Bone Marrow Transplant, 2004 Feb, 33(3), 303 - 10 Gram-negative bacteraemia (GNB) after 428 unrelated donor bone marrow transplants (UD-BMT): risk factors, prophylaxis, therapy and outcome; Mitchell AE et al.; Gram-negative infection is an important cause of morbidity and mortality after unrelated donor-bone marrow transplantation (UD-BMT) . We performed a retrospective case-control study to examine the risk factors, prophylaxis, therapy and outcome of Gram-negative bacteraemia (GNB) in 428 patients undergoing UD-BMT . The incidence of GNB was 3.6% in children and 19% in adults . Of the adults, 11% developed GNB >60 days post UD-BMT . Predisposing risk factors for GNB included 'high-risk' disease status, chronic graft-versus-host disease and use of systemic steroids . Fever, a raised C-reactive protein (CRP) and hypotension were common findings at presentation . Patients were routinely given prophylactic ciprofloxacin: resistance to this antibiotic was seen in 33% of isolates . We identified an age-matched control group undergoing UD-BMT over the same time period as the study group . Gram-positive bacteraemia was significantly more common in cases than controls . Mortality from GNB was 17% in children and 24% in adults . We conclude that GNB is a common complication of UD-BMT with a high associated mortality . Patients should be educated further to present rapidly with symptoms suggestive of infection. Antimicrob Agents Chemother, 2003 Dec, 47(12), 3945 - 9 Molecular epidemiology of orf513-bearing class 1 integrons in multiresistant clinical isolates from Argentinean hospitals; Arduino SM et al.; The spread of orf513-bearing class 1 integrons is associated with bla(CTX-M-2) in gram-negative clinical isolates in Argentina, with In35 being the most frequently found integron (74%) . Among 65 isolates without bla(CTX-M-2), only one harbored a novel orf513-bearing class 1 integron with the dfrA3b gene . The finding of orf513 not associated with class 1 integrons in two gram-positive strains indicates the widespread occurrence of this putative site-specific recombinase. J Vet Med A Physiol Pathol Clin Med, 2003 Sep, 50(7), 370 - 4 The cytology of the external ear canal in the normal dog and cat; Tater KC et al.; This study was conducted on 50 normal dogs and 52 normal cats in order to characterize the normal cytological findings in specimens taken from the vertical ear canal by swabbing . Yeast were detected in 96% of the dogs and 83% of the cats . Gram-positive cocci were found in 42% of the dogs and 71% of the cats . Rods were not seen . In dogs, the median numbers per high-power microscopic field (400x magnification) for yeast, cocci and keratinocytes were 0.2, 0 and 3.9, respectively . In cats, the median numbers were 0.2, 0.3 and 8, respectively . Nucleated keratinocytes were occasionally observed in both species, and should not be mistaken for a pathological process (parakeratotic hyperkeratosis). J Mol Biol, 2003 Nov 28, 334(3), 541 - 9 Structure of bacteriocin AS-48: from soluble state to membrane bound state; Sanchez-Barrena MJ et al.; The bacteriocin AS-48 is a membrane-interacting peptide, which displays a broad anti-microbial spectrum against Gram-positive and Gram-negative bacteria . The NMR structure of AS-48 at pH 3 has been solved . The analysis of this structure suggests that the mechanism of AS-48 anti-bacterial activity involves the accumulation of positively charged molecules at the membrane surface leading to a disruption of the membrane potential . Here, we report the high-resolution crystal structure of AS-48 and sedimentation equilibrium experiments showing that this bacteriocin is able to adopt different oligomeric structures according to the physicochemical environment . The analysis of these structures suggests a mechanism for molecular function of AS-48 involving a transition from a water-soluble form to a membrane-bound state upon membrane binding. Transpl Infect Dis, 2003 Sep, 5(3), 132 - 9 Infectious complications following nonmyeloablative allogeneic hematopoietic stem cell transplantation; Busca A et al.; Nonmyeloablative hematopoietic stem cell transplantation (NST) has been explored in hematological malignancies and solid tumors in an attempt to minimize treatment-related toxicity . Whether this approach is associated with reduced risk of infectious complications is unclear . The aim of the current study was to evaluate the infectious complications in a series of 32 consecutive adult patients who received NST at our institution . Peripheral blood stem cell grafts (n=30) or marrow grafts (n=2) were infused from human leukocyte antibody (HLA)-matched sibling (n=30), partially matched related (n=1), or unrelated (n=1) donors . Neutropenia developed in two-thirds of patients and lasted 16 days . Acute graft-versus-host disease (GVHD) grade II to IV was observed in 25% of patients, whereas 35% of patients had signs of extensive chronic GVHD . Twenty-two patients (69%) had at least one significant infectious episode . Bacteremia occurred in 19% of patients (n=5 gram-positive, n=1 gram-negative microorganisms) . Cytomegalovirus (CMV) infection was observed in 10 out of 28 (36%) evaluable patients; 4 of these had recurrent or persistent CMV antigenemia requiring a second-line treatment, but eventually the viremia cleared . No patients experienced CMV disease . Fungal infections were documented in five (16%) patients, comprising invasive fungal infections in two cases and mucosal fungal infections in three . Four patients died of transplant-related causes, and three of these died before day +100 . Infection was considered the primary cause of death in one patient (pulmonary aspergillosis) and contributed to death in another two . The actuarial probability of nonrelapse mortality at 100 days was 10% (95% confidence interval, 3-26%) . Our preliminary results suggest that NST is associated to a low incidence of bacteremia or fungal and viral infections . Whether these findings would translate into an improved overall survival needs to be confirmed in larger prospective studies. Annu Rev Genet, 2003, 37, 409 - 33 Structure, diversity, and evolution of protein toxins from spore-forming entomopathogenic bacteria; de Maagd RA et al.; Gram-positive spore-forming entomopathogenic bacteria can utilize a large variety of protein toxins to help them invade, infect, and finally kill their hosts, through their action on the insect midgut . These toxins belong to a number of homology groups containing a diversity of protein structures and modes of action . In many cases, the toxins consist of unique folds or novel combinations of domains having known protein folds . Some of the toxins display a similar structure and mode of action to certain toxins of mammalian pathogens, suggesting a common evolutionary origin . Most of these toxins are produced in large amounts during sporulation and have the remarkable feature that they are localized in parasporal crystals . Localization of multiple toxin-encoding genes on plasmids together with mobilizable elements enables bacteria to shuffle their armory of toxins . Recombination between toxin genes and sequence divergence has resulted in a wide range of host specificities. Eur J Intern Med, 2003 Nov, 14(7), 411 - 414 The value of the Von Reyn and the Duke diagnostic criteria for infective endocarditis in internal medicine practice . A study of 38 cases; Andres E et al.; BACKGROUND: The aim of our study was to compare the value of the Von Reyn and the Duke diagnostic criteria for infective endocarditis (IE) in internal medicine practice . METHODS: We determined the sensitivity and negative predictive value of these two sets of diagnostic criteria in 38 patients with established infective endocarditis who were followed in two departments of general internal medicine . The patients essentially had subacute endocarditis of the aortic valve (79%) with several systemic manifestations (100%) . Microbial documentation included mainly gram-positive cocci (60%) . RESULTS: With transthoracic echocardiography (TTE), the sensitivities of the Duke and the Von Reyn criteria were 65% and 35%, respectively, while with transesophageal echocardiography (TEE), these sensitivities were 75% and 35%, respectively . With TTE and TEE, the negative predictive values were 100% for the Duke diagnostic criteria versus only 71% for the Von Reyn criteria . CONCLUSIONS: This study confirms that the Duke diagnostic criteria are useful in practice for the management of patients with infective endocarditis followed in internal medicine. Microbiol Immunol, 2003, 47(10), 701 - 7 A silkworm larvae plasma test for detecting peptidoglycan in cerebrospinal fluid is useful for the diagnosis of bacterial meningitis; Inada K et al.; The silkworm larvae plasma (SLP) test has been established based on a cascade reaction triggered by either peptidoglycan or (1, 3)-beta-D-glucan to produce melanin . We applied this test to the diagnosis of bacterial meningitis . Cerebrospinal fluid (CSF) obtained from patients with bacterial meningitis due to gram-positive bacteria, gram-negative bacteria, or fungi, showed positive reactions to the test . In contrast, CSF from patients with viral meningitis or noninfectious illnesses gave negative reactions . Therefore, this test seems to be useful for diagnosis of bacterial and fungal meningitis . When this test was used together with two types of limulus tests, an endotoxin-specific test, and a conventional test, meningitis was further characterized as gram-positive, gram-negative or fungal meningitis . The SLP test requires a computerized instrument for quantitative colorimetric measurement . A qualitative alternative of this test also can be accomplished by visually observing the darkening color . Thus, this method can be applied for simple and rapid diagnosis of meningitis. Prog Nucleic Acid Res Mol Biol, 2003, 75, 113 - 37 DNA-protein interactions during the initiation and termination of plasmid pT181 rolling-circle replication; Khan SA; Initiation of DNA replication requires the generation of a primer at the origin of replication that can be utilized by a DNA polymerase for DNA synthesis . This can be accomplished by several means, including the synthesis of an RNA primer by a DNA primase or RNA polymerase, by nicking of one strand of the DNA to generate a free 3'-OH end that can be used as a primer, and by the utilization of the OH group present in an amino acid such as serine within an initiation protein as a primer . Furthermore, some single-stranded DNA genomes can utilize a snap-back 3'-OH end generated due to self-complementarity as a primer for DNA replication . The different modes of initiation require the generation of highly organized DNA-protein complexes at the origin that trigger the initiation of replication . A large majority of small, multicopy plasmids of Gram-positive bacteria and some of Gram-negative bacteria replicate by a rolling-circle (RC) mechanism (for previous reviews, see Refs.) . More than 200 rolling-circle replicating (RCR) plasmids have so far been identified and, based on sequence homologies in their replication regions, can be grouped into approximately seven families (Refs., and This review will focus on plasmids of the pT181 family that replicate by an RC mechanism . So far, approximately 25 plasmids have been identified as belonging to this family based on the sequence homology in their double-strand origins (dsos) and the genes encoding the initiator (Rep) proteins . This review will highlight our current understanding of the structural features of the origins of replication, and the DNA-protein and protein-protein interactions that result in the generation of a replication-initiation complex that triggers replication . It will discuss the molecular events that result in the precise termination of replication once the leading-strand DNA synthesis has been completed . This review will also discuss the various biochemical activities of the initiator proteins encoded by the plasmids of the pT181 family and the mechanism of inactivation of the Rep activity after supporting one round of leading-strand replication . Finally, the review will outline the mechanism of replication of the lagging strand of the pT181 plasmid as well as the limited information that is available on the role of host proteins in pT181 leading- and lagging-strand replication. Nucleic Acids Res, 2003 Nov 15, 31(22), 6619 - 23 Silencing of Toll pathway components by direct injection of double-stranded RNA into Drosophila adult flies; Goto A et al.; Double-stranded RNA (dsRNA) gene interference is an efficient method to silence gene expression in a sequence-specific manner . Here we show that the direct injection of dsRNA can be used in adult Drosophila flies to disrupt function of endogenous genes in vivo . As a proof of principle, we have used this method to silence components of a major signaling cascade, the Toll pathway, which controls fruit fly resistance to fungal and Gram-positive bacterial infections . We demonstrate that the knockout is efficient only if dsRNA is injected in 4- or more day-old flies and that it lasts for at least 1 week . Furthermore, we report dsRNA-based epistatic gene analysis via injection of a mixture of two dsRNAs and propose that injection of dsRNA represents a powerful method for rapid functional analysis of genes in Drosophila melanogaster adults, particularly of those whose mutations are lethal during development. Appl Environ Microbiol, 2003 Nov, 69(11), 6793 - 800 Microbial community dynamics associated with rhizosphere carbon flow; Butler JL et al.; Root-deposited photosynthate (rhizodeposition) is an important source of readily available carbon (C) for microbes in the vicinity of growing roots . Plant nutrient availability is controlled, to a large extent, by the cycling of this and other organic materials through the soil microbial community . Currently, our understanding of microbial community dynamics associated with rhizodeposition is limited . We used a (13)C pulse-chase labeling procedure to examine the incorporation of rhizodeposition into individual phospholipid fatty acids (PLFAs) in the bulk and rhizosphere soils of greenhouse-grown annual ryegrass (Lolium multiflorum Lam . var . Gulf) . Labeling took place during a growth stage in transition between active root growth and rapid shoot growth on one set of plants (labeling period 1) and 9 days later during the rapid shoot growth stage on another set of