Gram positive

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 skin and skin structure infections: linezolid and quinupristin/dalfopristin.

Eur J Immunol, 2003 Aug, 33(8), 2327 - 36
CRP-ductin, the mouse homologue of gp-340/deleted in malignant brain tumors 1 (DMBT1), binds gram-positive and gram-negative bacteria and interacts with lung surfactant protein D; Madsen J et al.; CRP-ductin is a protein expressed mainly by mucosal epithelial cells in the mouse . Sequence homologies indicate that CRP-ductin is the mouse homologue of human gp-340, a glycoprotein that agglutinates microorganisms and binds the lung mucosal collectin surfactant protein-D (SP-D) . Here we report that purified CRP-ductin binds human SP-D in a calcium-dependent manner and that the binding is not inhibited by maltose . The same properties have previously been observed for gp-340 binding of SP-D . CRP-ductin also showed calcium-dependent binding to both gram-positive and -negative bacteria . A polyclonal antibody raised against gp-340 reacted specifically with CRP-ductin in Western blots . Immunoreactivity to CRP-ductin was found in the exocrine pancreas, in epithelial cells throughout the gastrointestinal tract and in the parotid ducts . A panel of RNA preparations from mouse tissues was screened for CRP-ductin and SP-D expression by reverse transcription-PCR . The pancreas was the main site of synthesis of CRP-ductin, but transcripts were also readily amplified from salivary gland, the gastrointestinal tract, liver, testis, uterus and lung . Lung was the main site of synthesis of SP-D, but transcripts were also amplified from uterus, salivary gland, thymus, thyroid gland, pancreas and testis . We conclude that CRP-ductin is the mouse homologue of human gp-340 and that its capacity to bind SP-D as well as gram-negative and gram-positive bacteria suggests a role in mucosal immune defense.

Clin Infect Dis, 2003 Aug 1, 37(3), 382 - 9 Epub 2003 Jul 22.
Vancomycin versus placebo for treating persistent fever in patients with neutropenic cancer receiving piperacillin-tazobactam monotherapy; Cometta A et al.; This prospective, double-blind trial assessed whether the addition of a glycopeptide would be able to reduce the time to defervescence in neutropenic patients with cancer who had persistent fever 48-60 h after the initiation of empirical piperacillin-tazobactam monotherapy . Of 763 eligible patients, 165 with persistent fever were randomized to receive piperacillin-tazobactam therapy plus either vancomycin therapy or placebo . Defervescence was observed in 82 (95%) of 86 patients in the vancomycin group and in 73 (92%) of 79 patients in the placebo group (P=.52) . The distributions of the time to defervescence were not statistically significant between the 2 groups (estimated hazard ratio, 1.03; 95% confidence interval, 0.75-1.43; P=.75) . The number of additional episodes of gram-positive bacteremia and the percentage of patients for whom amphotericin B was empirically added to their therapy regimen were also similar in both groups . This study failed to demonstrate that the empirical addition of vancomycin therapy to the treatment regimen is of benefit to persistently febrile neutropenic patients with cancer.

Appl Microbiol Biotechnol, 2003 Aug, 62(2-3), 256 - 62 Epub 2003 Mar 20.
Roles of tert-butyl formate, tert-butyl alcohol and acetone in the regulation of methyl tert-butyl ether degradation by Mycobacterium austroafricanum IFP 2012; Francois A et al.; Mycobacterium austroafricanum IFP 2012 is a Gram-positive strain able to grow on methyl tert-butyl ether (MTBE) as a sole carbon and energy source . The effect of two downstream metabolites of MTBE, tert-butyl formate (TBF) and tert-butyl alcohol (TBA) on MTBE degradation was investigated using resting cells . The addition of low concentrations of TBF decreased the MTBE degradation rate by about 30% . In contrast, the addition of TBA did not have a significant effect on MTBE degradation rate, even at high concentrations; and it was also shown that TBA degradation occurred only once MTBE was exhausted . At neutral pH, TBF hydrolysis involved mainly an esterase-type activity regulated by the presence of TBA . The TBF degradation rate was about four times lower than the MTBE degradation rate . Furthermore, acetone was identified as an intermediate during TBA degradation . An acetone mono-oxygenase activity, inhibited by methimazole but not by acetylene, was suggested . It was different from the MTBE/TBA mono-oxygenase and, thus, acetone did not appear to compete with MTBE and TBA for the same enzyme . These new results show that the metabolic regulation of the early steps of MTBE degradation by M . austroafricanum IFP 2012 is complex, involving inhibition and competition phenomena.

Drugs Today (Barc), 2000 Apr, 36(4), 245 - 53
Clinical results in the treatment of respiratory infections with moxifloxacin; Lode H et al.; Respiratory infections are a common source of morbidity and mortality, with pneumonia being the number one cause of death from infectious disease in Western industrialized countries . Initial antibiotic therapy of upper and lower respiratory infections is often empiric, being directed at the pathogens that are most likely to be present . Leading pathogens in respiratory infections are S . pneumoniae, H . influenzae and M . catarrhalis, which have developed considerable resistance problems against previous standard antibiotics like beta-lactams, macrolides and tetracyclines in the last decade . Newly developed quinolones such as moxifloxacin combine enhanced in vitro activity against Gram-positive bacteria with maintenance of activity against Gram-negative organisms . Three comparative, prospective, randomized, double-blind studies in the treatment of community acquired sinusitis, AECB and CAP demonstrated equal or higher efficacy of moxifloxacin in comparison to standard antibiotic therapies.

Int J Food Microbiol, 2003 Aug 25, 85(3), 269 - 79
Potential for broad applications of flow cytometry and fluorescence techniques in microbiological and somatic cell analyses of milk; Gunasekera TS et al.; Monitoring the quality and safety of milk requires careful analysis of microbial and somatic cell loading . Our aim was to demonstrate proof of the principle that flow cytometry (FCM), coupled with fluorescence techniques for distinguishing between cell types, could potentially be employed in a wide variety of biological assays relevant to the dairy industry . To this end, we studied raw milk samples and ultraheat-treated milk, into which known numbers of bacteria or mouse cells were inoculated . For bacterial analyses, protein and lipids were removed, whereas only centrifugal lipid clearing was needed for somatic cell analyses . Cleared samples were stained with fluorescent dyes or with bacterial-specific fluorescent-labeled oligonucleotides and analyzed by FCM . A fluoresceinated peptide nucleic acid probe enabled efficient enumeration of bacteria in milk . Dual staining of samples with fluorescent dyes that indicate live (5-cyanol-2,3-ditolyl tetrazolium chloride, CTC or SYTO 9) or damaged cells (oxonol or propidium iodide, PI) enabled determination of viable bacteria in milk . Gram-positive and -negative bacteria were distinguished using hexidium iodide and SYTO 13 in dual staining of cleared milk samples . An FCM-based method gave a good correlation (r=0.88) with total microscopic counts of somatic cells in raw milk . The FCM method also correlated strongly (r=0.98) with the standard Fossomatic method for somatic cell detection . We conclude that FCM, coupled with fluorescence staining techniques, offers potentially diverse and rapid approaches to biological safety and quality testing in the dairy industry . Potential application of flow cytometers to a broad range of assays for milk biological quality should make this instrumentation more attractive and cost effective to the dairy industry and indeed the broader food industry.

Toxicon, 2003 Jun, 41(8), 1033 - 8
Structural, enzymatic and biological properties of new PLA(2) isoform from Crotalus durissus terrificus venom; Toyama MH et al.; We isolated a new PLA(2) from the Crotalus durissus terrificus venom that designated F15, which showed allosteric behavior with a V(max) of 8.5nmol/min/mg and a K(m) of 38.5 mM . The incubated heparin salt of this isolated F15 act a positive allosteric effector by increasing the V(max) to 10.2 nmol/min/mg, with decreasing the V(max) value to 20.5 mM . The crotapotin, on the other hand acts as a negative allosteric effector by increasing the V(max) values to 58.4 mM . F15 also showed high calcium dependence for its catalysis similar to that found for other PLA(2) enzymes isolated from these snake venoms . The replacement of calcium by other divalent ions such Mg(2+), Mn(2+), Cd(2+), Sn(2+) and Cu(2+) resulted in lower enzymatic activity . The optimum pH and temperature for the enzyme was 8.5 and 18 degrees C, respectively . F15 alone showed moderate neurotoxic activity in isolated mouse phrenic nerve diaphragm in comparison to other strong myotoxic PLA(2) such as bothropstoxin-I (BThtx-I), but this activity was highly neurotoxic in a chick biventrer cervis preparation, whereas BthTx-I did not reveal this high neurotoxicity . This new protein showed a high bactericidal effect against both Gram-negative and Gram-positive bacterial strains . F15 contained 122 amino acid residues, with a primary structure of: HLLQFNKMIKFETRKNAVPFYAFYGCYCGWGGQRRPKDATDRCCFVHDCCYGKLTKCNTKWDIYRYSLKSGYITCGKGTWCKEQICECDRVAAECLRRSLSTYKNEYMFYPKSRCRRPSETC . Its molecular mass and isoeletric point were 14.5 kDa and 8.85, both estimated by two dimensional electrophoresis . The amino acid sequence of the F15 revealed high sequence homology with F16 and F17 . F15 and the other PLA(2)s revealed highly conserved amino acid sequences principally for calcium binding loop and active site helix . F15 also showed a high homology with the lysine-rich region of myotoxic PLA(2).

J Mol Microbiol Biotechnol, 2003, 5(4), 206 - 15
Transcription regulators potentially controlled by HPr kinase/phosphorylase in Gram-negative bacteria; Boel G et al.; Phosphorylation and dephosphorylation at Ser-46 in HPr, a phosphocarrier protein of the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) is controlled by the bifunctional HPr kinase/phosphorylase (HprK/P) . In Gram-positive bacteria, P-Ser-HPr controls (1) sugar uptake via the PTS; (2) catabolite control protein A (CcpA)-mediated carbon catabolite repression, and (3) inducer exclusion . Genome sequencing revealed that HprK/P is absent from Gram-negative enteric bacteria, but present in many other proteobacteria . These organisms also possess (1) HPr, the substrate for HprK/P; (2) enzyme I, which phosphorylates HPr at His-15, and (3) one or several enzymes IIA, which receive the phosphoryl group from P approximately His-HPr . The genes encoding the PTS proteins are often organized in an operon with HPRK . However, most of these organisms miss CcpA and a functional PTS, as enzymes IIB and membrane-integrated enzymes IIC seem to be absent . HprK/P and the rudimentary PTS phosphorylation cascade in Gram-negative bacteria must therefore carry out functions different from those in Gram-positive organisms . The gene organization in many HprK/P-containing Gram-negative bacteria as well as some preliminary experiments suggest that HprK/P might control transcription regulators implicated in cell adhesion and virulence . In alpha-proteobacteria, HPRK is located downstream of genes encoding a two-component system of the EnvZ/OmpR family . In several other proteobacteria, HPRK is organized in an operon together with genes from the RPON region of ESCHERICHIA COLI (RPON encodes a sigma54) . We propose that HprK/P might control the phosphorylation state of HPr and EIIAs, which in turn could control the transcription regulators .

Diagn Microbiol Infect Dis, 2003 Jul, 46(3), 181 - 7
Cloning and sequencing an unknown gene of Tropheryma whipplei and development of two LightCycler PCR assays; Maibach RC et al.; Whipple's disease is a rare infectious illness that can affect any organ system in the body . It is caused by Tropheryma whipplei, a Gram-positive rod-shaped bacterium with a high G + C content, classified within the actinobacteria . For decades, laboratory detection has been based on microscopy and the periodic acid-Schiff (PAS) staining of biopsies . Recently, PCR has become a useful tool to detect T . whipplei DNA in various clinical specimens . However, a positive PCR result does not confirm Whipple's disease as it has been shown that asymptomatic persons can harbor T . whipplei DNA . Since there is not yet much known about the genome of T . whipplei, genome-walking represents a convenient method to determine unknown gene sequences . Starting from a RAPD fragment we have sequenced and cloned an open reading frame (ORF) of 843 bp . Two real-time PCR assays targeting the ORF fragment and the 16S rRNA gene, respectively, were developed . Compared to a conventional 16S rRNA PCR system the ORF LightCycler assay proved to be very specific (100%) but not sufficiently sensitive (62.4%) . In contrast, the 16S rRNA LightCycler assay showed a sensitivity of 95.7% and a specificity of 97.8% . Thus, the 16S rRNA gene assay but not that targeting the new ORF is a suitable alternative to conventional PCR methods.

Clin Ther, 2003 Jun, 25(6), 1846 - 71
Hospital resource use and cost of treatment with linezolid versus teicoplanin for treatment of serious gram-positive bacterial infections among hospitalized patients from South America and Mexico: results from a multicenter trial; Lopez H et al.; BACKGROUND: Linezolid is a novel oxazolidinone antibiotic that is effective for the treatment of gram-positive bacterial infections . The oral formulation has the potential to reduce length of stay (LOS) when used as a substitute for parenteral glycopeptide antibiotics . In a recent multinational trial comparing linezolid (i.v . followed by oral administration) with teicoplanin (i.v . alone or switched to i.m . administration), linezolid was found to have better efficacy (P = 0.005) and similar safety for treating serious gram-positive infections . OBJECTIVE: The purpose of this study was to compare hospital resource use (primarily LOS) and cost of treatment between linezolid and teicoplanin for hospitalized patients with serious gram-positive infections in South America and Mexico using data from the multinational trial . METHODS: In a multinational, Phase IIIb, open-label, comparator-controlled trial, data were collected from hospitalized patients in centers in 6 South America can countries and Mexico with suspected or confirmed serious gram-positive infections . Patients were randomly assigned to receive i.v . linezolid 600 mg BID (for the entire treatment period {7-28 days} or switched to oral linezolid 600 mg BID) or i.v . teicoplanin (for the entire treatment period or switched to i.m . teicoplanin) dosed per approved prescription information . Data on direct medical resource utilization were collected for each patient, including duration and doses of study medication, location of hospitalization and LOS, comedications, tests and procedures, and outpatient service usage . Unit costs for the medical resources were obtained from secondary sources . RESULTS: A total of 203 patients (97 treated with linezolid and 106 treated with teicoplanin) were enrolled from these 7 countries . The unadjusted results showed that compared with teicoplanin, patients treated with linezolid had a 3.1-day shorter mean i.v . antibiotic treatment duration (P < 0.001), a 2.0- to 2.2-day shorter median and mean LOS (P = 0.03), and a 311 US dollars lower mean total cost of treatment (P = NS) . After controlling for age, race, sex, site of infection, inpatient location when the antibiotic treatment started, number of historical and current comorbidities, and whether the patient had a diagnosis of systemic inflammatory response syndrome or sepsis, the multivariate adjusted results were similar to the unadjusted results . The linezolid group had a 1.6-day shorter adjusted LOS or 66% greater odds of early discharge (P = 0.049) and a 335 US dollars lower adjusted mean total cost of treatment (P = NS) . CONCLUSION: Linezolid was associated with shorter LOS and duration of IV antibiotic treatment than teicoplanin for serious gram-positive infections in the population studied . Linezolid therapy has the potential to reduce the total cost of treatment.

Arch Microbiol, 2003 Sep, 180(3), 211 - 7 Epub 2003 Jul 10.
In situ growth of the novel SM1 euryarchaeon from a string-of-pearls-like microbial community in its cold biotope, its physical separation and insights into its structure and physiology; Moissl C et al.; Recently, a unique archaeal/bacterial community that grows in a macroscopically visible string-of-pearls-like structure in cold (~10 degrees C), sulfurous marsh water was discovered . Here, a new technique is described that allows the fast and reliable growth of these string-of-pearls-like microbial communities in larger quantities on polyethylene nets in nature . The microbial net population, estimated to consist of about 10,000 single pearls, can be harvested once a week and the archaeal cells selectively separated by density gradient centrifugation . As in native pearls, the archaeal cell fraction obtained consisted of a single type of coccoid cells only, 0.6 micro m in diameter . This novel type of euryarchaea has been tentatively named SM1 euryarchaeon . Electron microscopy and immuno-fluorescence in situ hybridization (immuno-FISH) revealed that about 100 pili-like fibers, up to 3 micro m in length, emanate radially from the surface of each cell . The SM1 euryarchaeal cells exhibited a viability of about 90% . The optimal conditions for viability were temperatures between -2 degrees C and 20 degrees C, pH 5-9, and low salt conditions; cell viability was independent of oxygen partial pressures . The cultures stained gram-positive, the cell wall was sensitive to SDS, EDTA and Proteinase K treatment . The cells did not exhibit the typical fluorescence for methanogens and did not contain coenzyme F(420) . The G+C-content was 34.5 mol%.

Biomaterials, 2003 Oct, 24(23), 4161 - 72
A highly sensitive cell assay for validation of purification regimes of alginates; Leinfelder U et al.; Among the hydrogels used for microencapsulation of cells and tissues, alginate has been and will continue to be one of the most important biomaterials . A mandatory requirement for clinical immunoisolated transplantations is the reproducible production of biocompatible alginate . As shown here for alginates extracted from freshly collected algal stipes, the current assays used for validation of the quality of the alginate are not sufficient to screen for impurities arising from spores of gram-positive bacteria (and related contaminants) . To assess the quality of alginate, we have developed a cell assay based on the induction of apoptosis in Jurkat cells . This assay allows in combination with the "modified mixed lymphocyte" assay a rapid and sensitive screening for any fibrosis-inducing impurities in alginate samples (even during the purification regime) as demonstrated by transplantation experiments performed in parallel with BB rats (exhibiting an elevated macrophage activity).The results clearly demonstrate that the quality of the input algal material is of key relevance for the production of transplantation-grade alginate.

Zhongguo Wei Zhong Bing Ji Jiu Yi Xue, 2003 Jan, 15(1), 32 - 4
{Efficacy and nephrotoxicity of vancomycin in the treatment of Gram positive infections}; Du B et al.; OBJECTIVE: To evaluate the efficacy and nephrotoxicity of vancomycin in the treatment of Gram positive infections . METHODS: A retrospective study of 84 patients with Gram positive infections who were treated with vancomycin in intensive care unit of Peking union medical college hospital . RESULTS: Clinical effective rate was 90% (76/84), and bacterial eradication rate was also 90% (76/84) . By univariate analysis, lower respiratory tract infection (LRTI) and longer duration of infection prior to vancomycin treatment were associated with poor clinical response, while LRTI and higher total dose of vancomycin were associated with poor bacteriologic response . The prevalence of nephrotoxicity ranged from 11% (9/84) to 14% (12/84) . There was no significant difference in terms of nephrotoxicity prevalence whichever criteria of nephrotoxicity was applied . Nephrotoxicity could be reversed, either during or after treatment, in 22% to 44% of patients . The development of nephrotoxicity was associated with LRTI and poor bacteriological response . CONCLUSION: Vancomycin is a reliable and safe antibiotic in the treatment of Gram positive infections.

J AOAC Int, 2003 May-Jun, 86(3), 449 - 52
Screening method for detecting cross-contamination residues of tiamulin in swine feeds; Rodriguez-Comesana M et al.; A method was developed for the determination of tiamulin (TML), 14-deoxy-14-{(2-diethylaminoethyl)mercaptoacetoxy}mutilin hydrogen fumarate, a semisynthetic derivative of the naturally occurring antibiotic pleuromutilin produced by the fungus Pleurotus mutilis . This drug, with high activity against Gram-positive bacteria, some Gram-negative bacteria, and several strains of mycoplasms is administered to animals in food, drinking water, or by injection; however, its chemical structure causes problems in analysis of feeds . Although the molecule is charged below pH 8, attempts to analyze TML-containing extracts on ion-exchange columns or other polar stationary phases have failed . Additionally, TML shows no fluorescence activity and only poor UV activity . The present method consists of organic solvent extraction followed by liquid chromatography with UV detection . A low wavelength (208 nm) was used for detection . Limits of detection and quantitation, as well as data for recovery and repeatability obtained during characterization of the method, are described . The applicability of the optimized method was tested by analyzing commercial blank feeds processed after TML-medicated feeds.

Forensic Sci Int, 2003 Jul 8, 134(2-3), 169 - 71
Fatal subarachnoid hemorrhage complicating actinomycotic meningitis; Koda Y et al.; Actinomycosis is caused by Gram-positive Actinomyces species that are part of the normal oral flora with low virulence . We describe a rare case of sudden death of a 48-year-old man with actinomycotic basilar meningitis that was complicated by fatal subarachnoid hemorrhage . Autopsy revealed meningitis at the basilar region of the brain, and histological examination revealed characteristic bacterial aggregates with extensive leukocyte infiltration and severe vasculitis of arteries of this region . Rupture of an artery by severe arteritis was thought to be the cause of the subarachnoid hemorrhage . The probable primary source of infection was found in the left lung . To the best of our knowledge, the complication of subarachnoid hemorrhage has not been reported previously in actinomycotic meningitis.

J Vasc Surg, 2003 Jul, 38(1), 83 - 7
Femoral endarteritis associated with percutaneous suture closure: new technology, challenging complications; Whitton Hollis H Jr et al.; OBJECTIVE: Use of percutaneous suture closure devices after catheter-based interventions is increasing . We recently have seen several severe femoral arterial wall infections after use of such devices . The purpose of this study was to examine the incidence, comorbid associations, and management of femoral arterial infections associated with percutaneous suture closure devices . METHODS: We retrospectively reviewed all infectious complications that occurred after 2223 consecutive cardiac catheterization procedures performed over 12 months in a university-affiliated community teaching hospital . Outcome variables included demographics, procedural details, infection, type of arterial reconstruction required, mortality, and limb loss . RESULTS: During this study, 822 patients received percutaneous suture devices . Infection developed in 6 patients (0.7%) . The incidence of diabetes in the population undergoing percutaneous suture closure was 219 of 822 patients (26.6%) . Three comorbid conditions, noted in multiple patients with infectious complications, included diabetes mellitus, obesity, and placement of a percutaneous suture closure device within the past 6 months . Invasive femoral endarteritis developed in 4 patients . Gram-positive cocci predominated in 4 patients . In 1 patient with polymicrobial infection catastrophic complications developed, including multiple anastomotic ruptures and hemorrhage . A new method of repair that incorporated double-thickness everted saphenous vein was used in 2 patients, and safe arterial closure was achieved . There was 1 late fatality on postoperative day 36 . Limb salvage was achieved in all patients . CONCLUSIONS: Femoral endarteritis complicating percutaneous suture closure is a challenging new problem for vascular surgeons and can result in catastrophic complications . Customary techniques that use saphenous vein patch or interposition grafting are not adequate in all circumstances . Successful outcome requires operative exploration in patients with suspected infection . Removal of the percutaneous suture closure device and debridement to normal arterial wall is recommended in all patients with suspected femoral endarteritis, based on positive intraoperative Gram stains or abnormal appearance of the adjacent femoral artery . Early success with an autologous bolstered repair is reported . Caution is advised when considering the use of a percutaneous suture closure device in patients with comorbid conditions including diabetes, obesity, and previously implanted devices.

J Exp Zoolog A Comp Exp Biol, 2003 Jul 1, 298(1), 16 - 22
Effects of acid stress in adult Rana pipiens; Brodkin M et al.; The decline in frog populations is a well-recognized worldwide phenomenon and infectious disease has been implicated as a major cause in the global decline of amphibian populations . Rana pipiens are disappearing from many habitats where they used to flourish, and environmental acidification has been considered as a possible contributor to this disappearance . We present a model that integrates the results of several experiments on the effects of acid exposure on natural resistance and mortality of adult Rana pipiens . These studies suggest that different components of the natural defense mechanisms of these frogs have different acid sensitivities . We have shown previously that exposure to pH 5.5 leads to a reduction in splenic white blood cell number, viability, and to colonization of the spleen with both Gram positive and Gram negative bacteria . In this paper we show that exposure to pH 6.0 did not affect the number or viability of splenic white blood cells but did result in colonization of the spleen by bacteria . We also show that cold exposure by itself does not cause a systemic bacterial infection in adult Rana pipiens, but acid stress following cold exposure does . The data presented in this paper provide empirical evidence to support the hypothesis that acid stress may be a contributor to the decline of Rana pipiens in the northeastern region of the United States .

