining both standard and novel approaches . Challenges of industrial-scale manufacturing of therapeutics were considered in the development of this process. J Am Vet Med Assoc, 2004 May 15, 224(10), 1634 - 9 Efficacy of ceftiofur hydrochloride sterile suspension administered parenterally for the treatment of acute postpartum metritis in dairy cows; Chenault JR et al.; OBJECTIVE: To evaluate the efficacy of ceftiofur hydrochloride sterile suspension administered parenterally for treatment of acute postpartum metritis (APM) in dairy cows . DESIGN: Multilocation, randomized block, field trial . ANIMALS: 406 cows in the first 14 days postpartum . PROCEDURE: Cows with rectal temperatures > or = 39.5 degrees C (103.1 degrees F) without clinical signs of respiratory or gastrointestinal tract disease and with a fetid vaginal discharge were allocated randomly in blocks of 3 to 3 treatment groups: sterile saline (0.9% NaCl) solution administered at a dosage of 2 mL/45.4 kg (2 mL/100 lb), SC or IM, once daily for 5 days (control); or ceftiofur hydrochloride administered at a dosage of 1.1 or 2.2 mg of ceftiofur equivalents (CE)/kg (0.5 or 1 mg/lb, respectively), SC or IM, once daily for 5 days . Cows were evaluated on days 6, 10, and 14, and clinical cure or failure to cure was determined . Clinical cure was defined as no additional antimicrobial treatment administered, rectal temperature < 39.5 degrees C, and absence of a fetid vaginal discharge . RESULTS: On day 14, clinical cure rates were 77%, 65%, and 62% for the 2.2 mg of CE/kg, 1.1 mg of CE/kg, and control groups, respectively . No significant differences were detected in clinical cure rates between control and treatment groups on day 10 or 6 . CONCLUSIONS AND CLINICAL RELEVANCE: Ceftiofur hydrochloride administered at a dosage of 2.2 mg of CE/kg, SC or IM, once daily for 5 days was efficacious for treatment of APM in dairy cows. Am J Infect Control, 2004 May, 32(3), 161 - 9 Survey of oral care practices in US intensive care units; Binkley C et al.; BACKGROUND: Research has shown that oral care involving toothbrushes and topical antimicrobials improves the oral health of medically compromised patients and may reduce the incidence of nosocomial infections including pneumonia . This survey research was undertaken to determine the type and frequency of oral care in intensive care units (ICUs) in the United States and the attitudes, beliefs, and knowledge of health care workers . METHODS: A randomly selected survey of 102 ICUs within the continental United States participated with 556 respondents; 97% of respondents were registered nurses . MEASUREMENTS: Frequency and type of oral care provided, attitudes and beliefs, and knowledge and training in oral care were measured . RESULTS: Ninety-two percent of respondents perceived oral care to be a high priority . The primary methods of oral care involved the use of foam swabs, moisturizers, and mouthwash . Toothbrushes and toothpaste were used infrequently by almost 80% of respondents . The majority of nurses indicated a need for research-proven oral care standards and desired to learn more . CONCLUSIONS: In this random sample of ICUs, oral care methods were not consistent with current research and oral care protocols . The translation of oral care research into practice in the ICU may improve the quality of care and decrease the incidence of ventilator-associated pneumonia. Caries Res, 2004 May-Jun, 38(3), 286 - 93 Sugar alcohols: what is the evidence for caries-preventive and caries-therapeutic effects? Van Loveren C. The most widely used sugar alcohols are: xylitol, sorbitol, mannitol, maltitol, lactitol and the products Lycasin and Palatinit . It is often claimed that xylitol is superior to the other sugar alcohols for caries control . This paper examines clinical studies on the caries-preventive and therapeutic effects of sugar alcohols with emphasis on sorbitol and xylitol . It is concluded that chewing sugar-free gum 3 or more times daily for prolonged periods of time may reduce caries incidence irrespective of the type of sugar alcohol used . It may be sufficient to do this only on school days . Sucking xylitol-containing candies or tablets may have a similar effect as chewing xylitol chewing gum . Clinical trials suggest greater caries reductions from chewing gums sweetened with xylitol than from gums sweetened with sorbitol . However, the superiority of xylitol was not confirmed in 2 out of 4 clinical trials comparing the caries-preventive effect of xylitol- with sorbitol-sweetened gums . The caries-preventive effects of polyol-containing gums and candies seem to be based on stimulation of the salivary flow, although an antimicrobial effect cannot be excluded . There is no evidence for a caries-therapeutic effect of xylitol . These conclusions are in line with those of recent reviews and with the conclusions of the Scientific Committee on Medicinal Products and Medical Devices of the EU Commission . Caries Res, 2004 May-Jun, 38(3), 247 - 53 Salivary proteins: protective and diagnostic value in cariology? Van Nieuw Amerongen A, Bolscher JG, Veerman EC. Saliva is essential for a lifelong conservation of the dentition . Various functions of saliva are implicated in the maintenance of oral health and the protection of our teeth: (i) The tooth surface is continuously protected against wear by a film of salivary mucins and proline-rich glycoprotein . (ii) The early pellicle proteins, proline-rich proteins and statherin, promote remineralization of the enamel by attracting calcium ions . (iii) Demineralization is retarded by the pellicle proteins, in concert with calcium and phosphate ions in saliva and in the plaque fluid . (iv) Several salivary (glyco)proteins prevent the adherence of oral microorganisms to the enamel pellicle and inhibit their growth . (v) The salivary bicarbonate/carbonate buffer system is responsible for rapid neutralization of acids . An overview is presented on the major antimicrobial systems in human saliva . Not only the well-known major salivary glycoproteins, including mucins, proline-rich glycoprotein and immunoglobulins, but also a number of minor salivary (glyco)proteins, including agglutinin, lactoferrin, cystatins and lysozyme, are involved in the first line of defense in the oral cavity . Besides, small cationic antimicrobial peptides, e.g . defensins, cathelicidin and the histatins, have come into focus . These are potentially suited as templates for the design of a new generation of antibiotics, since they kill a broad spectrum of microorganisms, while hardly evoking resistance, in contrast to the classical antibiotics . Caries Res, 2004 May-Jun, 38(3), 223 - 9 Antimicrobials in future caries control? A review with special reference to chlorhexidine treatment; Twetman S; The aim of this paper was to examine recent evidence for the effect of the antibacterial approach to prevent and control caries with special reference to the use of chlorhexidine (CHX) . Existing information from the mid 1990s provided limited evidence for the effectiveness of CHX gels, rinses and toothpaste in preventing caries in permanent teeth of children and adolescents . An updated literature search on CHX intervention in controlled clinical trials from 1995 to May 2003 unveiled 22 studies covering over 4,500 patients with clinical caries as end point . The vast majority (n = 21) were dealing with CHX-containing varnishes . Since the studies exhibited disparities in design, diagnosis and intervention, the findings were subgrouped with respect to caries type and localization . According to the ranking system of the Swedish Council on Technology Assessment in Health Care, the evidence for an anticaries effect of CHX varnishes was rated as inconclusive for caries-active schoolchildren and adolescents with regular fluoride exposure . Regarding fissure caries, a preventive effect of CHX varnishes was demonstrated in 4 studies out of 5 when compared to no treatment in children with low fluoride exposure . The evidence for arresting root caries in dry-mouth patients and frail elderly subjects was inconclusive . In conclusion, the evidence from the recent literature was inconclusive for the use of CHX varnishes for caries prevention in risk groups . MMWR Morb Mortal