plants (labeling period 2) . Temporal differences in microbial community composition were more apparent than spatial differences, with a greater relative abundance of PLFAs from gram-positive organisms (i15:0 and a15:0) in the second labeling period . Although more abundant, gram-positive organisms appeared to be less actively utilizing rhizodeposited C in labeling period 2 than in labeling period 1 . Gram-negative bacteria associated with the 16:1omega5 PLFA were more active in utilizing (13)C-labeled rhizodeposits in the second labeling period than in the first labeling period . In both labeling periods, however, the fungal PLFA 18:2omega6,9 was the most highly labeled . These results demonstrate the effectiveness of using (13)C labeling and PLFA analysis to examine the microbial dynamics associated with rhizosphere C cycling by focusing on the members actively involved. J Biol Chem, 2004 Jan 30, 279(5), 3516 - 24 Epub 2003 Nov 03. Gram-positive DsbE proteins function differently from Gram-negative DsbE homologs . A structure to function analysis of DsbE from Mycobacterium tuberculosis; Goulding CW et al.; Mycobacterium tuberculosis, a Gram-positive bacterium, encodes a secreted Dsb-like protein annotated as Mtb DsbE (Rv2878c, also known as MPT53) . Because Dsb proteins in Escherichia coli and other bacteria seem to catalyze proper folding during protein secretion and because folding of secreted proteins is thought to be coupled to disulfide oxidoreduction, the function of Mtb DsbE may be to ensure that secreted proteins are in their correctly folded states . We have determined the crystal structure of Mtb DsbE to 1.1 A resolution, which reveals a thioredoxin-like domain with a typical CXXC active site . These cysteines are in their reduced state . Biochemical characterization of Mtb DsbE reveals that this disulfide oxidoreductase is an oxidant, unlike Gram-negative bacteria DsbE proteins, which have been shown to be weak reductants . In addition, the pK(a) value of the active site, solvent-exposed cysteine is approximately 2 pH units lower than that of Gram-negative DsbE homologs . Finally, the reduced form of Mtb DsbE is more stable than the oxidized form, and Mtb DsbE is able to oxidatively fold hirudin . Structural and biochemical analysis implies that Mtb DsbE functions differently from Gram-negative DsbE homologs, and we discuss its possible functional role in the bacterium. Am J Ophthalmol, 2003 Nov, 136(5), 791 - 6 Acute endophthalmitis following intravitreal triamcinolone acetonide injection; Moshfeghi DM et al.; PURPOSE: To report the clinical features, causative organisms, management, and visual acuity outcomes of eight eyes of eight patients who developed acute postoperative endophthalmitis following intravitreal injection of triamcinolone acetonide (IVTA) . DESIGN: Retrospective, multicenter, interventional, case series . METHODS: A retrospective, interventional, case series of all patients with acute postoperative endophthalmitis following IVTA at seven academic clinical centers between March 2001 and July 2002 . RESULTS: A total of 922 IVTAs were performed . Eight eyes of eight patients with acute postoperative endophthalmitis were identified in the 6 weeks following IVTA for an incidence of 0.87% (95% confidence interval of 0.38% to 1.70%) . The median time to presentation was 7.5 days (range, 1-15 days) after IVTA . The most common clinical findings were iritis (n = 8), vitritis (n = 8), hypopyon (n = 8), pain (n = 7), red eye (n = 6), and decreased vision (n = 5) . The median presenting visual acuity was 20/1127 (range, 20/60 to light perception) . Initial treatment consisted of vitreous tap and injection of antibiotics (n = 6) or pars plana vitrectomy and injection of intravitreal antibiotics (n = 2) . Intraocular cultures yielded identification in seven patients . One demonstrated intracellular gram-positive cocci in chains with numerous polymorphonuclear cells on gram stain . The median postinfection vision was 20/400 (range, 20/40 to no light perception) . Three patients ended up with no light perception visual acuity, including enucleation (n = 1) and phthisis (n = 1) . CONCLUSIONS: Acute postoperative endophthalmitis following IVTA occurs rapidly and can result in severe loss of vision. New Microbiol, 2003 Oct, 26(4), 329 - 37 Infectious agents in tissues from spontaneous abortions in the first trimester of pregnancy; Penta M et al.; Some evidence suggests that intrauterine infection plays a major role in the pathogenesis of early pregnancy loss, but the implication and prevalence of microrganisms in the aetiology of spontaneous abortion during the first trimester of pregnancy has not yet been well established . In this study, we analysed the tissues relative to the product of conception from abortions during the first trimester (51 spontaneous abortions and 56 voluntary pregnancy interruptions) in women attending the Gynecological Sciences Perinatology and Puericulture Department of "Policlinico Umberto I" . Specimens were investigated by cultural methods for the presence of yeasts, gram positive, gram negative bacteria, and genital mycoplasma . By molecular diagnostic procedures, DNA sequences of Chlamydia trachomatis, herpes simplex viruses, adenovirus, human papillomaviruses and human polyomaviruses BK and JC were searched . None of these agents could be found in voluntary pregnancy interruption samples, with the exception of 3.6% of specimens positive for adenovirus, whereas spontaneous abortion tissues were positive for at least one microrganism by 31.5% . Data analysis showed the occurrence of both monomicrobial and polymicrobial infections. Environ Sci Technol, 2003 Oct 15, 37(20), 4678 - 84 Electron spin resonance study of chromium(V) formation and decomposition by basalt-inhabiting bacteria; Kalabegishvili TL et al.; Bacterial reduction of Cr(VI) to Cr(III) compounds may produce reactive intermediates Cr(V) and Cr(IV), which can affect the mobility and toxicity of chromium in environments . To address this important subject, we conducted an electron spin resonance (ESR) study to understand the kinetics of the formation and decomposition of Cr(V) during Cr(VI) reduction by different gram-positive Cr(VI)-tolerant bacteria, which were isolated from polluted basalts from the United States of America and the Republic of Georgia . Results from our batch experiments show that during Cr(VI) reduction, the macromolecules at the cell wall of these bacteria could act as an electron donor to Cr(VI) to form a stable square-pyramidal Cr(V) complexes, which were reduced further probably via a one-electron transfer pathway to form Cr(IV) and Cr(III) compounds . The Cr(V) peak at the ESR spectrum possessed superhyperfine splitting characteristic of the Cr(V) complexes with diol-containing molecules . It appears that the kinetics of Cr(V) formation and decomposition depended on the bacterial growth phase and on the species . Both formation and decomposition of Cr(V) occurred more quickly when Cr(VI) was added at the exponential phase . In comparison with other gram-positive bacteria from the republic of Georgia, the formation and decomposition of Cr(V) in Arthrobacter species from the Unites States was significantly slower. Appl Microbiol Biotechnol, 2004 Feb, 63(5), 510 - 9 Epub 2003 Oct 31. Lincomycin, cultivation of producing strains and biosynthesis; Spizek J et al.; Lincomycin and its derivatives are antibiotics exhibiting biological activity against Gram-positive bacteria . The semi-synthetic chlorinated lincomycin derivative is used in clinical practice . The chemical structure of lincosamide antibiotics, cultivation of producing strains and analytical procedures used for separation and isolation of these compounds are described in this review . Biosynthesis of lincomycin and related compounds and its genetic control are briefly discussed. Leukemia, 2004 Jan, 18(1), 72 - 7 Infectious complications in pediatric acute myeloid leukemia: analysis of the prospective multi-institutional clinical trial AML-BFM 93; Lehrnbecher T et al.; Infections still remain a major cause of therapy-associated morbidity and mortality in children with acute myeloid leukemia (AML) . To improve supportive care measurements, detailed information on frequency and characteristic features of infectious complications is needed . We retrospectively analyzed the medical charts of 304 children, treated in 30 hospitals according to the multi-institutional clinical trial AML-BFM 93 . Overall, 855 infectious complications occurred in 304 patients (fever without identifiable source (n=523; 61.2%), clinically (n=57; 6.7%) and microbiologically documented infections (n=275; 32.1%)) . Neutropenia was present in 74.1% of the infectious episodes . In all, 20 patients died of infection-associated complications (15/276 (5.4%) patients without and 5/28 (17.9%) with Down syndrome), most of them during early induction therapy (n=11) . Blood stream infections occurred in 228 episodes (Gram-positive (n=202) and Gram-negative (n=42) pathogens) . Invasive fungal infection was probable or proven in 15 patients . In 113 out of the 855 infectious episodes (13.3%), pneumonia was radiologically diagnosed . Better strategies of supportive care might help to improve overall survival in children undergoing chemotherapy for AML . Therefore, children with AML should be treated in specialized pediatric centers, and there should be a very low threshold to readmit patients, in particular patients with pulmonary symptoms. Braz J Med Biol Res, 2003 Nov, 36(11), 1495 - 9 Epub 2003 Oct 22. Thiol-independent activity of a cholesterol-binding enterohemolysin produced by enteropathogenic Escherichia coli; Figueiredo PM et al.; Enterohemolysin produced by Escherichia coli associated with infant diarrhea showed characteristics similar to those of thiol-activated hemolysins produced by Gram-positive bacteria, including inactivation by cholesterol, lytic activity towards eukaryotic cells and thermoinstability . However, enterohemolysin activity was not inactivated by oxidation or by SH group-blocking agents (1 mM HgCl2, 1 mM iodoacetic acid) and the hemolysin (100 microg/ml) was not lethal to mice, in contrast to the lethality of the thiol-activated hemolysin family to animals . Earlier reports showed that intravenous injection of partially purified streptolysin O preparations (0.2 microg) was rapidly lethal to mice . These results suggest that E . coli enterohemolysin is not a thiol-activated hemolysin, despite its ability to bind cholesterol, probably due to the absence of free thiol-group(s) that characterize the active form of the thiol-activated hemolysin molecule. Curr Opin Microbiol, 2003 Oct, 6(5), 519 - 27 Genomic analysis of secretion systems; Pallen MJ et al.; Secretion of proteins into the extracellular environment is important to almost all bacteria, and in particular mediates interactions between pathogenic or symbiotic bacteria with their eukaryotic hosts . The accumulation of bacterial genome sequence data in the past few years has provided great insights into the distribution and function of these secretion systems . Three systems are responsible for secretion of proteins across the bacterial cytoplasmic membrane: Sec, SRP and Tat . Many novel examples of systems for transport across the Gram-negative bacterial cell envelope have been discovered through genome sequencing and surveys, including many novel type III secretion systems and autotransporters . Similarly, genomic data mining has revealed many new potential secretion substrates and identified unsuspected domains in secretion-associated proteins . Interestingly, genomic analyses have also hinted at the existence of a dedicated protein secretion system in Gram-positive bacteria, targeting members of the WXG100/ESAT-6 family of proteins, and have revealed an unexpectedly wide distribution of sortase-driven protein-targeting systems. J Hosp Infect, 2003 Nov, 55(3), 226 - 31 Validity of the four European test strains of prEN 12054 for the determination of comprehensive bactericidal activity of an alcohol-based hand rub; Kampf G et al.; A comprehensive bactericidal activity of an alcohol-based hand rub is essential for prevention of cross-transmission by the hands of healthcare workers . In Europe, however, only four test organisms are used to determine bactericidal activity according to prEN 12054 . The susceptibility of the various bacterial species against the commonly used alcohols is thought to be similar, but so far this has never been studied . We therefore evaluated the bactericidal activity of an alcohol-based hand rub (Sterillium) within 30 s in compliance with prEN 12054 and in a time kill test against 13 Gram-positive, 18 Gram-negative bacteria and 14 antibiotic-resistant bacterial pathogens . Each strain was evaluated in quadruplicate . Counts of the four test bacteria of prEN 12054 were reduced by factors exceeding 10(5) within 30 s . In the time kill test, all 13 Gram-positive and all 18 Gram-negative bacteria were reduced more than 10(5)-fold within 30 s, not only against the ATCC test strains but also against corresponding clinical isolates . Comparable reductions were also observed against all 14 emerging bacterial pathogens . The four European test bacteria were found to be sufficient to determine a comprehensive bactericidal activity of a propanol-based hand rub. Intensive Care Med, 2003 Dec, 29(12), 2157 - 61 Epub 2003 Oct 16. Lipopolysaccharide-binding protein: a possible diagnostic marker for Gram-negative bacteremia in neutropenic cancer patients; Oude Nijhuis CS et al.; OBJECTIVE: Cancer patients with febrile neutropenia after chemotherapy have a variable risk of bacterial infection . Especially Gram-negative bacteremia is associated with high mortality and/or morbidity . Early diagnosis of patients with Gram-negative bacteremia at the onset of febrile neutropenia is potentially useful in tailoring therapy . DESIGN AND SETTING: Prospective study at the Department of Pediatric Oncology and Internal Medicine of a university hospital . PATIENTS: Were analyzed 66 febrile neutropenic episodes in 57 adults and children . Patients were divided into four groups: those with Gram-negative bacteremia, Gram-positive bacteremia, clinical sepsis, or fever of unknown origin . MEASUREMENTS AND RESULTS: Plasma lipopolysaccharide-binding protein (LBP) and C-reactive protein (CRP) concentrations were determined . LBP at the onset of febrile neutropenia was significantly