Res Microbiol, 2003 Jun, 154(5), 337 - 44
Bacterial components induce cytokine and intercellular adhesion molecules-1 and activate transcription factors in dermal fibroblasts; Perfetto B et al.; This study investigated the effect of various structural components of Gram-positive (lipotheichoic acid and protein A) and Gram-negative (porins and lipopolysaccharide) bacteria on human dermal fibroblasts . Fibroblasts are important effector cells which have a potential role in augmenting the inflammatory response in various diseases . In this study we present a profile of TNF-alpha, IL-6 and IL-8, the expression of intercellular adhesion molecules (ICAM-1) and the activation of transcriptional nuclear factor NF-kB and AP-1 in human dermal fibroblasts stimulated by bacterial surface components . Compared to the controls, increased ICAM-1, IL-6 and IL-8 gene expression after stimulation of LPS and porins at 2 and 4 h was more evident than that obtained following stimulation of LTA and PA . Gene expression was also associated with the production of cytokine proteins in culture supernatants . TNF-alpha gene expression remained undetectable . Moreover, LPS and porin treatments determined IkBalpha phosphorylation and degradation in human dermal fibroblasts and the subsequent activation of nuclear factors NF-kB and AP-1 . These data suggest the importance of such stimuli in the first step of the inflammatory process, as well as the important role played by fibroblasts in skin inflammatory disease.

Physiol Behav, 2003 Jul, 79(2), 173 - 82
Pharmacokinetics and feeding responses to muramyl dipeptide in rats; Fosset S et al.; N-acetyl-muramyl-L-alanine-D-isoglutamine or muramyl dipeptide (MDP) is the minimally active subunit of bacterial peptidoglycan . During a systemic infection, the involvement of MDP has been demonstrated in food intake depression by the macrophage hydrolysis of Gram-positive bacteria . Under normal conditions, mammals are constantly exposed to the release of endogenous MDP from degraded gut flora and that of exogenous MDP from the diet . However, MDP digestion and absorption in the gastrointestinal tract are not fully understood, and their physiological significance needs to be clarified . After gavage (1.5 mg/kg), very low levels of MDP were found in the systemic circulation of rats and feeding patterns were not altered . In contrast, after the intraperitoneal injection of a similar dose, a depression in food intake was observed . The rats reduced their meal frequency and constant feeding rate, showing signs of satiety . The behavioral satiety sequence (BSS) was modified by behavioral changes, similar to those which appear during sickness, such as an increase in resting and a reduction in grooming . Our data suggest that the hypophagic effect of MDP may result from satiety and sickness behavior.

Zhonghua Liu Xing Bing Xue Za Zhi, 2003 Apr, 24(4), 296 - 9
{Application of multiplex semi-nested polymerase chain reaction in detection of pathogens in cerebrospinal fluid}; Yan ZY et al.; OBJECTIVE: To establish a new method of multiplex semi-nested polymerase chain reaction (PCR) to detect pathogens in cerebrospinal fluid (CSF) . METHODS: According to the analysis of the conservative and variable regions in bacterial 16S rRNA genes, we designed universal primers for all bacteria and specific primers for most gram-positive and gram-negative bacteria . All primers were added into the same reaction systems successively of a two-step PCR assay to amplify the different bacterial DNA in CSF, and the results were compared with common culture method with sensitivity and the specificity both detected at the same time . RESULTS: Both gram-positive and gram-negative bacteria amplified DNA fragment about 1,032 bp after first-step amplification with universal primers . In the second step, specific fragments of 336 bp and 127 bp were amplified in gram-positive and gram-negative bacteria respectively besides fragments of 1,032 bp; The detection limit for E . coli was 8 cfu/ml . The comparison of 62 CSF samples detected by both multiplex semi-PCR and conventional culture method revealed sensitivity, specificity, positive and negative values of 93.8%, 95.7%, 88.2%, and 97.8% respectively for PCR . CONCLUSION: The result suggested that the multiplex semi-nested PCR we established was sensitive, specific and rapid method for clinical laboratory to detect pathogens in CSF.

Arch Microbiol, 2003 Aug, 180(2), 88 - 100 Epub 2003 Jun 19.
A transporter of Escherichia coli specific for L- and D-methionine is the prototype for a new family within the ABC superfamily; Zhang Z et al.; An ABC-type transporter in Escherichia coli that transports both L- and D-methionine, but not other natural amino acids, was identified . This system is the first functionally characterized member of a novel family of bacterial permeases within the ABC superfamily . This family was designated the methionine uptake transporter (MUT) family (TC #3.A.1.23) . The proteins that comprise the transporters of this family were analyzed phylogenetically, revealing the probable existence of several sequence-divergent primordial paralogues, no more than two of which have been transmitted to any currently sequenced organism . In addition, MetJ, the pleiotropic methionine repressor protein, was shown to negatively control expression of the operon encoding the ABC-type methionine uptake system . The identification of MetJ binding sites (in gram-negative bacteria) or S-boxes (in gram-positive bacteria) in the promoter regions of several MUT transporter-encoding operons suggests that many MUT family members transport organic sulfur compounds.

Infect Immun, 2003 Jul, 71(7), 3979 - 87
Borrelia burgdorferi-induced tolerance as a model of persistence via immunosuppression; Diterich I et al.; If left untreated, infection with Borrelia burgdorferi sensu lato may lead to chronic Lyme borreliosis . It is still unknown how this pathogen manages to persist in the host in the presence of competent immune cells . It was recently reported that Borrelia suppresses the host's immune response, thus perhaps preventing the elimination of the pathogen (I . Diterich, L . Harter, D . Hassler, A . Wendel, and T . Hartung, Infect . Immun . 69:687-694, 2001) . Here, we further characterize Borrelia-induced immunomodulation in order to develop a model of this anergy . We observed that the different Borrelia preparations that we tested, i.e., live, heat-inactivated, and sonicated Borrelia, could desensitize human blood monocytes, as shown by attenuated cytokine release upon restimulation with any of the different preparations . Next, we investigated whether these Borrelia-specific stimuli render monocytes tolerant, i.e . hyporesponsive, towards another Toll-like receptor 2 (TLR2) agonist, such as lipoteichoic acid from gram-positive bacteria, or towards the TLR4 agonist lipopolysaccharide . Cross-tolerance towards all tested stimuli was induced . Furthermore, using primary bone marrow cells from TLR2-deficient mice and from mice with a nonfunctional TLR4 (strain C3H/HeJ), we demonstrated that the TLR2 was required for tolerance induction by Borrelia, and using neutralizing antibodies, we identified interleukin-10 as the key mediator involved . Although peripheral blood mononuclear cells tolerized by Borrelia exhibited reduced TLR2 and TLR4 mRNA levels, the expression of the respective proteins on monocytes was not decreased, ruling out the possibility that tolerance to Borrelia is attributed to a reduced TLR2 expression . In summary, we characterized tolerance induced by B . burgdorferi, describing a model of desensitization which might mirror the immunosuppression recently attributed to the persistence of Borrelia in immunocompetent hosts.

Biol Blood Marrow Transplant, 2003 Jun, 9(6), 373 - 82
Nonmyeloablative bone marrow transplantation: Infectious complications in 65 recipients of HLA-identical and mismatched transplants; Daly A et al.; Infections are a common complication of allogeneic bone marrow transplantation and the leading cause of transplantation-related mortality . It had been hypothesized that transplantation following nonmyeloablative preparative regimens would result in fewer infections by causing less mucosal injury, less graft-versus-host disease, and allowing earlier immune reconstitution . We have retrospectively reviewed the infectious complications of 65 consecutive patients with advanced hematologic malignancies who underwent bone marrow transplantation using a novel preparative regimen consisting of cyclophosphamide, thymic irradiation, and in vivo T-cell depletion . Cytomegalovirus (CMV) infection occurred in 52% of cases in which the donor or recipient had evidence of prior CMV exposure . Using a strategy of preemptive therapy and secondary prophylaxis with ganciclovir, no CMV disease occurred . Infections with gram-positive bacteria predominated over the first 100 days after bone marrow transplantation . Thereafter, the relative proportion of gram-negative infections increased without a significant increase in episodes of neutropenia . The rate of bacterial infections was not influenced by relapse of the underlying malignancy . Seven patients developed infections with Aspergillus species, which was the most common infectious cause of death in these patients . Infections with viruses other than CMV (n=10) and with protozoan organisms (n=2) also occurred . The use of HLA-mismatched donors, the occurrence of grade II-IV acute graft-versus-host disease, and treatment with corticosteroids did not influence the risk of CMV or bacterial or fungal infections in patients who underwent transplantation following this preparative regimen . Overall, the incidence and spectrum of infections in this series was similar to the reported incidence of infections following conventional myeloablative allogeneic stem cell transplantation . We conclude that a quantitative T-cell deficiency in these extensively T-cell depleted patients may be a risk factor for infection, even in the absence of graft-versus-host disease.

J Bacteriol, 2003 Jul, 185(13), 3828 - 41
Identification of pyrene-induced proteins in Mycobacterium sp . strain 6PY1: evidence for two ring-hydroxylating dioxygenases; Krivobok S et al.; In this study, the enzymes involved in polycyclic aromatic hydrocarbon (PAH) degradation were investigated in the pyrene-degrading Mycobacterium sp . strain 6PY1 . {(14)C}pyrene mineralization experiments showed that bacteria grown with either pyrene or phenanthrene produced high levels of pyrene-catabolic activity but that acetate-grown cells had no activity . As a means of identifying specific catabolic enzymes, protein extracts from bacteria grown on pyrene or on other carbon sources were analyzed by two-dimensional gel electrophoresis . Pyrene-induced proteins were tentatively identified by peptide sequence analysis . Half of them resembled enzymes known to be involved in phenanthrene degradation, with closest similarity to the corresponding enzymes from Nocardioides sp . strain KP7 . The genes encoding the terminal components of two distinct ring-hydroxylating dioxygenases were cloned . Sequence analysis revealed that the two enzymes, designated Pdo1 and Pdo2, belong to a subfamily of dioxygenases found exclusively in gram-positive bacteria . When overproduced in Escherichia coli, Pdo1 and Pdo2 showed distinctive selectivities towards PAH substrates, with the former enzyme catalyzing the dihydroxylation of both pyrene and phenanthrene and the latter preferentially oxidizing phenanthrene . The catalytic activity of the Pdo2 enzyme was dramatically enhanced when electron carrier proteins of the phenanthrene dioxygenase from strain KP7 were coexpressed in recombinant cells . The Pdo2 enzyme was purified as a brown protein consisting of two types of subunits with M(r)s of about 52,000 and 20,000 . Immunoblot analysis of cell extracts from strain 6PY1 revealed that Pdo1 was present in cells grown on benzoate, phenanthrene, or pyrene and absent in acetate-grown cells . In contrast, Pdo2 could be detected only in PAH-grown cells . These results indicated that the two enzymes were differentially regulated depending on the carbon source used for growth.

Br J Ophthalmol, 2003 Jul, 87(7), 834 - 8
Bacterial keratitis: predisposing factors, clinical and microbiological review of 300 cases; Bourcier T et al.; AIM: To identify predisposing factors and to define clinical and microbiological characteristics of bacterial keratitis in current practice . METHODS: A retrospective analysis of the hospital records of patients presenting with bacterial keratitis and treated at the Quinze-Vingts National Center of Ophthalmology, Paris, France, was performed during a 20 month period . A bacterial keratitis was defined as a suppurative corneal infiltrate and overlying epithelial defect associated with presence of bacteria on corneal scraping and/or that was cured with antibiotic therapy . Risk factors, clinical and microbiological data were collected . RESULTS: 300 cases (291 patients) of presumed bacterial keratitis were included . Potential predisposing factors, usually multiple, were identified in 90.6% of cases . Contact lens wear was the main risk factor (50.3%) . Trauma or a history of keratopathy was found in 15% and 21% of cases, respectively . An organism was identified in 201 eyes (68%) . 83% of the infections involved Gram positive bacteria, 17% involved Gram negative bacteria, and 2% were polymicrobial . Gram negative bacteria were associated with severe anterior chamber inflammation (p=0.004), as well as greater surface of infiltrates (p=0.01) . 99% of ulcers resolved with treatment, but only 60% of patients had visual acuity better than the level at admission, and 5% had very poor visual outcome . CONCLUSIONS: Contact lens wear is the most important risk factor . Most community acquired bacterial ulcers resolve with appropriate treatment.

Arch Insect Biochem Physiol, 2003 Jul, 53(3), 125 - 33
Cloning and expression of gallerimycin, an antifungal peptide expressed in immune response of greater wax moth larvae, Galleria mellonella; Schuhmann B et al.; A novel defensin-like peptide was identified in the greater wax moth, Galleria mellonella . It was discovered in a haemocyte cDNA bank enriched with transcripts upregulated after immune challenge via subtractive hybridisation and suppressive PCR . The deduced amino acid sequence of the defensin-like peptide exhibits similarities to the antifungal peptides drosomycin from Drosophila melanogaster and heliomicin from Heliothis virescens . Therefore, it has been termed gallerimycin . Upregulation of gallerimycin after stimulation of the immune system by LPS-injection was demonstrated by quantitative real-time PCR . A full-size cDNA was cloned and overexpressed in Escherichia coli Origami cells in order to obtain a functional peptide with disulfide bridges . The recombinant peptide was active against the entomopathogenic fungus Metarhizium anisopliae, but not against yeast, gram-negative and gram-positive bacteria .

Transplantation, 2003 Jun 15, 75(11), 1898 - 900
Disseminated encephalitozoon cuniculi infection in a Mexican kidney transplant recipient; Gamboa-Dominguez A et al.; BACKGROUND: No cases of Encephalitozoon cuniculi infection have been reported in transplant patients . METHODS: A 42-year-old man received a renal transplant 8 months earlier because of terminal glomerulonephritis and was admitted with cough, fever, diarrhea, abdominal pain, and colon wall thickening . While under rapamycin (2 g/day), cyclosporine A (4.4 mg/kg/day), and prednisone (100 mg/day) therapy, he developed Banff grade IB graft rejection and was treated with methylprednisolone (1 g/day) for 3 days and oral prednisone (60 mg/d) . RESULTS: Microbiologic studies were inconclusive, and biopsy specimens of ileum, colon, liver, and the grafted kidney revealed numerous gram-positive microsporidia spores . Parasitophorous vacuoles containing various developing stages of Encephalitozoon were seen . Immunofluorescence studies identified the etiologic agent as E . cuniculi . Albendazole therapy resulted in clinical improvement but no eradication after 10 months of follow-up . CONCLUSIONS: This report describes what is, to the authors' knowledge, the first case of disseminated E . cuniculi infection in a kidney transplant human immunodeficiency virus-negative patient from Mexico.

J Exp Med, 2003 Jun 16, 197(12), 1745 - 54
Stimulus-dependent deacylation of bacterial lipopolysaccharide by dendritic cells; Lu M et al.; We describe here a previously unrecognized property of dendritic cells (DCs), the ability to deacylate the lipid A moiety of gram-negative bacterial LPSs . Both immature DCs of the XS52 cell line and bone marrow-derived DCs produce acyloxyacyl hydrolase, an enzyme that detoxifies LPS by selectively removing the secondary acyl chains from lipid A . Acyloxyacyl hydrolase expression decreased when DCs were incubated with IL-4, IL-1 beta, TNF alpha, and an agonistic CD40 antibody (maturation cocktail), and increased after treatment with LPS, CpG oligodeoxynucleotides, or a gram-positive bacterium (Micococcus luteus) . Maturation cocktail treatment also diminished, whereas LPS treatment enhanced or maintained the cells' ability to kill Escherichia coli, deacylate LPS, and degrade bacterial protein . Enzymatic deacylation of LPS is an intrinsic, regulated mechanism by which DCs may modulate host responses to this potent bacterial agonist.

Int J Syst Evol Microbiol, 2003 May, 53(Pt 3), 871 - 4
Amycolatopsis keratiniphila sp . nov., a novel keratinolytic soil actinomycete from Kuwait; Al-Musallam AA et al.; A keratinolytic, nocardioform actinomycete, strain D2(T), was isolated from Kuwait marsh soil by the bait technique using animal wool . Strain D2(T) was an aerobic, Gram-positive organism that produced light-grey aerial mycelium but no specific spore chains . 16S rDNA sequence analyses and chemotaxonomic markers were consistent with the classification of strain D2(T) in the genus Amycolatopsis, i.e . it had meso-diaminopimelic acid in its peptidoglycan, arabinose and galactose as its diagnostic sugars, the polar lipids phosphatidylinositol, phosphatidylethanolamine, hydroxyphosphatidylethanolamine and diphosphatidylglycerol, menaquinone MK-9(H4) and an iso-/anteiso-branched fatty acid pattern combined with 10-methyl-branched and 2-hydroxy-branched fatty acids . Amycolatopsis japonica was the closest phylogenetic neighbour of strain D2(T), showing 99.4% 16S rDNA sequence similarity with the novel strain . A . japonica and strain D2(T) could be clearly separated from each other on the basis of their low DNA-DNA reassociation value (55.9%) . These data, together with its distinct physiological traits, led to the conclusion that strain D2(T) represented a novel species within the genus Amycolatopsis, for which the name Amycolatopsis keratiniphila (type strain D2(T) = DSM 44409(T) = NRRL B24117(T)) is proposed.

Int J Syst Evol Microbiol, 2003 May, 53(Pt 3), 811 - 4
Allofustis seminis gen . nov., sp . nov., a novel Gram-positive, catalase-negative, rod-shaped bacterium from pig semen; Collins MD et al.; An unknown Gram-positive, catalase-negative, facultatively anaerobic, non-spore-forming, rod-shaped bacterium originating from semen of a pig was characterized using phenotypic, molecular chemical and molecular phylogenetic methods . Chemical studies revealed the presence of a directly cross-linked cell wall murein based on L-lysine and a DNA G + C content of 39 mol% . Comparative 16S rRNA gene sequencing showed that the unidentified rod-shaped organism formed a hitherto unknown subline related, albeit loosely, to Alkalibacterium olivapovliticus, Alloiococcus otitis, Dolosigranulum pigrum and related organisms, in the low-G + C-content Gram-positive bacteria . However, sequence divergence values of > 11% from these recognized taxa clearly indicated that the novel bacterium represents a separate genus . Based on phenotypic and phylogenetic considerations, it is proposed that the unknown bacterium from pig semen be classified as a new genus and species, Allofustis seminis gen . nov., sp . nov . The type strain is strain 01-570-1(T) (= CCUG 45438(T) = CIP 107425(T)).

Am J Clin Oncol, 2003 Jun, 26(3), 285 - 8
Cefepime versus ticarcillin and clavulanate potassium and aztreonam for febrile neutropenia therapy in high-dose chemotherapy patients; Fleming DR et al.; An open labeled randomized trial comparing the efficacy and cost of empirically applied cefepime (C) as monotherapy versus combination therapy consisting of ticarcillin and clavulanate potassium and aztreonam (T/A) was performed in febrile neutropenic patients following high-dose chemotherapy (HDC) +/- radiation, with or without peripheral blood stem cell support . Over a 28-month period, 126 patients were screened and included in the study . Using afebrile status following 3 days of therapy as a primary endpoint, both regimens produced comparable clinical response rates (C = 55% vs . T/A = 61%) . Also, the use of vancomycin for resistant gram-positive infections and alteration of gram-negative infection coverage was similar in both groups (C = 40% vs . T/A = 47% and C = 29% vs . T/A = 24%) . Both treatment groups had similar needs for empirical antifungal therapy (C = 25% vs . T/A = 22%) . There was a postrandomization difference between the two groups in that the "C" group had a significantly higher number of allogeneic transplants and non-stem-cell-supported patients, whereas the "T/A" group had a significantly greater number of autologous peripheral blood stem cell patients (p < 0.0001) . Despite this difference, the C group had a significantly lower cost ratio than the T/A group (p = 0.016) . In conclusion, we have shown that C treatment of febrile neutropenic patients following HDC results in similar efficacy and lower cost when compared to T/A, despite the inclusion of higher risk patients in the C group.

Nihon Kokyuki Gakkai Zasshi, 2003 Apr, 41(4), 276 - 81
{A case of pulmonary actinomycosis with a unique finding in the chest MR image}; Sito T et al.; A 57-year old man, who was complaining of a productive cough and right shoulder pain, was admitted to our hospital because of an irregularly shaped mass located at rt . S1 on a chest radiograph . Bronchoscopy revealed no evidence suggesting lung cancer or any specific infection, either pathologically or microbiologically . CT-guided biopsy revealed changes resembling lymphocytic or plasmocytic interstitial pneumonitis with thickening of the alveolar septum and with accumulations of mononuclear cells and plasma cells, indicating the proliferation of bronchus-associated lymphoid tissue (BALT system) . Since no definitive diagnosis was considered possible, a right upper lobectomy was performed . Histopathologic examination of tissue from the right upper lobe revealed sulfur granules and branching Gram-positive filamentous bacteria, and the condition was pathologically diagnosed as pulmonary actinomycosis . In the center of the mass lesion, the patient's chest MRI showed a very small area with a low signal intensity in T1- and a high signal in T2-weighted images, which suggested an accumulation of fluid in the actinomycotic abscess . As detailed MR findings in this condition have not been well described in the literature, the MRI evidence seen in this case may be useful for the diagnosis of actinomycosis.

Microbiol Mol Biol Rev, 2003 Jun, 67(2), 277 - 301, table of contents
Conjugative plasmid transfer in gram-positive bacteria; Grohmann E et al.; Conjugative transfer of bacterial plasmids is the most efficient way of horizontal gene spread, and it is therefore considered one of the major reasons for the increase in the number of bacteria exhibiting multiple-antibiotic resistance . Thus, conjugation and spread of antibiotic resistance represents a severe problem in antibiotic treatment, especially of immunosuppressed patients and in intensive care units . While conjugation in gram-negative bacteria has been studied in great detail over the last decades, the transfer mechanisms of antibiotic resistance plasmids in gram-positive bacteria remained obscure . In the last few years, the entire nucleotide sequences of several large conjugative plasmids from gram-positive bacteria have been determined . Sequence analyses and data bank comparisons of their putative transfer (tra) regions have revealed significant similarities to tra regions of plasmids from gram-negative bacteria with regard to the respective DNA relaxases and their targets, the origins of transfer (oriT), and putative nucleoside triphosphatases NTP-ases with homologies to type IV secretion systems . In contrast, a single gene encoding a septal DNA translocator protein is involved in plasmid transfer between micelle-forming streptomycetes . Based on these clues, we propose the existence of two fundamentally different plasmid-mediated conjugative mechanisms in gram-positive microorganisms, namely, the mechanism taking place in unicellular gram-positive bacteria, which is functionally similar to that in gram-negative bacteria, and a second type that occurs in multicellular gram-positive bacteria, which seems to be characterized by double-stranded DNA transfer.

J Clin Microbiol, 2003 Jun, 41(6), 2788 - 90
Tuboovarian abscess caused by Atopobium vaginae following transvaginal oocyte recovery; Geissdorfer W et al.; A 39-year-old woman with tubarian sterility fell ill with acute pelvic inflammatory disease 2 months after transvaginal oocyte recovery . Laparotomy revealed a large tuboovarian abscess, from which Atopobium vaginae, an anaerobic gram-positive coccoid bacterium of hitherto unknown clinical significance, was isolated . The microbial etiology and the risk of pelvic infections following transvaginal punctures are discussed.