Wkly Rep, 2004 May 21, 53(19), 407 - 10 Fatal cases of Rocky Mountain spotted fever in family clusters--three states, 2003; Centers for Disease Control and Prevention (CDC); Rocky Mountain spotted fever (RMSF), a tickborne infection caused by Rickettsia rickettsii and characterized by a rash, has a case-fatality rate as high as 30% in certain untreated patients . Even with treatment, hospitalization rates of 72% and case-fatality rates of 4% have been reported . This report summarizes the clinical course of three fatal cases of RMSF in children and related illness in family members during the summer of 2003 . These cases underscore the importance of 1) prompt diagnosis and appropriate antimicrobial therapy in patients with RMSF to prevent deaths and 2) consideration of RMSF as a diagnosis in family members and contacts who have febrile illness and share environmental exposures with the patient. Vet Ther, 2004 Spring, 5(1), 60 - 74 Pharmacokinetics and lung tissue concentrations of tulathromycin, a new triamilide antibiotic, in cattle; Nowakowski MA et al.; The pharmacokinetics of the new triamilide antibiotic tulathromycin was investigated in two cattle studies . Following a single subcutaneous injection, the drug was rapidly absorbed and bioavailability was excellent . High and persistent levels of the drug in lung tissue were observed as well . These attributes are advantageous for an antimicrobial drug indicated for the treatment of bacterial and mycoplasmal respiratory diseases in cattle. J Med Microbiol, 2004 Jun, 53(Pt 6), 535 - 8 Helicobacter pylori: antibiotic resistance and eradication rates in Suffolk, UK, 1991-2001; Cameron EA et al.; Helicobacter pylori infection causes a number of gastrointestinal diseases and its current treatment is based on multidrug regimes including acid suppression and antimicrobials . The success of these regimes is determined by a number of factors including antibiotic resistance, which varies widely but is an increasing problem . Local data are important in establishing the most cost-effective eradication regime . Data have been collected prospectively on antibiotic resistance at Ipswich Hospital (Suffolk, UK) in all consecutive isolates of H . pylori from 1991 to 2001 . The success of regimes consisting of a proton pump inhibitor, amoxycillin and metronidazole (PPI/A/M) has also been evaluated in patients found positive on serological testing in primary care using urea breath testing . Overall, metronidazole resistance was found in 31.7 % of isolates and clarithromycin resistance in 5.3 % . A significant increase in metronidazole resistance from 29.1 to 37.0 % (P = 0.022) and a decrease in clarithromycin resistance from 10.3 to 3.8 % (P = 0.014) was seen over the study period . Metronidazole resistance was significantly more common in women (P < 0.001) and young patients (P < 0.001) . Eradication with PPI/A/M was successful in 89.9 % of patients and did not change significantly over the study period . Eradication rates were lower in young patients (P < 0.001) . Whilst metronidazole resistance is increasing in Suffolk, this does not seem to have a significant effect on eradication rates . Metronidazole-based regimes are still effective first-line treatments in most patients. J Inorg Biochem, 2004 Jun, 98(6), 1151 - 9 Equilibrium characterization of the As(III)-cysteine and the As(III)-glutathione systems in aqueous solution; Rey NA et al.; Some arsenic compounds were the first antimicrobial agents specifically synthesized for the treatment of infectious diseases such as syphilis and trypanosomiasis . More recently, arsenic trioxide has been shown to be efficient in the treatment of acute promyelocytic leukemia . The exact mechanism of action has not been elucidated yet, but it seems to be related to arsenic binding to vicinal thiol groups of regulatory proteins . Glutathione is the major intracellular thiol and plays important roles in the cellular defense and metabolism . This paper reports on a study of the interactions between arsenic(III) and either cysteine or glutathione in aqueous solution . The behavior observed for the As(III)-glutathione system is very similar to that of As(III)-cysteine . In both cases, the formation of two complexes in aqueous solution was evidenced by NMR and electronic spectroscopies and by potentiometry . The formation constants of the cysteine complexes {As(H(-1)Cys)(3)}, log K = 29.84(6), and {As(H(-2)Cys)(OH)(2)}(-), log K = 12.01(9), and of the glutathione complexes {As(H(-2)GS)(3)}(3-), log K = 32.0(6), and {As(H(-3)GS)(OH)(2)}(2-), log K = 10(3) were calculated from potentiometric and spectroscopic data . In both cases, the {As(HL)(3)} species, in which the amine groups are protonated, predominate from acidic to neutral media, and the {As(L)(OH)(2)} species appear in basic medium (the charges were omitted for the sake of simplicity) . Spectroscopic data clearly show that the arsenite-binding site in both complexes is the sulfur atom of cysteine . In the {As(L)(OH)(2)} species, the coordination sphere is completed by two hydroxyl groups . In both cases, arsenic probably adopts a trigonal pyramidal geometry . Above pH 10, the formation of {As(OH)(2)O}(-) excludes the thiolates from arsenic coordination sites . At physiological pH, almost 80% of the ligand is present as {As(HL)(3)}. Bioorg Med Chem Lett, 2004 Jun 21, 14(12), 3165 - 8 Structure-based design, synthesis, and antimicrobial activity of purine derived SAH/MTA nucleosidase inhibitors; Tedder ME et al.; The structure-based design, synthesis, and biological activity of novel inhibitors of S-adenosyl homocysteine/methylthioadenosine (SAH/MTA) nucleosidase are described . Using 6-substituted purine and deaza purines as the core scaffolds, a systematic and structure guided series of modifications provided low nM inhibitors with broad-spectrum antimicrobial activity. Bioorg Med Chem Lett, 2004 Jun 21, 14(12), 3069 - 72 The synthesis and antimicrobial evaluation of a new series of isoxazolinyl oxazolidinones; Weidner-Wells MA et al.; A series of oxazolidinone antibacterial agents containing a 5-substituted isoxazol-3-yl moiety were synthesized via a nitrile oxide {3+2} dipolar cycloaddition reaction . These compounds were screened against a panel of susceptible and resistant Gram-positive organisms . Several analogs from this series were comparable to or more potent than linezolid in vitro. Br J Dermatol, 2004 May, 150(5), 904 - 9 Involvement of granulysin-producing T cells in the development of superficial microbial folliculitis; Oono T et al.; BACKGROUND: Granulysin is a recently identified antimicrobial protein expressed on cytotoxic T cells, natural killer (NK) cells and NKT cells . It has been shown that granulysin contributes to the defence mechanisms against mycobacterial infection . Superficial microbial folliculitis is a common skin disease . In a previous report, we showed that, as a first line of defence, alpha-defensin, a human neutrophil peptide, and beta-defensin (human beta-defensin-2) were expressed in infiltrating neutrophils and in lesional epidermal keratinocytes, respectively, in superficial folliculitis . As we also observed many infiltrating lymphocytes in lesional dermis, we hypothesized that infiltrating lymphocytes may possess antimicrobial substances, such as granulysin, and play a role in the defence mechanism as a second line of defence . OBJECTIVES: Seven specimens of superficial microbial folliculitis diagnosed clinically and histologically were examined by means of immunohistochemistry . To identify the phenotype of cells expressing granulysin, confocal laser microscopic examination was performed . RESULTS: A dense lymphoid cell infiltrate was observed in pustules, in the perivascular regions . A large number of these lymphoid cells were positive for granulysin . Phenotypically, cells consisted of CD3+ T cells, CD8+ T cells and UCHL-1+ T cells . CD20+ cells and CD56+ cells were not observed . Microscopic examination with a confocal laser showed that the lymphocytes producing granulysin were CD3+ and CD4+ T cells but not CD8+ T cells . CONCLUSIONS: We showed that many granulysin-bearing T cells infiltrated affected follicles and perilesional dermis in superficial microbial folliculitis . However, few