higher in patients with Gram-negative bacteremia than those with fever of unknown origin and those with Gram-positive bacteremia . Using a cutoff value for LBP proved to have much greater sensitivity, specificity, and positive and negative predictive value for Gram-negative bacteremia than the best cutoff value for CRP . CONCLUSIONS: An initial high LBP level might predict Gram-negative bacteremia in cancer patients with febrile neutropenia . These results may have potential clinical impact by allowing therapy to be initiated for these patients at a very early stage. Shock, 2003 Nov, 20(5), 415 - 9 Modulation of the lipopolysaccharide receptor complex (CD14, TLR4, MD-2) and toll-like receptor 2 in systemic inflammatory response syndrome-positive patients with and without infection: relationship to tolerance; Calvano JE et al.; The lipopolysaccharide (LPS) receptor complex consists of two interacting receptors (CD14 and TLR4) and an associated protein (MD-2) . When engaged by LPS, as in gram-negative infection, this complex transduces a signal detected by MyD88 and passed onward by a cascade of the IRAKs, TRAF6, and NIK, resulting in activation of NF-kappaB . A similar cascade, mediated by TLR2, occurs with ligands derived from gram-positive bacteria . In vitro studies of human monocytes have shown that TLR4 mRNA is paradoxically upregulated in response to "tolerizing" doses of LPS . This study evaluated changes in vivo of blood monocyte CD14, TLR4, TLR2, and MD-2 mRNA by reverse transcription followed by real-time polymerase chain reaction in surgical intensive care unit patients and in normal controls . In addition cell-surface receptor expression of TLR2, TLR4, and CD14 was assessed by flow cytometry in patients and normal controls . Inflammation-induced acute tolerance to LPS was evaluated by ex vivo whole blood tumor necrosis factor alpha production and was significantly reduced in patients compared with controls, confirming LPS hyporesponsiveness . Monocyte mRNA and cell-surface receptor expression of TLR4 were increased 2.4-fold (P < 0.05) and 1.7-fold (P <.002), respectively, in patients compared with normal controls . Monocyte TLR2 mRNA, MD-2 mRNA and CD14 and TLR2 cell-surface expression were not significantly changed compared with controls . The present study suggests that the acute inflammatory condition associated with peripheral cellular LPS hyporesponsiveness is neither specific to prior infectious challenge nor can be ascribed to significant alterations in expression of the cell-surface LPS binding complex proteins. Shock, 2003 Nov, 20(5), 402 - 14 Peptidoglycan and lipoteichoic acid in gram-positive bacterial sepsis: receptors, signal transduction, biological effects, and synergism; Wang JE et al.; In sepsis and multiple organ dysfunction syndrome (MODS) caused by gram-negative bacteria, lipopolysaccharide (LPS) initiates the early signaling events leading to the deleterious inflammatory response . However, it has become clear that LPS can not reproduce all of the clinical features of sepsis, which emphasize the roles of other contributing factors . Gram-positive bacteria, which lack LPS, are today responsible for a substantial part of the incidents of sepsis with MODS . The major wall components of gram-positive bacteria, peptidoglycan and lipoteichoic acid, are thought to contribute to the development of sepsis and MODS . In this review, the literature underlying our current understanding of how peptidoglycan and lipoteichoic acid activate inflammatory responses will be presented, with a focus on recent advances in this field. J Vet Diagn Invest, 2003 Sep, 15(5), 473 - 5 Valvular endocarditis associated with Helcococcus ovis infection in a bovine; Post KW et al.; A 12-month-old Angus bull calf with a history of fever and lethargy of several weeks' duration was necropsied . Macroscopic findings included general dehydration, congestion, and edema within the craniodorsal lobes of the lung, multifocal ecchymotic hemorrhages on the dorsal epaxial and gluteal muscles, bloody ingesta within the gastrointestinal tract, and a 4- x 4- x 5-cm irregular plaque located on the right atrioventricular heart valve . Microscopically, there were focally extensive pulmonary alveolar infiltrates of histiocytes and neutrophils, large numbers of necrotic hypereosinophilic hepatocytes located within the centrilobular and midzonal regions of the liver, and, within the plaque from the right atrioventricular valve, a large mass formed by abundant laminated fibrin that contained numerous small multifocal aggregates of gram-positive cocci . This report describes the first apparent isolation of Helcococcus ovis from cattle. Planta Med, 2003 Aug, 69(8), 745 - 9 New eremophilane sesquiterpenes from Cacalia ainsliaeflora; Mao M et al.; One known eremophilanolide, 3beta-angeloyloxy-8beta,10beta-dihydroxy-6beta-methoxyeremophilenolide (1) and seven new eremophiane sesquiterpenes, 3beta-angeloyloxy-6beta-ethoxy-8beta,10beta-dihydroxyeremophilenolide (2), 3beta,6beta-diangeloyloxy-10alpha-hydroxy-8alpha-methoxyeremophilenolide, 6beta-angeloyloxy-8beta,10beta-dihydroxy-3-oxo-eremophilenolide, 6beta,8beta,10beta-trihydroxy-3-oxo-eremophilenolide, 3beta-angeloyloxy-6beta,10beta-dihydroxy-eremophila-7(11),8(9)-dien-8,12-olide (6), 3beta-angeloyloxy-10beta-hydroxy-6beta-methoxy-eremophila-7(11),8(9)-dien-8,12-olide, 3beta-angeloyloxy-8-oxo-eremophila-6.9-dien-12-oic acid ethyl ester, were isolated from the roots of Cacalia ainsliaeflora . Their structures were elucidated by spectroscopic methods . Compounds 1, 2 and 6 were assayed against both Gram-positive and Gram-negative bacteria . No positive activities were observed. Curr Pharm Biotechnol, 2003 Aug, 4(4), 248 - 59 The methylerythritol phosphate pathway and its significance as a novel drug target; Testa CA et al.; Isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) are the precursors for all isoprenoid compounds . Two pathways are found in Nature for their biosynthesis . The mevalonate (MVA) pathway is found in eukaryotes, algae, archae and some gram-positive bacteria . Gram-negative bacteria, plants and some gram-positive bacteria utilize the methyl erythritol phosphate (MEP) pathway . The distribution and the orthogonal nature of the pathways make the MEP pathway an attractive new target for antibiotics and herbicides . The MEP pathway is essential for bacterial viability . Inhibitors to the MEP pathway represent a "dual-use technology" because potential targets include potential biological warfare agents in addition to common human pathogens . The CDC has three categories designated for Biological Diseases/Agents . Three of the six entities designated as the highest priority (Category A) are organisms that utilize, or appear to utilize, the MEP pathway . Among the 12 second highest priority agents (Category B) listed, 8 are organisms that appear to utilize the MEP pathway . Common human pathogens that can be targeted include the organisms responsible for peptic ulcers, tuberculosis, malaria, food safety threats, and sexually transmitted diseases . There is so far only one inhibitor reported that specifically blocks the MEP pathway and is being investigated clinically . This compound, fosmidomycin, has been shown to be somewhat effective in treating Plasmodium falciparum, the parasite responsible for malaria . We foresee that new MEP pathway inhibitors will open up an entirely new class of antibiotics . An MEP pathway intermediate has also been shown to be the most potent gammadelta T cell activator. Syst Appl Microbiol, 2003 Sep, 26(3), 357 - 66 Planococcus rifietensis sp . nov, isolated from algal mat collected from a sulfurous spring in Campania (Italy); Romano I et al.; The taxomony of strain M8, isolated from algal mat formed at the origin of a sulfurous spring in Rifieto (Savignano Irpino, Campania, Italy), was investigated in a polyphasic approach . The morphological, physiological and genetic characteristics were compared with of Planococcus and Planomicrobium species . The isolate grew optimally at pH 9.0, 1.8 M NaCl at 37 degrees C . The cells were Gram-positive cocci that form pairs, tetrads and aggregates of several cells . The isolate was aerobic/microaerophilic and accumulated glycine-betaine, as a major osmolyte, with minor components glutamate and an unknown compound . M8 was able to hydrolyse X-Glc (5-bromo-4-chloro-3-indoyl beta-d-glucopyranoside) . The polar lipid profile consisted of phosphatidylglycerol and diphosphatidylglycerol as major components, and phosphocholine as a minor compound . MK8 was the only quinone found and the fatty acid composition was dominated by branched acids, mainly aiC15:0 . The G+C content of DNA was 47.9% and its phylogenetic position was established by 16S rRNA gene sequencing as a member of the genus Planococcus . The DNA/DNA similarity of M8 to the type species Planococcus citreus was less than 55% . For this reason and for physiological and chemotaxonomic features, it is proposed to create a new species Planococcus rifietensis sp . nov. Pediatr Infect Dis J, 2003 Sep, 22(9 Suppl), S193 - 200 Safety and tolerability of linezolid in children; Saiman L et al.; BACKGROUND: Linezolid, an oxazolidinone, is effective in the treatment of adults and children with community-acquired and nosocomial pneumonia and uncomplicated and complicated skin and skin structure infections (SSSIs), including infections caused by Gram-positive resistant pathogens . Because of the increasing use of linezolid, it is important to review the common adverse events (AEs) associated with its use in children with the use of data from clinical trials . OBJECTIVE: The safety and tolerability of linezolid in pediatric patients with Gram-positive infections were determined in four pediatric clinical studies . Study I included pediatric patients with community-acquired pneumonia; Study II included otitis media; Study III included SSSIs; and Study IV included complicated SSSIs, nosocomial pneumonia and bacteremia . METHODS: Studies I and II had no comparator arm . Study III was randomized and compared linezolid with cefadroxil . Study IV also was randomized and compared linezolid with vancomycin . Patients <12 years of age received linezolid 10 mg/kg; patients age 12 years and older received 600 mg (intravenous/oral) . Dosing frequency (two to three times daily) varied depending on age and clinical diagnosis . The primary safety endpoints were AEs, drug-related AEs, serious AEs and selected laboratory tests . RESULTS: In the 4 studies 958 patients were included in the intent-to-treat analysis . In the linezolid vs . cefadroxil study (Study III), the most common AEs in patients treated with linezolid were diarrhea (7.8%), headache (6.5%) and upper respiratory tract infection (3.7%) . In the linezolid vs . vancomycin study (Study IV), the most common AEs in the linezolid group were fever (14.1%), diarrhea (10.8%) and vomiting (9.4%) . The most common drug-related AEs for linezolid in all 4 studies were diarrhea, vomiting, loose stools and nausea . None of these common AEs or drug-related AEs occurred more frequently in patients treated with linezolid than in those in the comparator group . CONCLUSIONS: Linezolid was safe and well-tolerated in pediatric patients with community-acquired pneumonia, otitis media, SSSIs and infections caused by Gram-positive resistant pathogens. Pediatr Infect Dis J, 2003 Sep, 22(9 Suppl), S186 - 92 Hematologic effects of linezolid in young children; Meissner HC et al.; BACKGROUND: Linezolid is an effective and well-tolerated antibiotic for the treatment of Gram-positive infections, including hospital and community-acquired pneumonia and complicated and uncomplicated skin and skin structure infections . In adults linezolid treatment for >/=2 weeks has been associated with reversible hematopoietic suppression, primarily thrombocytopenia . OBJECTIVE: To evaluate the occurrence of hematologic effects in children with Gram-positive infections in an open label study of linezolid vs . vancomycin . METHODS: Detailed analyses of hematologic data, including reported hematologic adverse events, complete blood counts, reticulocyte index (RI) and iron studies (serum iron and transferrin saturation), were conducted in both groups at baseline and during and after treatment with the use of an intent-to-treat analysis . RESULTS: Three hundred sixteen patients (median age, 1.65 yr) randomized 2:1 to linezolid (n = 215) or vancomycin (n = 101) were treated . Total treatment durations were similar in the vancomycin group (12.2 +/- 6.4 days; median, 11.0 days) and the linezolid group (11.3 +/- 5.0 days; median, 11.0 days) (P = 0.20) . No significant differences were noted in drug-related hematologic events, such as thrombocytopenia (linezolid, 1.9% vs . vancomycin, 0%; P = 0.170), anemia (linezolid, 1.4% vs . vancomycin, 1.0%; P = 0.771) or neutropenia (linezolid, 0% vs . vancomycin, 0%) . Hemoglobin values also were similar between treatment groups when assessed by shifts from baseline to lowest recorded value . Frequency of occurrence of any substantially abnormal value for hemoglobin (15.7% vs . 12.4%), platelets (12.9% vs . 13.4%) and neutrophils (5.9% vs . 4.3%) were similar in the linezolid and vancomycin groups . No clinically relevant changes in RI or iron studies were noted between treatment groups, and parallel increases in RI occurred with both linezolid and vancomycin . CONCLUSIONS: No significant differences in hematologic profiles between linezolid and vancomycin occurred in this pediatric population. Pediatr Infect Dis J, 2003 Sep, 22(9 Suppl), S172 - 7 Linezolid for the treatment of complicated skin and skin structure infections in children; Yogev R et al.; BACKGROUND: Gram-positive pathogens are a major cause of complicated skin and skin structure infections (CSSSIs) in children . Many pathogens are developing decreased susceptibility to currently used antibiotics, increasing the need for new therapies . Linezolid is well-tolerated and effective in the treatment of these infections in adults . OBJECTIVE: To evaluate the clinical efficacy and safety of iv/oral linezolid and iv vancomycin in children with Gram-positive CSSSIs . METHODS: Hospitalized children <12 years of age were randomized (2:1 ratio) to receive either linezolid 10 mg/kg iv every 8 h (with the option to change treatment to oral linezolid suspension 10 mg/kg every 8 h) or iv vancomycin 10 to 15 mg/kg every 6 to 24 h (according to age) . Clinical response, tolerance and safety were evaluated at follow-up . The results of a subset analysis of patients with CSSSIs are presented here . RESULTS: One hundred twenty intent-to-treat patients (linezolid 80, vancomycin 40) with CSSSI were included in this analysis . Clinical cure rates for clinically evaluable patients with CSSSI did not differ between treatment groups (linezolid, 93.2% vs . vancomycin, 90.0%; P = 0.594) . Significantly fewer linezolid-treated patients experienced drug-related adverse events than did vancomycin-treated patients (23% vs . 