Appl Environ Microbiol, 2003 Jun, 69(6), 3103 - 9
Characterization of the microbial community in indoor environments: a chemical-analytical approach; Sebastian A et al.; An integrated procedure is presented whereby gas chromatography-ion trap mass spectrometry is used to determine chemical markers of gram-negative bacterial lipopolysaccharide (3-hydroxy fatty acids with 10 to 18 carbon atoms), gram-positive bacteria (branched-chain fatty acids with 15 and 17 carbon atoms), bacterial peptidoglycan (muramic acid), and fungal biomass (ergosterol) in samples of settled house dust . A hydrolysate of (13)C-labeled cyanobacterial cells is used as an internal standard for the first three markers . These analyses require two dust samples, one for 3-OH fatty acids, branched-chain fatty acids, and muramic acid and another for ergosterol . The method may be used to characterize microbial communities in environmental samples.

Pediatrics, 2003 Jun, 111(6 Pt 1), 1411 - 5
Platelet count and sepsis in very low birth weight neonates: is there an organism-specific response?
Guida JD, Kunig AM, Leef KH, McKenzie SE, Paul DA.
OBJECTIVE: Thrombocytopenia is commonly observed in very low birth weight (VLBW) neonates with sepsis . Specific platelet responses to different infectious agents have not been extensively characterized . The objectives of this study were to examine platelet counts and platelet indices in preterm neonates with culture-proven sepsis to determine if there are organism-specific platelet responses . STUDY DESIGN: We analyzed a cohort of all VLBW neonates (birthweight <1500 g) born over a 4-year period and admitted to a single level III neonatal intensive care unit (N = 943) . Thrombocytopenia was defined as a platelet count <100,000/mm(3) . Platelet count, nadir, duration of thrombocytopenia, and mean platelet volume (MPV) were examined during episodes of culture-proven sepsis . Analysis of variance, Kruskal-Wallis, Mann-Whitney U, and chi(2) tests were used to compare groups, and data are expressed as mean +/- standard deviation . RESULTS: Sepsis was diagnosed in 154 (16%) of 943 patients in the study population . Of the sepsis episodes, 54% were associated with thrombocytopenia and 61% with an elevation in MPV . Infections were grouped by organism type: Gram-positive bacteria (117/154, 76%), Gram-negative bacteria (24/154, 16%), and fungi (13/154, 8%) . When compared with patients with Gram-positive sepsis, those with Gram-negative or fungal sepsis had a significantly lower initial platelet count, a lower platelet nadir, a higher incidence of thrombocytopenia, and a greater duration of thrombocytopenia . The decrease in platelet count from baseline was also significantly less in the Gram-positive infections than in the fungal infections . Although there was an overall increase in MPV from baseline, there were no differences between groups . CONCLUSIONS: In our population of VLBW infants, sepsis is frequently associated with thrombocytopenia and an elevation in MPV . However, fungal and Gram-negative pathogens are associated with a lower platelet count and more prolonged thrombocytopenia compared with Gram-positive pathogens . We conclude that common pathogens causing sepsis have different effects on platelet kinetics.

J Biol Chem, 2003 Jul 18, 278(29), 26319 - 22 Epub 2003 May 30.
Functional diversity of the Drosophila PGRP-LC gene cluster in the response to lipopolysaccharide and peptidoglycan; Werner T et al.; The peptidoglycan recognition protein PGRP-LC is a major activator of the imd/Relish pathway in the Drosophila immune response . Three transcripts are generated by alternative splicing of the complex PGRP-LC gene . The encoded transmembrane proteins share an identical intracellular part, but each has a separate extracellular PGRP-domain: x, y, or a . Here we show that two of these isoforms play unique roles in the response to different microorganisms . Using RNA interference in Drosophila mbn-2 cells, we found that PGRP-LCx is the only isoform required to mediate signals from Gram-positive bacteria and purified bacterial peptidoglycan . By contrast, the recognition of Gram-negative bacteria and bacterial lipopolysaccharide requires both PGRP-LCa and LCx . The third isoform, LCy, is expressed at lower levels and may be partially redundant . Two additional PGRP domains in the gene cluster, z and w, are both included in a single transcript of a separate gene, PGRP-LF . Suppression of this transcript does not block the response to any of the microorganisms tested.

Biochem Soc Trans, 2003 Jun, 31(Pt 3), 659 - 63
Evolutionary relationships, but functional differences, between the Drosophila and human Toll-like receptor families; Gangloff M et al.; The Toll receptor was first found to function in the dorsoventral patterning pathway of Drosophila embryos . It is activated by a specific protein ligand, Spatzle, generated at ventral positions in the early embryo . Drosophila Toll (dToll) also functions in innate immune responses to Gram-positive bacteria and fungi, and Spatzle is required for this response . We have shown that Spatzle is necessary and sufficient for activation of the dToll pathway, and that it probably acts by cross-linking two molecules of Toll to form homodimers . In the present paper, we contrast this mode of regulation with that proposed for the vertebrate Toll-like receptor family, which mediate analogous responses to pathogen pattern antigens . In contrast with dToll, these receptors appear to be activated by direct exposure to pathogen patterns, such as peptidoglycan and lipopolysaccharide . We discuss the evolutionary basis of this functional divergence of the vertebrate and invertebrate Toll-like receptors.

Biochem Soc Trans, 2003 Jun, 31(Pt 3), 648 - 51
Toll and Toll-like receptors in Drosophila; Bilak H et al.; The Drosophila Toll receptor controls the immune response to Gram-positive bacteria and fungi by activating a signalling pathway partially conserved throughout evolution . The Drosophila genome encodes eight additional Toll-related receptors, most of which appear to carry out developmental rather than immune functions . One exception may be Toll-9, which shares structural and functional similarities with mammalian TLRs.

ALTEX, 2003, 20(2), 59 - 63
Comparison of the reactivity of human and rabbit blood towards pyrogenic stimuli; Schindler S et al.; A comparison between humans and rabbits was performed based on stimulation of whole blood with well-known pyrogens from Gram-negative and Gram-positive bacteria, such as lipopolysaccharide (LPS) and lipoteichoic acid (LTA), respectively . The reactivity was measured as release of IL-1 beta and IL-8 by ELISA . The reactivity of the two species towards LPS was found to be similar, whereas their reactivity towards LTA differed considerably . Differences between the levels of IL-1 beta and IL-8 release were observed in both species . This finding suggests that the In vitro Pyrogen Test (IPT) which uses human blood to detect contaminations, e.g . of injectable drugs, might predict the human reaction to the contamination better than the "gold standard" rabbit pyrogen test.

J Nat Prod, 2003 May, 66(5), 699 - 702
Sesquiterpene lactones from Anthemis altissima and their anti-Helicobacter pylori activity; Konstantinopoulou M et al.; Seven sesquiterpene lactones, (-) sivasinolide (1), a new naturally occurring eudesmanolide (altissin, 2), desacetyl-beta-cyclopyrethrosin (3), tatridin-A (4), 1-epi-tatridin B (5), 1alpha,10beta-epoxy-6-hydroxy-1,10H-inunolide (6), and spiciformin (7), were isolated from Anthemis altissima . Also isolated were 10 known flavonoids, namely, apigenin (8), kaempferol 4'-methyl ether (9), quercetin (10), quercetin 3-methyl ether (11), isorhamnetin (12), rhamnetin (13), 6-hydroxyquercetin 3,6,4'-trimethyl ether (14), isoquercetrin (15), taxifolin (16), and eriodictyol (17), and one phenolic acid, chlorogenic acid (18) . The structure and the stereochemistry of compound 2 were deduced by spectroscopic methods . The in vitro activity of the sesquiterpene lactones (1-5) against Helicobacter pylori, as well as against three Gram-positive and three Gram-negative bacteria growing aerobically, was tested using the microdilution method . Compounds 8-18 have also been tested against H . pylori.

J Biochem (Tokyo), 2003 Mar, 133(3), 271 - 7
SRCL/CL-P1 recognizes GalNAc and a carcinoma-associated antigen, Tn antigen; Yoshida T et al.; SRCL /CL-P1 was recently identified as a scavenger receptor with a C-type lectin domain, which was expressed in vascular endothelial cells and could bind to Gram-positive and Gram-negative bacteria, yeast and oxidized LDL . We found that SRCL was expressed in some but not all nurse-like cells examined . Furthermore, to characterize the C-type lectin domain of SRCL, the secreted form of the C-type lectin domain (LEC-AP) of SRCL, which was fused to the signal sequence of IgG and alkaline phosphatase, was expressed in 293/EBNA-1 cells and the culture medium was used for the in vitro binding assay . LEC-AP specifically bound to GalNAc-conjugated gel in a Ca(2+)-dependent manner, and this binding was inhibited by free GalNAc, L-, D-fucose, D-galactose, lactose, and especially T antigen and Tn antigen . Furthermore, we examined whether or not SRCL could take up saccharide-conjugated particles . 293/EBNA-1 cells stably expressing SRCL were found to take up GalNAc but not mannose-conjugated particles on confocal microscopy . The binding of GalNAc-conjugated particles to these cells was quantitatively measured by comparing the x-means of individual cell populations . An approximately 2.1-fold increase in immunofluorescence intensity was observed for the SRCL transfectants compared to control vector transfectants . Our results provide a basis for understanding the scavenger function of SRCL as to carbohydrate-containing ligands.

J Vet Pharmacol Ther, 2003 Jun, 26(3), 193 - 8
Efficacy of intramammary treatment with procaine penicillin G vs . procaine penicillin G plus neomycin in bovine clinical mastitis caused by penicillin-susceptible, gram-positive bacteria--a double blind field study; Taponen S et al.; The efficacy of intramammary treatments containing procaine penicillin G alone (treatment A) or a combination of procaine penicillin G and neomycin (treatment B) was compared in treating clinical bovine mastitis caused by gram-positive bacteria susceptible in vitro to penicillin G . Both treatments were supplemented with a single intramuscular injection of procaine penicillin G on the first day of treatment . The study was carried out using a double blind design on commercial dairy farms in Southern Finland . A total of 56 quarters were treated with treatment A and 61 with treatment B . The cure rates for both treatments were equal, which suggests that the use of the penicillin G-aminoglycoside combination does not increase the efficacy of the treatment over that achieved by using penicillin G alone in bovine clinical mastitis caused by penicillin-susceptible, gram-positive bacteria.

Biol Cell, 2003 Jan-Feb, 95(1), 53 - 7
Molecular characterization of a defensin in the IZD-MB-0503 cell line derived from immunocytes of the insect Mamestra brassicae (Lepidoptera); Mandrioli M et al.; The induction of anti-microbial peptides against Gram positive and negative bacteria in the IZD-MB-0503 cell line from the lepidopteran Mamestra brassicae is demonstrated, while no anti-fungal activity is detected . The identification of a defensin-like molecule active against Gram positive bacteria is described for the first time in Lepidoptera . This molecule shows between 43% and 59% homology with group A defensins from other dipteran and hymenopteran species.

J Food Prot, 2003 May, 66(5), 847 - 53
In vitro assessment of the cytotoxicity of nisin, pediocin, and selected colicins on simian virus 40-transfected human colon and Vero monkey kidney cells with trypan blue staining viability assays; Murinda SE et al.; Gram-positive bacterial bacteriocins (nisin and pediocin) and gram-negative bacterial bacteriocins (colicins {Col} E1, E3, E6, E7, and K) were evaluated for cytotoxicity against cultured simian virus 40-transfected human colon (SV40-HC) and Vero monkey kidney (Vero) cells . Bacteriocin-treated cells were assessed for viability by trypan blue staining . Monolayers of SV40-HC and Vero cells were cultured in tissue culture plates (35 degrees C, 10% CO2 in humidified air) with the use of Dulbecco's modified Eagle's medium supplemented with 10% (vol/vol) calf serum . Actively growing cells in the log phase (ca . 10(4) cells per ml) were treated with individual partially purified bacteriocin preparations at 170, 350, and 700 activity units per ml . Duplicate culture plates for each bacteriocin treatment and untreated controls were withdrawn after 16, 32, and 48 h of incubation . Cells were dissociated with trypsin and treated with trypan blue and were then counted in a hemocytometer with the use of a phase-contrast microscope . Viability assays indicated dose-dependent toxicity for some bacteriocins . Nisin, pediocin, and Col E6 were the most cytotoxic bacteriocins; SV40-HC cells demonstrated greater sensitivity than Vero cells did . Some bacteriocins can be toxic to mammalian cells; therefore, bacteriocins intended for use as biopreservatives must be evaluated for toxicity to mammalian cells and for other toxicities . Col E1, Col E3, Col E7, and Col K demonstrated little toxicity at the activities tested, indicating that they are safe and thus have potential for use as food biopreservatives.

Int J Food Microbiol, 2003 Jun 25, 83(3), 281 - 93
The effect of abrupt osmotic shifts on the lag phase duration of foodborne bacteria; Mellefont LA et al.; The effects of osmotic environment and inoculum history on lag times were examined . Abrupt osmotic shifts of cultures were found to induce lag phases in a variety of foodborne bacteria . Relative lag times (RLT; the ratio of lag time to generation time) were used to differentiate the effects of the shift from those of the outgrowth environment . In general, osmotic downshifts induced larger RLTs than equivalent upshifts . An observed reduction in RLT at very low a(w), however, was unexpected . For an osmotic downshift, differences were observed in the RLT response of the Gram-negative and -positive strains tested . RLTs were usually extended for Gram-negative organisms as conditions became less favourable for growth . In comparison, RLT remained relatively unaffected for Gram-positive organisms . The observations reported in this study demonstrate that lag time can be understood in terms of the amount of work to be done to adjust to new environmental conditions and the rate at which that work is done, and are consistent with known strategies for osmoregulation employed by the various organisms studied.

Drugs Today (Barc), 2001 Sep, 37(9), 617 - 628
Telithromycin (HMR 3647): The first ketolide antibiotic; Xiong YQ et al.; Telithromycin, the first ketolide antibiotic to undergo clinical development, has been specifically designed to treat community-acquired respiratory tract infections, including those caused by resistant pathogens . Like macrolides, telithromycin inhibits protein synthesis by acting mainly on the 50S ribosomal subunit . The defining structural characteristic is a keto function in place of the C3-cladinose moiety, which greatly improves acid stability and confers a lack of induction of MLSb resistance . Telithromycin provides potent activity against a wide spectrum of Gram-positive and Gram-negative organisms, including erythromycin-resistant pneumococci and atypical/intracellular organisms . Preliminary results from clinical trials have demonstrated that telithromycin may provide a convenient and effective compact treatment option for select respiratory tract infections such as community-acquired pneumonia, acute bacteria exacerbations of chronic bronchitis, acute sinusitis and tonsillitis/pharyngitis . (c) 2001 Prous Science . All rights reserved.

J Biol Chem, 2003 Jul 18, 278(29), 26410 - 7 Epub 2003 May 09.
Biosynthesis of dTDP-3-acetamido-3,6-dideoxy-alpha-D-galactose in Aneurinibacillus thermoaerophilus L420-91T; Pfoestl A et al.; The glycan chain of the S-layer protein of Aneurinibacillus thermoaerophilus L420-91T (DSM 10154) consists of d-rhamnose and 3-acetamido-3,6-dideoxy-d-galactose (d-Fucp3NAc) . Thymidine diphosphate-activated d-Fucp3NAc serves as precursor for the assembly of structural polysaccharides in Gram-positive and Gram-negative organisms . The biosynthesis of dTDP-3-acetamido-3,6-dideoxy-alpha-d-galactose (dTDP-d-Fucp3NAc) involves five enzymes . The first two steps of the reaction are catalyzed by enzymes that are part of the well studied dTDP-l-rhamnose biosynthetic pathway, namely d-glucose-1-phosphate thymidyltransferase (RmlA) and dTDP-d-glucose-4,6-dehydratase (RmlB) . The enzymes catalyzing the last three synthesis reactions have not been characterized biochemically so far . These steps include an isomerase, a transaminase, and a transacetylase . We identified all five genes involved by chromosome walking in the Gram-positive organism A . thermoaerophilus L420-91T and overexpressed the three new enzymes heterologously in Escherichia coli . The activities of these enzymes were monitored by reverse phase high performance liquid chromatography, and the intermediate products formed were characterized by 1H and 13C nuclear magnetic resonance spectroscopy analysis . Alignment of the newly identified proteins with known sequences revealed that the elucidated pathway in this Gram-positive organism may also be valid in the biosynthesis of the O-antigen of lipopolysaccharides of Gram-negative organisms . The key enzyme in the biosynthesis of dTDP-d-Fucp3NAc has been identified as an isomerase, which converts the 4-keto educt into the 3-keto product, with concomitant epimerization at C-4 to produce a 6-deoxy-d-xylo configuration . This is the first report of the functional characterization of the biosynthesis of dTDP-d-Fucp3NAc and description of a novel type of isomerase capable of synthesizing dTDP-6-deoxy-d-xylohex-3-ulose from dTDP-6-deoxy-d-xylohex-4-ulose.

BMC Microbiol . 2003 May 09;3(1):10.
Genomic homogeneity between Mycobacterium avium subsp . avium and Mycobacterium avium subsp . paratuberculosis belies their divergent growth rates; Bannantine JP et al.; BACKGROUND: Mycobacterium avium subspecies avium (M . avium) is frequently encountered in the environment, but also causes infections in animals and immunocompromised patients . In contrast, Mycobacterium avium subspecies paratuberculosis (M . paratuberculosis) is a slow-growing organism that is the causative agent of Johne's disease in cattle and chronic granulomatous infections in a variety of other ruminant hosts . Yet we show that despite their divergent phenotypes and the diseases they present, the genomes of M . avium and M . paratuberculosis share greater than 97% nucleotide identity over large (25 kb) genomic regions analyzed in this study . RESULTS: To characterize genome similarity between these two subspecies as well as attempt to understand their different growth rates, we designed oligonucleotide primers from M . avium sequence to amplify 15 minimally overlapping fragments of M . paratuberculosis genomic DNA encompassing the chromosomal origin of replication . These strategies resulted in the successful amplification and sequencing of a contiguous 11-kb fragment containing the putative Mycobacterium paratuberculosis origin of replication (oriC) . This fragment contained 11 predicted open reading frames that showed a conserved gene order in the oriC locus when compared with several other Gram-positive bacteria . In addition, a GC skew analysis identified the origin of chromosomal replication which lies between the genes dnaA and dnaN . The presence of multiple DnaA boxes and the ATP-binding site in dnaA were also found in M . paratuberculosis . The strong nucleotide identity of M . avium and M . paratuberculosis in the region surrounding the origin of chromosomal replication led us to compare other areas of these genomes . A DNA homology matrix of 2 million nucleotides from each genome revealed strong synteny with only a few sequences present in one genome but absent in the other . Finally, the 16s rRNA gene from these two subspecies is 100% identical . CONCLUSIONS: We present for the first time, a description of the oriC region in M . paratuberculosis . In addition, genomic comparisons between these two mycobacterial subspecies suggest that differences in the oriC region may not be significant enough to account for the diverse bacterial replication rates . Finally, the few genetic differences present outside the origin of chromosomal replication in each genome may be responsible for the diverse growth rates or phenotypes observed between the avium and paratuberculosis subspecies.

Am J Respir Crit Care Med, 2003 Jul 15, 168(2), 158 - 64 Epub 2003 May 08.
Toll-like receptor-mediated tumor necrosis factor and interleukin-10 production differ during systemic inflammation; Adib-Conquy M et al.; Major trauma is associated with a decreased capacity of patients' leukocytes to produce proinflammatory cytokines on in vitro stimulation . We studied leukocytes from 48 patients with trauma and showed that this hyporeactivity was restricted to gram-negative bacteria, Escherichia coli endotoxin, and unmethylated bacterial DNA, whereas Leptospira interrogans endotoxin-induced tumor necrosis factor production was similar to that observed with healthy donors . As well, tumor necrosis factor and interleukin-6 production in response to gram-positive bacteria was not altered . The expression of toll-like receptor (TLR) 2 was not reduced on patients' monocytes as compared with healthy control subjects, whereas that of TLR4 was reduced . However, the hyporeactivity to gram-negative bacteria and E . coli endotoxin cannot be fully explained by the downregulation of TLR4 . Indeed, unlike proinflammatory cytokines, after stimulation with these microbial products the release of antiinflammatory cytokines was increased as compared with healthy control subjects . The increased interleukin-10 production was analyzed in terms of intracellular signaling in peripheral blood mononuclear cells from patients with trauma: our results suggest the involvement of p38 mitogen-activated protein kinase, Sp-1 transcription factor, heterotrimeric Gi protein, and phosphatidylinositol-3'-kinase . In conclusion, the immunodysregulation described for patients with trauma is not a generalized phenomenon but depends on the stimulus and the signaling pathway.

Mar Pollut Bull, 2003 May, 46(5), 573 - 86
Changes in zooxanthellae density, morphology, and mitotic index in hermatypic corals and anemones exposed to cyanide; Cervino JM et al.; Sodium cyanide (NaCN) is widely used for the capture of reef fish throughout Southeast Asia and causes extensive fish mortality, but the effect of NaCN on reef corals remains debated . To document the impact of cyanide exposure on corals, the species Acropora millepora, Goniopora sp., Favites abdita, Trachyphyllia geoffrio, Plerogyra sp., Heliofungia actinformis, Euphyllia divisa, and Scarophyton sp., and the sea anemone Aiptasia pallida were exposed to varying concentrations of cyanide for varying time periods . Corals were exposed to 50, 100, 300, and 600 mg/l of cyanide ion (CN(-)) for 1-2 min (in seawater, the CN(-) forms hydrocyanic acid) . These concentrations are much lower than those reportedly used by fish collectors . Exposed corals and anemones immediately retracted their tentacles and mesenterial filaments, and discharged copious amounts of mucus containing zooxanthellae . Gel electrophoreses techniques found changes in protein expression in both zooxanthellae and host tissue . Corals and anemones exposed to cyanide showed an immediate increase in mitotic cell division of their zooxenthellae, and a decrease in zooxanthellae density . In contrast, zooxanthellae cell division and density remained constant in controls . Histopathological changes included gastrodermal disruption, mesogleal degradation, and increased mucus in coral tissues . Zooxanthellae showed pigment loss, swelling, and deformation . Mortality occurred at all exposure levels . Exposed specimens experienced an increase in the ratio of gram-negative to gram-positive bacteria on the coral surface . The results demonstrate that exposure cyanide causes mortality to corals and anemones, even when applied at lower levels than that used by fish collectors . Even brief exposure to cyanide caused slow-acting and long-term damage to corals and their zooxanthellae.

J Immunol, 2003 May 15, 170(10), 5203 - 9
Cumulative Toll-like receptor activation in human macrophages treated with whole bacteria; Nau GJ et al.; Purified components from bacteria selectively activate Toll-like receptors (TLR), leading to shared and unique responses in innate immune cells . Whole bacteria contain agonists for multiple TLR and induce a common macrophage activation program of transcription . It is not known, however, whether the stimulation of specific TLR by whole bacteria results in differential activation of the innate immune system . We evaluated gene expression data from human macrophages and found a unique gene expression profile induced by Gram-negative bacteria . In contrast, Gram-positive bacteria evoked few specific alterations in gene expression . LPS, a TLR4-specific ligand, was sufficient to elicit the distinct expression profile observed with Gram-negative bacteria . TLR4 activation regulated gene expression by both an IFN-dependent and an IFN-independent mechanism, illustrated by I-TAC and IL-12 p70, respectively . IL-12 p70 was produced by cells in whole blood exposed to Gram-negative bacteria, demonstrating faithful reproduction of the macrophage response in mixed populations of cells and identifying a potential diagnostic marker of infection . Our results show that the macrophage response to bacteria is dominated by the accumulated input from multiple TLR . For macrophages exposed to Gram-negative bacteria, gene expression changes encompass those induced by Gram-positive bacteria plus a distinct TLR4 response . This distinct TLR4 response may provide the basis to diagnose clinical Gram-negative infections.