granulysin-positive lymphoid cells were observed in sterile pustular lesions . Our observations indicated that adaptive immunity such as granulysin, a lymphocyte-produced antimicrobial protein, may play an important role in the cutaneous defence mechanism. Bioconjug Chem, 2004 May-Jun, 15(3), 530 - 5 Promotion of peptide antimicrobial activity by fatty acid conjugation; Chu-Kung AF et al.; Three peptides, YGAA{KKAAKAA}(2) (AKK), KLFKRHLKWKII (SC4), and YG{AKAKAAKA}(2) (KAK), were conjugated with lauric acid and tested for the effect on their structure, antibacterial activity, and eukaryotic cell toxicity . The conjugated AKK and SC4 peptides showed increased antimicrobial activity relative to unconjugated peptides, but the conjugated KAK peptide did not . The circular dichroism spectrum of AKK showed a significantly larger increase in its alpha-helical content in the conjugated form than peptide KAK in a solution containing phosphatidylethanolamine/phosphotidylglycerol vesicles, which mimics bacterial membranes . The KAK and AKK peptides and their corresponding fatty acid conjugates showed little change in their structure in the presence of phosphatidylcholine vesicles, which mimic the cell membrane of eukaryotic cells . The hemolytic activity of the KAK and AKK peptides and conjugates was low . However, the SC4 fatty acid conjugate showed a large increase in hemolytic activity and a corresponding increase in helical content in the presence of phosphatidylcholine vesicles . These results support the model of antimicrobial peptide hemolytic and antimicrobial activity being linked to changes in secondary structure as the peptides interact with lipid membranes . Fatty acid conjugation may improve the usefulness of peptides as antimicrobial agents by enhancing their ability to form secondary structures upon interacting with the bacterial membranes. Aust Vet J, 2004 Apr, 82(4), 223 - 7 Survey for papillomatous digital dermatitis in Australian dairy cattle; Milinovich GJ et al.; OBJECTIVE: To determine whether Treponema-associated papillomatous digital dermatitis (PDD) occurs in Australian dairy cattle . DESIGN: Mail-out questionnaire and histological and bacteriological examination of biopsy tissue from suspect PDD lesions . PROCEDURE: The questionnaire was mailed to 375 veterinarians to evaluate their knowledge of PDD, determine if they had observed the disease in Australian dairy cattle, and to request biopsy material from suspicious cases . Biopsies were examined for histological and bacteriological evidence of PDD, including for the presence of spirochaetes . RESULTS: Eighty-eight replies to the questionnaire were received (23.5%) . Of 52 respondents who were aware of PDD as a possible cause of lameness, 26 reported observing the condition in Australian cattle . Of 32 respondents who were unaware of the condition, 6 reported observing lesions that might have been PDD . The majority of reports of PDD-like lesions came from the southern Australian states, the condition occurring during periods of high rainfall and proving responsive to topical or parenteral application of antimicrobials . Biopsies from five erosive lesions showed histological similarity to PDD whereas biopsies from five proliferative lesions were consistent with chronic inflammation, fibroma or cutaneous papilloma . The presence of spirochaetes was not demonstrated in any of the lesions by histological or bacteriological methods . CONCLUSION: Anecdotal reports and analysis of biopsy material confirm that a condition similar to PDD does occur sporadically in dairy cattle in southern Australia . However, this condition has so far not been shown to be associated with the presence of spirochaetes in the lesions. Immunogenetics, 2004 Jun, 56(3), 170 - 7 Epub 2004 May 18. Bioinformatic discovery and initial characterisation of nine novel antimicrobial peptide genes in the chicken; Lynn DJ et al.; Antimicrobial peptides (AMPs) are essential components of innate immunity in a range of species from Drosophila to humans and are generally thought to act by disrupting the membrane integrity of microbes . In order to discover novel AMPs in the chicken, we have implemented a bioinformatic approach that involves the clustering of more than 420,000 chicken expressed sequence tags (ESTs) . Similarity searching of proteins-predicted to be encoded by these EST clusters-for homology to known AMPs has resulted in the in silico identification of full-length sequences for seven novel gallinacins (Gal-4 to Gal-10), a novel cathelicidin and a novel liver-expressed antimicrobial peptide 2 (LEAP-2) in the chicken . Differential gene expression of these novel genes has been demonstrated across a panel of chicken tissues . An evolutionary analysis of the gallinacin family has detected sites-primarily in the mature AMP-that are under positive selection in these molecules . The functional implications of these results are discussed. Pediatr Surg Int, 2004 Jun, 20(6), 402 - 7 Epub 2004 May 18. Advances in the treatment of oesophageal atresia over three decades: the 1970s and the 1990s; Orford J et al.; Oesophageal atresia management has evolved alongside the development of paediatric surgery . An analysis of a 30-year prospective collection of oesophageal atresia cases treated at the Royal Alexandra Hospital for Children, NSW, Australia is presented . There has been a dramatic change in the surgical approach . Fewer infants are undergoing operations of cervical oesophagostomy, gastrostomy and oesophageal replacement . More, including fragile infants, are surviving with a repaired native oesophagus . Fewer infants are suffering morbidity secondary to anastomotic leak . The progression in treatment has been enabled by improved neonatal support due to advances in neonatology, neonatal anaesthesia, nutritional support and antimicrobial therapy . Oesophageal atresia treatment and outcome has changed markedly over three decades . Cooperative multi-centre database development is now required to provide data in order to further refine treatment for clinical challenges that remain. Proc Natl Acad Sci U S A, 2004 May 25, 101(21), 8233 - 8 Epub 2004 May 17. A gene cluster for secondary metabolism in oat: implications for the evolution of metabolic diversity in plants; Qi X et al.; The evolution of the ability to synthesize specialized metabolites is likely to have been key for survival and diversification of different plant species . Oats (Avena spp.) produce antimicrobial triterpenoids (avenacins) that protect against disease . The oat beta-amyrin synthase gene AsbAS1, which encodes the first committed enzyme in the avenacin biosynthetic pathway, is clearly distinct from other plant beta-amyrin synthases . Here we show that AsbAS1 has arisen by duplication and divergence of a cycloartenol synthase-like gene, and that its properties have been refined since the divergence of oats and wheat . Strikingly, we have also found that AsbAS1 is clustered with other genes required for distinct steps in avenacin biosynthesis in a region of the genome that is not conserved in other cereals . Because the components of this gene cluster are required for at least four clearly distinct enzymatic processes (2,3-oxidosqualene cyclization, beta-amyrin oxidation, glycosylation, and acylation), it is unlikely that the cluster has arisen as a consequence of duplication of a common ancestor . Although clusters of paralogous genes are common in plants (e.g., gene clusters for rRNA and specific disease resistance), reports of clusters of genes that do not share sequence relatedness and whose products contribute to a single selectable function are rare {Gierl, A . & Frey, M . (2001) Planta 213, 493-498} . Taken together, our evidence has important implications for the generation of metabolic diversity in plants. Arch Dermatol, 2004 May, 140(5), 563 - 6 Acaricidal activity of Melaleuca alternifolia (tea tree) oil: in vitro sensitivity of sarcoptes scabiei var hominis to terpinen-4-ol; Walton SF et al.; OBJECTIVE: To compare the acaricidal activity of Melaleuca alternifolia (tea tree) oil (TTO) and some of its individual active components on the itch mite Sarcoptes scabiei var hominis . DESIGN: In vitro acaricide sensitivity assessment . SETTING: The