48%; P = 0.006) . The percentages of patients with laboratory abnormalities, including selected hematologic parameters, were generally low and similar between the treatment groups . CONCLUSIONS: Linezolid given iv or orally was well-tolerated and safe . It was as effective as vancomycin in treating children with Gram-positive CSSSIs. Pediatr Infect Dis J, 2003 Sep, 22(9 Suppl), S164 - 71 Linezolid for the treatment of children with bacteremia or nosocomial pneumonia caused by resistant gram-positive bacterial pathogens; Jantausch BA et al.; BACKGROUND: Nosocomial infections, particularly hospital-acquired pneumonia (HAP) and bacteremia, are an increasing concern in pediatric hospitals and pediatric intensive care units . Gram-positive pathogens are a leading cause of these infections in children . Linezolid is well-tolerated and as effective as vancomycin in the treatment of these infections in adults . OBJECTIVE: To evaluate the clinical effectiveness and safety of iv/oral linezolid and iv vancomycin in children with resistant Gram-positive HAP or bacteremia . METHODS: Hospitalized children <12 years of age were randomized 2:1 to linezolid or vancomycin . Patients received linezolid 10 mg/kg iv every 8 h with the option to change treatment to oral linezolid suspension 10 mg/kg every 8 h or iv vancomycin 10 to 15 mg/kg every 6 to 24 h . Clinical response was evaluated at follow-up . Results from an analysis of patients with HAP or bacteremia are presented . RESULTS: Thirty-nine patients (linezolid, 23; vancomycin, 16) with HAP and 113 patients with bacteremia (linezolid, 81; vancomycin, 32) were included in the intent-to-treat group . Clinical cure rates for clinically evaluable patients with HAP did not differ between treatment groups (linezolid, 90.0% and vancomycin, 100%; P = 0.305) . No significant difference was seen in clinical cure rates in the clinically evaluable population between the linezolid and vancomycin groups for patients with catheter-related bacteremia (84.8 and 80.0%, respectively; P = 0.716) or patients with bacteremia of unknown source (79.2 and 69.2%, respectively; P = 0.501) . In this subset fewer linezolid-treated patients had drug-related adverse events than did vancomycin-treated patients (19.4% vs . 28.3%; P = 0.230) . Similar percentages of patients with laboratory abnormalities, including selected hematologic parameters, were seen in both treatment groups . CONCLUSIONS: Intravenous/oral linezolid was well-tolerated and as effective as vancomycin in treating children with resistant Gram-positive HAP or bacteremia. Pediatr Infect Dis J, 2003 Sep, 22(9 Suppl), S153 - 7 Linezolid pharmacokinetics in pediatric patients: an overview; Jungbluth GL et al.; BACKGROUND: There are a number of physiologic and developmental differences between children and adults that can influence the absorption, distribution, metabolism and elimination of a drug . Therefore it is important to determine the specific pharmacokinetic characteristics for individual drugs in pediatric patients so that appropriate age-specific dosage regimens can be developed and evaluated in clinical trials . This review summarizes the pharmacokinetic parameters of linezolid in pediatric patients and the rationale for the approved dosing recommendations for this population . METHODS: The pharmacokinetics of linezolid in pediatric patients has been evaluated in 4 clinical trials, including >180 patients ranging in age from preterm newborn infants up to 18 years of age . In all of these studies, patients received a single intravenous dose of linezolid . Plasma linezolid concentrations have been determined by validated high performance liquid chromatography (adult studies) or liquid chromatography/mass spectrometry/mass spectrometry (pediatric studies) methods . RESULTS: The pharmacokinetics of linezolid, especially elimination clearance, is age-dependent . Children younger than 12 years of age have a smaller area under the drug concentration-time curve, a faster clearance and a shorter elimination half-life than adults . Although clearance rates in newborn infants are similar to those in adults, clearance increases rapidly during the first week of life, becoming 2- to 3-fold higher than in adults by the seventh day of life . The clearance of linezolid decreases gradually among young children, becoming similar to adult values by adolescence . The pharmacokinetics of linezolid in children age 12 years and older is not significantly different from that of adults . CONCLUSIONS: Because of the higher clearance and lower area under the drug concentration-time curve, a shorter dosing interval for linezolid is required for children younger than 12 years of age to produce adequate drug exposure against target Gram-positive pathogens. J Long Term Eff Med Implants, 2003, 13(3), 139 - 54 Management and prevention of tetanus; Edlich RF et al.; The World Health Organization was committed to eliminating neonatal tetanus by 1995 . Three years after this date, the infection killed over 400,000 babies a year, even though a safe, effective vaccine had been available for most of this century . The frequency of tetanus in the developing world epitomizes the healthcare disparity between the developed and the developing world . Consequently, the priority of the medical profession must be prevention, with the development of simpler immunization schedules with longer protection . Consequently, the purpose of this collective review is to provide an overview to the management of tetanus as well as to review the immunization strategy that will prevent this potentially deadly illness . Tetanus is caused by Cloistridium tetani, which is an obligate anaerobic, gram-positive rod that is motile and readily forms endospores . Although C . tetani is located everywhere, the disease is encountered largely in underdeveloped, overcrowded, and economically disadvantaged countries . C . tetani is widespread in the feces of domestic animals and humans, while spores of C . tetani are abundant in soil and in the environment surrounding the habitation of humans and animals . Tetanus usually follows deep penetrating wounds where anaerobic bacterial growth is facilitated . Three basic forms of tetanus may be distinguished: local, cephalic, and generalized . At least 80% of the cases are the generalized form . In the adult patient, the most characteristic sign of generalized tetanus is lockjaw, or trismus . The diagnosis of tetanus is most frequently made on clinical manifestations, rather than on bacteriologic findings . The three objectives of management of tetanus are: (1) to provide supportive care until the tetanospasmin that is fixed in tissue has been metabolized; (2) to neutralize circulating toxin; and (3) to remove the source of tetanospasmin . Because there is essentially no immunity to tetanus toxoid, the only effective way to control tetanus is by prophylactic immunization. Presse Med, 2003 Sep 6, 32(28), 1317 - 8 {Diagnosis by blood smear of a fulminant pneumococcemia complicating a myeloma}; Avenas S et al.; INTRODUCTION: During a pneumococcal septicemia, the direct revelation of bacteria on a blood smear is rare and has been mainly described in splenectomised patients . OBSERVATION: A 62 year-old non-splenectomised man suffering from myeloma died, a few hours after his admission to emergency department, from a fulminant pneumococcemia, the diagnosis of which had been rapidly evoked on the examination of a blood smear showing numerous Gram positive diplococci . COMMENTS: This observation highlights the potential severity of pneumococcal infections in patients suffering from altered humoral immunity and the diagnostic interest, in this case, of a systematic routine blood smear permitting rapid initiation of appropriate treatment. Eur J Cardiothorac Surg, 2003 Oct, 24(4), 601 - 7 Bacterial colonization of the donor lower airways is a predictor of poor outcome in lung transplantation; Avlonitis VS et al.; OBJECTIVE: At the time of lung transplant, we routinely perform bronchoalveolar lavage (BAL) of the donor lungs on the recipient operating table immediately before implantation, for bacterial and fungal cultures . We sought to determine whether the results correlate with the outcome . METHODS: We retrospectively analysed 115 consecutive cadaveric lung transplants (single lung: 42; bilateral lung: 63; heart-lung: 10) performed over 4 years . RESULTS: Fifty-three (46%) grafts had positive BAL (bacteria: 33; fungus: 10; mixed: 10) and 62 (54%) were negative . Recipients with donor BAL culture positive for bacteria had lower mean oxygenation index in the first 6 h compared with those with negative bacterial culture (36.5+/-14.73 vs . 44.1+/-16.79 kPa) (P=0.019) . They also had longer median intensive treatment unit stay (2.5 vs . 1.5 days) (P=0.035), and median time of mechanical ventilation (37.5 vs . 23.0 h) (P=0.008), as well as inferior 6-month, 1-year, 2-year and 4-year cumulative survival (79, 77, 74, 60% vs . 93, 92, 88, 79% respectively) (P=0.04) . There was no difference in the above parameters between recipients with Gram-negative (n=18) and recipients with Gram-positive bacteria (n=19) in the donor BAL . Incidence of acute rejection within the first 2 weeks and time of onset of bronchiolitis obliterans syndrome (BOS) were similar in the bacteria-positive and bacteria-negative groups . Recipients with donor BAL positive for fungi alone had similar outcome with the negatives . There was no difference in the donor oxygenation index and age, recipient age, transplant type and ischaemic time between compared groups . There was a significant difference in the median length of donor mechanical ventilation between donors with Gram-positive and donors with Gram-negative bacteria in the BAL (24 vs . 48 h) (P=0.01), as well as between donors with fungi alone in the BAL and donors with negative BAL (67 vs . 48 h) (P=0.04) . CONCLUSIONS: Donor lungs with lower airways colonized with bacteria result in inferior recipient outcome . Bacterial colonization of the donor lower airways could therefore be used as a marker of donor lung injury, but evidence from a prospective study is necessary. Int J Syst Evol Microbiol, 2003 Sep, 53(Pt 5), 1609 - 14 Flow cytometric sorting, phylogenetic analysis and in situ detection of Oscillospira guillermondii, a large, morphologically conspicuous but uncultured ruminal bacterium; Yanagita K et al.; Flow cytometric sorting based on its large cell size enabled an enriched fraction of Oscillospira guillermondii cells to be obtained from the rumen contents of a sheep . Phylogenetic analysis based on cloned 16S rDNA sequences indicated that the bacterium is a member of the low-G+C Gram-positive bacterial cluster . Sporobacter termitidis and Papillibacter cinnamivorans were the most closely related known species, with sequence similarities of only 86.3-88.1 % . Fluorescently labelled 16S rRNA-targeted oligonucleotide probes specific for Oscillospira were designed and applied to the rumen sample from which the enriched fraction was obtained . The probes hybridized specifically with the large, morphologically conspicuous Oscillospira cells. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2003 Sep, 96(3), 283 - 7 Fungal load and candidiasis in Sjögren's syndrome; Radfar L et al.; OBJECTIVE: We sought to investigate the prevalence of Candida carriage and the relationships between salivary flow rates and oral Candida load in patients with Sjogren's syndrome (SS) . METHODS: The oral Candida load of patients with SS was evaluated by culturing oral rinse (swish and spit) samples . Culture, Gram stain, and wet-mount test results were reported . RESULTS: One hundred three patients (96 women) met European criteria for SS (91 with primary SS and 12 with secondary SS) . The mean age (95% confidence interval) was 55 years (range, 51-57 years) . Oral rinse cultures were positive in 77% of subjects . The total stimulated salivary flow rate was inversely correlated with oral Candida load (r = -0.47; P </=.0001) . The oral rinse samples yielded gram-positive results in 38% of patients with SS, and the Fungi-Fluor assay (wet mount) results were positive in 49% . CONCLUSIONS: The prevalence of Candida carriage varies according to the methods used to determine the presence of the organism and is similar to that reported in the literature . A low stimulated salivary flow rate-not a low unstimulated flow rate-was associated with Candida carriage. Zhongguo Wei Zhong Bing Ji Jiu Yi Xue, 2003 Sep, 15(9), 523 - 5 {Organism distribution and drug resistance in 7 cases of severe acute respiratory syndrome death patients with secondary bacteria infection}; Wang JB et al.; OBJECTIVE: To study the organism distribution and drug resistance of seven cases of severe acute respiratory syndrome (SARS) death patients with secondary bacterial infection . METHODS: Thirty strains of bacteria from seven cases of SARS patients with secondary bacterial infection were classified and drug resistance was analyzed . RESULTS: Seven cases of SARS death patients were all infected secondly and 5 cases were polyinfection . Twenty-four (31.5 percent) of 76 examed samples were positive . There were 30 strains of bacteria isolated from seven cases of SARS death patients with secondary bacterial infection . There were 9 strains of Gram negative bacteria (GNB), 8 strains of Gram positive cocci (GPC), and 13 strains of Fungi . The sensitive rate of vancomycin to GPC was 100.0 percent . The sensitive rate of imipenem, piperacillin/tazobactan was 100.0 percent, 44.5 percent, respectively . The sensitive rate of fluconazole to fungi was 92.4 percent . CONCLUSION: SARS patients are consitive to be infected secondary bacteria . Secondary bacteria infection is one of important reason of death. Diagn Microbiol Infect Dis, 2003 Sep, 47(1), 341 - 4 Lymphocutaneous nocardiosis due to Nocardia brasiliensis; Maraki S et al.; Nocardia species are