Appl Environ Microbiol, 2003 May, 69(5), 2928 - 35
An improved protocol for quantification of freshwater Actinobacteria by fluorescence in situ hybridization; Sekar R et al.; We tested a previously described protocol for fluorescence in situ hybridization of marine bacterioplankton with horseradish peroxidase-labeled rRNA-targeted oligonucleotide probes and catalyzed reporter deposition (CARD-FISH) in plankton samples from different lakes . The fraction of Bacteria detected by CARD-FISH was significantly lower than after FISH with fluorescently monolabeled probes . In particular, the abundances of aquatic Actinobacteria were significantly underestimated . We thus developed a combined fixation and permeabilization protocol for CARD-FISH of freshwater samples . Enzymatic pretreatment of fixed cells was optimized for the controlled digestion of gram-positive cell walls without causing overall cell loss . Incubations with high concentrations of lysozyme (10 mg ml(-1)) followed by achromopeptidase (60 U ml(-1)) successfully permeabilized cell walls of Actinobacteria for subsequent CARD-FISH both in enrichment cultures and environmental samples . Between 72 and >99% (mean, 86%) of all Bacteria could be visualized with the improved assay in surface waters of four lakes . For freshwater samples, our method is thus superior to the CARD-FISH protocol for marine Bacteria (mean, 55%) and to FISH with directly fluorochrome labeled probes (mean, 67%) . Actinobacterial abundances in the studied systems, as detected by the optimized protocol, ranged from 32 to >55% (mean, 45%) . Our findings confirm that members of this lineage are among the numerically most important Bacteria of freshwater picoplankton.

Chemosphere, 2003 Jul, 52(1), 185 - 93
Separation of the strength and selectivity of the microbiological effect of synthetic dyes by spectral mapping technique; Oros G et al.; The growth inhibitory effect of 30 synthetic dyes on 22 bacteria (test organisms) belonging to various taxonomic groups was determined . The strength (potency) and selectivity of the biological effect were separated by the spectral mapping technique, reducing the dimensionality of the selectivity maps to two by the nonlinear mapping technique . The relationship between biological effect and physicochemical parameters of dyes was elucidated by stepwise regression analysis . It has been established that the strength of the effect of anthracene and trityl derivatives was higher than that of azobenzene dyes and significantly depended on the hydrophobicity of the compound . The selectivity of the effect also depended on hydrophobicity and on the nonpolar unsaturated surface area of the dyes . Gram negative and Gram positive bacteria differed in the strength and selectivity of their response to dyes indicating the marked impact of the taxonomical position on the response . Contrary to other multivariate mathematical statistical methods biological activity may be divided by SPM into potency and selectivity values, therefore, application of the technique in future QSAR studies is highly recommended.

Int J Syst Evol Microbiol, 2003 Mar, 53(Pt 2), 569 - 76
Conexibacter woesei gen . nov., sp . nov., a novel representative of a deep evolutionary line of descent within the class Actinobacteria; Monciardini P et al.; A novel Gram-positive bacterial strain was isolated from forest soil . According to its 16S rRNA sequence, this strain is a deep-rooting member of the class Actinobacteria . The 16S rRNA sequence is most closely related (approximately 94% identity) to clones of uncultured bacteria detected in different terrestrial environments, while showing only a remote relationship (approximately 90% identity or less) to sequences of cultured species . Cells of the first cultured representative of this phylogenetic cluster are small, short rods that are motile by peritrichous flagella, catalase- and oxidase-positive and grow under aerobic conditions . In liquid culture, flagella from different cells can aggregate to form networks, clearly visible under the light microscope . The peptidoglycan contains meso-diaminopimelic acid and is directly cross-linked (type A1gamma) . Mycolic acids are not present . The polar lipids are phosphatidylinositol and an unidentified phospholipid . Menaquinone MK-7(H4) was detected as the predominant isoprenoid quinone . Oleic, 14-methylpentadecanoic, hexadecanoic and omega6c-heptadecenoic acids are the predominant components of the cellular fatty acid profile . The DNA G + C content is 71 mol% . The distinct phylogenetic position and the unusual combination of chemotaxonomic characteristics justify the proposal of a new genus and species, Conexibacter woesei gen . nov., sp . nov., with the type strain ID131577T (=DSM 14684T =JCM 11494T).

Int J Syst Evol Microbiol, 2003 Mar, 53(Pt 2), 547 - 53
Phylogenetic analysis of some Sporomusa sub-branch members isolated from human clinical specimens: description of Megasphaera micronuciformis sp . nov; Marchandin H et al.; Two unknown, Gram-negative, catalase-negative and strictly anaerobic cocci were isolated from two independent human samples (strains AIP 49.01 and AIP 412.00T) . Comparative 16S rRNA gene sequencing demonstrated that these two organisms displayed 99.8% sequence identity and that they are members of the Sporomusa sub-branch of the low-G + C Gram-positive bacteria . The most closely related 16S rDNA sequences were from Megasphaera sp . oral clone BU057 (99.8%) and from isolates of Megasphaera cerevisiae and Megasphaera elsdenii (94.5 and 93.8%, respectively) . Phylogenetic analysis based on 16S rDNA sequences showed that these two strains were most closely related to M . elsdenii and belonged to the Megasphaera genus . Differences from previously described Megasphaera species in terms of size, biochemical tests (particularly the analysis of metabolic end products), gas production and DNA G+C content indicated that the two strains studied represent a novel species of anaerobic Gram-negative cocci . The name Megasphaera micronuciformis sp . nov . is proposed for these two isolates . It is also proposed that the uncultured organism previously deposited as Megasphaera sp . oral clone BU057 should be named 'Candidatus Megasphaera micronuciformis' . The type strain of Megasphaera micronuciformis is AIP 412.00T (=CIP 107280T =CCUG 45952T).

Int J Syst Evol Microbiol, 2003 Mar, 53(Pt 2), 485 - 90
Solirubrobacter pauli gen . nov., sp . nov., a mesophilic bacterium within the Rubrobacteridae related to common soil clones; Singleton DR et al.; A novel bacterium, strain B33D1T, isolated from agricultural soil, was characterized taxonomically and phylogenetically . Strain B33D1T was a Gram-positive, aerobic rod of medium length that formed long chains on a common laboratory medium . However, B33D1T grew poorly on the surface of agar plates and was sensitive to desiccation . The optimal growth temperature was 30 degrees C (range 19-38 degrees C) . The organism grew well on a variety of sugars and was capable of utilizing a few amino acids as sole carbon sources . Phylogenetically, the most closely related described species to strain B33D1T was Rubrobacter xylanophilus, which possessed 86% 16S rRNA sequence similarity . However, a number of 16S rRNA gene clones derived from soil samples possessed up to 93% sequence similarity . These results placed strain B33D1T within the subclass Rubrobacteridae of the phylum Actinobacteria . The novel genus and species Solirubrobacter pauli gen . nov., sp . nov . is proposed, with strain B33D1T (=ATCC BAA-492T =DSM 14954T) as the type strain.

Tunis Med, 2003 Jan, 81(1), 63 - 6
{Endometrial actinomycosis . Two case reports}; Chatti S et al.; Actinomyces is an aerobic, Gram positive bacteria saprophyte of the genital tract . The endometrial involvement is extremely rare . The authors report two cases of pelvic actinomycosis in a 58 and 55 year old women, the second using intrauterine device for 11 years.

Biochemistry, 2003 Apr 29, 42(16), 4717 - 26
S-Ribosylhomocysteinase (LuxS) is a mononuclear iron protein; Zhu J et al.; S-Ribosylhomocysteinase (LuxS) catalyzes the cleavage of the thioether linkage of S-ribosylhomocysteine (SRH) to produce L-homocysteine and 4,5-dihydroxy-2,3-pentanedione (DHPD) . This is a key step in the biosynthetic pathway of the type II autoinducer (AI-2) in both Gram-positive and Gram-negative bacteria . Previous studies demonstrated that LuxS contains a divalent metal cofactor, which has been proposed to be a Zn(2+) ion . To gain insight into the catalytic mechanism of this unusual reaction and the function of the metal cofactor, we developed an efficient expression and purification system to produce LuxS enriched in either Fe(2+), Co(2+), or Zn(2+) . Comparison of the catalytic properties and stability of the metal-substituted LuxS with those of the native enzyme revealed that the native metal ion is Fe(2+) . The electronic absorption spectrum of the Co(II)-substituted LuxS underwent dramatic catalysis-dependent changes, suggesting the direct involvement of the metal ion in catalysis . Site-directed mutagenesis studies showed that Glu-57 and Cys-84 are essential for catalysis, most likely acting as general acids/bases . Reaction in D(2)O resulted in the incorporation of deuterium at the C-1, C-2, and C-5 positions of the diketone product . These data suggest a catalytic mechanism in which the metal ion catalyzes an intramolecular redox reaction, shifting the carbonyl group from the C-1 position to the C-3 position of the ribose . Subsequent beta-elimination at the C-4 and C-5 positions releases homocysteine as a free thiol.

Proc Natl Acad Sci U S A, 2003 Apr 29, 100(9), 5052 - 6 Epub 2003 Apr 17.
The riboswitch-mediated control of sulfur metabolism in bacteria; Epshtein V et al.; Many operons in Gram-positive bacteria that are involved in methionine (Met) and cysteine (Cys) biosynthesis possess an evolutionarily conserved regulatory leader sequence (S-box) that positively controls these genes in response to methionine starvation . Here, we demonstrate that a feed-back regulation mechanism utilizes S-adenosyl-methionine as an effector . S-adenosyl-methionine directly and specifically binds to the nascent S-box RNA, causing an intrinsic terminator to form and interrupt transcription prematurely . The S-box leader RNA thus expands the family of newly discovered riboswitches, i.e., natural regulatory RNA aptamers that seem to sense small molecules ranging from amino acid derivatives to vitamins.

J Clin Microbiol, 2003 Apr, 41(4), 1363 - 9
16S ribosomal DNA sequence-based analysis of clinically significant gram-positive anaerobic cocci; Song Y et al.; Sequence analysis of the 16S rRNA gene represents a highly accurate and versatile method for bacterial classification and identification, even when the species in question is notoriously difficult to identify by phenotypic means . However, its use for identification based on public sequence databases is not without limitation due to the presence of ambiguous data in the databases . In this study, we evaluated the utility of 16S ribosomal DNA sequencing as a means of identifying clinically important gram-positive anaerobic cocci (GPAC) by sequencing 13 type strains of established GPAC species and 156 clinical isolates that had been studied only by phenotypic tests . Among the 13 type strains of GPAC species we tested, only 4 gave a "perfect" match with their corresponding sequences in GenBank, whereas the other 9 had lower sequence similarities (<98%) . This indicates that data in the public database may be inaccurate at times . Based on the sequences of the 13 type strains obtained in this study, 84% (131 of 156) of the clinical isolates were accurately identified to species level, with the remaining 25 clinical strains revealing nine unique sequences that may represent eight novel species . This finding is in contrast to the phenotypic identification results, by which only 56% of isolates were correctly identified to species level.

N Engl J Med, 2003 Apr 17, 348(16), 1546 - 54
The epidemiology of sepsis in the United States from 1979 through 2000; Martin GS et al.; BACKGROUND: Sepsis represents a substantial health care burden, and there is limited epidemiologic information about the demography of sepsis or about the temporal changes in its incidence and outcome . We investigated the epidemiology of sepsis in the United States, with specific examination of race and sex, causative organisms, the disposition of patients, and the incidence and outcome . METHODS: We analyzed the occurrence of sepsis from 1979 through 2000 using a nationally representative sample of all nonfederal acute care hospitals in the United States . Data on new cases were obtained from hospital discharge records coded according to the International Classification of Diseases, Ninth Revision, Clinical Modification . RESULTS: Review of discharge data on approximately 750 million hospitalizations in the United States over the 22-year period identified 10,319,418 cases of sepsis . Sepsis was more common among men than among women (mean annual relative risk, 1.28 {95 percent confidence interval, 1.24 to 1.32}) and among nonwhite persons than among white persons (mean annual relative risk, 1.90 {95 percent confidence interval, 1.81 to 2.00}) . Between 1979 and 2000, there was an annualized increase in the incidence of sepsis of 8.7 percent, from about 164,000 cases (82.7 per 100,000 population) to nearly 660,000 cases (240.4 per 100,000 population) . The rate of sepsis due to fungal organisms increased by 207 percent, with gram-positive bacteria becoming the predominant pathogens after 1987 . The total in-hospital mortality rate fell from 27.8 percent during the period from 1979 through 1984 to 17.9 percent during the period from 1995 through 2000, yet the total number of deaths continued to increase . Mortality was highest among black men . Organ failure contributed cumulatively to mortality, with temporal improvements in survival among patients with fewer than three failing organs . The average length of the hospital stay decreased, and the rate of discharge to nonacute care medical facilities increased . CONCLUSIONS: The incidence of sepsis and the number of sepsis-related deaths are increasing, although the overall mortality rate among patients with sepsis is declining . There are also disparities among races and between men and women in the incidence of sepsis . Gram-positive bacteria and fungal organisms are increasingly common causes of sepsis .

FEMS Microbiol Rev, 2003 Apr, 27(1), 65 - 74
Ionophore resistance of ruminal bacteria and its potential impact on human health; Russell JB et al.; In recent years, there has been a debate concerning the causes of antibiotic resistance and the steps that should be taken . Beef cattle in feedlots are routinely fed a class of antibiotics known as ionophores, and these compounds increase feed efficiency by as much as 10% . Some groups have argued that ionophore resistance poses the same public health threat as conventional antibiotics, but humans are not given ionophores to combat bacterial infection . Many ruminal bacteria are ionophore-resistant, but until recently the mechanism of this resistance was not well defined . Ionophores are highly lipophilic polyethers that accumulate in cell membranes and catalyze rapid ion movement . When sensitive bacteria counteract futile ion flux with membrane ATPases and transporters, they are eventually de-energized . Aerobic bacteria and mammalian enzymes can degrade ionophores, but these pathways are oxygen-dependent and not functional in anaerobic environments like the rumen or lower GI tract . Gram-positive ruminal bacteria are in many cases more sensitive to ionophores than Gram-negative species, but this model of resistance is not always clear-cut . Some Gram-negative ruminal bacteria are initially ionophore-sensitive, and even Gram-positive bacteria can adapt . Ionophore resistance appears to be mediated by extracellular polysaccharides (glycocalyx) that exclude ionophores from the cell membrane . Because cattle not receiving ionophores have large populations of resistant bacteria, it appears that this trait is due to a physiological selection rather than a mutation per se . Genes responsible for ionophore resistance in ruminal bacteria have not been identified, but there is little evidence that ionophore resistance can be spread from one bacterium to another . Given these observations, use of ionophores in animal feed is not likely to have a significant impact on the transfer of antibiotic resistance from animals to man.

Nat Immunol, 2003 May, 4(5), 478 - 84
The Drosophila immune system detects bacteria through specific peptidoglycan recognition; Leulier F et al.; The Drosophila immune system discriminates between different classes of infectious microbes and responds with pathogen-specific defense reactions through selective activation of the Toll and the immune deficiency (Imd) signaling pathways . The Toll pathway mediates most defenses against Gram-positive bacteria and fungi, whereas the Imd pathway is required to resist infection by Gram-negative bacteria . The bacterial components recognized by these pathways remain to be defined . Here we report that Gram-negative diaminopimelic acid-type peptidoglycan is the most potent inducer of the Imd pathway and that the Toll pathway is predominantly activated by Gram-positive lysine-type peptidoglycan . Thus, the ability of Drosophila to discriminate between Gram-positive and Gram-negative bacteria relies on the recognition of specific forms of peptidoglycan.

J Microbiol Methods, 2003 Jun, 53(3), 295 - 307
Detection of fatty acids from intact microorganisms by molecular beam static secondary ion mass spectrometry; Ingram JC et al.; We report the use of a surface analysis approach, static secondary ion mass spectrometry (SIMS) equipped with a molecular (ReO(4)(-)) ion primary beam, to analyze the surface of intact microbial cells . SIMS spectra of 28 microorganisms were compared to fatty acid profiles determined by gas chromatographic analysis of transesterfied fatty acids extracted from the same organisms . The results indicate that surface bombardment using the molecular primary beam cleaved the ester linkage characteristic of bacteria at the glycerophosphate backbone of the phospholipid components of the cell membrane . This cleavage enables direct detection of the fatty acid conjugate base of intact microorganisms by static SIMS . The limit of detection for this approach is approximately 10(7) bacterial cells/cm(2) . Multivariate statistical methods were applied in a graded approach to the SIMS microbial data . The results showed that the full data set could initially be statistically grouped based upon major differences in biochemical composition of the cell wall . The gram-positive bacteria were further statistically analyzed, followed by final analysis of a specific bacterial genus that was successfully grouped by species . Additionally, the use of SIMS to detect microbes on mineral surfaces is demonstrated by an analysis of Shewanella oneidensis on crushed hematite . The results of this study provide evidence for the potential of static SIMS to rapidly detect bacterial species based on ion fragments originating from cell membrane lipids directly from sample surfaces.

J Am Soc Mass Spectrom, 2003 Apr, 14(4), 342 - 51
Experimental factors affecting the quality and reproducibility of MALDI TOF mass spectra obtained from whole bacteria cells; Williams TL et al.; Numerous experimental factors are shown to significantly influence the spectra obtained when bacteria are analyzed by MALDI TOF/MS . Detailed investigation of the instrument parameters and sample preparation are all shown to influence the spectra . Of these, the preanalysis sample preparation steps incorporate the most important elements influencing the quality and reproducibility of the spectra . Some of the most important sample preparation factors include the method employed for sterilization, the type of matrix, the matrix solvent and concentration of cells in the matrix, as well as the type and concentration of acid added to the matrix . The effects of these parameters, as well as other aspects of sample preparation and the effects of several instrumental parameters on spectra are presented . Optimization and control of all experimental variables leads to a stable protocol for analysis of bacteria . The protocol employs a Nd:Yag laser and describes both sample handling and instrument conditions which consistently yield reproducible MALDI TOF mass spectra with greater than 25 peaks from both gram-positive and gram-negative bacteria.

J Med Microbiol, 2003 Apr, 52(Pt 4), 309 - 13
Specific PCR detection of Peptostreptococcus magnus; Riggio MP et al.; Peptostreptococcus magnus is the most pathogenic and one of the most common Gram-positive anaerobic cocci found in human clinical specimens . The organism has been isolated in pure culture from a range of serious infections, including meningitis and endocarditis . However, isolation of Peptostreptococcus magnus from the oral cavity has rarely been attempted . Identification of Peptostreptococcus magnus in clinical specimens is reliant upon microbiological culture and biochemical methods, which often give ambiguous results . The aim of this study was to develop a PCR assay for the specific detection of Peptostreptococcus magnus in oral clinical specimens . PCR primers specific for Peptostreptococcus magnus DNA were derived by comparison of 16S rRNA gene sequences and selection of primers that demonstrated specificity at their 3' ends for Peptostreptococcus magnus . PCR positivity for Peptostreptococcus magnus DNA was indicated by the amplification of a 553 bp product . The PCR assay was then used to attempt detection of Peptostreptococcus magnus DNA in subgingival plaque samples from adult periodontitis patients and pus aspirates from subjects with acute dento-alveolar abscesses . The PCR assay was demonstrated to be highly specific for Peptostreptococcus magnus DNA, since no PCR products were obtained when genomic DNA from a wide range of other oral bacteria, including closely related Peptostreptococcus species, was used in the PCR assay . Confirmation of specific amplification of Peptostreptococcus magnus DNA was obtained by digestion of PCR products with the restriction endonuclease RsaI, which gives a unique restriction profile for Peptostreptococcus magnus . Of the 33 subgingival plaque samples analysed, 2 (6 %) were positive for Peptostreptococcus magnus DNA . None of the 60 pus aspirates analysed was positive for Peptostreptococcus magnus DNA . It is concluded that Peptostreptococcus magnus is not a major pathogen in adult periodontitis or dento-alveolar abscesses . The PCR assay provides a more rapid, specific and sensitive alternative to conventional methods for identification of Peptostreptococcus magnus in clinical specimens.

Mol Microbiol, 2003 Apr, 48(2), 349 - 60
A Streptomyces coelicolor functional orthologue of Escherichia coli RNase E shows shuffling of catalytic and PNPase-binding domains; Lee K et al.; Previous work has detected an RNase E-like endoribonucleolytic activity in cell extracts obtained from Streptomyces . Here, we identify a Streptomyces coelicolor gene, rns, encoding a 140 kDa protein (RNase ES) that shows endoribonucleolytic cleavage specificity characteristic of RNase E, confers viability on and allows propagation of Escherichia coli cells lacking RNase E and accomplishes RNase E-like regulation of plasmid copy number in E . coli . However, notwithstanding its complementation of rne-deleted E . coli, RNase ES did not accurately process 9S rRNA from E . coli . Additionally, whereas RNase E is normally required for E . coli survival, rns is not an essential gene in S . coelicolor . Deletion analysis mapped the catalytic domain of RNase ES near its centre and showed that regions located near the RNase ES termini interact with an S . coelicolor homologue of polynucleotide phosphorylase (PNPase) - a major component of E . coli RNase E-based degradosomes . The interacting arginine- and proline-rich segments resemble the C-terminally located degradosome scaffold region of E . coli RNase E . Our results indicate that RNase ES is a structurally shuffled RNase E homologue showing evolutionary conservation of functional RNase E-like enzymatic activity, and suggest the existence of degradosome-like complexes in Gram-positive bacteria.

Sheng Wu Gong Cheng Xue Bao, 2002 Nov, 18(6), 662 - 6
{Cloning of the replication origin (oriC) of Streptoverticillum caespitosus and transformation of Streptomyces lividans ZX7}; Ma W et al.; Streptoverticillum caespitosus ATCC27422 is a producing strain of mitomycin A for cancer therapy . Taking the advantage of the conserved sequences of genes flanking the oriC of high G + C Gram-positive bacteria, a 1.3 kb DNA fragment containing oriC and its flanking region was cloned by PCR . Nuleotide sequence comparisons revealed that the cloned fragment is more than 80% identical to the same region of S . coelicolor . There are 22 DnaA-boxes in the oriC region, and the conserved sequence of DnaA-box is TTGTCCACA . The plasmid containing the oriC of S . caespitosus was constructed (pMJ9), and it was able to transform the protoplast of Streptomyces lividans ZX7 at the frequency of 3.2 x 10(2) transformants/micrograms plasmid DNA . The colony and mycelia's morphology of the transformants are normal . The constructed plasmid can exist stable in the host as a low copy extra-chromosome replicon . The high rate of the homology and the cross genus replication initiation activity suggests close relationship between Streptomyces and Streptoverticillum in the evolution . While the maximum likelihood phylogenetic tree based upon the oriC of S . caespitosus and several Streptomyces spp . revealed that S . caespitosus differed extensively from the Streptomyces spp . This result supports the separation of Streptoverticillum from Streptomyces.

Eur J Immunol, 2003 Apr, 33(4), 1127 - 38
Cooperation between toll-like receptor 2 and 4 in the brain of mice challenged with cell wall components derived from gram-negative and gram-positive bacteria; Laflamme N et al.; In this study we investigated whether induction of toll-like receptor 2 (TLR2) amplifies the effect of a cell wall component derived from gram-positive bacteria, namely peptidoglycan (PGN) . Mice received a first systemic lipopolysaccharide (LPS) injection to pre-induce TLR2 in various regions of the brain, and 6 h later, a second administration of either LPS or PGN . The data show a robust transcriptional activation of TLR2, TNF-alpha and monocyte chemotactic protein-1 (MCP-1) in microglial cells of mice challenged twice with LPS, whereas PGN essentially abolished this response . TLR4 plays a critical role in this process, because C3H/HeJ mice no longer responded to LPS but exhibited a normal reaction to PGN . Conversely, a robust signal for genes encoding innate immune proteins was found in the brain of TLR2-deficient mice challenged with LPS . However, the second LPS bolus failed to trigger TNF-alpha and IL-12 in TLR2-deficient mice, while the same treatment caused a strong induction of these genes in the cerebral tissue of wild-type littermates . The present data provide evidence that cooperation exists between TLR4 and TLR2 . While TLR4 is absolutely necessary to engage the innate immune response in the brain, TLR2 participates in the regulation of genes encoding TNF-alpha and IL-12 during severe endotoxemia . Such collaboration between TLR4 and TLR2 may be determinant for the transfer from the innate to the adaptive immunity within the CNS of infected animals.