Menzies School of Health Research laboratory, located near the Infectious Diseases Ward of the Royal Darwin Hospital, Australia, where patients are admitted and treated for crusted scabies . PARTICIPANTS: Scabies mites (S scabiei var hominis) were collected from a 20-year-old Aboriginal woman admitted to the Royal Darwin Hospital with crusted scabies.Interventions Within 3 hours of collection, scabies mites were placed in continuous direct contact with the TTO products and control acaricides and were observed at regular intervals . MAIN OUTCOME MEASURES: Percentage of mites dead at regular observation intervals between 5 minutes and 24 hours during continuous exposure to the TTO products and acaricides . RESULTS: The 5% TTO and active component terpinen-4-ol were highly effective in reducing mite survival times . Statistically significant differences in mite survival curves were observed for 5% TTO, 2.1% terpinen-4-ol, 5% permethrin, and ivermectin (100 microg/g of Emulsifying Ointment British Pharmacopoeia 88) . In vivo effectiveness was also observed . CONCLUSIONS: Documentation of resistance against antiectoparasitic compounds is increasing . Reported S scabiei treatment failures with lindane, crotamiton, and benzyl benzoate, as well as likely emerging resistance to 5% permethrin and oral ivermectin, are of concern and advocate for the identification and development of novel acaricidal drugs . Tea tree oil is a membrane-active biocide extracted from the tree M alternifolia . It is a principal antimicrobial in a wide range of pharmaceuticals sold in Australia, with the main active component being oxygenated terpenoids . The results suggest that TTO has a potential role as a new topical acaricide and confirm terpinen-4-ol as the primary active component. FEBS Lett, 2004 May 21, 566(1-3), 301 - 6 The cyclic antimicrobial peptide RTD-1 induces stabilized lipid-peptide domains more efficiently than its open-chain analogue; Abuja PM et al.; The effects of a mammalian cyclic antimicrobial peptide, rhesus theta defensin 1 (RTD-1) and its open chain analogue (oRTD-1), on the phase behaviour and structure of model membrane systems (dipalmitoyl phosphatidylcholine, DPPC and dipalmitoyl phosphatidylglycerol, DPPG) were studied . The increased selectivity of RTD-1 for anionic DPPG over zwitterionic DPPC was shown by differential scanning calorimetry . RTD-1, at a molar peptide-lipid ratio of 1:100, induced considerable changes in the phase behaviour of DPPG, but not of DPPC . The main transition temperature, Tm, was unchanged, but additional phase transitions appeared above Tm . oRTD-1 induced similar effects . However, the effects were not observable below a peptide:lipid molar ratio of 1:50, which correlates with the weaker biological activity of oRTD-1 . Small- and wide-angle X-ray scattering revealed for DPPG the appearance of additional structural features induced by RTD-1 above Tm, which were interpreted as correlated lamellar structures, with increased order of the fatty acyl side chains of the lipid . It is proposed that after initial electrostatic interaction of the cationic rim of the peptide with the anionic DPPG headgroups, leading to stabilized lipid-peptide clusters, the hydrophobic face of the peptide assists in its interaction with the fatty acyl side chains eventually leading to membrane disruption. Int J Cancer, 2004 Jul 10, 110(5), 695 - 700 Promoter polymorphisms of tumor necrosis factor-alpha are associated with risk of gastric mucosa-associated lymphoid tissue lymphoma; Wu MS et al.; Genes involved in regulating antimicrobial immunity and inflammation may modulate the risk of Helicobacter pylori-associated diseases . IL-1 and TNF-alpha are major cytokines detected in H . pylori-infected tissues . We aimed to determine the role of gene polymorphisms for these cytokines and their receptors in 2 distinct H . pylori-related gastric malignancies, adenocarcinoma (GAC) and maltoma . Genotyping for IL-1beta (-31 C/T, -511 C/T), TNF-alpha (-238 G/A, -308 G/A, -857 C/T, -863 C/A, -1031 T/C), TNFR1 (-383 A/C) and TNFR2 (196 G/T) was undertaken for 70 patients with maltoma and 204 patients with noncardia GAC and compared to 210 unrelated healthy controls . Genotype frequencies showed no differences among patients with GAC or maltoma and controls for IL-1beta, TNFR1 or TNFR2 . The TNF-alpha -857 T variant was significantly underrepresented in maltoma compared to controls (6.4% vs . 14.3%, p = 0.018), conferring a 3-fold decrease in risk (OR = 0.33, 95% CI 0.15-0.75) . Comparison of allele frequencies between GAC and controls failed to show any statistical significance for TNF-alpha polymorphisms . We concluded that TNF-alpha -857 T itself or a neighboring gene may modify the risk of maltoma . The differences in genetic background as well as divergent clinicopathologic features between GAC and maltoma support the notion that fundamental mechanistic differences exist in these 2 well-defined H . pylori-related malignancies . Klin Mikrobiol Infekc Lek, 2004 Apr, 10(2), 73 - 9 {Antimicrobial prophylaxis in surgery}; Stastnik M; Antimicrobial prophylaxis is an important factor influencing the risk of infection at the spot of surgical interventions (SSI) . SSIs are the most important nosocomial infections of hospitalized surgical patients; they are responsible for increases of 10 to 20 % in the total costs of treatment . The efficacy of antimicrobial prophylaxis hinges on four basic factors . The first is a correctly indicated prophylaxis (in surgical operations with a confirmed reduction of SSI risk after prophylaxis and/or in cases of surgical operations, where a possible early or organ SSI could have tragic consequences) . The second factor is the choice of the best possible antimicrobial for a specific indication . The third factor is the best possible time for the administration of prophylaxis (in most indications at the beginning of anaesthesia) . The fourth factor influencing the efficacy of prophylaxis is its administration for only the absolutely minimum time period necessary (in most indications best is a single administration, possibly including a second peroperative ATB dose) . The high rate of errors in the actual practice of prophylaxis and the confirmed efficacy of implementing local recommendations indicate that it is absolutely necessary to define national and local recommendations for antimicrobial prophylaxis, to ensure that surgeons adhere to these recommendations and to initiate SSI surveillance in the Czech Republic. Med Klin (Munich), 2004 May 15, 99(5), 251 - 60 {Infectious diseases-Part II: New agents, resistances, and treatment strategies}; Salzberger B et al.; A number of new antimicrobial agents have been licensed in the last years, most prominently a number of new antiviral and antimycotic agents . The development of new antibacterial agents has slowed down, new agents are mainly targeted at a small but rapidly growing number of patients infected with bacteria resistant to the current antibiotics.The rates of resistance against antibiotics are rapidly rising, especially for gram-positive cocci and, in addition, for gram-negative nosocomial agents . Rising rates of resistances are further present in chronic viral infections, e . g., HIV infection . Instruments to monitor and minimize the rates of resistances are necessary.Important changes are to be found in a number of infectious complications . For patients with sepsis a new therapeutic principle, drocretogrin, has been found to reduce mortality . Additionally, low-dose corticosteroids in selected patients, close control of blood sugar and other interventions have shown to be effective . Corticosteroids have proven to be effective as well in the reduction of complications in adults with bacterial meningitis, most pronounced in pneumococcal meningitis . With new drugs licensed for the treatment of malaria and changes in the epidemiology, the guidelines for therapy and prevention have been reformulated. Clin Rev Allergy Immunol, 2004 Apr, 26(2), 115 - 25 Toll-like receptors and immune response in allergic disease; Gangloff SC et al.; Allergic reactions are dominated by the preferential development of specific Th2 responses against innocuous antigens in atopic individuals . This can