Gram-positive bacteria responsible for systemic or cutaneous infections in humans . Nocardia brasiliensis is the most common infective agent in the cutaneous form of nocardiosis . We describe a case of a previously healthy man, who presented with lymphocutaneous Nocardia brasiliensis infection, and was successfully treated with trimethoprim-sulfamethoxazole . The identification of the isolate was confirmed by nucleotide sequence analysis of the 16S rRNA gene. Mol Gen Mikrobiol Virusol, 2003, (3), 3 - 8 {The pleiotropic function of the phosphoenolpyruvate-dependent phosphotransferase system in bacteria . Communication II}; Gershanovich VN; The structure and function of regulators and anti-terminators is under discussion in gram-positive bacteria . The regulators of lichen and levan operons (LiR and LevR) as well as the implementation of both gram-positive and negative regulations of operons by them are in the focus of attention . Po-independent termination is regarded by the example of the regulatory activity for the utilization systems of glucose (GlcT) beta-glucosides (LicT), sucrose (low-efficiency system SacY-SacX) and of glycerin (GlcP) . Changes in the functional activity of the above systems, which are dependent on a condition of anti-terminators (phosphorylated or dephosphorylated forms and an ability to demirelize etc.) are regarded from the viewpoint of a possibility of occurrence of catabolic repression. Braz Dent J, 2003, 14(2), 95 - 8 Tissue reactions to a component of root canal system bacteria: lipoteichoic acid; Costa ED et al.; Lipoteichoic acid (LTA), present in Gram-positive microorganisms, has physiochemical characteristics that allow it to act as an immunogen . Due to polymicrobial characteristics of root canal infections, LTA can participate in the development of periapical disease . The reaction of the rat subcutaneous tissue to Teflon tube implants, filled with Fibrinol soaked in lipoteichoic acid (concentration of 150 microg/ml), was observed . Lipoteichoic acid provoked an inflammatory tissue reaction. J Chemother, 2003 Aug, 15(4), 323 - 8 Treatment of gram-positive surgical sepsis: role of the oxazolidinones; Colizza S et al.; Surgical sepsis is still too frequent (the thirteenth cause of death in the U.S.) with an estimated cost of 5-10 billion dollars . Since the early 1990s antibiotic resistance has become a serious public health problem, with a relevant increase in nosocomial gram-positive infections . The oxazolidinones, a new class of antibiotics acting on bacterial synthesis at a very early stage, were first commercialized in 1987, and linezolid was the first antibiotic in this class registered for clinical use . This new molecule has been shown to be effective in a variety of aerobic and anaerobic infections (both nosocomial and community-acquired), especially those due to gram-positive agents . Linezolid has complete bioavailability in parenteral and oral administration, is well tolerated and shows little toxicity, thus favoring a shortened hospital stay, improving the patient's quality of life and reducing social costs . Oxazolidinones may be considered the first choice in the treatment of resistant gram-positive infections. Curr Gastroenterol Rep, 2003 Oct, 5(5), 379 - 85 Whipple's disease; Fenollar F et al.; Whipple's disease is an infectious disease caused by a gram-positive bacterium, Tropheryma whipplei . The first case was reported in 1907 by GH Whipple . Its classic symptoms are diarrhea and arthralgias, but symptoms can be various . Cardiac or central nervous system involvement, not always associated with digestive symptoms, may also be observed . For a long time, diagnosis has been based on duodenal biopsy, which is positive using periodic acid-Schiff staining . However, for patients without digestive symptoms, results can be negative, leading to a delay in diagnosis . For 10 years, a tool based on polymerase chain reaction targeting the 16S rDNA sequence has been used . In vitro culture of the bacterium, achieved 3 years ago, has allowed new perspectives for diagnosis and treatment . The natural evolution of the disease without treatment is always fatal . Current treatment is based on administration of trimethoprim-sulfamethoxazole for at least 1 year. J Clin Microbiol, 2003 Sep, 41(9), 4134 - 40 Ribosomal DNA sequencing for identification of aerobic gram-positive rods in the clinical laboratory (an 18-month evaluation); Bosshard PP et al.; We have evaluated over a period of 18 months the use of 16S ribosomal DNA (rDNA) sequence analysis as a means of identifying aerobic gram-positive rods in the clinical laboratory . Two collections of strains were studied: (i) 37 clinical strains of gram-positive rods well identified by phenotypic tests, and (ii) 136 clinical isolates difficult to identify by standard microbiological investigations, i.e., identification at the species level was impossible . Results of molecular analyses were compared with those of conventional phenotypic identification procedures . Good overall agreement between phenotypic and molecular identification procedures was found for the collection of 37 clinical strains well identified by conventional means . For the 136 clinical strains which were difficult to identify by standard microbiological investigations, phenotypic characterization identified 71 of 136 (52.2%) isolates at the genus level; 65 of 136 (47.8%) isolates could not be discriminated at any taxonomic level . In comparison, 16S rDNA sequencing identified 89 of 136 (65.4%) isolates at the species level, 43 of 136 (31.6%) isolates at the genus level, and 4 of 136 (2.9%) isolates at the family level . We conclude that (i) rDNA sequencing is an effective means for the identification of aerobic gram-positive rods which are difficult to identify by conventional techniques, and (ii) molecular identification procedures are not required for isolates well identified by phenotypic investigations. FASEB J, 2003 Sep, 17(12), 1727 - 9 Epub 2003 Jul 03. Airway epithelia regulate expression of human beta-defensin 2 through Toll-like receptor 2; Wang X et al.; The goal of this study is to investigate whether TLR2 mediates hBD2 induction through NF-kappaB in response to bacterial components in the human airway epithelia . We showed that hTLR2 is expressed in the airway epithelial cells by immunohistochemical staining and RT-PCR . The biology of hTLR2 in this context was studied initially in 293 cells transfected with a plasmid expressing hTLR2 together with an hBD2 promoter-driven luciferase reporter (hBD2-promoter-LUC) . Upon incubation with lipoteichoic acid (LTA), the major cell wall component of gram-positive bacteria, luciferase activity was greatly increased compared with mock stimulation . Analysis of mutation constructs of the hBD2 promoter revealed that NF-kappaB sites are important for hTLR2-mediated hBD2 up-regulation upon LTA stimulation . When hBD2-promoter-LUC was transfected into primary human airway epithelia cells (EC), the luciferase activity was greatly increased upon LTA stimulation compared with mock stimulation . The hBD2 promoter mutation constructs were also tested in EC, which confirmed the studies in 293 cells . When a plasmid expressing a dominant-negative mutant of hTLR2 was co-transfected with hBD2-promoter-LUC into EC, LTA could not stimulate hBD2 expression . These data provide convincing evidence that up-regulation of hBD2 can be induced through hTLR2-mediated NFkappaB/IkappaB pathway in the human airway epithelial cells. Biochem J, 2003 Dec 1, 376(Pt 2), 433 - 40 Structural and biochemical characterization of a new type of lectin isolated from carp eggs; Galliano M et al.; A previously unidentified glycoprotein present in the eggs of the carp ( Cyprinus carpio ) was isolated and structurally characterized . The protein binds to a Sepharose 4B matrix and can be eluted with 0.4 M N -acetylglucosamine . The protein has an apparent molecular mass of 26686.3 Da . On the basis of gel-filtration chromatography, the protein appears to be present in solution as a monomer . The sequence of its 238 amino acids, the position of its four disulphide bridges and the composition of its single N-linked carbohydrate chain were determined . The lectin shows a very low agglutinating activity for human A-type erythrocytes and interacts with both Gram-positive and -negative bacteria . These latter interactions are inhibited by N -acetylglucosamine . A database search shows that its amino acid sequence is similar to that of the members of an invertebrate lectin family that includes tachylectin-1 . Tachylectin-1 is present in the amoebocytes of the horseshoe crab, Tachypleus tridentatus, and plays a role in the innate defence system of this species . Homologous genes are also present in other fish, having 85% identity with a gene expressed in the oocytes of the crucian carp ( Carassius auratus gibelio ) and 78% identity with a gene in the cDNA library of the zebrafish ( Danio rerio ). Virology, 2003 Sep 1, 313(2), 401 - 14 Comparative analysis of bacterial viruses Bam35, infecting a gram-positive host, and PRD1, infecting gram-negative hosts, demonstrates a viral lineage; Ravantti JJ et al.; Extra- and intracellular viruses in the biosphere outnumber their cellular hosts by at least one order of magnitude . How is this enormous domain of viruses organized? Sampling of the virosphere has been scarce and focused on viruses infecting humans, cultivated plants, and animals as well as those infecting well-studied bacteria . It has been relatively easy to cluster closely related viruses based on their genome sequences . However, it has been impossible to establish long-range evolutionary relationships as sequence homology diminishes . Recent advances in the evaluation of virus architecture by high-resolution structural analysis and elucidation of viral functions have allowed new opportunities for establishment of possible long-range phylogenic relationships-virus lineages . Here, we use a genomic approach to investigate a proposed virus lineage formed by bacteriophage PRD1, infecting gram-negative bacteria, and human adenovirus . The new member of this proposed lineage, bacteriophage Bam35, is morphologically indistinguishable from PRD1 . It infects gram-positive hosts that evolutionarily separated from gram-negative bacteria more than one billion years ago . For example, it can be inferred from structural analysis of the coat protein sequence that the fold is very similar to that of PRD1 . This and other observations made here support the idea that a common early ancestor for Bam35, PRD1, and adenoviruses existed. Biochem J, 2003 Nov 15, 376(Pt 1), 15 - 33 Polyester synthases: natural catalysts for plastics; Rehm BH; Polyhydroxyalkanoates (PHAs) are biopolyesters composed of hydroxy fatty acids, which represent a complex class of storage polyesters . They are synthesized by a wide range of different Gram-positive and Gram-negative bacteria, as well as by some Archaea, and are deposited as insoluble cytoplasmic inclusions . Polyester synthases are the key enzymes of polyester biosynthesis and catalyse the conversion of (R)-hydroxyacyl-CoA thioesters to polyesters with the concomitant release of CoA . These soluble enzymes turn into amphipathic enzymes upon covalent catalysis of polyester-chain formation . A self-assembly process is initiated resulting in the formation of insoluble cytoplasmic inclusions with a phospholipid monolayer and covalently attached polyester synthases at the surface . Surface-attached polyester synthases show a marked increase in enzyme activity . These polyester synthases have only recently been biochemically characterized . An overview of these recent findings is provided . At present, 59 polyester synthase structural genes from 45 different bacteria have been cloned and the nucleotide sequences have been obtained . The multiple alignment of the primary structures of these polyester synthases show an overall identity of 8-96% with only eight strictly conserved amino acid residues . Polyester synthases can been assigned to four classes based on their substrate specificity and subunit composition . The current knowledge on the organization of the polyester synthase genes, and other genes encoding proteins related to PHA metabolism, is compiled . In addition, the primary structures of the 59 PHA synthases are aligned and analysed with respect to highly conserved amino acids, and biochemical features of polyester synthases are described . The proposed catalytic mechanism based on similarities to alpha/beta-hydrolases and mutational analysis is discussed . Different threading algorithms suggest that polyester synthases belong to the alpha/beta-hydrolase superfamily, with a conserved cysteine residue as catalytic nucleophile . This review provides a survey of the known biochemical features of these unique enzymes and their proposed catalytic mechanism. Biochem Biophys Res Commun, 2003 Sep 19, 309(2), 449 - 56 Arisostatins A induces apoptosis through the activation of caspase-3 and reactive oxygen species generation in AMC-HN-4 cells; Kim YH et al.; A microbial secondary metabolite, arisostatins A (As-A), was originally discovered as a substance carrying the antibiotic activity against Gram-positive bacteria and shown to possess potent anti-tumor properties . The mechanism by which arisostatins A initiates apoptosis remains poorly understood . In the present report we investigated the effect of arisostatins A on activation of the apoptotic pathway in HN-4 cells . Arisostatins A was shown to be responsible for the inhibition of HN-4 cell growth by inducing apoptosis . Treatment with 4 microM arisostatins A for 24h produced morphological features of apoptosis and DNA fragmentation in HN-4 cells . Arisostatins A caused dose-dependent apoptosis and DNA fragmentation of HN-4 cells used as a model . Treatment with caspase inhibitor significantly reduced the arisostatins A-induced caspase 3 activation . In addition, arisostatins A-induced apoptosis was associated with the generation of reactive oxygen species (ROS), which was prevented by an antioxidant NAC (N-acetyl-cysteine) . These data indicate that cytotoxic effect of arisostatins A on HN-4 cells is attributable to the induced apoptosis and that arisostatins A-induced apoptosis is mediated by caspase-3 activation pathway, loss of mitochondrial transmembrane potential (DeltaPsi(m)), and release of cytochrome c into cytosol. Microbiology, 2003 Sep, 149(Pt 9), 2687 - 