Hepatology, 2003 Apr, 37(4), 897 - 901
Spontaneous bacterial peritonitis in asymptomatic outpatients with cirrhotic ascites; Evans LT et al.; The prevalence and natural history of spontaneous bacterial peritonitis in asymptomatic patients with ascites secondary to cirrhosis is unknown . From a prospectively recorded database, we reviewed the clinical and laboratory features of all outpatients with cirrhotic ascites undergoing paracentesis between July 1994 and December 2000 . The prevalence of spontaneous bacterial peritonitis in the population of 427 cirrhotic outpatients as defined by neutrocytic ascites (absolute neutrophil count >or=250 cells/mm(3)) was 3.5% . Of the 15 patients with neutrocytic ascites, 6 were culture positive (1.4%) and 9 culture negative (2.1%) . Eight other patients (1.9%) had bacterascites . The organisms cultured from ascitic fluid in these asymptomatic patients with culture positive neutrocytic ascites and bacterascites were predominantly gram positive . No patient developed hepatorenal syndrome, and 1-year survival of 67% was better than historical data from hospitalized patients with spontaneous bacterial peritonitis . Moreover, patients who did not receive antibiotics for neutrocytic ascites fared no worse than patients who did receive antibiotics . In conclusion, spontaneous bacterial peritonitis in outpatients with cirrhotic ascites is less frequent, occurs in patients with less advanced liver disease, and may have a better outcome than its counterpart in hospitalized patients . In addition, the organisms cultured from ascitic fluid in outpatients are predominantly gram positive . A reassessment of diagnostic criteria for spontaneous bacterial peritonitis in outpatients may be required.

Environ Microbiol, 2003 Apr, 5(4), 321 - 6
Reproduction and metabolism at - 10 degrees C of bacteria isolated from Siberian permafrost; Bakermans C et al.; We report the isolation and properties of several species of bacteria from Siberian permafrost . Half of the isolates were spore-forming bacteria unable to grow or metabolize at subzero temperatures . Other Gram-positive isolates metabolized, but never exhibited any growth at - 10 degrees C . One Gram-negative isolate metabolized and grew at - 10 degrees C, with a measured doubling time of 39 days . Metabolic studies of several isolates suggested that as temperature decreased below + 4 degrees C, the partitioning of energy changes with much more energy being used for cell maintenance as the temperature decreases . In addition, cells grown at - 10 degrees C exhibited major morphological changes at the ultrastructural level.

J Perinat Med, 2003, 31(1), 60 - 8
Early markers of late-onset sepsis in premature neonates: clinical, hematological and cytokine profile; Gonzalez BE et al.; BACKGROUND: Late-onset sepsis in the NICU is a major problem associated with high morbidity and mortality . OBJECTIVE: To determine if clinical characteristics, hematological parameters and serial measurements of serum IL-6 and IL-8 can detect late-onset sepsis in premature neonates prior to positive blood cultures . DESIGN: The study was done in 2 phases . The first phase (S1) was a retrospective evaluation of clinical signs and timing of blood culture positivity in all neonates with late-onset cultures proven sepsis from 1991-1998 . The second phase (S2) was a prospective study that enrolled infants > or = 72 hours old, suspected of sepsis based on the presence of criteria identified in S1 . At that time (day 0), blood was drawn for a CBC with differential, blood culture, IL-6 and IL-8 levels; cytokine levels were repeated on day 1 . Infants with positive cultures were diagnosed as confirmed sepsis; those with negative cultures, as no sepsis . RESULTS: S1: Of the 48 episodes of culture proven, late-onset sepsis, 54% of the blood cultures were positive by 24 hours and 90% by 48 hours . The most common presenting signs were desaturations (50%) and increased gastric residuals (33%); I/T ratio > 0.16 differentiated between gram-positive, negative and fungal infections (p = 0.007) . S2: 27 infants were enrolled . Eight (mean {SEM} gestational age of 28.2 {0.94} weeks; birth weight of 1.15 {0.11} kg) had positive blood cultures; 19 (gestational age of 27.7 {0.9} weeks; birth weight of 1.06 {0.13} kg) had no sepsis . Infants with sepsis were more likely to have apnea/bradycardia (p = 0.002); no differences in hematological profile, as compared to those with no sepsis . Seven (88%) infants had positive blood cultures by 48 hours . Median values of IL-6 (pg/ml) were higher in infants with sepsis vs . those with no sepsis on days 0 {40 vs . 13} (p = 0.03) and 1 {24 vs . 9} (p < 0.001) . IL-8 levels were not significantly different . CONCLUSIONS: In both S1 and S2, a majority of the blood cultures were positive by 48 hours . IL-6 levels on days 0 and 1 were significantly higher in infants with confirmed sepsis, prior to the blood culture being positive . IL-6 levels may be useful in the initiation as well as early termination of antibiotic therapy in late-onset neonatal sepsis.

Int J Syst Evol Microbiol, 2003 Jan, 53(Pt 1), 105 - 12
Detection and characterization of Pasteuria 16S rRNA gene sequences from nematodes and soils; Duan YP et al.; Various bacterial species in the genus Pasteuria have great potential as biocontrol agents against plant-parasitic nematodes, although study of this important genus is hampered by the current inability to cultivate Pasteuria species outside their host . To aid in the study of this genus, an extensive 16S rRNA gene sequence phylogeny was constructed and this information was used to develop cultivation-independent methods for detection of Pasteuria in soils and nematodes . Thirty new clones of Pasteuria 16S rRNA genes were obtained directly from nematodes and soil samples . These were sequenced and used to construct an extensive phylogeny of this genus . These sequences were divided into two deeply branching clades within the low-G + C, Gram-positive division; some sequences appear to represent novel species within the genus Pasteuria . In addition, a surprising degree of 16S rRNA gene sequence diversity was observed within what had previously been designated a single strain of Pasteuria penetrans (P-20) . PCR primers specific to Pasteuria 16S rRNA for detection of Pasteuria in soils were also designed and evaluated . Detection limits for soil DNA were 100-10,000 Pasteuria endospores (g soil)(-1).

Int J Syst Evol Microbiol, 2003 Jan, 53(Pt 1), 99 - 103
Xylanimonas cellulosilytica gen . nov., sp . nov., a xylanolytic bacterium isolated from a decayed tree (Ulmus nigra); Rivas R et al.; A bacterial strain, designated XIL07T, isolated from a decayed tree, Ulmus nigra, in Salamanca (Spain) produced abundant cellulases and xylanases . The micro-organism was Gram-positive, aerobic, coccoid and non-motile . Growth was observed on many carbohydrates, including cellulose and xylan as the sole carbon sources . No growth was observed with acetate, citrate, gluconate, inositol, malate or mannitol as carbon sources . The strain showed very weak catalase activity . HPLC analysis of menaquinones revealed two peaks: the main peak corresponded with MK-9(H4) and the smaller one with MK-8(H4) . The major fatty acid found was anteiso-C15:0 (12-methyl tetradecanoic acid) . Mycolic acids were absent . The polar lipids detected were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides . Peptidoglycan type was A4alpha, L-Lys-D-Asp . The cell-wall sugars detected were galactose and rhamnose . The complete 16S rDNA sequence of strain XIL07T was obtained and phylogenetic analysis based on the neighbour-joining method indicated that this bacterium belongs to the high-G + C-content Gram-positive bacteria and that the closest related genera are Promicromonospora and Cellulosimicrobium . The DNA G + C content was 73 mol% . According to the data obtained in this work, this bacterium belongs to a new genus in the family Promicromonosporaceae and the name Xylanimonas cellulosilytica gen . nov., sp . nov . is proposed; the type strain is strain XIL07T (=LMG 20990T =CECT 5975T).

Mol Gen Mikrobiol Virusol, 2003, (1), 14 - 26
{Pleiotropic function of phosphoenolpyruvate-dependent phosphotransferase system in bacteria . Report 1}; Gershanovich VN; Modern data (collected mainly in the 1998-2001 studies) about the transport of carbohydrates in bacteria, about the regulation of utilization of sugars via the glycolytic pathway as well as about the regulation of transformation of pyruvat into the products of secondary metabolism and of tricarboxylic acid cycle are presented in the survey . Issues, related with the regulation of synthesis of enzymes involved in the last mentioned process, are discussed in detail . Besides, the key pathways pertaining to the regulation of synthesis and activity of adenylate cyclase; elimination of the inductor in the gram-negative bacteria and entry of phage lambda DNA into E . coli are described . As for the gram-positive bacteria, properties of their main components (involved in catabolic repression), i.e . HPr, K/P, CcpA, CCpB and CcpC, cre, are presented . The mechanisms of catabolic repressions and of catabolic activation in bacteria are in the focus of attention . Finally, issues related with the structural organization of PTS as well as molecular-and-biological aspects of the interaction of proteins of the mentioned system are considered in the survey.

J Antimicrob Chemother, 2003 Apr, 51(4), 971 - 5 Epub 2003 Feb 25.
Teicoplanin therapeutic drug monitoring in critically ill patients: a retrospective study emphasizing the importance of a loading dose; Pea F et al.; Data obtained as part of our routine drug monitoring of teicoplanin therapy (therapeutic drug monitoring, TDM) in adult critically ill patients being treated for suspected or documented Gram-positive multiresistant infections were assessed, retrospectively . Data were available for 202 patients (146 male, 56 female; age 58 +/- 16 years) with a total number of 829 teicoplanin trough plasma levels (C(min)) assessed . The percentage of patients with adequate teicoplanin concentrations (C(min) >/= 10 mg/L) during the treatment period substantially increased from 3.2% on day 2, to 35%, 70%, 90% and approximately 95% on days 4, 7, 11 and 15, respectively . The findings suggest that optimal teicoplanin therapy was achieved only after at least 4, and probably 7, days of therapy in most cases, mainly because of a failure to use an appropriate loading dose . Among the possible causes for the reluctance to use a loading dose, concern over the potential nephrotoxicity of teicoplanin was a major factor . We conclude that loading doses of teicoplanin (6 mg/kg every 12 h for at least three doses) must be considered mandatory in all patients, regardless of their renal function, to enable optimal drug concentrations to be achieved early in the treatment period . Subsequently, TDM is important to ensure that dose regimens are optimized to the individual requirements of the patients.

FEMS Microbiol Lett, 2003 Mar 14, 220(1), 15 - 20
Tylosin-resistant bacteria cultivated from agricultural soil; Onan LJ et al.; In this study we analyzed the numbers and types of cultivable tylosin-resistant bacteria from six agricultural soils that differed with respect to their association with subtherapeutic antibiotic use . The proportion of tylosin-resistant bacteria to the total number of bacteria cultivated was significantly higher (7.2-16.5%) at three sites affected by subtherapeutic antibiotic use compared to three sites unaffected by subtherapeutic antibiotic use (0.7-2.5%) . We also detected differences in the types of cultivable tylosin-resistant bacteria . At a site affected by subtherapeutic antibiotic use, we detected tylosin-resistant bacteria from the alpha- and beta-subdivisions of Proteobacteria . In contrast, at a site unaffected by subtherapeutic use, we detected only Streptomyces-like (high-G+C Gram-positive) tylosin-resistant bacteria . Our results suggest a link between subtherapeutic use of antibiotics and the numbers and types of antibiotic-resistant bacteria in nearby soil . However, other factors, such as soil type and temporal variation, may have also contributed to the differences observed.

Proc Natl Acad Sci U S A, 2003 Apr 1, 100(7), 4316 - 21 Epub 2003 Mar 17.
Cloning and engineering of the cinnamycin biosynthetic gene cluster from Streptomyces cinnamoneus cinnamoneus DSM 40005; Widdick DA et al.; Lantibiotics are ribosomally synthesized oligopeptide antibiotics that contain lanthionine bridges derived by the posttranslational modification of amino acid residues . Here, we describe the cinnamycin biosynthetic gene cluster (cin) from Streptomyces cinnamoneus cinnamoneus DSM 40005, the first, to our knowledge, lantibiotic gene cluster from a high G+C bacterium to be cloned and sequenced . The cin cluster contains many genes not found in lantibiotic clusters from low G+C Gram-positive bacteria, including a Streptomyces antibiotic regulatory protein regulatory gene, and lacks others found in such clusters, such as a LanT-type transporter and a LanP-type protease . Transfer of the cin cluster to Streptomyces lividans resulted in heterologous production of cinnamycin . Furthermore, modification of the cinnamycin structural gene (cinA) led to production of two naturally occurring lantibiotics, duramycin and duramycin B, closely resembling cinnamycin, whereas attempts to make a more widely diverged derivative, duramycin C, failed to generate biologically active material . These results provide a basis for future attempts to construct extensive libraries of cinnamycin variants.

Biochem J, 2003 Jun 1, 372(Pt 2), 577 - 85
Thermotoga neapolitana adenylate kinase is highly active at 30 degrees C; Vieille C et al.; The adenylate kinase (AK) gene from Thermotoga neapolitana, a hyperthermophilic bacterium, was cloned and overexpressed in Escherichia coli, and the recombinant enzyme was biochemically characterized . The T . neapolitana AK (TNAK) sequence indicates that this enzyme belongs to the long bacterial AKs . TNAK contains the four cysteine residues that bind Zn(2+) in all Gram-positive AKs and in a few other Zn(2+)-containing bacterial AKs . Atomic emission spectroscopy and titration data indicate a content of 1 mol of Zn(2+)/mol of recombinant TNAK . The EDTA-treated enzyme has a melting temperature (T (m)=93.5 degrees C) 6.2 degrees C below that of the holoenzyme (99.7 degrees C), identifying Zn(2+) as a stabilizing feature in TNAK . TNAK is a monomeric enzyme with a molecular mass of approx . 25 kDa . TNAK displays V (max) and K (m) values at 30 degrees C identical with those of the E . coli AK at 30 degrees C, and displays very high activity at 80 degrees C, with a specific activity above 8000 units/mg . The unusually high activity of TNAK at 30 degrees C makes it an interesting model to test the role of enzyme flexibility in activity.

J Bacteriol, 2003 Mar, 185(6), 1976 - 86
Sequence of the 165-kilobase catabolic plasmid pAO1 from Arthrobacter nicotinovorans and identification of a pAO1-dependent nicotine uptake system; Igloi GL et al.; The 165-kb catabolic plasmid pAO1 enables the gram-positive soil bacterium Arthrobacter nicotinovorans to grow on the tobacco alkaloid L-nicotine . The 165,137-nucleotide sequence, with an overall G+C content of 59.7%, revealed, besides genes and open reading frames (ORFs) for nicotine degradation, a complete set of ORFs for enzymes essential for the biosynthesis of the molybdenum dinucleotide cofactor, as well as ORFs related to uptake and utilization of carbohydrates, sarcosine, and amino acids . Of the 165 ORFs, approximately 50% were related to metabolic functions . pAO1 conferred to A . nicotinovorans the ability to take up L-{(14)C}nicotine from the medium, with an K(m) of 5.6 +/- 2.2 micro M . ORFs of putative nicotine transporters formed a cluster with the gene of the D-nicotine-specific 6-hydroxy-D-nicotine oxidase . ORFs related to replication, chromosome partitioning, and natural transformation functions (dprA) were identified on pAO1 . Few ORFs showed similarity to known conjugation-promoting proteins, but pAO1 could be transferred by conjugation to a pAO1-negative strain at a rate of 10(-2) to 10(-3) per donor . ORFs with no known function represented approximately 35% of the pAO1 sequence . The positions of insertion sequence elements and composite transposons, corroborated by the G+C content of the pAO1 sequence, suggest a modular composition of the plasmid.

Gene Expr Patterns, 2002 Dec, 2(3-4), 311 - 7
Tissue and stage-specific expression of the Tolls in Drosophila embryos; Kambris Z et al.; The Drosophila transmembrane receptor Toll plays a key role in specifying the dorsoventral axis of the embryo . At later stages of development, it controls the immune response of the fly to fungal and Gram-positive bacterial infections . The Drosophila genome has a total of nine Toll-like genes, including the previously characterized Toll (Toll-1) and 18-wheeler (Toll-2) . Here we describe the embryonic expression patterns of the seven Toll-like genes Toll-3 through Toll-9 . We find that these genes have distinct expression domains and that their expression is dynamically changing throughout embryonic development . This complex and tissue-specific regulation of Toll-like gene expression strongly suggests a role in embryonic development for most Drosophila Tolls . The evolving picture on the Toll family members in Drosophila contrasts with that of mammalian Toll-like receptors, which are predominantly expressed in immune responsive cells where their activation occurs via microbial structural determinants.

Histochem Cell Biol, 2003 Feb, 119(2), 103 - 8 Epub 2003 Jan 18.
Toll-like receptor 2 is expressed by alveolar epithelial cells type II and macrophages in the human lung; Droemann D et al.; The ability of the host to recognize pulmonary invasion by pathogenic organisms and establish an appropriate host response to infection requires innate immune defense mechanisms . Early bacterial clearance in the lung is mediated by alveolar macrophages (AM) and polymorphonuclear neutrophils . Additionally alveolar epithelial cells type II (AEC-II) may act as immunoregulatory cells . The toll-like receptors (TLR) are part of this innate immune defense, recognizing conserved patterns on microorganisms . Toll-like receptor 2 (TLR2) is crucial in detecting components of gram-positive bacteria and mycobacteria . Signals initiated by the interaction of TLR2 with bacterial components direct the subsequent inflammatory response . The detection of TLR2 mRNA in human lung tissue prompted us to localize the expression of mRNA and protein at the cellular level using a novel method for tissue fixation . We utilized HOPE-fixed lung specimen sections for targeting mRNA by in situ hybridization and protein by immunohistochemistry using the monoclonal antibody TL2.1 . In normal lung areas the expression of TLR2 mRNA and protein was found to be located in cells resembling AEC-II and AM . Expression of mRNA was verified by RT-PCR and DNA sequencing . These results indicate a potential mechanism of increased immunosurveillance at the alveolar level controlling the localized infection.

J Matern Fetal Neonatal Med, 2002 Nov, 12(5), 304 - 12
Evidence of participation of soluble CD14 in the host response to microbial invasion of the amniotic cavity and intra-amniotic inflammation in term and preterm gestations; Espinoza J et al.; OBJECTIVE: Endotoxin has been implicated in the mechanism responsible for the setting of infection in preterm labor . To exert its biological effects, endotoxin binds to a circulating protein known as lipopolysaccharide binding protein (LBP) and presents endotoxin monomers to CD14, which may be a membrane-bound receptor or a soluble molecule . The endotoxin-LBP-CD14 complex interacts with Toll-like receptor 4 and other regulatory proteins leading to cellular activation and an inflammatory response . The purpose of this study was to determine whether microbial invasion of the amniotic cavity (MIAC)/intra-amniotic inflammation (both preterm and term) and parturition at term are associated with changes in the amniotic fluid and umbilical plasma soluble concentrations of CD14 (sCD14) . STUDY DESIGN: Amniotic fluid was retrieved by amniocentesis from 88 patients in the following groups: group 1, preterm labor with intact membranes with MIAC/intra-amniotic inflammation (n = 18) and without these conditions (n = 26); group 2, term gestations not in labor without MIAC/intra-amniotic inflammation (n = 11), in labor without MIAC/intra-amniotic inflammation (n = 12) and in labor with MIAC/intra-amniotic inflammation (n = 13); and group 3, patients who underwent genetic amniocentesis at mid-trimester (n = 8) . A sample of cord blood was obtained after delivery in all patients except those in group 3 . sCD14 was assayed with a sensitive and specific immunoassay . Non-parametric statistics were used for analysis . A p value of < 0.05 was considered significant . RESULTS: sCD14 was detectable in 97% (85/88) of the amniotic fluid samples . Amniotic fluid sCD14 concentrations were lower in patients at term than in the mid-trimester of pregnancy (mid-trimester: median 482 ng/ml, range 258-838 ng/ml vs . term no labor: median 7 ng/ml, range 2-274 ng/ml, p = 0.01) . Among patients with preterm labor with intact membranes, the median amniotic fluid sCD14 level of patients with MIAC/intra-amniotic inflammation was higher than in patients without these conditions (median 1568 ng/ml, range 98-5887 ng/ml vs . median 645 ng/ml, range 0-3961 ng/ml, respectively; p = 0.01) . Among women at term in labor, those with MIAC/intra-amniotic inflammation had a higher median amniotic fluid sCD14 concentration than those without these conditions (median 85 ng/ml, range 2-1113 ng/ml vs . median 17 ng/ml, range 0-186 ng/ml; p = 0.01) . MIAC/intra-amniotic inflammation in women with preterm labor with intact membranes was associated with a higher median umbilical venous plasma sCD14 concentration (median 744 ng/ml, range 0-3620 ng/ml vs . median 0 ng/ml, range 0-2060 ng/ml; p = 0.04) . sCD14 was undetectable in plasma from umbilical cords of all neonates born to women at term . An increase in amniotic fluid concentration of sCD14 was observed in cases of intrauterine infection, not only by gram-negative bacteria, but also gram-positive bacteria and Ureaplasma spp . CONCLUSION: sCD14 is a physiological constituent of amniotic fluid, and its concentrations at term are lower than in the mid-trimester . Intrauterine infection/inflammation is associated with a higher median amniotic fluid sCD14 concentration in both preterm and term parturition . Neonates born from mothers with preterm labor with intact membranes and MIAC/intra-amniotic inflammation had a higher median concentration of sCD14 in umbilical cord plasma than those without these conditions . sCD14 concentrations are increased in the amniotic fluid and umbilical cord blood even in the absence of a microbiologically proven gram-negative infection . CD14 appears to participate in the host response to intrauterine infection even in cases involving genital mycoplasmas.

Mol Pharmacol, 2003 Mar, 63(3), 617 - 23
Erythromycin, roxithromycin, and clarithromycin: use of slow-binding kinetics to compare their in vitro interaction with a bacterial ribosomal complex active in peptide bond formation; Dinos GP et al.; In a cell-free system derived from Escherichia coli, it is shown that clarithromycin and roxithromycin, like their parent compound erythromycin, do not inhibit the puromycin reaction (i.e., the peptide bond formation between puromycin and AcPhe-tRNA bound at the P-site of 70S ribosomes programmed with heteropolymeric mRNA) . Nevertheless, all three antibiotics compete for binding on the ribosome with tylosin, a 16-membered ring macrolide that behaves as a slow-binding, slowly reversible inhibitor of peptidyltransferase . The mutually exclusive binding of these macrolides to ribosomes is also corroborated by the fact that they protect overlapping sites in domain V of 23S rRNA from chemical modification by dimethyl sulfate . From this competition effect, detailed kinetic analysis revealed that roxithromycin or clarithromycin (A), like erythromycin, reacts rapidly with AcPhe-tRNA.MF-mRNA x 70S ribosomal complex (C) to form the encounter complex CA which is then slowly isomerized to a more tight complex, termed C*A . The value of the overall dissociation constant, K, encompassing both steps of macrolide interaction with complex C, is 36 nM for erythromycin, 20 nM for roxithromycin, and 8 nM for clarithromycin . Because the off-rate constant of C*A complex does not significantly differ among the three macrolides, the superiority of clarithromycin as an inhibitor of translation in E . coli cells and many Gram-positive bacteria may be correlated with its greater rate of association with ribosomes.