reflect alterations in innate immune mechanisms . Toll-like receptors (TLRs) have evolved as key molecules in innate and adaptive immunity . Their activation by structurally distinct exogenous or endogenous ligands present at the cell microenvironment plays a critical role in antimicrobial defense . The global view is that TLR activation induces antigen-presenting cells to produce cytokines that favor Th1-type immune responses, suggesting that it might prevent the development of deleterious Th2 responses in allergy . On the basis of epidemiological studies and recent data, it has been established that TLRs play a role in the development of Th2 responses . However, more information is needed to fully understand the mechanism of TLR involvement and the implication of immune cells that express TLRs in the Th1/Th2 cytokine profiles . Several TLRs, such as TLR9, TLR7, and TLR8, can be considered as good target candidates . Some TLR ligands, such as CpG DNA, are effective adjuvants, strong inducers of both IL-5 and eosinophilia downregulation . They are also potential links to allergen epitopes that could provide new allergen-specific immunotherapy regimens for the treatment of allergic disorders. FEMS Immunol Med Microbiol, 2004 Jun 1, 41(2), 141 - 8 Anti-complement effects of lactoferrin-derived peptides; Samuelsen O et al.; Lactoferrin is an important biological molecule with many functions such as modulation of the inflammatory response, iron metabolism and antimicrobial defense . One effect of lactoferrin is the inhibition of the classical complement pathway . This study reports that antimicrobial peptides derived from the N-terminal region from both human and bovine lactoferrin, lactoferricin H and lactoferricin B, respectively, inhibit the classical complement pathway . No inhibitory effect of these peptides was observed on the alternative complement pathway in an AP50 assay . However, lactoferricin B reduced the inhibitory properties of serum against Escherichia coli in a concentration dependent manner . These results suggest that the N-terminal region of lactoferrin is the important part in the inhibition of complement activation and that these peptides possess other important properties than their antimicrobial effect. Burns, 2004 Jun, 30(4), 329 - 33 Effects of cytokines and heat shock on defensin levels of cultured keratinocytes; Bick RJ et al.; Burns have been associated with high levels of circulating pro-inflammatory cytokines which promote systemic inflammatory response syndrome (SIRS), immunosuppression and sepsis for which no effective treatment is currently available . Defensins, a family of cationic naturally occurring antimicrobial peptides, are considered important components of the innate immune system and enhance adaptive immunity . This study examines the effects of pro-inflammatory cytokines, interleukin-1beta (IL-1beta), gamma-interferon (IFNgamma) and tumor necrosis factor-alpha (TNFalpha) on human beta-defensin-2 (HBD-2) levels in cultured keratinocytes . We also examined the effects of heat shock at 42 degrees C . The results demonstrate that only TNFalpha shows significant induction of HBD-2 but this induction was not sustained in the long-term . In addition, endogenous levels of defensin were significantly reduced by exposure to heat shock . The keratinocytes also responded to IL-1beta by becoming hypertrophic . These results indicate that stress-related, pro-inflammatory cytokines can induce keratinocytes to synthesize HBD-2, while heat shock appears to reduce its production . These experiments give us further insight into the role of natural antimicrobial peptides under conditions of stress. Can Vet J, 2004 Apr, 45(4), 309 - 11 Availability and estimates of veterinary antimicrobial use in British Columbia; Fraser E et al.; The amount of antimicrobial use is a significant selection pressure that alters the frequency of antimicrobial resistance . This paper summarizes attempts to estimate the weight of antimicrobial purchases in British Columbia for use in animals . The data reported here do not capture all sources of veterinary antimicrobial use in British Columbia . This paper highlights how information deficits on veterinary drug use complicate the development of an evidence-based policy framework for combating antimicrobial resistance. Tumori, 2004 Jan-Feb, 90(1), 132 - 5 The development of pulmonary adenocarcinoma in a patient with Job's syndrome, a rare immunodeficiency condition; Oztop I et al.; The hyperimmunoglobulin E (HIE) (Job's) syndrome often has it onset in childhood and is characterized by markedly elevated serum IgE levels, chronic dermatitis and recurrent pyogenic infections . Lymphoid malignancies have most commonly been associated with this syndrome while the first case in the literature of carcinoma associated with HIE syndrome was a squamous cell carcinoma of the vulva, described by Clark et al . in 1998 . We observed a male patient with Job's syndrome diagnosed at age three who presented with bone pain and a metastatic epithelial tumor of the bone revealed by biopsy . Diagnostic procedures aimed at detecting the primary site showed multiple mediastinal lymph nodes with lung and liver metastases on computed tomography scans and an extradural spinal metastasis at the upper thoracic level on magnetic resonance imaging . Although the patient refused a bronchoscopic procedure, a diagnosis of pulmonary adenocarcinoma was established on the basis of sputum cytology and the clinical aspects of tumor extent . Intravenous corticosteroids and palliative radiotherapy were given for the spinal metastasis . Palliative chemotherapy could not be started because of the patient's poor performance status as well as nosocomial fungal pneumonia and pseudomonal urogenital infection with bacteremia . Despite the antifungal and broad-spectrum antimicrobial treatments, the patient died of pseudomonal sepsis. Hepatogastroenterology, 2004 May-Jun, 51(57), 822 - 4 Living-related liver transplantation in a patient with end-stage hepatolithiasis and a biliary-bronchial fistula; Hirohashi K et al.; Liver transplantation in patients with end-stage hepatolithiasis is complicated by the high incidence of the suppurative cholangitis and systemic infection . A 43-year-old Korean-Japanese woman with hepatolithiasis, biliary cirrhosis, suppurative cholangitis, and biliary-bronchial fistula underwent living-related liver transplantation (LRLT) using a right lobe graft of her sister . The risk of infection was minimized by preoperative percutaneous transhepatic biliary drainage initiated 2 months before transplantation . The native liver was resected en bloc with the extrahepatic bile ducts and the infected section of the right hemidiaphragm . Opportunistic infection was prevented by limiting antimicrobial therapy to the interval from preoperative day 3 to postoperative day 4 . Immunosuppressive agents were given below standard dose . The postoperative course following LRLT was uncomplicated, and hepatic function was good . Careful management of infection and adequate graft size are essential for successful LRLT in patients with end-stage hepatolithiasis. Arch Environ Health, 2003 Jul, 58(7), 433 - 41 Chronic sinusitis: defective T-cells responding to superantigens, treated by reduction of fungi in the nose and air; Dennis DP; In this study, the author used endoscopic sinus photography to study the effects of reduction of fungi in the nose, and in environmental air, on the sinus mucosa of 639 patients diagnosed with chronic rhinosinusitis . Sinus mucosal photographs were taken before and after reduction of fungal load in the nose and air, to determine if there was an optimum environmental air fungal load associated with sinus mucosal recovery to normal appearance . Systemic symptoms associated with fungal exposure, which resolved when fungus was removed from the patient and the environmental air and reappeared with recurrent environmental fungal exposure, are also discussed and are termed systemic fungal symptoms . Interventions consisted of nasal fungal load reduction with normal saline nasal irrigations and antimicrobial nasal sprays, and environmental air fungal load reduction with high-efficiency particulate air (HEPA) filtration in combination