96 Glucose and trehalose PTS permeases of Spiroplasma citri probably share a single IIA domain, enabling the spiroplasma to adapt quickly to carbohydrate changes in its environment; Andre A et al.; Spiroplasma citri is a plant-pathogenic mollicute phylogenetically related to Gram-positive bacteria . Spiroplasma cells are restricted to the phloem sieve tubes and are transmitted from plant to plant by the leafhopper vector Circulifer haematoceps . In the plant sieve tubes, S . citri grows on glucose and fructose, whereas in the leafhopper haemolymph the spiroplasma must grow on trehalose, the major sugar in insects . Previous studies in this laboratory have shown that fructose utilization was a key factor of spiroplasmal pathogenicity . To further study the implication of sugar metabolism in the interactions of S . citri with its plant host and its leafhopper vector, genes encoding permease enzymes II (EII(Glc) and EII(Tre)) of the S . citri phosphoenolpyruvate : glucose and phosphoenolpyruvate : trehalose phosphotransferase systems (PTS) were characterized . Mapping studies revealed that the EII(Glc) complex was split into two distinct polypeptides, IIA(Glc) and IICB(Glc), encoded by two separate genes, crr and ptsG, respectively . As expected, S . citri polypeptides IIA(Glc) and IICB(Glc) were more phylogenetically related to their counterparts from Gram-positive than to those from Gram-negative bacteria . The trehalose operon consisted of three genes treR, treP and treA, encoding a transcriptional regulator, the PTS permease (EII(Tre)) and the amylase, respectively . However, in contrast to the fructose-PTS permease, which is encoded as a single polypeptide (IIABC(Fru)) containing the three domains A, B and C, the trehalose-PTS permease (IIBC(Tre)) lacks its own IIA domain . No trehalose-specific IIA could be identified in the spiroplasmal genome, suggesting that the IIBC(Tre) permease probably functions with the IIA(Glc) domain . In agreement with this statement, yeast two-hybrid system experiments revealed that the IIA(Glc) domain interacted not only with IIB(Glc) but also with the IIB(Tre) domain . The results are discussed with respect to the ability of the spiroplasma to adapt from the phloem sap of the host plant to the haemolymph and salivary gland cells of the insect vector. J Bacteriol, 2003 Sep, 185(18), 5473 - 82 Biochemical characterization of a beta-galactosidase with a low temperature optimum obtained from an Antarctic arthrobacter isolate; Coker JA et al.; A psychrophilic gram-positive isolate was obtained from Antarctic Dry Valley soil . It utilized lactose, had a rod-coccus cycle, and contained lysine as the diamino acid in its cell wall . Consistent with these physiological traits, the 16S ribosomal DNA sequence showed that it was phylogenetically related to other Arthrobacter species . A gene (bgaS) encoding a family 2 beta-galactosidase was cloned from this organism into an Escherichia coli host . Preliminary results showed that the enzyme was cold active (optimal activity at 18 degrees C and 50% activity remaining at 0 degrees C) and heat labile (inactivated within 10 min at 37 degrees C) . To enable rapid purification, vectors were constructed adding histidine residues to the BgaS enzyme and its E . coli LacZ counterpart, which was purified for comparison . The His tag additions reduced the specific activities of both beta-galactosidases but did not alter the other characteristics of the enzymes . Kinetic studies using o-nitrophenyl-beta-D-galactopyranoside showed that BgaS with and without a His tag had greater catalytic activity at and below 20 degrees C than the comparable LacZ beta-galactosidases . The BgaS heat lability was investigated by ultracentrifugation, where the active enzyme was a homotetramer at 4 degrees C but dissociated into inactive monomers at 25 degrees C . Comparisons of family 2 beta-galactosidase amino acid compositions and modeling studies with the LacZ structure did not mimic suggested trends for conferring enzyme flexibility at low temperatures, consistent with the changes affecting thermal adaptation being localized and subtle . Mutation studies of the BgaS enzyme should aid our understanding of such specific, localized changes affecting enzyme thermal properties. Biodegradation, 2003 Aug, 14(4), 241 - 7 Growth-substrate dependent dechlorination of 1,2-dichloroethane by a homoacetogenic bacterium; De Wildeman S et al.; A rod shaped, gram positive, non sporulating Acetobacterium strain was isolated that dechlorinated 1,2dichloroethane (1,2-DCA) to ethene at a dechlorination rate of up to 2 nmol Cl- min(-1) mg(-1) of protein in the exponential growth phase with formate (40 mM) as the substrate . Although with other growth substrates such as pyruvate, lactate, H2/CO2, and ethanol higher biomass productions were obtained, the dechlorination rate with these substrates was more than 10-fold lower compared with formate growing cells . Neither cell extracts nor autoclaved cells of the isolated Acetobacterium strain mediated the dechlorination of 1,2-DCA at significant rates . The addition of 1,2-DCA to the media did not result in increased cell production . No significant differences in corrinoid concentrations could be measured in cells growing on several growth-substrates . However, these measurements indicated that differences in corrinoid structure might cause the different dechlorination activity . The Acetobacterium sp . strain gradually lost its dechlorination ability during about 10 transfers in pure culture, probably due to undefined nutritional requirements . 16S rDNA analysis of the isolate revealed a 99.7% similarity with Acetobacterium wieringae . However, the type strains of A . wieringae and A . woodii did not dechlorinate 1,2-DCA. Cancer, 2003 Sep 1, 98(5), 1039 - 47 Treatment of febrile neutropenic patients with cancer who require hospitalization: a prospective randomized study comparing imipenem and cefepime; Raad II et al.; BACKGROUND: The objective of the current study was to compare the efficacy and safety of imipenem and cefepime in the treatment of adult patients with cancer who had fever and neutropenia requiring hospitalization according to Infectious Disease Society of America criteria . METHODS: In the current prospective randomized clinical trial at a university-affiliated tertiary cancer center, adult patients with cancer who had fever (> or = 38.3 degrees C or > or = 38.0 degrees C for > 2 hours) and neutropenia (< or = 500/mm(3) or < 1000/mm(3) but declining) requiring hospitalization were randomized to receive either cefepime or imipenem . Vancomycin or amikacin was added on suspicion of gram-positive or gram-negative bacterial infection, respectively . RESULTS: Patients who received an imipenem regimen or a cefepime regimen were comparable in terms of age, gender, underlying malignancy, prior transplantation, degree and trend of neutropenia, and presence of central venous catheters (P > or = 0.3) . An intent-to-treat analysis showed a 68% response rate to the imipenem regimen, compared with a 75% response rate to the cefepime regimen (P = 0.2) . The rates of antibiotic-related adverse events and superinfections also were comparable (P = 0.6) . There was no difference in response among patients who received imipenem or cefepime alone compared with patients who also received vancomycin or amikacin (P = 1.0) . Leukemia was the only independent risk factor associated with a poor outcome (odds ratio, 4.6; 95% confidence interval, 1.9-10.7; P < 0.0001) . CONCLUSIONS: Imipenem and cefepime had similar efficacy and safety profiles in the treatment of adult cancer patients with fever and neutropenia who required hospitalization . The addition of either vancomycin or amikacin may not be necessary . Mol Microbiol, 2003 Sep, 49(5), 1201 - 12 Specific targeting of a DNA-binding protein to the SPP1 procapsid by interaction with the portal oligomer; Stiege AC et al.; The icosahedral procapsid of tailed bacteriophages is composed of a large number of identical subunits and of minor proteins found in a few copies . Proteins present in a very low copy number are targeted to the viral procapsid by an unknown mechanism . Bacteriophage SPP1 procapsids and mature virions contain two copies of gp7 on average . Gp7 forms stable complexes with the SPP1 portal protein gp6 . Deletion of the gp6 carboxyl-terminus and the mutation Y467-->C localized in the same region prevent gp6-gp7 complex formation . Gp7 binds double-stranded and single-stranded DNA . Gp6 competes for this interaction, and purified gp6-gp7 complexes do not bind DNA . Procapsid structures assembled in the absence of gp6 or carrying the mutant gp6 Y467-->C lack gp7 . The gp6-gp7 interaction thus targets gp7 to the procapsid where the portal protein is localized asymmetrically at a single vertex of the icosahedral structure . The interaction between the two proteins is disrupted during viral assembly . Proteins homologous to gp6 and gp7 are coded by contiguous genes in a variety of phage genomes from Gram-positive bacteria, suggesting that the gp6-gp7 complex is widespread in this group of phages . Transient association with the portal protein, an essential component of tailed bacteriophages and herpes viruses, provides a novel strategy to target minor proteins to the virion structure that might be operative in a large number of viruses. Neurochem Res, 2003 Sep, 28(9), 1359 - 67 Nocardia asteroides culture filtrates cause dopamine depletion and cytotoxicity in PC12 cells; Camp DM et al.; Experimental infection of BALB/c mice with the gram-positive bacterium Nocardia asteroides produces marked loss of nigrostriatal dopamine neurons, resulting in striatal dopamine depletion . To investigate the mechanism(s) responsible for this neuronal loss, we evaluated the influence of N . asteroides cell-free culture filtrates on rat pheochromocytoma PC12 cells, an in vitro model for dopamine neurons . Changes in cell viability and cell numbers were minimal after 24 h, but increased with longer incubation . In contrast, dopamine depletion occurred after 30 min incubation, and was greater with GUH-2 filtrate than with filtrate from the less virulent strain 10905 . Incubation with the culture filtrate decreased viability in neuroblastoma and glioma cell lines, indicating that cytotoxic effects were not limited to dopaminergic cells . These findings suggest that the loss of nigrostriatal dopamine neurons and concomitant striatal dopamine depletion in Nocardia-infected mice may be due, at least in part, to the neurotoxicity of nocardial secretory products. Mem Inst Oswaldo Cruz, 2003 Jun, 98(4), 529 - 32 Epub 2003 Aug 18. Bloodstream infections in late-stage acquired immunodeficiency syndrome patients evaluated by a lysis centrifugation system; Rosas RC et al.; Opportunistic infections, which affect acquired immunodeficiency syndrome (Aids) patients, are frequently disseminated and may cause bloodstream infections (BSI) . The aim of this study was to evaluate the main causes of BSI in Aids patients with advanced stage of the disease, with special emphasis on the identification of fungemia . During a 21 months period, all patients with Aids (CD4 < 200) and febrile syndrome admitted to 3 university hospitals were systematically evaluated . For each patient presenting fever, a pair of blood cultures was collected and processed by using a commercial lysis-centrifugation system . One hundred and eleven patients (75 males) with a mean age of 36 years (median 33 years) and mean CD4 count of 64 cells/ml were included . Among the 111 patients evaluated we documented 54 episodes of BSI, including 46 patients with truly systemic infections and 8 episodes considered as contaminants . BSI were caused by gram-positive bacteria (43%), fungi (20%), gram-negative bacteria (15%), mycobacteria (15%), and mixed flora (7%) . The crude mortality rate of our patients was 39%, being 50% for patients with BSI and 31% for the others . In conclusion, BSI are a common related to systemic infections on Aids patients with advanced stage of disease and is associated with a high rate of mortality. Immunity, 2003 Aug, 19(2), 269 - 79 Recognition of pneumococcal peptidoglycan: an expanded, pivotal role for LPS binding protein; Weber JR et al.; Lipopolysaccharide binding protein (LBP) has a well-established role in LPS-induced immune responses . Here, we report that LBP also plays an essential role in the innate immune response to Gram-positive pneumococci, specifically to their major inflammatory component, pneumococcal cell wall (PCW) . LBP was present in the CSF of patients with meningitis, and LBP-deficient mice failed to develop meningeal inflammation . LBP enhanced PCW-induced cell signaling and TNF-alpha release . LBP bound specifically to PCW multimers, indicating novel lipid-independent binding capability for LBP . We propose the iterative anionic groups along the glycan backbone of the cell wall are a crucial structure for recognition by LBP . Such a function for LBP expands its role to Gram-positive infections. Chem Commun (Camb), 2003 Aug 7, (15), 1842 - 3 A family of macrocyclic antibiotics with a mixed peptide-peptoid beta-hairpin backbone conformation; Shankaramma SC et al.; Macrocyclic peptidomimetics having a mixed peptide-peptoid backbone have been synthesized and shown to possess antibiotic activity against gram-positive and -negative bacteria with a low hemolytic activity against human erythrocytes; one is shown to adopt a regular beta-hairpin conformation by NMR in aqueous solution. Eur J Biochem, 2003 Sep, 270(17), 3593 - 602 A thermoacidophilic endoglucanase (CelB) from Alicyclobacillus acidocaldarius displays high sequence similarity to arabinofuranosidases belonging to family 51 of glycoside hydrolases; Eckert K et al.; A 100-kDa protein with endoglucanase activity was purified from Triton X-100 extract of cells of the thermoacidophilic Gram-positive bacterium Alicyclobacillus acidocaldarius . The enzyme exhibited activity towards carboxy methyl cellulose and oat spelt xylan with pH and temperature optima of 4 and 80 degrees C, respectively . Cloning and nucleotide sequence analysis of the corresponding gene (celB) revealed an ORF encoding a preprotein of 959 amino acids which is consistent with an extracellular localization . Purified recombinant CelB and a variant lacking the C-terminal 203 amino acid residues (CelBtrunc) displayed similar enzymatic properties as the wild-type protein . Analysis of product