Lancet, 2003 Feb 22, 361(9358), 637 - 44
Sequencing and analysis of the genome of the Whipple's disease bacterium Tropheryma whipplei; Bentley SD et al.; BACKGROUND: Whipple's disease is a rare multisystem chronic infection, involving the intestinal tract as well as various other organs . The causative agent, Tropheryma whipplei, is a Gram-positive bacterium about which little is known . Our aim was to investigate the biology of this organism by generating and analysing the complete DNA sequence of its genome . METHODS: We isolated and propagated T whipplei strain TW08/27 from the cerebrospinal fluid of a patient diagnosed with Whipple's disease . We generated the complete sequence of the genome by the whole genome shotgun method, and analysed it with a combination of automatic and manual bioinformatic techniques . FINDINGS: Sequencing revealed a condensed 925938 bp genome with a lack of key biosynthetic pathways and a reduced capacity for energy metabolism . A family of large surface proteins was identified, some associated with large amounts of non-coding repetitive DNA, and an unexpected degree of sequence variation . INTERPRETATION: The genome reduction and lack of metabolic capabilities point to a host-restricted lifestyle for the organism . The sequence variation indicates both known and novel mechanisms for the elaboration and variation of surface structures, and suggests that immune evasion and host interaction play an important part in the lifestyle of this persistent bacterial pathogen.

J Pharmacol Exp Ther, 2003 Mar, 304(3), 1093 - 102
Inhibition of endotoxin response by e5564, a novel Toll-like receptor 4-directed endotoxin antagonist; Mullarkey M et al.; Alpha-D-glucopyranose,3-O-decyl-2-deoxy-6-O-{2-deoxy-3-O-{(3R)-3-methoxydecyl}-6-O-methyl-2-{{(11Z)-1-oxo-11-octadecenyl}amino}-4-O-phosphono-beta-D-glucopyranosyl}-2-{(1,3-dioxotetradecyl)amino}-1-(dihydrogen phosphate), tetrasodium salt (E5564) is a second-generation synthetic lipodisaccharide designed to antagonize the toxic effects of endotoxin, a major immunostimulatory component of the outer cell membrane of Gram negative bacteria . In vitro, E5564 dose dependently (nanomolar concentrations) inhibited lipopolysaccharide (LPS)-mediated activation of primary cultures of human myeloid cells and mouse tissue culture macrophage cell lines as well as human or animal whole blood as measured by production of tumor necrosis factor-alpha and other cytokines . E5564 also blocked the ability of Gram negative bacteria to stimulate human cytokine production in whole blood . In vivo, E5564 blocked induction of LPS-induced cytokines and LPS or bacterial-induced lethality in primed mice . E5564 was devoid of agonistic activity when tested both in vitro and in vivo and has no antagonistic activity against Gram positive-mediated cellular activation at concentrations up to 1 microM . E5564 blocked LPS-mediated activation of nuclear factor-kappaB in toll-like receptor 4/MD-2-transfected cells . In a mouse macrophage cell line, activity of E5564 was independent of serum, suggesting that E5564 exerts its activity through the cell surface receptor(s) for LPS, without the need for serum LPS transfer proteins . Similar to (6-O-{2-deoxy-6-O-methyl-4-O-phosphono-3-O-{(R)-3-Z-dodec-5-endoyloxydecl}-2-{3-oxo-tetradecanoylamino}-beta-O-phosphono-alpha-D-glucopyranose tetrasodium salt (E5531), another lipid A-like antagonist, E5564 associates with plasma lipoproteins, causing low concentrations of E5564 to be quantitatively inactivated in a dose- and time-dependent manner . However, compared with E5531, E5564 is a more potent inhibitor of cytokine generation, and higher doses retain activity for durations likely sufficient to permit clinical application . These results indicate that E5564 is a potent antagonist of LPS and lacks agonistic activity in human and animal model systems, making it a potentially effective therapeutic agent for treatment of disease states caused by endotoxin.

Acta Pol Pharm, 2002 Sep-Oct, 59(5), 347 - 51
Application of topological indices for prediction of the biological activity of selected alkoxyphenols; Pyka A; The topological indices based on adjacency and distance matrices, and electrotopological states were calculated for selected meta and para alkoxyphenols . The toxicities of alkoxyphenols on gram-positive M . pyogenes var . aurens and gram-negative S . typhosa bacteria were calculated on the basis of linear or parabolic equations with one topological index or hydrophobic constants . It was affirmed, that structural descriptors describe better the toxic properties of studied alkoxyphenols than the hydrophobic constants.

Ginekol Pol, 2002 Sep, 73(9), 779 - 82
{Hydronephrosis in course of actinomycosis of female reproductive organs}; Iwaszko R et al.; OBJECTIVES: Pelvic actinomycosis is a chronic suppurative granulomatous disease caused by an anaerobic Gram positive germ, Actinomyces . The most difficult task was to obtain the diagnosis in a patient with an intrauterine device and poor general health, signs of infection and a pelvic syndrome . RESULTS: This is a report of a case of hydronephrosis in course of actinomycosis of female reproductive organs in a patient aged 46 years with the intrauterine contraceptive device admitted to The Department of Gynecology & Obstetric in Hospital of Slupsk . Symptoms were presented as an acute abdomen associated with painful epigastric and mass in right adnexa . A pre-operative diagnosis of the right hydronephrosis, on ovarian tumor and uterus myoma were detected which lead to an extensive and difficult surgery . The pathologic process infiltrated the retroperitoneal space simulated sarcoma or lymphoma . The disease was serious and required hysterectomy with salpingo-oophorectomy and intra-abdominal drainage . Actinomycosis was confirmed by the postoperative histopathologic examination, and the patient was successfully treated with penicillin . The patient was completely free of symptoms two months within the operation . CONCLUSIONS: In patients who have intrauterine contraceptive devices or who have had them removed recently, abdominal pain, recurrent vaginal bleeding or adnexal masses should prompt a thorough search for potentially pathogenic actinomyces in the genital tract.

Angew Chem Int Ed Engl, 2003 Feb 17, 42(7), 730 - 65
Vancomycin assembly: nature's way; Hubbard BK et al.; Antibiotics are precious resources in the fight to combat bacterial infections caused by pathogenic organisms . Vancomycin is one of the antibiotics of last resort in the treatment of life-threatening infections by gram-positive bacteria . The rules by which nature assembles the glycopeptide (vancomycin) and lipoglycopeptide (teicoplanin) antibiotics are becoming elucidated and verified: first amino acids are synthesized, then joined together and cross-linked . This knowledge opens up approaches for reprogramming strategies at the level of altered monomers, swapped assembly lines, and different post-assembly tailoring enzymes.

Surg Infect (Larchmt), 2000, 1(1), 15 - 21
Pyogenic liver abscess; Zibari GB et al.; BACKGROUND: A current assessment of liver abscesses should allow for better understanding of the pathogenesis of the disease and improve the effectiveness of diagnosis and treatment . Amebic liver abscess occurs more commonly than pyogenic liver abscess on a worldwide basis . However, in the United States, pyogenic liver abscess predominates . The purpose of our study was to evaluate the etiology, management, morbidity, and mortality of all patients admitted to our medical center with diagnoses of pyogenic liver abscess between 1983 and 1996 . METHODS: A retrospective chart review was performed on all patients admitted to our medical center, Louisiana State University Medical Center, Shreveport, with diagnoses of pyogenic liver abscess . RESULTS: Twenty patients were admitted with diagnoses of pyogenic liver abscess . The subjects were 65% (13/20) male and 65% (13/20) African-American and had an average age of 52 years . The most common presenting symptoms were fever and pain . The most common physical finding was right upper-quadrant tenderness . The most common etiologies of pyogenic liver abscesses were cryptogenic, trauma, and biliary, while portal vein was the source for only 10% of the cases . The right lobe of the liver was involved in 95% of the cases, and 70% of these liver abscesses were solitary . Computed tomography (CT)-and ultrasound-guided percutaneous drainage were performed in 85% (17/20) of patients with liver abscesses . One patient was treated by open drainage, three patients were treated with antibiotics alone, and three patients did not respond to aspiration and catheter placement, which subsequently required open drainage . The culture results were as follows: 50% were gram-negative organisms, 25% were gram-positive organisms, 10% were anaerobic organisms, and 15% of the abscess were sterile . Sixty percent of the positive abscess cultures were polymicrobial . CONCLUSIONS: CT scan- and ultrasound-guided percutaneous drainage of pyogenic liver abscesses were safe and effective methods of treatment . The right lobe of the liver was involved in 95% of cases . Although no one species predominated, gram-negative bacteria were the most common organism cultured, and 60% of the abscesses were polymicrobial . There was no in-house mortality in this review.

Clin Infect Dis, 2003 Mar 1, 36(5), e69 - 70 Epub 2003 Feb 18.
Tsukamurella tyrosinosolvens intravascular catheter infection identified using 16S ribosomal DNA sequencing; Sheridan EA et al.; Cultures of blood from a hemodialysis line repeatedly yielded a gram-positive rod . The organism was identified as Tsukamurella tyrosinosolvens by 16S ribosomal DNA sequencing, and the patient was treated successfully by removal of the line.

Surg Infect (Larchmt), 2001 Winter, 2(4), 255 - 63; discussion 264-5
Tertiary peritonitis (recurrent diffuse or localized disease) is not an independent predictor of mortality in surgical patients with intraabdominal infection; Evans HL et al.; BACKGROUND: It is well documented that tertiary peritonitis is associated with different microbiological flora and worse outcomes than secondary peritonitis . It is unknown, however, if these differences can be explained simply by the nosocomial nature of tertiary peritonitis and underlying severity of illness . METHODS: We reviewed all episodes of intraabdominal infection on the inpatient surgical services at a university hospital over a 46-month period . Univariate analysis and logistic regression were used to compare 91 episodes of secondary peritonitis that progressed to tertiary peritonitis (recurrent diffuse or localized intraabdominal infection) to all episodes of secondary peritonitis (n = 453) to identify predictors for developing tertiary peritonitis . Logistic regression was also used to identify predictors of mortality among patients with secondary (n = 473) or tertiary peritonitis (n = 129) . RESULTS: Of 602 episodes of intraabdominal infection identified, there were 473 episodes of secondary peritonitis, including 20 patients who died within seven days of diagnosis . A total of 129 episodes of tertiary peritonitis were identified, of which 91 were preceded by a single episode of secondary peritonitis, and 38 were preceded by an episode of secondary peritonitis and at least one prior episode of tertiary peritonitis . Tertiary peritonitis was associated with a high APACHE II score (14.9 +/- 0.7), pancreatic or small bowel source, drainage only at initial intervention, gram-positive and fungal pathogens, and a high mortality rate (19%) . Increasing APACHE II score (OR 1.07, 95% CI 1.03-1.16, p = 0.0009) independently predicted progression from secondary to tertiary peritonitis while increasing age (OR 0.98, 95% CI 0.97-0.99, p = 0.01) and appendiceal source (OR 0.12, 95% CI 0.02-0.68, p = 0.02) predicted non-progression to tertiary peritonitis . Independent predictors of mortality in this population included increasing age (OR 1.06, 95% CI 1.03-1.1, p < 0.001), increasing APACHE II score (OR 1.18, 95% CI 1.11-1.3, p < 0.001), and four comorbidities: cerebrovascular disease (OR 4.3, 95% CI 1.4-13.1, p = 0.01), malignant disease (OR 2.9, 95% CI 1.3-6.5, p = 0.01), hemodialysis dependency (OR 3.8, 95% CI 1.3-11.2, p = 0.02), and liver disease (OR 4.2, 95% CI 1.6-15.1, p = 0.03) . Tertiary peritonitis was not an independent predictor of mortality . CONCLUSIONS: We were unable to demonstrate, when compared to secondary peritonitis, that tertiary peritonitis is a significant independent predictor of mortality when other variables are taken into account . This suggests that the high mortality associated with tertiary peritonitis is more a function of the patient population in which it occurs than the severity of the pathologic process itself.

J Periodontol, 2003 Jan, 74(1), 85 - 9
Interactions of Porphyromonas gingivalis with host cells: implications for cardiovascular diseases; Kuramitsu HK et al.; BACKGROUND: Recent epidemiological studies have suggested a contribution of periodontitis in atherosclerotic diseases . Two mechanisms have been proposed to explain such a connection involving general inflammatory responses and/or specific effects of periodontal bacteria on host tissues . METHODS: The role of the periodontopathogen Porphyromonas gingivalis as a potential contributor to atherosclerosis has been investigated in model systems using human umbilical vein endothelial cells (HUVEC) and murine J774 macrophage cell cultures . RESULTS: P . gingivalis 381 was demonstrated to induce foam cell formation in J774 macrophage cell cultures in the presence of low-density lipoproteins . The active bacterial component involved in this process appears to be lipopolysaccharide . This effect was not limited to these organisms as several other Gram-positive and Gram-negative oral bacteria exhibited the same property . In addition, in a more specific manner, P . gingivalis induced monocyte chemoattractant protein-1 secretion in HUVEC cultures . CONCLUSIONS: The fimbriae of strain 381 are important, but are not required, for this inductive effect . Taken together, these results suggest a potential role for P . gingivalis in several steps involved in atherosclerotic lesion formation.

Heart, 2003 Mar, 89(3), 258 - 62
Blood culture negative endocarditis: analysis of 63 cases presenting over 25 years; Lamas CC et al.; OBJECTIVE: To analyse cases of blood culture negative endocarditis (BCNE) seen at St Thomas' Hospital, London, between 1975 and 2000 . METHODS: Data on all episodes of endocarditis with negative blood cultures seen at St Thomas' Hospital between 1975 and 2000 were collected prospectively and analysed retrospectively . RESULTS: Sixty three patients with BCNE were seen during the study period: 48 (76%) with native and 15 (24%) prosthetic valve infection . BCNE accounted for 12.2% of the 516 cases of endocarditis seen at St Thomas' Hospital . The diagnosis of endocarditis was clinically definite by the Duke criteria in only 21% (7 of 34) of cases of pathologically proven native valve endocarditis but in 62% (21 of 34) of cases by the St Thomas' modifications of the criteria . Comparable figures for the 11 cases of pathologically proven prosthetic valve endocarditis were 45% and 73% . Despite negative blood cultures a causative organism was identified in 31 (49%) of the 63 cases: in 15 by serology (8 Coxiella burnetii, 6 Bartonella species, and 1 Chlamydia psittaci); in 9 cases by culture of the excised valve; in 3 by microscopy of the excised valve, on which large numbers of Gram positive cocci were seen although the culture was sterile; and in the other 4 by isolation from a site other than the excised valve (2 respiratory specimens, 1 from the pacemaker tip, and 1 from an excised embolus) . In addition 5 of the 6 cases of Bartonella infection were confirmed by polymerase chain reaction study of the excised valve . Two thirds of the 32 patients for whom no pathogen was identified had received antibiotics before blood was cultured . Thus truly "negative" endocarditis was very uncommon (6% of the cases) . CONCLUSION: If blood cultures are negative in definite or suspected endocarditis, serum should be analysed for Bartonella, Coxiella, and Chlamydia species antibodies, and the excised valve or (rarely) embolus should be analysed by microscopy, culture, histology, and relevant polymerase chain reaction . Other specimens may be relevant . The Duke criteria performed poorly in BCNE; St Thomas' additional minor criteria gave more definite diagnoses.

Pathol Int, 2003 Feb, 53(2), 121 - 5
Primary sclerosing lipogranuloma with broad necrosis of the scrotum; Terada T et al.; A-25-year-old man was admitted because of a painless tumor of the scrotum . The patient denied a history of exogenous material injection and trauma in the scrotum . Physical and radiological examination revealed a mass in the scrotum, and blood laboratory tests showed no significant findings except for mild eosinophilia (5.6%) . Resection of the mass was performed . The mass was isolated and located in the subcutaneous tissue of the scrotum . The mass was rectangular and symmetrical, and measured 65 x 45 x 15 mm . Histologically, the mass was composed of adipose tissue with fibrosis . Many epithelioid granulomas with multinucleated giant cells of foreign body and Langhans' types and heavy infiltrates of lymphocytes and eosinophils were recognized . Characteristically, the lesion showed broad coagulative and lytic necrosis . Congestion and edema suggestive of ischemia were seen in some areas . Special stains for acid-fast bacteria, gram-positive bacteria and fungi failed to detect any microorganisms . Polymerase chain reaction for mycobacterium tuberculosis revealed no reaction products . Immunohistochemically, the majority of lymphocytes were CD45RO-positive T cells, and S-100 protein-positive cells and CD68-positive macrophages were scattered in small amounts . The appearances were typical for sclerosing lipogranuloma except for the necrosis . Although the pathological mechanism of the broad necrosis is unclear, the necrosis might be the result of ischemia . Our case suggests that primary sclerosing lipogranuloma of the scrotum might show broad necrosis, and that T-cell-mediated immune response might play a part in the formation of lipogranuloma.

APMIS, 2002 Nov, 110(11), 753 - 70
The role of bacteriolysis in the pathophysiology of inflammation, infection and post-infectious sequelae; Ginsburg I; The literature dealing with the biochemical basis of bacteriolysis and its role in inflammation, infection and in post-infectious sequelae is reviewed and discussed . Bacteriolysis is an event that may occur when normal microbial multiplication is altered due to an uncontrolled activation of a series of autolytic cell-wall breaking enzymes (muramidases) . While a low-level bacteriolysis sometimes occurs physiologically, due to "mistakes" in cell separation, a pronounced cell wall breakdown may occur following bacteriolysis induced either by beta-lactam antibiotics or by a large variety of bacteriolysis-inducing cationic peptides . These include spermine, spermidine, bactericidal peptides defensins, bacterial permeability increasing peptides from neutrophils, cationic proteins from eosinophils, lysozyme, myeloperoxidase, lactoferrin, the highly cationic proteinases elastase and cathepsins, PLA2, and certain synthetic polyamino acids . The cationic agents probably function by deregulating lipoteichoic acid (LTA) in Gram-positive bacteria and phospholipids in Gram-negative bacteria, the presumed regulators of the autolytic enzyme systems (muramidases) . When bacteriolysis occurs in vivo, cell-wall- and -membrane-associated lipopolysaccharide (LPS (endotoxin)), lipoteichoic acid (LTA) and peptidoglycan (PPG), are released . These highly phlogistic agents can act on macrophages, either individually or in synergy, to induce the generation and release of reactive oxygen and nitrogen species, cytotoxic cytokines, hydrolases, proteinases, and also to activate the coagulation and complement cascades . All these agents and processes are involved in the pathophysiology of septic shock and multiple organ failure resulting from severe microbial infections . Bacteriolysis induced in in vitro models, either by polycations or by beta-lactams, could be effectively inhibited by sulfated polysaccharides, by D-amino acids as well as by certain anti-bacteriolytic antibiotics . However, within phagocytic cells in inflammatory sites, bacteriolysis tends to be strongly inhibited presumably due to the inactivation by oxidants and proteinases of the bacterial muramidases . This might results in a long persistence of non-biodegradable cell-wall components causing granulomatous inflammation . However, persistence of microbial cell walls in vivo may also boost innate immunity against infections and against tumor-cell proliferation . Therapeutic strategies to cope with the deleterious effects of bacteriolysis in vivo include combinations of autolysin inhibitors with combinations of certain anti-inflammatory agents . These might inhibit the synergistic tissue- and- organ-damaging "cross talks" which lead to septic shock and to additional post-infectious sequelae.

Plasmid, 2003 Jan, 49(1), 44 - 52
Characterization of the cryptic plasmid pBGR1 from Bartonella grahamii and construction of a versatile Escherichia coli-Bartonella spp . shuttle cloning vector; Seubert A et al.; We report herein the isolation and molecular characterization of pBGR1, the first native plasmid isolated from the genus Bartonella . Cloning and sequencing revealed a 2725-base pair (bp) cryptic plasmid comprising two open reading frames of considerable length, which were designated rep and mob . The regions containing rep and mob are separated by 140-bp inverted repeat sequences and display a difference in G + C content from one another . A 1435-bp SacI-BclI fragment containing the rep gene is sufficient to mediate replication in the species Bartonella henselae and Bartonella tribocorum, while this replicon does not appear to be functional in Escherichia coli . The Rep protein of 190 amino acids (aa) shares homology to putative replication proteins of cryptic plasmids of Gram-negative origin, which form a subgroup of the rolling-circle replication proteins of the pSN2 plasmid superfamily of Gram-positive bacteria . The Mob protein of 333 aa is related to mobilization proteins of several cryptic plasmids and is associated with a conserved recombination site A . The tra functions of RP4 can mobilize pBGR1 derivatives in a mob-dependent manner . Mobilizable pBGR1-based E . coli-Bartonella spp . shuttle vectors were constructed and were shown to be maintained in B . tribocorum during in vivo passage in a rat model in the absence of antibiotic selection . The small size and stability of these shuttle cloning vectors should render them particularly valuable for genetic studies in Bartonella spp .

Syst Appl Microbiol, 2002 Dec, 25(4), 498 - 506
Allisonella histaminiformans gen . nov., sp . nov . A novel bacterium that produces histamine, utilizes histidine as its sole energy source, and could play a role in bovine and equine laminitis; Garner MR et al.; When cattle and horses are fed large amounts of grain, histamine can accumulate in the gastrointestinal tract, and this accumulation can cause an acute inflammation of the hooves (laminitis) . When ruminal fluid from dairy cattle fed grain supplements was serially diluted in anaerobic MRS medium containing histidine (50 mM), histamine was detected at dilutions as high as 10(-7) . The histidine enrichments were then transferred successively in an anaerobic, carbonate-based medium (50 mM histidine) without glucose . The histamine producing bacteria could not be isolated from the rumens of cattle fed hay; however, histamine producing bacteria could be isolated the feces of cattle fed grain and the cecum of a horse . All of the histamine producing isolates had the same ovoid morphology . The cells stained Gram-negative and were resistant to the ionophore, monensin (25 microM) . The doubling time was 110 min, and the yield was 1.5 mg cell protein per mmol histidine . The G+C content was 46.8% . Lysine was the only other amino acid used, but lysine did not allow growth if histidine was absent . Because carbohydrate and organic acid utilization was not detected, it appeared that the isolates used histidine decarboxylation as their sole mechanism of energy derivation . 16s rRNA gene sequencing indicated that the isolates were most closely related to low G+C Gram-positive bacteria (firmicutes), but similarities were < or = 94% . Because the most closely related bacteria (Dialister pneumonsintes, Megasphaera elsdenii and Selenomonas ruminantium) did not produce histamine from histidine, we propose that these histamine producing bacteria be assigned to a new genus, Allisonella, as Allisonella histaminiformans gen . nov., sp . nov . The type strain is MR2 (ATCC BAA610, DSM 15230).

Rev Esp Quimioter, 2002 Sep, 15(3), 257 - 63
Immediate vs . delayed imipenem treatment in cancer patients with profound neutropenia induced by high-dose chemotherapy: results of a randomized study; Garcia-Saenz JA et al.; A study was carried out to compare the use of prophylactic imipenem administered at the onset of profound neutropenia (immediate) with therapeutic imipenem administered at the onset of neutropenic fever (delayed) in cancer patients treated with high-dose chemotherapy . A total of 65 patients who were scheduled to receive two cycles of high-dose cyclophosphamide, etoposide, cisplatin (CEP) chemotherapy were randomized to receive imipenem either at presentation of neutropenia (immediate imipenem arm, prophylactic arm) or at commencement of neutropenic fever (delayed imipenem arm, therapeutic arm) . Treatment was crossed over when the second CEP chemotherapy cycle was received . Of the 65 patients, 41 received the two planned cycles and 24 received only the first . Compared with the delayed imipenem arm, the immediate imipenem arm was associated with lower fever incidence (86.3% vs . 100%, p=0.0142) and Gram-negative bacteria infection {4/51 (7.8%) vs . 14/55 (25.5%), OR=0.24, p =0.031} . There were fewer episodes of pneumonia (2% vs . 12.7%), septic shock (0% vs . 3.6%) and deaths from infection (0% vs . 3.6%), but these differences did not reach statistical significance . With regard to delayed imipenem, for every seven patients with immediate imipenem, one episode of febrile neutropenia was avoided; for every six patients administered immediate imipenem, one case of Gram-negative infection was avoided; and for every nine patients administered immediate imipenem, one episode of pneumonia was avoided . There were no differences in the incidence of Gram-positive infections nor in the length of hospitalization between the two treatment arms . In conclusion, compared to its conventional delayed use, immediate imipenem significantly reduces the frequency of febrile neutropenia and Gram-negative infections in patients with high-dose chemotherapy.