with ionizers or evaporation of a solution of botanical extract . Main outcome measures were obtained with environmental air 1-hr gravity-plate fungal colony counts, laser air particle counts, and endoscopic sinus photography . Blood levels of immunoglobulins IgG and IgE for 7 common molds were also determined . After intervention, 94% of patients who used antimicrobial nasal sprays and who reduced their environmental fungal air count to 0-4 colonies per 1-hr agar gravity-plate exposure (n = 365) exhibited normal sinus mucosa by endoscopic exam . Environmental air fungal counts that exceeded 4 colonies resulted in sinus mucosal abnormalities ranging from edema, to pus and/or nasal polyps at higher counts . Neutralization of allergy, and/or surgery, were used as appropriate following implementation of environmental measures . On the basis of these observations, as well as detailed clinical experience and a review of the current literature, the author hypothesizes that the pathogenesis of chronic rhinosinusitis, allergic fungal sinusitis, and systemic fungal symptoms is a genetic defect at the variable beta chain helper T-cell receptor (TCR Vbeta) site which requires the presence of an antigen (fungus) . Chronic sinusitis patients who have recurring exposure to environmental air that contains fungal concentrations in excess of 4 colonies per 1-hr agar plate exposure appear to have an increased risk of persistent chronic sinusitis and/or systemic symptoms, regardless of the medical treatment provided. Harefuah, 2004 Feb, 143(2), 121 - 5, 166 {Bacteriophages as antibacterial agents}; Shasha SM et al.; Bacteriophages are viruses that only infect bacteria . They have played an important role in the development of molecular biology and have been used as anti-bacterial agents . Since their independent discovery by Twort and d'Herelle, they have been extensively used to prevent and treat bacterial infections, mainly in Eastern Europe and the former Soviet Union . In western countries this method has been sporadically employed on humans and domesticated animals . However, the discovery and widespread use of antibiotics, coupled with doubts about the efficacy of phage therapy, led to an eclipse in the use of phage in medicine . The emergence of antibiotic resistant bacteria, especially strains that are multiply resistant, has resulted in a renewed interest in alternatives to conventional drugs . One of the possible replacements for antibiotics is the use of bacteriophages as antimicrobial agents . This brief review aims to describe the history of bacteriophage and early clinical studies on their use in bacterial disease prophylaxis and therapy, and discuss the advantages and disadvantages of bacteriophage in this regard. J Infect Dis, 2004 Jun 1, 189(11), 2129 - 38 Epub 2004 May 10. Sphingosine 1-phosphate induces antimicrobial activity both in vitro and in vivo; Garg SK et al.; Sphingosine 1-phosphate (S1P), a polar sphingolipid metabolite, is involved in a wide spectrum of biological processes, including Ca(++) mobilization, cell growth, differentiation, motility, and cytoskeleton organization . Here, we show a novel role of S1P in the induction of antimicrobial activity in human macrophages that leads to the intracellular killing of nonpathogenic Mycobacterium smegmatis and pathogenic M . tuberculosis . Such activity is mediated by host phospholipase D, which favors the acidification of mycobacteria-containing phagosomes . Moreover, when it was intravenously injected in mycobacteria-infected mice, S1P reduced mycobacterial growth and pulmonary tissue damage . These results identify S1P as a novel regulator of the host antimicrobial effector pathways. J Hosp Infect, 2004 May, 57(1), 44 - 51 The procedures of hygiene to control hospital-acquired diarrhoea in paediatric wards: a multicentre audit; Jusot JF et al.; An audit was carried out in paediatric wards to study the compliance of healthcare workers (HCWs) to the procedures recommended for the control of hospital-acquired diarrhoea . Thirty-two paediatric wards in the southeast of France participated on a voluntary basis in this prospective observational study after completing a self-administered questionnaire recording measures of hygiene . All the observations were made by the same investigator and focused on preventive procedures: use of single room, handwashing, hand disinfection, overclothing, single-use gloves and masks . Two hundred and seventy patient-HCW contacts were observed, including mainly diapering, temperature measurement, collection of blood sample and catheter care . The isolation of patients in a single room and use of gowns by HCWs were significantly associated with diarrhoea . Whereas handwashing before care was performed by HCWs in more than 95% of all the procedures, the compliance in the use of disposable gloves by HCWs was only of 39.4% for technical procedures (including those with potential exposure to blood) and 20.3% for diapering or temperature measurement . A substantial agreement between reported and observed measures of hygiene was observed for handwashing before contact and hand disinfection with antimicrobial soap before contact . In contrast, this agreement was moderate for use of single room, handwashing after contact, overclothing and wearing disposable gloves after a diaper change . Despite the excellent compliance of HCWs to handwashing, clearer recommendations for the indication and use of disinfectants and disposable gloves are urgently needed. Immunity, 2004 May, 20(5), 637 - 49 Monomeric and polymeric gram-negative peptidoglycan but not purified LPS stimulate the Drosophila IMD pathway; Kaneko T et al.; Insects depend solely upon innate immune responses to survive infection . These responses include the activation of extracellular protease cascades, leading to melanization and clotting, and intracellular signal transduction pathways inducing antimicrobial peptide gene expression . In Drosophila, the IMD pathway is required for antimicrobial gene expression in response to gram-negative bacteria . The exact molecular component(s) from these bacteria that activate the IMD pathway remain controversial . We found that highly purified LPS did not stimulate the IMD pathway . However, lipid A, the active portion of LPS in mammals, activated melanization in the silkworm Bombyx morii . On the other hand, the IMD pathway was remarkably sensitive to polymeric and monomeric gram-negative peptidoglycan . Recognition of peptidoglycan required the stem-peptide sequence specific to gram-negative peptidoglycan and the receptor PGRP-LC . Recognition of monomeric and polymeric peptidoglycan required different PGRP-LC splice isoforms, while lipid A recognition required an unidentified soluble factor in the hemolymph of Bombyx morii. Curr Infect Dis Rep, 2004 Jun, 6(3), 187 - 190 Nebulization of Antibiotics in Management of Sinusitis; Vaughan WC; Chronic sinusitis with recurrent acute infections continues to be a significant medical problem . Even after aggressive medical and surgical management, some patients continue to have recurrent infections . These infections are often multidrug-resistant . Topical delivery of medications into body cavities has been practiced for decades . Recently, the use of prescription antibiotic, antifungal, and anti-inflammatory topical medications has increased for sinus patients . This article examines emerging data on nebulized antimicrobials for patients with sinusitis. Cent Eur J Public Health, 2004 Mar, 12 Suppl, S57 - 9 Biological activity of 2-hydroxythiobenzanilides and related compounds; Kubicova L et al.; Thiobenzanilides substituted in thioacyl moiety with one or more hydroxy groups are interesting for their biological effects depending on the substitution pattern . New findings in mechanisms of action of 2-hydroxybenzanilides insert 2-hydroxybenzanilides and their analogues, e.g . substituted thiobenzanilides, among interesting compounds in the development of new potential antimicrobial drugs . The present review paper with 32 references links up with our previous communications which reviewed biological activity of 2-hydroxybenzanilides and related compounds, and includes the research of mono-, di-, and trihydroxythiobenzanilides carried out in