formation suggested an endo mode of action . Remarkable stability was observed at pH values between 1 and 7 and 60% of activity were retained after incubation for 1 h at 80 degrees C . CelB displayed homology to members of glycoside hydrolase family 51, being only the second entry with activity typical of an endoglucanase but lacking activity on p-nitrophenyl-alpha-l-arabinofuranoside (pNPAraf) . Highest sequence similarity was found towards the other endoglucanase F from Fibrobacter succinogenes (EGF), forming a distinct group in the phylogenetic tree of this family . Analysis of the amino acid composition of the catalytic domains demonstrated that CelB contains fewer charged amino acids than its neutrophilic counterparts, which is in line with adaptation to low pH . Wild-type and full-length recombinant CelB were soluble only in Triton X-100 . In contrast, CelBtrunc was completely water soluble, suggesting a role of the C-terminal region in cell association . This C-terminal hydrophobic region displayed local sequence similarities to an alpha-amylase from the same organism. Proc Natl Acad Sci U S A, 2003 Sep 2, 100(18), 10320 - 5 Epub 2003 Aug 13. Fatty acid biosynthesis in Mycobacterium tuberculosis: lateral gene transfer, adaptive evolution, and gene duplication; Kinsella RJ et al.; Mycobacterium tuberculosis is a high GC Gram-positive member of the actinobacteria . The mycobacterial cell wall is composed of a complex assortment of lipids and is the interface between the bacterium and its environment . The biosynthesis of fatty acids plays an essential role in the formation of cell wall components, in particular mycolic acids, which have been targeted by many of the drugs used to treat M . tuberculosis infection . M . tuberculosis has approximately 250 genes involved in fatty acid metabolism, a much higher proportion than in any other organism . In silico methods have been used to compare the genome of M . tuberculosis CDC1551 to a database of 58 complete bacterial genomes . The resulting alignments were scanned for genes specifically involved in fatty acid biosynthetic pathway I . Phylogenetic analysis of these alignments was used to investigate horizontal gene transfer, gene duplication, and adaptive evolution . It was found that of the eight gene families examined, five of the phylogenies reconstructed suggest that the actinobacteria have a closer relationship with the alpha-proteobacteria than expected . This is either due to either an ancient transfer of genes or deep paralogy and subsequent retention of the genes in unrelated lineages . Additionally, adaptive evolution and gene duplication have been an influence in the evolution of the pathway . This study provides a key insight into how M . tuberculosis has developed its unique fatty acid synthetic abilities. Nucleic Acids Res . 2003 Aug 15;31(16):e95. Restriction site tagged (RST) microarrays: a novel technique to study the species composition of complex microbial systems; Zabarovsky ER et al.; We have developed a new type of microarray, restriction site tagged (RST), for example NotI, microarrays . In this approach only sequences surrounding specific restriction sites (i.e . NotI linking clones) were used for generating microarrays . DNA was labeled using a new procedure, NotI representation, where only sequences surrounding NotI sites were labeled . Due to these modifications, the sensitivity of RST microarrays increases several hundred-fold compared to that of ordinary genomic microarrays . In a pilot experiment we have produced NotI microarrays from Gram-positive and Gram-negative bacteria and have shown that even closely related Escherichia coli strains can be easily discriminated using this technique . For example, two E.coli strains, K12 and R2, differ by less than 0.1% in their 16S rRNA sequences and thus the 16S rRNA sequence would not easily discriminate between these strains . However, these strains showed distinctly different hybridization patterns with NotI microarrays . The same technique can be adapted to other restriction enzymes as well . This type of microarray opens the possibility not only for studies of the normal flora of the gut but also for any problem where quantitative and qualitative analysis of microbial (or large viral) genomes is needed. Acta Odontol Latinoam, 1999, 12(2), 63 - 74 Correlation between bacterial counts in saliva and subgingival plaque; Testa M et al.; The aim of this study was to examine the correlation between the number and type of bacteria from periodontal pockets more than 4 mm deep and saliva in 26 patients . Periodontal pocket samples were taken with paper points and transferred to 0.1 ml of enriched thioglicollate broth . Saliva samples were collected simultaneously in aseptic flasks . Both samples were processed within the first hour . They were inoculated in Schaedler agar plus 5 micrograms/ml vitamin K and 5% blood, TSBV agar and MGB agar to perform colony counts and identification . Spirochete counts per microscopic field were obtained by direct light microscopy of Gram-stained preparations . The results show a fair to good correlation between both samples for anaerobic, pigmented gram-negative rods, anaerobic non-pigmented gram-negative rods, spirochetes, facultative gram-negative rods other than Actinobacillus actinomycetemcomitans, anaerobic, gram-positive cocci and anaerobic gram-positive rods (Spearman correlation coefficient 0.51 to 0.96) . The correlation coefficient values for A.a., facultative gram-positive rods, facultative gram-positive cocci and facultative gram-negative cocci were lower than 0.21 . There were no significant differences between the counts in both samples for all the bacterial groups (Student's t test, p > 0.1) . We may conclude that, under the experimental conditions of the present study, saliva samples and periodontal pocket samples are equally useful to detect subgingival organisms associated with periodontal disease in the oral cavity . Saliva samples were useful to evaluate risk and periodontal therapy in individual patients or groups. J Immunol, 2003 Aug 15, 171(4), 1647 - 51 Cutting edge: SR-PSOX/CXC chemokine ligand 16 mediates bacterial phagocytosis by APCs through its chemokine domain; Shimaoka T et al.; SR-PSOX and CXC chemokine ligand (CXCL)16, which were originally identified as a scavenger receptor and a transmembrane-type chemokine, respectively, are indicated to be identical . In this study, we demonstrate that membrane-bound SR-PSOX/CXCL16 mediates adhesion and phagocytosis of both Gram-negative and Gram-positive bacteria . Importantly, our prepared anti-SR-PSOX mAb, which suppressed chemotactic activity of SR-PSOX, significantly inhibited bacterial phagocytosis by human APCs including dendritic cells . Various scavenger receptor ligands inhibited the bacterial phagocytosis of SR-PSOX . In addition, the recognition specificity for bacteria was determined by only the chemokine domain of SR-PSOX/CXCL16 . Thus, SR-PSOX/CXCL16 may play an important role in facilitating uptake of various pathogens and chemotaxis of T and NKT cells by APCs through its chemokine domain. Genome Res, 2003 Aug, 13(8), 1800 - 9 Tropheryma whipplei Twist: a human pathogenic Actinobacteria with a reduced genome; Raoult D et al.; The human pathogen Tropheryma whipplei is the only known reduced genome species (<1 Mb) within the Actinobacteria {high G+C Gram-positive bacteria} . We present the sequence of the 927303-bp circular genome of T . whipplei Twist strain, encoding 808 predicted protein-coding genes . Specific genome features include deficiencies in amino acid metabolisms, the lack of clear thioredoxin and thioredoxin reductase homologs, and a mutation in DNA gyrase predicting a resistance to quinolone antibiotics . Moreover, the alignment of the two available T . whipplei genome sequences (Twist vs . TW08/27) revealed a large chromosomal inversion the extremities of which are located within two paralogous genes . These genes belong to a large cell-surface protein family defined by the presence of a common repeat highly conserved at the nucleotide level . The repeats appear to trigger frequent genome rearrangements in T . whipplei, potentially resulting in the expression of different subsets of cell surface proteins . This might represent a new mechanism for evading host defenses . The T . whipplei genome sequence was also compared to other reduced bacterial genomes to examine the generality of previously detected features . The analysis of the genome sequence of this previously largely unknown human pathogen is now guiding the development of molecular diagnostic tools and more convenient culture conditions. Ann Thorac Surg, 2003 Aug, 76(2), 401 - 5; discussion 405-6 Twenty-six years of experience with the modified eloesser flap; Thourani VH et al.; BACKGROUND: Empyema thoracis is a common thoracic problem with a multitude of therapeutic options . The modified Eloesser flap (MEF) is one means of dealing with this problem in selected complicated patients . The purpose of this study is to report our 26-year experience with the MEF . METHODS: A review of 78 patients who had a MEF from 1975 to 2001 was performed . RESULTS: There were 52 males (67%) and 26 females (33%) . Mean age was 59 +/- 14 years . The overall length of stay was 26 +/- 27 days, while mean postoperative length of stay was 16 +/- 17 days . Microbiology of the empyema cavity revealed a predominance of gram-positive organisms . Before a modified Eloesser flap, all patients failed initial conservative interventions and 23 patients (29%) failed surgical interventions . Operative indications were as follows: parapneumonic effusions, 35 patients (45%); postresectional, 23 patients (29%); tuberculosis related, 7 patients (9%); malignant effusion, 4 patients (5%); esophageal fistulas, 4 patients (5%); abdominal sepsis, 3 patients (4%); and hemothorax secondary to trauma, 2 patients (3%) . The inverted-U incision was performed in all patients . Average rib resection was 3 +/- 1 ribs . There were no intraoperative complications and adequate drainage was achieved in all patients . Thirty-day morbidity/mortality was 4 patients (5%): 3 died of sepsis and 1 died of metabolic encephalopathy; although long-term follow-up (mean: 109 +/- 141 months) revealed no additional morbidity related to the MEF . CONCLUSIONS: We demonstrate that MEF can be performed as a safe, definitive surgical procedure for the treatment of chronic empyema thoracis . The MEF remains an important option in the surgical treatment of chronic, complicated empyema thoracis. Am J Kidney Dis, 2003 Aug, 42(2), 350 - 4 Depression and its association with peritonitis in long-term peritoneal dialysis patients; Troidle L et al.; BACKGROUND: Depression is the most common psychological disorder among patients with end-stage renal disease and has been associated with mortality in patients maintained on hemodialysis therapy . Peritonitis is the leading cause of technique failure among long-term peritoneal dialysis (PD) patients . This prospective study is designed to examine the relationship between depression and peritonitis . METHODS: All patients on long-term PD therapy in our unit between January 1, 1997, and January 31, 2002, completed a Beck Depression Inventory (BDI) assessment at 6-month intervals . BDI scores were analyzed 2 ways . First, patients were placed into either group I (BDI score < 10) or group II (BDI score > or = 11) and were reclassified based on subsequent scores . Second, multivariable analysis was performed looking at initial BDI score as a risk factor for peritonitis, adjusting for age older than 65 years, diabetes, coronary artery disease, and race . RESULTS: One hundred sixty-two patients were enrolled, and 281 individual BDI assessments were completed . There was a significantly greater incidence of diabetes and coronary artery disease in group II . Rates for overall and gram-positive peritonitis were significantly greater in group II patients compared with group I patients . Using Cox regression, only BDI score of 11 or greater was associated with the development of peritonitis (hazard ratio, 2.7; 95% confidence interval, 1.2 to 6.0) . CONCLUSION: There is an association between BDI score of 11 or greater and the development of peritonitis . Whether treatment of depression can impact on the rate of peritonitis remains to be examined. Curr Opin Immunol, 2003 Aug, 15(4), 408 - 15 Innate sensors for Gram-positive bacteria; Weber JR et al.; More than half of invasive bacterial infections are Gram-positive in origin . This class of bacteria has neither endotoxins nor an outer membrane, yet it generates some of the most powerful inflammatory responses known in medicine . Some recent seminal studies go a long way toward settling the controversies that surround the process by which Gram-positive bacterial surfaces trigger the human immune system . Although the components of the cell wall are now chemically defined in exquisite detail and the interaction with the toll-like receptor 2 pathway has been discovered, it is only very recently that definitive studies combining these advanced biochemical and cell biological tools have been carried out . It is these breakthrough studies that have finally confirmed the paradigm of innate sensors for Gram-positive bacteria. Phytochemistry, 2003 Aug, 63(8), 945 - 52 Polypropionate lactones of deoxysugars glycosides from slime mold Lycogala epidendrum; Rezanka T et al.; Two novel polypropionate lactone glycosides (1 and 2, i.e . lycogalinosides A and B) were isolated from the slime mold Lycogala epidendrum . Their structures, including the absolute configurations of the hydroxyl and methyls groups, were determined by means of extensive spectroscopic data such as mass, IR, UV, and 1D and 2D NMR spectra and chemical degradation followed by spectroscopic and chromatographic analysis . Compounds 1 and 2 are unique in structure containing a 2-deoxy-alpha-L-fucopyranosyl-(1-4)-6-deoxy-beta-D-gulopyranosyl unit and a beta-D-olivopyranosyl-(1-4)-beta-D-fucopyranosyl unit, respectively, and showed growth inhibitory activities against Gram-positive bacteria. An Med Interna, 2003 Jul, 20(7), 347 - 50 {Early hospital mortality due to infectious diseases}; Vallejo M et al.; BACKGROUND: Infectious diseases are an important health problem . Early hospital mortality (EHM) (first 48 hours after hospital admission) give us information about the etiology and the focus of infection . This study was designed