J Vet Diagn Invest, 2003 Jan, 15(1), 57 - 9
Encephalitozoon cuniculi placentitis and abortion in a quarterhorse mare; Patterson-Kane JC et al.; Encephalitozoon cuniculi is a microsporidial parasite, which has rarely been reported to cause placentitis in animals . A late-term aborted fetus and placenta from a Quarterhorse were presented to the Livestock Disease Diagnostic Center, University of Kentucky, for diagnostic examination . There was a necrotizing placentitis, with distension of many chorionic epithelial cells by intracytoplasmic vacuoles containing 1-2-microm-diameter, elongated, gram-positive organisms . The organisms were identified as E . cuniculi by electron microscopy and by polymerase chain reaction using primers to microsporidial ribosomal DNA . Joints of the fetus were swollen, with gross and microscopic lesions of synovitis; however, E . cuniculi DNA was not detected.

Biotechnol Appl Biochem, 2003 Feb, 37(Pt 1), 21 - 6
A new antibiotic, TH818, and its properties; Rachev R et al.; Antibiotic TH818 was isolated and purified from a culture broth of Streptomyces fulvoviolaceus 818 by extraction and reversed-phase liquid chromatography . The antibiotic TH818 possesses high molecular mass and unique structure, constructed from carbohydrates, amino acids and lipids . The hydrolysis experiments showed the presence of six fatty acids (hexadecanoic, hexadecenoic, octadecanoic, iso-nonadecanoic, docosanoic and pentacosanoic acids), four sugars (glucose, galactose, N -acetylglucosamine and N -acetylgalactosamine), and seven amino acids (threonine, alanine, glutamic acid, glycine, serine and two unidentified) . TH818 has a broad spectrum of anti-microbial activity against Gram-positive and Gram-negative bacteria, yeasts and fungi.

Scand J Infect Dis, 2002, 34(11), 863 - 6
Cerebral mycotic aneurysm complicating Stomatococcus mucilaginosus infective endocarditis; Perez-Vega C et al.; Stomatococcus mucilaginosus is a gram-positive coccus present in the normal flora of the mouth and upper respiratory tract of humans . Although traditionally believed to be an organism of low virulence, S . mucilaginosus has recently been recognized as an emerging opportunistic pathogen, especially in patients with chronic immunosuppressive diseases . This report describes the case of a patient with known mitral valve prolapse, who presented with a mycotic cerebral aneurysm revealing a non-nosocomial spontaneous S . mucilaginosus endocarditis . The spectrum of infections due to this opportunistic pathogen is reviewed, with particular focus on assessing the clinical characteristics and prognosis of S . mucilaginosus infective endocarditis.

Crit Care Med, 2003 Feb, 31(2), 390 - 4
Bacteremic sepsis in intensive care: temporal trends in incidence, organ dysfunction, and prognosis; Hugonnet S et al.; OBJECTIVE: To determine whether changes have occurred at our center in the incidence, patterns of organ dysfunctions, prognostic factors, and case-fatality rate of bacteremic sepsis . DESIGN: Retrospective comparative study (1984-1988 vs . 1994-1997) . SETTING: University hospital surgical intensive care unit . PATIENTS: A total of 372 critically ill patients with sepsis and positive blood cultures . INTERVENTION: None . MEASUREMENTS AND MAIN RESULTS: In the 1984-1988 study, 176 patients developed bacteremic sepsis (3.2 per 100 admissions), with a fatality rate of 35% at 28 days, whereas in the 1994-1997 study, 196 patients had bacteremic sepsis (4.3 per 100 admissions), with a 28-day fatality rate of 37% . The frequency of primary bacteremia increased from 21% to 47% (p <.001), paralleled by an increase in the proportional frequency of Gram-positive microorganisms . In 1984-1988, 69% (n = 120) of patients had at least one organ dysfunction, compared with 80% (n = 156) in 1994-1997 (p =.02) . The prevalence of pulmonary and cardiac dysfunction increased . The case- fatality rate of septic shock remained high (69% vs . 68%) . For both cohorts, the two strongest predictors of mortality remained the Acute Physiology and Chronic Health Evaluation II score at the onset of sepsis and the number of evolving organ dysfunctions . CONCLUSION: The fatality rate of bacteremic sepsis remained constant over the study period, despite an increased incidence of bacteremia and associated organ dysfunction . Continued efforts need to be directed toward the prevention of bacteremic sepsis, given the magnitude and poor prognosis of this syndrome.

J Clin Microbiol, 2003 Feb, 41(2), 908 - 10
Nocardia africana isolated from a feline mycetoma; Hattori Y et al.; Nocardia africana was isolated from a subcutaneous nodule of a cat . The isolate developed orange, wrinkled colonies . The bacteria were rod shaped to coccoid (1 by 5 microm) and gram positive . Analysis of the 16S ribosomal DNAs of the isolate and a reference strain of N . africana showed 100% homology.

J Clin Microbiol, 2003 Feb, 41(2), 819 - 21
Optimum detection times for bacteria and yeast species with the BACTEC 9120 aerobic blood culture system: evaluation for a 5-year period in a Turkish university hospital; Durmaz G et al.; We tracked and documented the time of positivity of blood cultures by using the BACTEC 9120 (Becton Dickinson Diagnostic Instrument Systems) blood culture system over a 5-year study period . A 7-day protocol of the incubation period was selected, and a total of 11156 blood cultures were evaluated . The clinically significant microorganisms (32.95%) were isolated in 3676 specimens . Gram-positive and -negative bacterial isolation rates were found to be 41.07 and 44.88%, respectively . Yeasts were found in 14.03% of all pathogens . Both the false-positivity and -negativity rates were very low (0.1 and 0.3%, respectively) . The mean detection times for all of the pathogens were determined to be 19.45 h . Yeasts, nonfermentative gram-negative bacteria, and Brucella melitensis strains were isolated within 5 days . By taking these data into account, we decided to establish a 5-day-incubation protocol in our laboratory instead of the 7 days that are commonly used.

J Clin Microbiol, 2003 Feb, 41(2), 640 - 4
Characterization of some actinomyces-like isolates from human clinical sources: description of Varibaculum cambriensis gen nov, sp nov; Hall V et al.; Fifteen strains of an anaerobic, catalase-negative, gram-positive diphtheroid-shaped bacterium recovered from human sources were characterized by phenotypic and molecular chemical and molecular genetic methods . The unidentified bacterium showed some resemblance to Actinomyces species and related taxa, but biochemical testing, polyacrylamide gel electrophoresis analysis of whole-cell proteins, and amplified 16S ribosomal DNA restriction analysis indicated the strains were distinct from all currently named Actinomyces species and related taxa . Comparative 16S rRNA gene sequencing studies showed that the bacterium represents a hitherto-unknown phylogenetic line that is related to but distinct from Actinomyces, Actinobaculum, Arcanobacterium, and Mobiluncus: We propose, on the basis of phenotypic and phylogenetic evidence, that the unknown bacterium from human clinical specimens should be classified as a new genus and species, Varibaculum cambriensis gen . nov., sp . nov . The type strain of Varibaculum cambriensis sp . nov . is CCUG 44998(T) = CIP 107344(T).

Jpn J Antibiot, 2000 Jun, 53 Suppl B, 82 - 90
{Clinical studies of azithromycin, a new macrolide antibiotic, for infections in the field of surgery}; Nakayama I; The efficacy of a newly developed macrolide antibiotic, azithromycin, for infections in the field of surgery, was investigated clinically by means of collaborative studies conducted in 17 major institutes and their affiliated hospitals throughout Japan . The following results were obtained . Clinical assessment: Azithromycin was administered at a dose of 250 mg or 500 mg once a day for 3 days . Clinical efficacy was evaluated in 170 patients . These subjects consisted of 81 with superficial purulent diseases, 12 with mastitis, 25 with periproctal abscess, 42 with superficial secondary infection due to trauma, burn and operative wound, 5 with cholecystitis or cholangitis, and 5 with other infections . The clinical efficacy rate was 96.3% (78/81) for superficial purulent diseases, 83.3% (10/12) for mastitis, 84.0%(21/25) for periproctal abscess, and 76.2%(32/42) for superficial secondary infection due to trauma, burn and operative wound . The overall clinical efficacy rate was 88.8%(151/170) respectively . The bacteriological eradication rate was 87.9%(116/132) for gram-positive bacteria, 85.0%(34/40) for gram-negative bacteria, and 100%(63/63) for anaerobic strains of casual bacteria, which were isolated from 140 patients . The overall bacteriological eradication rate was 90.6%(213/235) respectively . Adverse effects were observed in 6 of 170 patients in whom they were evaluated . They consisted of gastrointestinal symptoms in 5 patients and exanthema in 1 . Abnormal changes in clinical laboratory test values were observed in 5 patients, and consisted of eosinophilia in 1, elevations of S-GOT and S-GPT in 1, elevations of S-GOT, S-GPT and gamma-GTP in 1, elevation of S-GPT in 1, and elevations of AL-P and gamma-GTP in 1 . These results suggest that azithromycin is very useful for surgical infections in the field of surgery.

Novartis Found Symp, 2002, 248, 171 - 6; discussion 176-80, 277-82
Mechanisms by which gram-positive bacteria and tobacco smoke stimulate mucin induction through the epidermal growth factor receptor (EGFR); Basbaum C et al.; Mucin, the major macromolecular component of mucus, is generally considered to be a protective substance . When overproduced in a variety of lung diseases, however, mucin gives rise to clinical problems such as airway obstruction and recurrent infection . Our approach to identifying drug targets for the control of mucin overproduction is the analysis of cellular signalling pathways linking stimuli in the diseased lung to mucin transcription . Here we show that mucin transcription in response to both gram-positive bacteria and tobacco smoke is mediated through activation of the epidermal growth factor receptor (EGFR) . The mode of activation of EGFR in response to bacterial lipoteichoic acid involves cleavage of the transmembrane ligand HBEGF by ADAM 10, whereas the activation of EGFR in response to smoke involves cleavage of amphiregulin by ADAM 17.

Clin Infect Dis, 2003 Feb 15, 36(4), 462 - 7 Epub 2003 Jan 28.
Tetracycline therapy: update; Roberts MC; Tetracyclines have been used for treatment of a wide variety of gram-positive and gram-negative bacterial infections since the 1950s . In addition to being effective against traditional bacteria, tetracyclines have been used to treat infections due to intracellular chlamydiae, mycoplasmas, rickettsiae, and protozoan parasites and a variety of noninfectious conditions . They are important for treatment of and prophylaxis against infections with bacteria that could be used in biological weapons . Bacterial resistance to tetracycline was identified shortly after the introduction of therapy . At present, tetracycline resistance in bacteria can occur by acquisition of >or=1 of the 36 different genes, by mutations to host efflux pumps or in their 16S rRNA sequences, or by alteration in the permeability of the cell . In contrast, tetracycline resistance has not yet been described in protozoa or other eukaryotic organisms.

J Biol Chem, 2003 Apr 11, 278(15), 12873 - 80 Epub 2003 Feb 03.
The molecular basis of the expansive substrate specificity of the antibiotic resistance enzyme aminoglycoside acetyltransferase-6'-aminoglycoside phosphotransferase-2" . The role of ASP-99 as an active site base important for acetyl transfer; Boehr DD et al.; The most frequent determinant of aminoglycoside antibiotic resistance in Gram-positive bacterial pathogens is a bifunctional enzyme, aminoglycoside acetyltransferase-6'-aminoglycoside phosphotransferase-2" (AAC(6')- aminoglycoside phosphotransferase-2", capable of modifying a wide selection of clinically relevant antibiotics through its acetyltransferase and kinase activities . The aminoglycoside acetyltransferase domain of the enzyme, AAC(6')-Ie, is the only member of the large AAC(6') subclass known to modify fortimicin A and catalyze O-acetylation . We have demonstrated through solvent isotope, pH, and site-directed mutagenesis effects that Asp-99 is responsible for the distinct abilities of AAC(6')-Ie . Moreover, we have demonstrated that small planar molecules such as 1-(bromomethyl)phenanthrene can inactivate the enzyme through covalent modification of this residue . Thus, Asp-99 acts as an active site base in the molecular mechanism of AAC(6')-Ie . The prominent role of this residue in aminoglycoside modification can be exploited as an anchoring site for the development of compounds capable of reversing antibiotic resistance in vivo.

Proc Natl Acad Sci U S A, 2003 Feb 18, 100(4), 1541 - 6 Epub 2003 Jan 31.
PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin; Gust B et al.; Streptomycetes are high G+C Gram-positive, antibiotic-producing, mycelial soil bacteria . The 8.7-Mb Streptomyces coelicolor genome was previously sequenced by using an ordered library of Supercos-1 clones . Here, we describe an efficient procedure for creating precise gene replacements in the cosmid clones by using PCR targeting and lambda-Red-mediated recombination . The cloned Streptomyces genes are replaced with a cassette containing a selectable antibiotic resistance and oriT(RK2) for efficient transfer to Streptomyces by RP4-mediated intergeneric conjugation . Supercos-1 does not replicate in Streptomyces, but the clones readily undergo double-crossover recombination, thus creating gene replacements . The antibiotic resistance cassettes are flanked by yeast FLP recombinase target sequences for removal of the antibiotic resistance and oriT(RK2) to generate unmarked, nonpolar mutations . The technique has been used successfully by >20 researchers to mutate around 100 Streptomyces genes . As an example, we describe its application to the discovery of a gene involved in the production of geosmin, the ubiquitous odor of soil . The gene, Sco6073 (cyc2), codes for a protein with two sesquiterpene synthase domains, only one of which is required for geosmin biosynthesis, probably via a germacra-1 (10) E,5E-dien-11-ol intermediate generated by the sesquiterpene synthase from farnesyl pyrophosphate.

Curr Drug Targets, 2003 Feb, 4(2), 181 - 90
Fluoroquinolones: structure and target sites; Higgins PG et al.; The quinolones are a potent group of drugs that target the essential bacterial enzymes DNA gyrase and topoisomerase IV . DNA gyrase is the primary target of Gram negative organisms however, it is topoisomerase IV that is the primary target of Gram positive organisms . Within these enzymes is a highly conserved region centered round the active site where resistance mutations occur . These mutations are almost always identical, irrespective of organism . In spite of the homology of this region, amino acid sequence analysis shows that there are defined differences between the Gram groups, particularly in topoisomerase IV, and it is speculated that herein lies the origin of target preference . Since the first quinolone nalidixic acid was developed, the quinolones have undergone structural modifications, in particular the addition of a fluorine at position 6, to produce the fluoroquinolones . This has seen their potency and pharmakokinetic profile greatly increase . In vitro selection of resistance mutations has allowed the observation of how resistance is acquired and some of the modifications in newer fluoroquinolones have resulted in the shift of primary target from topoisomerase IV to gyrase with Gram positives . Curiously, purified topoisomerase IV is still more sensitive even if gyrase is the primary target . Gyrase remains the primary target for Gram negatives.

Expert Opin Investig Drugs, 2003 Feb, 12(2), 117 - 37
Novel agents for the treatment of resistant Gram-positive infections; Woodford N; Bacteria have proved themselves able to develop resistance to every antibiotic used clinically . Traditional agents used for treatment of serious infections caused by Gram-positive species have recently been supplemented with the introduction of linezolid, quinupristin-dalfopristin, several new quinolones and telithromycin . However, resistance to many of these agents has already been reported and, although each currently retains activity against the vast majority of clinical isolates of its target species, their long-term efficacy is uncertain . We must look to develop other compounds to replace and hopefully improve upon existing anti-Gram-positive agents . Daptomycin (a lipopeptide), oritavancin and dalbavancin (both second-generation glycopeptides) and ramoplanin (a glycolipodepsipeptide) are among the agents in advanced stages of development and, at present, many seem likely to proceed to licensing . In addition, it is encouraging that many agents active against novel bacterial targets have been discovered and are in earlier stages of development . In the next two decades, we should be optimistic that a regular flow of new anti-Gram-positive agents will enable us to offset the constant spectre of bacterial resistance.

Wei Sheng Wu Xue Bao, 1999 Aug, 39(4), 376 - 80
{Conjugational actinomycetes of plasmid RSF101 from Escherichia coli to the rare actinomyceces of Nocardia asteriodes and Streptoverticillum caespitosus}; Li W et al.; RSF1010 is a naturally occurring Escherichia coli broad host-range plasmid about 8.7 kb in size . It can be mobilized at high frequency between different gram-negative bacterial species when transfer functions are available in trans . Following the pioneering work of conjugational transfer of RSF1010 from E . coli to Streptomyces lividans and Mycobacterium smegmatis, the transfer of this plasmid by conjugation from E . coli S17.1 tp two gram-positive rare actinomycetes, Nocardia asteroides 3927 and Streptoverticillum caespitosus ATCC27422 was first time reported in this study . Southern blot analysis of the total DNA extracted from the actinomycetes' exconjugants proved that RSF1010 had been transferred from E . coli into the two new hosts and maintained staby in the exconjugants . Meanwhile, partial deletions of RSF1010 replicon loosing its antibiotics resistance makers were readily detected in E . coli . The implenmentation of this observation was discussed.

Wei Sheng Wu Xue Bao, 1999 Oct, 39(5), 396 - 401
{Characterization and phylogenetic analysis of the new species of the Leucobacter tropicalis sp . nov.}; Liu H et al.; A new aerobic, gram-positive, non-sporulating, rod-shaped organism is described . Strain F-8 was isolated from tropical soil, and has the following characteritics: the menaquinone contains a side chain with 11 isoprenyl units(MK-11); 2,4-diaminobutyric acid, glutamic acid, alanine, glycine and a small amount of gamma-aminobutyric acid present in the cell wall at a molar ratio of 0.77:1.0:1.66:0.95 . The major cellular sugar were rhamnose, galactose and glucose . The G + C content of DNA is 68 mol% . The 16S rRNA gene was amplified, cloned and sequenced, a phylogenetic tree was constructed on the 16S rRNA gene sequences . The tree clearly indicated that strain F-8 forming the same lineage with leucobacter komagatae, the level of binary sequence similarity between strain F-8 and L . komagatae is 96% . Based on the characteristics, allow the placement of strain F-8 into the genus Leucobacter, but F-8 showed some differences in biochemical and physical characters from the reported species L . komagae . A new species name, Leucobacter tropicalis sp . nov . was proposed for this isolate . The type strain is designated F-8.

Wei Sheng Wu Xue Bao, 2001 Dec, 41(6), 716 - 22
{Diversity of metabolites and their bio-activities in myxobacterium Sorangium cellulosum}; Li Y et al.; Different Sorangium cellulosum strains not only showed diversity in their cell and fruiting body morphologies, but also differences of bio-activities and components of the metabolites . All the Sorangial strains studied in this paper had no activity on Gram-negative bacteria, some were able to repress Gram-positive bacteria . However, all the strains were able to repress growth of fungi and tumor cells strongly and widely . Thin layer chromatography assay of metabolites showed multi-components in the metabolites, and most of them have abilities of repressing fungi and tumor cells in different degrees . Four strains were found to be able to produce compounds with activity of promoting polymerization of microtubule . Based on Rf value on TLC, the bio-active component produced by So33-1 strain was similar to Epothilone A, while that of So81 was much different . The results of this paper suggested that Sorangium cellulosum is a much beneficial resource for screening nature compounds with bio-activities against eukaryotes.

J Infect Dis, 2003 Jan 15, 187(2), 287 - 91 Epub 2002 Dec 19.
Lipopolysaccharide-binding protein serum levels in patients with severe sepsis due to gram-positive and fungal infections; Blairon L et al.; Lipopolysaccharide-binding protein (LBP) is increased in patients with severe gram-negative infections, but LBP serum levels have not been reported for in patients with gram-positive and fungal infections . LBP serum levels were determined in patients with severe sepsis secondary to gram-positive or fungal infections and were compared with LBP serum levels obtained from patients with gram-negative mixed infections and from healthy volunteers . Thirty-seven episodes of severe sepsis were analyzed among 24 patients . LBP serum levels were significantly increased in patients with severe sepsis (46.4+/-28.3 microg/mL), compared with that of healthy volunteers (5.7+/-1.9 microg/mL; P<.0001) . On the other hand, LBP serum levels obtained from patients with gram-negative infections (40.80+/-34.79 microg/mL) did not differ from those obtained from patients with gram-positive (35.55+/-23.95 microg/mL) or fungal (39.90+/-22.19 microg/mL) infections . These data suggest that LBP is an aspecific marker of sepsis, and the response was not clearly correlated with severity . Furthermore, in patients with multiple episodes of sepsis, LBP response seems to be of lesser magnitude after each subsequent episode of severe sepsis.

Enferm Infecc Microbiol Clin, 2003 Jan, 21(1), 24 - 9
{Incidence of bacteremia among histologic groups of hematologic malignancies}; Teira R et al.; INTRODUCTION: Subgroups having dissimilar prognoses are being identified among cancer patients with infection . Previous studies have suggested that these differences may be related to the histologic diagnosis, but this issue has not as yet been demonstrated . METHODS: We reviewed the medical records of all patients admitted with acute leukemia (AL) or lymphoma (ML) from 1988 to 1998 . Incidence of bacteremia was calculated for the following subgroups: acute lymphocytic leukemia (ALL), acute myelocytic leukemia (AML), AML following refractory anaemia with excess blasts (AML-RAEB), high-grade ML (HGML), intermediate-grade ML (IGML), low-grade ML (LGML) and indeterminate ML (IML) . Kaplan-Meier curves of time to the first positive blood culture were constructed and compared by means of log-rank test . RESULTS: In the period covered there were 244 new diagnoses of AL or ML: 62 AML, 32 ALL, 20 AML-RAEB, 78 HGML, 7 IGML, 37 LGML and 6 IML . At the end of the study period, 44 patients were alive, 147 were known to have died at a certain date and 53 had been formally lost to follow-up (most of them, transferred for hospice care) . Among 684 blood cultures, there were 51 contaminations and 155 significant isolates . Among the latter, gram-positive bacteria were isolated in 74 and gram-negative bacteria in 47; in 27 cases more than 1 bacterial species were recovered . Fungi were isolated in 7 cases . The incidence of bacteremia expressed as cases per 1000 patient-days was 5.80 for AML, 5.03 for AML-RAEB, 1.56 for ALL, 0.21 for HGML and 0.40 for the remaining ML . Time to the first positive blood culture was significantly shorter for AML than for any other group, and was shorted for ALL and AML-RAEB than for ML . CONCLUSION: Differences in the incidence of bacteremia were observed among histologically-defined groups of unselected patients with hematologic malignancies.

J Environ Qual, 2003 Jan-Feb, 32(1), 198 - 206
Stable carbon isotope ratio and composition of microbial fatty acids in tropical soils; Burke RA et al.; The soil microbial community plays a critical part in tropical ecosystem functioning through its role in the soil organic matter (SOM) cycle . This study evaluates the relative effects of soil type and land use on (i) soil microbial community structure and (ii) the contribution of SOM derived from the original forest vegetation to the functioning of pasture and sugarcane (Saccharum spp.) ecosystems . We used principal components analysis (PCA) of soil phospholipid fatty acid (PLFA) profiles to evaluate microbial community structure and PLFA stable carbon isotope ratios (delta13C) as indicators of the delta13C of microbial substrates . Soil type mainly determined the relative proportions of gram positive versus gram negative bacteria whereas land use primarily determined the relative proportion of fungi, protozoa, and actinomycetes versus other types of microorganisms . Comparison of a simple model to our PLFA delta13C data from land use chronosequences indicates that forest-derived SOM is actively cycled for appreciably longer times in sugarcane ecosystems developed on Andisols (mean turnover time = 50 yr) than in sugarcane ecosystems developed on an Oxisol (mean turnover time = 13 yr) . Our analyses indicate that soil chronosequence PLFA delta13C measurements can be useful indicators of the contribution that SOM derived from the original vegetation makes to continued ecosystem function under the new land use.