the last period. Am J Vet Res, 2004 May, 65(5), 610 - 5 Effect of danofloxacin and tilmicosin on body temperatures of beef calves with pneumonia experimentally induced by inoculation with Mannheimia haemolytica; Fajt VR et al.; OBJECTIVE: To examine effects of danofloxacin and tilmicosin on continuously recorded body temperature in beef calves with pneumonia experimentally induced by inoculation of Mannheimia haemolytica . ANIMALS: 41 Angus-cross heifers (body weight, 160 to 220 kg) without a recent history of respiratory tract disease or antimicrobial treatment, all from a single ranch . PROCEDURE: Radiotransmitters were implanted intravaginally in each calf . Pneumonia was induced intrabronchially by use of logarithmic-phase cultures of M . haemolytica . At 21 hours after inoculation, calves were treated with saline (0.9% NaCl) solution, danofloxacin, or tilmicosin . Body temperature was monitored from 66 hours before inoculation until 72 hours after treatment . Area under the curve (AUC) of the temperature-time plot and mean temperature were calculated for 3-hour intervals and compared among treatment groups . RESULTS: The AUCs for 3-hour intervals did not differ significantly among treatment groups for any of the time periods . Analysis of the mean temperature for 3-hour intervals revealed significantly higher temperatures at most time periods for saline-treated calves, compared with temperatures for antimicrobial-treated calves; however, we did not detect significant differences between the danofloxacin- and tilmicosin-treated calves . The circadian rhythm of temperatures before exposure was detected again approximately 48 hours after bacterial inoculation . CONCLUSIONS AND CLINICAL RELEVANCE: Danofloxacin and tilmicosin did not differ in their effect on mean body temperature for 3-hour intervals but significantly decreased body temperature, compared with body temperature in saline-treated calves . Normal daily variation in body temperature must be considered in the face of respiratory tract disease during clinical evaluation of feedlot cattle. Orthopade, 2004 Apr, 33(4), 397 - 404 {Special aspects of implant-associated infection in orthopedic surgery . From the pathophysiology to custom-tailored prevention strategies}; Schierholz JM et al.; One of the most important risk factors in orthopedic surgery is implant-associated infection . Adhesion and colonization mediated implant infections are extremely resistant to antibiotics and host defences and frequently persist until the biomaterial or foreign body is removed, which is standard therapy . Tissue damage caused by surgery and foreign body implantation increases the susceptibility to infections, activates host defences and stimulates the generation of inflammatory mediators including radicals that are further aggravated by bacterial activity and toxins . Nearly one third of implant-related infections can be prevented by strictly following established infection control guidelines . However, a significant number of implant-associated infections remains . The escape of bacteria from host defence and antibiotic therapy makes the development of infection-resistant materials as anti-microbial drug delivery systems feasible . This concept consists of the sustained delivery of antimicrobial drugs into the local microenvironment of implants avoiding systemic side effects exceeding usual systemic concentrations by magnitudes of order. Intensive Care Med, 2004 Aug, 30(8), 1557 - 63 Epub 2004 May 13. Variation of infected cell count in bronchoalveolar lavage and timing of ventilator-associated pneumonia; Dupont H et al.; OBJECTIVES: The aim of this study was to evaluate and compare the accuracy of the percentage of infected cells (%IC) in bronchoalveolar lavage (BAL) for ventilator-associated pneumonia (VAP) diagnosis according to its onset from the initiation of mechanical ventilation . PATIENTS: One hundred and eight patients admitted to a surgical ICU were retrospectively included (1999-2001) . A total of 171 cases of VAP were diagnosed on clinical, biological, chest X-ray and BAL results (threshold >/=10(4 )cfu/ml) . RESULTS: The %IC significantly decreased with the timing of VAP diagnosis: 12.2+/-12.1% for VAP occurring less than 7 days after the initiation of mechanical ventilation, 7.4+/-9.2% for VAP occurring between 7 and 15 days and 4.8+/-6.4% for VAP after 15 days ( p=0.0002), despite the same number of elements and proportion of polymorphonuclear neutrophils in BAL . In addition, a relationship between the %IC and the pathogen responsible for VAP was observed for P . aeruginosa {higher for VAP <7 days than for VAP 7-15 days ( p=0.01) and VAP >15 days ( p=0.006)} and S . aureus {lower for VAP >15 days than VAP 7-15 days ( p=0.04) and VAP <7 days ( p=0.04)} . Furthermore, the %IC in BAL was lower in patients undergoing antimicrobial therapy than in patients without antibiotics ( p=0.04) . Three factors were independently associated with the %IC: quantitative culture of BAL (beta=0.42, p<0.0001), ongoing antimicrobial therapy (beta= -0.21, p=0.003) and onset of VAP (beta= -0.17, p=0.01) . CONCLUSIONS: A relationship between the %IC in BAL, duration of ventilation, quantitative culture of BAL and ongoing antimicrobial therapy has been proved in this study . The %IC for VAP diagnosis may not be accurate in patients with ongoing antibiotics and late onset infections (>7 days). Inform Prim Care, 2004, 12(1), 19 - 26 Changing public attitudes to antibiotic prescribing: can the internet help? Madle G, Kostkova P, Mani-Saada J, Weinberg J, Williams P. INTRODUCTION: Antimicrobial resistance is a global problem with serious implications for modern medicine . Education of the public is essential for reducing patient pressure on GPs and subsequent inappropriate prescribing . Evaluation of educational interventions is necessary to assess their impact on public knowledge and attitudes . The aim of this study was to evaluate the effect of a health information website, part of the National electronic Library of Infection, on user knowledge and attitudes . METHOD: Questionnaires testing user knowledge and attitudes before and after using the website . RESULTS: There were significant improvements in knowledge about the use of antibiotics and antibiotic resistance . Expectations that antibiotics should be prescribed were significantly reduced after using the website . Health professionals showed a significantly greater knowledge about antibiotics and were less likely to expect antibiotics to be prescribed for acute otitis media than non-health professionals before using the website . There was no significant difference between the knowledge of these groups after using the website, but non-health professionals continued to have higher expectations of antibiotics being prescribed than health professionals . CONCLUSIONS: Health information websites can play a significant role in influencing public knowledge and attitudes . Further research is needed to investigate how people learn from these interventions and to determine their long-term impact on public attitudes and subsequent behaviour. Annu Rev Biophys Biomol Struct, 2004, 33, 95 - 118 Structure, molecular mechanisms, and evolutionary relationships in DNA topoisomerases; Corbett KD et al.; Topoisomerases are enzymes that use DNA strand scission, manipulation, and rejoining activities to directly modulate DNA topology . These actions provide a powerful means to effect changes in DNA supercoiling levels, and allow some topoisomerases to both unknot and decatenate chromosomes . Since their initial discovery over three decades ago, researchers have amassed a rich store of information on the cellular roles and regulation of topoisomerases, and have delineated general models for their chemical and physical mechanisms . Topoisomerases are now known to be necessary for the survival of cellular organisms and many viruses and are rich clinical targets for anticancer and antimicrobial treatments . In recent years, crystal structures have been obtained for each of the four types of topoisomerases in a number of distinct conformational and substrate-bound states . In addition, sophisticated biophysical methods have been utilized to study details of topoisomerase reaction dynamics