because no articles have been found about this subject . MATERIAL AND METHODS: We reviewed the medical records coded by the ICD-9-CM of all patients that suffered from EHM due to infectious diseases during the period 1992 to 1999 . RESULTS: Of all the patients analyzed, 0.7% died of EHM, and of theses, 6.9% were due to an infectious disease . Median age was 73.2 years; 56.1% were men . Index of comorbidity was higher than 1 in 59,9%, and 70,7% never has been admitted to the hospital before . At admission, fever was present in 43.9% . The illness severity was 60.9% sepsis, 24.4% severe sepsis, 13.4% septic shock and 1.2% multiorgan failure . Causes of death were respiratory (76.8%; pneumonia 58.5%) . Pneumonia was more frequent among aged 65 years and older (p = 0.03) . In 69.5% no microbiological techniques were performed with independence of the clinical severity or the presence or absence of fever . In 85.4% the casual agent was unidentified, but in the case of isolation, gram positive was the most frequent microorganism . CONCLUSIONS: Infections are an important cause of EHM, and community-acquired respiratory tract infection (mainly pneumonia) the most frequent cause of EHM . Patients were admitted to the hospital with sepsis in 60.9%, perhaps due to a diagnostic or therapeutic delay . Among aged 65 years and older, microbiological diagnostic procedures were rarely employed. Int J Syst Evol Microbiol, 2003 Jul, 53(Pt 4), 1201 - 5 Phylogenetic analysis identifies the 'megabacterium' of birds as a novel anamorphic ascomycetous yeast, Macrorhabdus ornithogaster gen . nov., sp . nov; Tomaszewski EK et al.; An organism commonly referred to as 'megabacterium' colonizes the gastric isthmus of many species of birds . It is weakly gram-positive and periodic acid-Schiff-positive and stains with silver stains . Previous studies have shown that it has a nucleus and a cell wall similar to those seen in fungi . Calcofluor white M2R staining suggests that the cell wall contains chitin, a eukaryote-specific substance, and rRNA in situ hybridization demonstrates that it is a eukaryote . To characterize this organism phylogenetically, DNA was extracted from purified cells . rDNA was readily amplified by PCR with pan-fungal DNA primer sets and primer sets derived from the newly determined sequence, but not with bacteria-specific primer sets . Specific primer sets amplified rDNA from isthmus scrapings from an infected bird, but not from a non-infected bird or other control DNA . The sequence was confirmed to derive from the purified organism by in situ rRNA hybridization using a specific probe . Phylogenetic analysis of sequences of the 18S rDNA and domain D1/D2 of 26S rDNA showed the organism to be a previously undescribed anamorphic ascomycetous yeast representing a new genus . The name Macrorhabdus ornithogaster gen . nov., sp . nov . is proposed for this organism . The type material is CBS 9251T (= NRRL Y-27487T). Int J Syst Evol Microbiol, 2003 Jul, 53(Pt 4), 985 - 94 Rhodoglobus vestalii gen . nov., sp . nov., a novel psychrophilic organism isolated from an Antarctic Dry Valley lake; Sheridan PP et al.; A novel, psychrophilic, gram-positive bacterium (designated strain LV3T) from a lake near the McMurdo Ice Shelf, Antarctica, has been isolated and characterized . This organism formed red-pigmented colonies, had an optimal growth temperature of 18 degrees C and grew on a variety of media between -2 and 21 degrees C . Scanning electron micrographs of strain LV3T that showed small rods with unusual bulbous protuberances during all phases of growth were of particular interest . The G + C content of the genomic DNA was approximately 62 mol% . The cell walls contained ornithine as the diamino acid . The major fatty acids were anteiso-C15:0, iso-C16:0 and anteiso-C17:0 . Cells grown at -2 degrees C contained significant amounts of anteiso-C15:1 . The major menaquinones found in strain LV3T were MK-11 and MK-12 . Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain LV3T was a member of the family Microbacteriaceae and related to, but distinct from, organisms belonging to the genera Agreia, Leifsonia and Subtercola . In addition, alignments of 16S rRNA sequences showed that the sequence of strain LV3T contained a 13 bp insertion that was found in only a few related sequences . Based on the low growth temperature, unusual cell shape, distinct 16S rRNA gene sequence and structure and cell-wall amino acid and menaquinone compositions, Rhodoglobus vestalii gen . nov., sp . nov . is proposed, with the type strain LV3T (= ATCC BAA-534T = CIP 107482T). Int J Syst Evol Microbiol, 2003 Jul, 53(Pt 4), 977 - 84 Leifsonia rubra sp . nov . and Leifsonia aurea sp . nov., psychrophiles from a pond in Antarctica; Reddy GS et al.; Two unique psychrophilic strains (CMS 76rT and CMS 81yT) were isolated from a cyanobacterial mat sample from a pond in Wright Valley, McMurdo, Antarctica . Both isolates were assigned to the genus Leifsonia, since they were gram-positive, curved rods, non-motile, catalase-positive, contained DL-2,4-diaminobutyric acid, menaquinone MK-11, phosphatidylglycerol and diphosphatidylglycerol, had a high content of anteiso- and iso-branched fatty acids and had a DNA G + C content of 64-66 mol% . In addition, both isolates were related to the five reported species of Leifsonia at a level of about 95-96% 16S rDNA sequence similarity and differed from one another by 2.5% . Strains CMS 76rT and CMS 81yT also differed from one another in many other phenotypic characteristics and exhibited only 30% relatedness at the DNA-DNA level, thus indicating that they represent two different species . Furthermore, these two isolates also showed many distinct differences with respect to the reported species of Leifsonia in terms of their phenotypic characteristics, biochemical properties, chemotaxonomic features, sensitivity to various antibiotics and 16S rDNA similarity, clearly indicating that strains CMS 76rT (= MTCC 4210T = DSM 15304T = CIP 107783T) and CMS 81yT (= MTCC 4657T = DSM 15303T = CIP 107785T) represent the type strains of two novel species of Leifsonia, for which the names Leifsonia rubra sp . nov . and Leifsonia aurea sp . nov . are proposed. Int J Syst Evol Microbiol, 2003 Jul, 53(Pt 4), 971 - 5 Alkaliphilus crotonatoxidans sp . nov., a strictly anaerobic, crotonate-dismutating bacterium isolated from a methanogenic environment; Cao X et al.; Two bacterial strains were isolated from methanogenic butyrate-oxidizing mixed cultures . The cells were straight to slightly curved, gram-positive rods that were motile by means of multiple flagella and formed endospores . Growth was observed in the temperature range 15-45 degrees C (optimum 37 degrees C) and pH range 5.5-9.0 (optimum pH 7.5) . The novel isolates were strictly anaerobic chemo-organotrophs capable of utilizing yeast extract, peptone, tryptone and a variety of sugars and organic acids, but not glucose . None of the accessory electron acceptors tested (elemental sulfur, thiosulfate or fumarate) improved growth, except crotonate, which was dismutated to butyrate and acetate . The G + C content of the DNA of one of the isolates, strain B11-2T, was 30.6 mol% . Phylogenetic analysis based on 16S rDNA sequence similarity between strain B11-2T and some other strictly anaerobic, spore-forming bacteria indicated that the novel isolates represented a species in cluster XI within the low-GC gram-positive bacteria, being most closely related to Alkaliphilus transvaalensis JCM 10712T . DNA-DNA relatedness between strain B11-2T and A . transvaalensis JCM 10712T was 21% . On the basis of physiological and molecular properties, and cellular fatty acid and cell wall compositions, the novel isolates are proposed to represent a novel species of the genus Alkaliphilus, for which the name Alkaliphilus crotonatoxidans is proposed (type strain B11-2T=AS 1.2897T=JCM 11672T). Zh Mikrobiol Epidemiol Immunobiol, 2003 May-Jun, (3), 109 - 13 {Bacteriocins: criteria, classification, characteristics, methods of detection}; Blinkova LP; The review on bacteriocins of Gram negative and Gram positive bacteria . Criteria making it possible to regard antagonistic substances as bateriocins or bacteriocin-like substances and on their classification are presented . Examples of bacteriocins naming depending on the taxonomic position of the producer culture are given . Information on the physico-chemical and biological properties of bacteriocins and their purification is presented as well as on detection tools of bacteriocins in microorganisms and evaluation of the producer activity of the bacteriological culture. Klin Monatsbl Augenheilkd, 2003 Jul, 220(7), 481 - 5 {Post-traumatic endophthalmitis after penetrating injury in Vietnam: risk factors, microbiological aspect and visual outcome}; Tran TP et al.; BACKGROUND: Post-traumatic endophthalmitis associated with penetrating injury represents a distinct kind of intraocular infection and is still a severe problem for diagnosis and treatment in developing countries . We report the incidence, microbiological presentation and visual outcome of post-traumatic endophthalmitis in South Vietnam . PATIENTS AND METHOD: In a retrospective study medical reports of 515 patients with penetrating injury treated at Eye Hospital in Ho Chi Minh city, Vietnam, from 1/1999 to 5/2000 were reviewed and analysed . The treatment was standardised: 1 . after vitreous-biopsy intravitreous injection of 0.1 mg/0.1 ml Gentamycine and 1 mg/0.1 ml Vancomycine; 2 . subconjunctival injection of 25 mg/0.5 ml Vancomycine and 20 mg/0.5 ml Gentamycine; 3 . topical and systemic antibiotic treatment due to microbiological presentation; 4 . systemic steroids treatment . Clinical parameters were evaluated for association with an increased risk of endophthalmitis . RESULTS: Out of 515 patients with penetrating injury 61 patients (11.8 %) with endophthalmitis were treated and evaluated . Mean time interval from trauma to diagnosis of endophthalmitis was 16.8 +/- 5.6 days . Follow-up period was 15.6 +/- 23.4 days . Visual acuity at the diagnosis of endophthalmitis in 96 % of all patients was only finger counting or light perception . Enucleation was needed in 14 % of patients, 29 % of patients developed phthisis bulbi . A visual acuity better than finger counting was reached in only 10 % of all patients with endophthalmitis . Cultures of the vitreous aspirate were positive in only 50 % of cases available and showed Gram-positive (51 %), Gram-negative bacteria (33 %) and fungi (16 %) . Risk factors found to be significant were: 1 . a purely corneal wound; 2 . wound length less than 5 mm; 3 . surgical primary repair more than 24 hour after trauma; 4 . inadequate antibiotic treatment and 5 . a rural injury setting . CONCLUSION: Post-traumatic endophthalmitis is still a major problem in Vietnam . Delay in primary wound closure or inadequate antibiotic treatment worsen the prognosis profoundly . Comprehensive prophylactic antibiotic treatment at the time of injury repair is in dispensable . In risk eyes particular attention should be paid to prophylaxis and signs of infection . Further prospective studies are necessary to develop optimal and adequate diagnostic and treatment options in Vietnam. Chemotherapy, 2003 Jul, 49(4), 209 - 11 Lung, pleural and colon actinomycosis in an immunocompromised patient: a rare form of presentation; Charalabopoulos K et al.; Actinomycosis is caused by gram-positive filamentous organisms of the genus Actinomyces, which may spread through trauma . Most commonly, it is a cervicofacial disease due to dental infection or a thoracic disease secondary to aspiration of foreign bodies . Primary abdominal infection usually follows some form of mucosal disruption . Any organ of the human body may be involved so that a wide range of symptoms may be present . We report a rare form of actinomycosis involving the lung, pleura and colon concomitantly in an immunocompromised patient . A fine needle aspiration from a lung lesion detected the characteristic sulfur granules, and a pleural effusion culture confirmed the diagnosis . Clinical manifestations and treatment are discussed . Actinomycetes are rarely opportunistic agents in immunocompromised patients; thus the disease deserves special attention in those patients . Eur J Pharm Sci, 2003 Jul, 19(4), 245 - 51 Endospores of B subtilis are pyrogenic and activate Mono Mac 6 cells: importance of the CD14 receptor; Moesby L et al.; The monocytic cell line Mono Mac 6 is sensitive to pyrogens and interleukin-6 secretion is induced after exposure to pyrogens . The aim of this study is to examine the pyrogenic activity and the interleukin-6-inducing capacity of the Gram-positive B . subtilis bacteria, endospores and isolated cell wall components . Furthermore the involvement of CD14 in activation of interleukin-6 release is investigated . All test substances are pyrogenic in the rabbit pyrogen test . The test substance is incubated with monocytic cells (Mono Mac 6) for 24 h and the secreted interleukin-6 is determined in a sandwich immunoassay . B . subtilis bacteria and endospores induce interleukin-6 in a dose-dependent manner . Endospores are less potent than bacteria . Lipoteichoic acid (LTA) isolated from B . subtilis induces interleukin-6 in a dose-dependent manner, whereas muramyl dipeptide (MDP) is unable to induce interleukin-6 . Lipopolysaccharides (LPS) dose-dependently induce interleukin-6 release, but the curve differs from that of LTA both in shape and offset . The interleukin-6 secretion induced by LPS, LTA and B . subtilis bacteria can be blocked by 73-85% by an antibody directed against CD14, whereas the antibody only blocks 25% of B . subtilis endospores-induced interleukin-6 release . The results might indicate that B . subtilis endospores use an additional pathway to CD14 to activate mononuclear cells. J Drugs Dermatol, 2003 Aug, 2(4), 378 - 83 Linezolid and quinupristin/dalfopristin: novel antibiotics for gram-positive infections of the skin; Schweiger ES et al.; With the continuing development of clinical drug resistance among bacteria, the need for new, effective agents to treat multi-drug-resistant Gram-positive infections remains important . With treatment options limited, it has become critical to identify antibiotics with novel mechanisms of activity . Several new drugs have emerged as possible therapeutic alternatives . This review focuses on agents newly introduced and FDA-approved for the treatment of