and enzymology . A synthesis of these approaches has provided researchers with new physical insights into how topoisomerases employ chemistry and allostery to direct the large-scale molecular motions needed to pass DNA strands through each other. Drugs, 2004, 64(10), 1091 - 124 QT prolongation with antimicrobial agents: understanding the significance; Owens RC Jr; Cardiac toxicity has been relatively uncommon within the antimicrobial class of drugs, but well described for antiarrhythmic agents and certain antihistamines . Macrolides, pentamidine and certain antimalarials were traditionally known to cause QT-interval prolongation, and now azole antifungals, fluoroquinolones and ketolides can be added to the list . Over time, advances in preclinical testing methods for QT-interval prolongation and a better understanding of its sequelae, most notably torsades de pointes (TdP), have occurred . This, combined with the fact that five drugs have been removed from the market over the last several years, in part because of QT-interval prolongation-related toxicity, has elevated the urgency surrounding early detection and characterisation methods for evaluating non-antiarrhythmic drug classes . With technological advances and accumulating literature regarding QT prolongation, it is currently difficult or overwhelming for the practising clinician to interpret these data for purposes of formulary review or for individual patient treatment decisions . Certain patients are susceptible to the effects of QT-prolonging drugs . For example, co-variates such as gender, age, electrolyte derangements, structural heart disease, end organ impairment and, perhaps most important, genetic predisposition, underlie most if not all cases of TdP . Between and within classes of drugs there are important differences that contribute to delayed repolarisation (e.g . intrinsic potency to inhibit certain cardiac ion currents or channels, and pharmacokinetics) . To this end, a risk stratification scheme may be useful to rank and compare the potential for cardiotoxicity of each drug . It appears that in most published cases of antimicrobial-associated TdP, multiple risk factors are present . Macrolides in general are associated with a greater potential than other antimicrobials for causing TdP from both a pharmacodynamic and pharmacokinetic perspective . The azole antifungal agents also can be viewed as drugs that must be weighed carefully before use since they also have both pharmacodynamic and pharmacokinetic characteristics that may trigger TdP . The fluoroquinolones appear less likely to be associated with TdP from a pharmacokinetic perspective since they do not rely on cytochrome P450 (CYP) metabolism nor do they inhibit CYP enzyme isoforms, with the exception of grepafloxacin and ciprofloxacin . Nonetheless, patient selection must be carefully made for all of these drugs . For clinicians, certain responsibilities are assumed when prescribing antimicrobial therapy: (i) appropriate use to minimise resistance; and (ii) appropriate patient and drug selection to minimise adverse event potential . Incorporating information learned regarding QT interval-related adverse effects into the drug selection process may serve to minimise collateral iatrogenic toxicity. Int J Tuberc Lung Dis, 2004 Feb, 8(2), 253 - 9 A microplate indicator-based method for determining the susceptibility of multidrug-resistant Mycobacterium tuberculosis to antimicrobial agents; Morcillo N et al.; SETTING: Reference Laboratory, Buenos Aires Province Tuberculosis Control Program, Dr Cetrangolo Hospital, Argentina . OBJECTIVE: To obtain a rapid, inexpensive method of determining minimal inhibitory concentrations (MIC) of several drugs acting on multidrug-resistant Mycobacterium tuberculosis (MDR-TB), a colorimetric, microplate-based assay (M-MTT) was developed . DESIGN: One hundred and one clinical isolates were studied . Drugs were placed in a microtiter plate, and several two fold dilutions were made in situ . Kanamycin, capreomycin, ethionamide, para-aminosalicylic acid and clarithromycin were tested in a range concentration of 8.0-0.25 microg/ml, cycloserine 60.0-1.9 microg/ml, clofazimine 3.0-0.10 microg/ml, levofloxacin 4.0-0.13 microg/ml and rifabutin 1.0-0.13 microg/ml . General indicator 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) at 5.0 mg/ml was used for visualising cellular growth and viability . The proportion method with Middlebrook 7H11 was used as the gold standard . RESULTS: MICs by M-MTT were obtained at on average 8 days and correlated with those obtained using the proportion method . In our conditions, the total cost of MIC determination for nine drugs was 5.00 US dollars . CONCLUSION: M-MTT could be used as a simple, rapid, low-cost technology to test the susceptibility of MDR-TB strains to several second-line and alternative drugs, with the objective of orienting future treatment regimens. Cell Tissue Res, 2004 Jun, 316(3), 391 - 401 Epub 2004 May 11. Earthworm leukocyte populations specifically harbor lysosomal enzymes that may respond to bacterial challenge; Engelmann P et al.; Earthworm leukocytes (coelomocytes) are responsible for innate cellular immune functions such as phagocytosis and encapsulation against parasites and pathogens . Microbial killing results from the combined action of the phagocytic process with humoral immune factors such as agglutinins (e.g., lectins), lysosomal enzymes (e.g., acid phosphatase, lysozyme), and various cytotoxic and antimicrobial molecules . There is also evidence of weak adaptive immune responses against foreign transplants . This study focused on aspects of the innate immune response . First, anti-human acid phosphatase (anti-AcP) polyclonal antibody characterized different acid hydrolase patterns in coelomocytes . Second, flow cytometry identified a strongly immunoreactive coelomocyte population . Third, ultrastructural and cytochemical analyses revealed acid phosphatase in discrete granules (lysosomes) of effector hyaline and granular coelomocytes but not in mature chloragocytes . Coelomocytes were exposed to bacteria to assess how phagocytosis influences: (a) the production of acid phosphatase using Western blot, and (b) release of acid phosphatase using ELISA from cell-free coelomic fluid . Fourth, after phagocytosis, acid phosphatase levels differed between controls and experimentals . Fifth, we found a 39-kDa molecule that reacted intensely with anti-AcP . Our results suggest that effector earthworm coelomocytes may not eliminate pathogens only by phagocytosis but also by extracellular lysis. Otolaryngol Head Neck Surg, 2004 May, 130(5 Suppl), S95 - 118 Clinical practice guideline: Otitis media with effusion; Rosenfeld RM et al.; The clinical practice guideline on otitis media with effusion (OME) provides evidence-based recommendations on diagnosing and managing OME in children . This is an update of the 1994 clinical practice guideline "Otitis Media With Effusion in Young Children," which was developed by the Agency for Healthcare Policy and Research (now the Agency for Healthcare Research and Quality) . In contrast to the earlier guideline, which was limited to children aged 1 to 3 years with no craniofacial or neurologic abnormalities or sensory deficits, the updated guideline applies to children aged 2 months through 12 years with or without developmental disabilities or underlying conditions that predispose to OME and its sequelae . The American Academy of Pediatrics, American Academy of Family Physicians, and American Academy of Otolaryngology-Head and Neck Surgery selected a subcommittee composed of experts in the fields of primary care, otolaryngology, infectious diseases, epidemiology, hearing, speech and language, and advanced practice nursing to revise the OME guideline . The subcommittee made a strong recommendation that clinicians use pneumatic otoscopy as the primary diagnostic method and distinguish OME from acute otitis media (AOM) . The subcommittee made recommendations that clinicians should (1) document the laterality, duration of effusion, and presence and severity of associated symptoms at each assessment of the child with OME; (2) distinguish the child with OME who is at risk for speech, language, or learning problems