Trends Genet, 2003 Feb, 19(2), 107 - 12
Transient genetic asymmetry and cell fate in a bacterium; Dworkin J; Certain species of Gram-positive bacteria can initiate a developmental program that results in the formation of two daughter cells with different fates . One cell develops into a spore and the other cell undergoes programmed lysis, with each process being mediated by a cascade of cell-type-specific transcription factors . An early and critical step in this developmental pathway is the formation of a division septum near one pole, creating two compartments of different sizes . But how is this morphological asymmetry translated into the transcriptional asymmetry of the two compartments? Recent results suggest that the chromosomal position of the genes encoding several key components of the transcriptional regulatory network has an important role in this process.

Crit Care Med, 2003 Jan, 31(1 Suppl), S85 - 93
Drotrecogin alfa (activated) (recombinant human activated protein C) for the treatment of severe sepsis; Bernard GR; OBJECTIVE: To review the data supporting drotrecogin alfa (activated) for severe sepsis treatment . DATA SOURCES: Published research and data from the Protein C Worldwide Evaluation in Severe Sepsis (PROWESS) trial . DATA EXTRACTION AND SYNTHESIS: The coagulation cascade and intense inflammation play a central role in the development of organ failure due to severe sepsis . Drotrecogin alfa (activated) has anti-inflammatory, antithrombotic, profibrinolytic, and other properties that may explain the beneficial results seen in both animal models and humans with severe sepsis . Drotrecogin alfa (activated) produces a robust reduction in the mortality rate of patients with severe sepsis that is evident across nearly every subgroup examined in the phase III clinical trial and has an acceptable safety profile with bleeding during infusion as the only significant risk associated with therapy . The relative risk reductions for mortality seen in Gram-negative, Gram-positive, pneumonia, abdominal sources, shock, and nonshock are similar to the intent-to-treat population, 19.4% . Treatment also increases days alive and free from mechanical ventilation and shock . CONCLUSIONS: Coagulopathy and systemic inflammation are almost universal in patients with severe sepsis . Treatment of this disorder with drotrecogin alfa (activated) directly addresses these derangements and substantially reduces morbidity and mortality rates with potential for bleeding during infusion as the only known risk.

Surg Infect (Larchmt), 2002 Autumn, 3(3), 251 - 257
In Vitro Assessment of Microbial Barrier Properties of Dermabond((R)) Topical Skin Adhesive; Bhende S et al.; BACKGROUND: Several different techniques are used for wound closure . Cyanoacrylate tissue adhesives are less invasive and easy to apply, and the cosmetic results are comparable to or better than the conventional methods for wound repair . The purpose of this study is to demonstrate that Dermabond((R)) Topical Skin Adhesive, 2-octylcyanoacrylate (a registered trademark of Ethicon) is an effective barrier against the penetration of microorganisms in an in vitro model . MATERIALS AND METHODS: Octylcyanoacrylate adhesive was evaluated in vitro as a barrier to microbial penetration using a strike through test . Agar media containing pH sensitive dye was used . The dye changed color in the presence of acidic microbial metabolic products . Octylcyanoacrylate adhesive was applied to the agar surface to form uniformly thick films . A total of 300 single layered films and an additional 300 triple layered films were evaluated . A 10-&mgr;L aliquot of inoculum containing at least 1 x 10(3) cfu was applied to the surface of each film . Plates were incubated at 37 degrees C for 72 h and were observed for growth and color change every 24 h during the incubation period . RESULTS: A series of binomial calculations were performed, varying the level of effectiveness of the test and the level of statistical confidence . Of the 600 test articles evaluated, 598 retained their patency at the end of 72 h . One individual test film was invalidated due to extrinsic contamination . The data presented indicated that octylcyanoacrylate tissue adhesive provided a barrier to microbial penetration with 95% confidence of 99% efficacy for 72 h in this in vitro model . CONCLUSION: The results of these in vitro experiments supported the hypothesis that octylcyanoacrylate tissue adhesive is an effective barrier to microbial penetration by gram-positive and gram-negative motile and nonmotile species.

Br J Haematol, 2003 Jan, 120(2), 209 - 16
Elderly haematological patients with chemotherapy-induced febrile neutropenia have similar rates of infection and outcome to younger adults: a prospective study of risk-adapted therapy; Garcia-Suarez J et al.; We prospectively evaluated 131 consecutive episodes of fever and chemotherapy-induced neutropenia in 85 adults with haematological malignancies to determine whether older patients (aged < 60 years) have different causes of fever and outcome than younger adults (aged < 60 years) . Patients were stratified into high-risk and low-risk groups according to previously published criteria . High-risk patients received ceftazidime plus amikacin and low-risk patients received ceftazidime alone . All patients were hospitalized until fever and neutropenia resolved . Ninety one high-risk episodes were documented: 56 occurring in older patients (mean age 69 years) and 35 in younger adults (mean age 45 years) . Non-Hodkgin's lymphoma and acute myeloid leukaemia were the most frequent underlying neoplasias in both age groups . Intensity of chemotherapy was similar in both age groups . Mean neutrophil count at entry, median duration of neutropenia, rate of documented infection, incidence of bacteraemia, response to therapy, overall mortality and infectious mortality were similar in the two high-risk age subgroups . The elderly subgroup had a trend to have more Gram-negative infections and the younger patients more Gram-positive infections . In addition, 40 low-risk episodes were registered: 29 in elderly patients (mean age 68 years) and 11 in younger patients (mean age 44 years) . Elderly low-risk patients had more concurrent diseases that younger ones (P = 0.124) . Mean neutrophil count at entry, median duration of severe neutropenia and rate of response were similar in the two age subgroups . All low-risk patients survived . In conclusion, elderly haematological cancer patients with febrile neutropenia show similar rates of infection and outcome to younger ones.

J Nat Prod, 2003 Jan, 66(1), 73 - 9
New bioactive rosigenin analogues and aromatic polyketide metabolites from the freshwater aquatic fungus Massarina tunicata; Oh H et al.; Four new rosigenin analogues (massarigenins A-D; 1-4) and two new aromatic polyketide-derived secondary metabolites (massarinins A and B; 6, 7) and have been isolated from the freshwater aquatic fungus Massarina tunicata . The structures of these compounds were determined primarily by analysis of NMR data, and that of compound 1 was verified by X-ray crystallography . The known compound 4-(2-hydroxybutynoxy)benzoic acid (11) was also obtained, and its absolute stereochemistry was assigned . Several of these metabolites showed antibiotic activity against Gram-positive bacteria.

J Food Prot, 2003 Jan, 66(1), 94 - 102
Resistance of bacterial isolates from poultry products to therapeutic veterinary antibiotics; Boothe DH et al.; Bacterial isolates from poultry products were tested for their susceptibility to 10 antibiotics commonly used in the therapeutic treatment of poultry . Bacteria were isolated from fresh whole broiler carcasses or from cut-up meat samples (breast with or without skin, wings, and thighs) that were either fresh or stored at 4 or 13 degrees C (temperatures relevant to poultry-processing facilities) . The Biolog system was used to identify isolates, and a broth dilution method was used to determine the antibiotic resistance properties of both these isolates and complementary cultures from the American Type Culture Collection . The antibiotics to which the most resistance was noted were penicillin G, sulfadimethoxine, and erythromycin; the antibiotic to which the least resistance was noted was enrofloxacin . Individual isolates exhibited resistances to as many as six antibiotics, with the most common resistance pattern involving the resistance of gram-negative bacteria to penicillin G, sulfadimethoxine, and erythromycin . Differences in resistance patterns were noted among 18 gram-positive and 7 gram-negative bacteria, and comparisons were made between species within the same genus . The data obtained in this study provide a useful reference for the species and resistance properties of bacteria found on various raw poultry products, either fresh or stored at temperatures and for times relevant to commercial processing, storage, and distribution . The results of this study show that resistance to antibiotics used for the therapeutic treatment of poultry occurs in bacteria in the processing environment.

Clin Infect Dis, 2003 Feb 1, 36(3), 313 - 8 Epub 2003 Jan 13.
Clinical experience with linezolid for the treatment of nocardia infection; Moylett EH et al.; Linezolid is an oxazolidinone that has activity against most gram-positive bacteria, including in vitro activity against all Nocardia species and strains . We describe 6 clinical cases of nocardiosis that were successfully treated with linezolid . Two patients had underlying X-linked chronic granulomatous disease, and 2 patients were receiving chronic corticosteroid therapy . Four of 6 patients had disseminated disease, and 2 of these 4 patients had multiple brain abscesses . Four patients primarily received monotherapy; for the fifth patient, linezolid was added to a failing multiple-drug regimen, and, for the sixth patient, it was used as part of combination therapy . All 6 patients were successfully treated, although 1 patient had a presumed relapse of central nervous system infection after premature discontinuation of the drug . Linezolid appears to be an effective alternative for the treatment of nocardiosis.

J Immunol, 2003 Feb 1, 170(3), 1399 - 405
Lipopolysaccharide (LPS)-binding protein mediates LPS detoxification by chylomicrons; Vreugdenhil AC et al.; Chylomicrons have been shown to protect against endotoxin-induced lethality . LPS-binding protein (LBP) is involved in the inactivation of bacterial toxin by lipoproteins . The current study examined the interaction among LBP, chylomicrons, and bacterial toxin . LBP was demonstrated to associate with chylomicrons and enhance the amount of LPS binding to chylomicrons in a dose-dependent fashion . In addition, LBP accelerated LPS binding to chylomicrons . This LBP-induced interaction of LPS with chylomicrons prevented endotoxin toxicity, as demonstrated by reduced cytokine secretion by PBMC . When postprandial circulating concentrations of chylomicrons were compared with circulating levels of low density lipoprotein, very low density lipoprotein, and high density lipoprotein, chylomicrons exceeded the other lipoproteins in LPS-inactivating capacity . Furthermore, highly purified lipoteichoic acid, an immunostimulatory component of Gram-positive bacteria, was detoxified by incubation with LBP and chylomicrons . In conclusion, our results indicate that LBP associates with chylomicrons and enables chylomicrons to rapidly bind bacterial toxin, thereby preventing cell activation . Besides a role in the detoxification of bacterial toxin present in the circulation, we believe that LBP-chylomicron complexes may be part of a local defense mechanism of the intestine against translocated bacterial toxin.

J Bacteriol, 2003 Feb, 185(3), 929 - 37
In vivo analysis of HPr reveals a fructose-specific phosphotransferase system that confers high-affinity uptake in Streptomyces coelicolor; Nothaft H et al.; HPr, the histidine-containing phosphocarrier protein of the bacterial phosphotransferase system (PTS), serves multiple functions in carbohydrate uptake and carbon source regulation in low-G+C-content gram-positive bacteria and in gram-negative bacteria . To assess the role of HPr in the high-G+C-content gram-positive organism Streptomyces coelicolor, the encoding gene, ptsH, was deleted . The ptsH mutant BAP1 was impaired in fructose utilization, while growth on other carbon sources was not affected . Uptake assays revealed that BAP1 could not transport appreciable amounts of fructose, while the wild type showed inducible high-affinity fructose transport with an apparent K(m) of 2 microM . Complementation and reconstitution experiments demonstrated that HPr is indispensable for a fructose-specific PTS activity . Investigation of the putative fruKA gene locus led to identification of the fructose-specific enzyme II permease encoded by the fruA gene . Synthesis of HPr was not specifically enhanced in fructose-grown cells and occurred also in the presence of non-PTS carbon sources . Transcriptional analysis of ptsH revealed two promoters that are carbon source regulated . In contrast to what happens in other bacteria, glucose repression of glycerol kinase was still operative in a ptsH background, which suggests that HPr is not involved in general carbon regulation . However, fructose repression of glycerol kinase was lost in BAP1, indicating that the fructose-PTS is required for transduction of the signal . This study provides the first molecular genetic evidence of a physiological role of the PTS in S . coelicolor.

J Microbiol Methods, 2003 Mar, 52(3), 395 - 8
A rapid method for RNA preparation from Gram-positive bacteria; Oh ET et al.; We have developed a rapid method for preparation of RNA from various Gram-positive bacteria . Unlike some methods, this method does not require lysozyme and proteinase K . Instead, we used glass beads to break cells more efficiently . Using this method, we successfully isolated the total RNA from various Gram-positive bacteria . This method is rapid, simple, and more economic when compared to the previously reported ones . Prepared RNA can be used for the transcriptional analysis of various Gram-positive bacteria.

Immunity, 2003 Jan, 18(1), 109 - 20
Intrinsic requirement for zinc finger transcription factor Gfi-1 in neutrophil differentiation; Hock H et al.; We report essential roles of zinc finger transcription factor Gfi-1 in myeloid development . Gene-targeted Gfi-1(-/-) mice lack normal neutrophils and are highly susceptible to abscess formation by gram-positive bacteria . Arrested, morphologically atypical, Gr1(+)Mac1(+) myeloid cells expand with age in the bone marrow . RNAs encoding primary but not secondary or tertiary neutrophil (granulocyte) granule proteins are expressed . The atypical Gr1(+)Mac1(+) cell population shares characteristics of both the neutrophil and macrophage lineages and exhibits phagocytosis and respiratory burst activity . Reexpression of Gfi-1 in sorted Gfi-1(-/-) progenitors ex vivo rescues neutrophil differentiation in response to G-CSF . Thus, Gfi-1 not only promotes differentiation of neutrophils but also antagonizes traits of the alternate monocyte/macrophage program.

J Infect Chemother, 2002 Dec, 8(4), 358 - 60
The first isolation of Nocardia nova from an HIV-1 infected individual in Japan; Koibuchi T et al.; Nocardia species are opportunistic pathogens of immunocompromised patients . We report a case of Nocardia nova infection complicating Pneumocystis carinii pneumonia (PCP) in an HIV-1 infected individual . A 27-year-old man with hemophilia A was admitted on October 17, 2000, with fever and dyspnea . CD4 cell counts were 5/microl on admission . Prophylaxis against PCP was administered by inhalation of pentamidine isethionate because he was allergic to sulfamethoxazole-trimethoprim (SMZ-TMP) . He was diagnosed with PCP from chest X-ray and bronchoalveolar lavage . The sputum obtained for culture on admission was positive for Gram-positive branching rods; the organism was later identified as Nocardia nova . He died from respiratory failure on November 7, 2000 . Although PCP might be a principal factor in respiratory failure, this case shows the need to consider pulmonary nocardiosis as a cause of respiratory illness in patients with advanced HIV-1 infection.

J Antibiot (Tokyo), 2002 Oct, 55(10), 855 - 62
GEX1 compounds, novel antitumor antibiotics related to herboxidiene, produced by Streptomyces sp . I . Taxonomy, production, isolation, physicochemical properties and biological activities; Sakai Y et al.; Six structurally related antitumor antibiotics named GEX1 compounds were isolated from a culture broth of Streptomyces sp . GEX1A was identified as a known herbicide, herboxidiene, structurally interested by the tetrahydropyran moiety and the side chain including a conjugated diene . GEX1Q1 to approximately Q5 were determined as novel compounds related to herboxidiene . All GEX1 compounds showed cytotoxicity with IC50 values of 0.0037 to approximately 0.99 microM against human tumor cell lines in vitro, but were not active against both gram-positive and -negative bacteria . Though GEX1A/herboxidiene exhibited antitumor activity in murine tumor-planted mouse models, both GEX1Q3 and GEX1Q5 did not.

Rev Med Suisse Romande, 2002 Nov, 122(11), 535 - 7
{Pelvic and abdominal actinomycosis . Case report and review of the literature}; Cirafici L et al.; Pelvic and abdominal actinomycosis is a chronic suppurating granulomatosis caused by a Gram positive gem, Actinomyces Israelii . Manifestations of this rare disease may mimic cancer, inflammatory bowel disease, or diverticulitis . These syndromes lead to surgical exeresis . The diagnosis is obtained from the pathology report . We report the case of a 56 years old woman with large bowel obstruction secondary to extensive pelvic and abdominal actinomycosis . A review of the literature shows that the abdominal-pelvic form has been increasing over the past 10 years secondary to the increased and prolonged use of the intrauterine device . Treatment of this condition consists of a combination of antibiotics and surgery to achieve complete recovery.

J Gastroenterol, 2002, 37(12), 1028 - 34
Risk factors for infections in cirrhotic patients with and without hepatocellular carcinoma; Yoneyama K et al.; BACKGROUND: Patients with liver cirrhosis (LC) frequently have complications with bacterial infections, and these infections increase the mortality rate . However, a detailed analysis of infections associated with LC patients has not yet been performed . METHODS: We analyzed 325 patients with LC with and without hepatocellular carcinoma (HCC) who were hospitalized between 1997 and 1999 . RESULTS: Infections developed in 70 (21.5%) patients and 48 (68.6%) of these developed infections during hospitalization . The mortality rate of 28.6% (20/70) in patients with infectious complications was higher than that of 12.5% (32/255) in patients without infectious complications . Forty (57.1%) of the 70 patients had infections of unknown causes; 11 (15.7%) had sepsis; 6 (8.6%) had intravenous hyperalimentation (IVH) infection; 3 (4.3%) each had spontaneous bacterial peritonitis (SBP), liver abscess, and cholecystitis; and 4 (5.7%) had other infections . Bacterial cultures of blood were prepared from 73 of the 325 patients (22.5%), and were positive in 22 of the 73 patients (30.1%) . Of these 22 culture-positive patients, 11 had sepsis, 6 had IVH infection, 2 had liver abscess, 1 had cholecystitis, 1 had pneumonia, and 1 had decubitus ulcer . Gram-positive bacterial strains were detected most frequently, in 16 of the 24 strains isolated . Univariate analysis revealed significant differences between the groups with and without infectious complications with regard to hepatitis B virus infection, Child-Pugh classification, ascites, esophageal varices, survival rate, total-bilirubin (T-Bil), albumin (Alb), lactate dehydrogenase (LDH), total cholesterol (T-chol), and prothrombin time (PT) . On multivariate analysis, the Alb level was selected as a significant independent factor contributing to the development of infections . CONCLUSIONS: Patients with advanced cirrhosis with low Alb levels should be carefully treated, and the administration of broad spectrum antibiotics covering gram-positive bacteria needs to be considered in the treatment of infections.

Ann Acad Med Singapore, 2002 Nov, 31(6), 802 - 4
Gemella empyema cured without antibiotics: a case report; Poulose V; INTRODUCTION: Gemella are gram-positive bacteria that are commensals of the upper respiratory tract in humans and infrequently known to cause infections . CLINICAL PICTURE: We report a case of thoracic empyema due to Gemella morbillorum in an elderly Chinese male who had been having symptoms for 3 months and no response to multiple courses of antibiotics . INTERVENTION: The collection was drained with ultrasound guidance . OUTCOME: Drainage of the empyema produced rapid resolution of symptoms and no antibiotics were started . CONCLUSIONS: Gemella is a very rare cause of empyema which usually responds well to treatment.

Afr J Med Med Sci, 2002 Jun, 31(2), 111 - 4
Contamination levels of in-use disinfectants in a teaching hospital in Lagos, Nigeria; Ogunsola FT et al.; In-use testing of the disinfectants; Hibitane (5% w/v Chlorhexidinegluconate), Hibiscrub (4% w/v Chlorhexidinegluconate), Savlon (3% w/v Chlorhexidine/Cetrimide), hydrogen peroxide (6% w/v hydrogen peroxide with stabilizer) and a common household bleach Jik (3.5% w/v sodium hypochlorite), was carried out over a two-month period at a university teaching hospital in Nigeria . Contamination levels were high with 82 (63.1%) of the 130 in-use disinfectants contaminated . However, a few of the stock solutions remained sterile . One hundred and thirty-four isolates were obtained of which 120 (91%) were gram-negative with Pseudomonas species being the commonest, constituting 67.2% of all the isolates . Gram-positive organisms made up the remaining 12 (9.0%) isolates . All the Pseudomonas spp . were resistant to gentamicin, ceftazidime, nalidixic acid and perfloxacin . Contributory factors for the high contamination levels were dilution of disinfectants with tap water, inadequate care of stock solution bottles and long storage of the diluted disinfectants in the wards.

J Lipid Res, 2003 Jan, 44(1), 1 - 10
The application of computational methods to explore the diversity and structure of bacterial fatty acid synthase; Zhang YM et al.; Acyl carrier protein (ACP) is a central element in the bacterial, type II dissociated fatty acid synthase (FAS II) system . ACP delivers the fatty acyl intermediates to a variety of enzymes with different biochemical functions and 3-dimensional (3-D) structures . Computational techniques have proved invaluable in guiding the experimental designs that have uncovered the recognition helix on ACP and the common features on its target enzymes responsible for specific protein*protein interactions . Escherichia coli has been the model organism for the study of FAS II, but the availability of complete genomic sequences of a growing number of bacteria coupled with computational bioinformatics has led to new discoveries on the mechanisms that regulate E . coli FAS II and allowed the differences between the E . coli paradigm and major groups of pathogens to be identified and experimentally addressed . Computational methods facilitated the discovery of the E . coli fatty acid synthesis transcriptional regulator, FabR, and led to the identification of novel bacterial FAS II proteins in Gram-positive pathogens, including enoyl-ACP reductases (FabK and FabL) and trans-2-cis-3-decenoyl-ACP isomerase FabM . As more genomic sequences and 3-D coordinates are added to the databases, the power and resolution of the computational approaches will increase to offer deeper insight into the structure, diversity and function of lipid metabolic pathways.

J Clin Microbiol, 2003 Jan, 41(1), 106 - 9
Identification of catalase-negative, non-beta-hemolytic, gram-positive cocci isolated from milk samples; Fortin M et al.; This study was undertaken in an effort to improve the identification scheme of catalase-negative, non-beta-hemolytic, gram-positive cocci isolated from milk samples obtained from cows . First, the sensitivity and specificity of the identification procedure currently in use in our laboratory were compared to the results obtained with API 20 STREP strips which were set as the gold standard . Second, a number of other identification tests, which could contribute to increase the sensitivity and specificity of the identification procedure of these microorganisms, were evaluated and selected . The data have shown that there is a necessity to review the identification procedure . Some modifications are suggested to laboratories doing milk sample analyses . A standardized procedure, using the CAMP test, esculin and sodium hippurate hydrolysis, the presence of the enzymes pyrolidonyl arylaminase and leucine aminopeptidase, and acid production from 1% inulin and raffinose broth, would not only improve the results of the identification process of gram-positive cocci isolated from milk samples but also ensure greater uniformity of the epidemiological data.

J Nutr, 2003 Jan, 133(1), 322S - 327S
Metabolic effects of infection on protein and energy status; Powanda MC et al.; A review of some of the seminal studies of metabolism during various infections indicates that similar patterns of metabolic alterations occur during these illnesses . This patterned metabolic array occurs whether the infection is caused by a gram-positive or a gram-negative bacterium, a rickettsia or a virus, or is respiratory or systemic . In all instances, the previously healthy host responds to infection with cytokine-mediated alterations that appear to occur in proportion to the infectious challenge and to the likelihood of death . These alterations also can occur in the vaccinated host, if the infectious challenge is sufficiently great . Because of their widespread occurrence and seemingly ingrained status, these metabolic alterations may be presumed to be of survival benefit to the host . Whether this patterned array of metabolic sequelae is of benefit to the host, or even to the species, its widespread and systematic occurrence allows it to be of value in assessing the safety and efficacy of vaccines and drugs to prevent or treat a wide variety of infections . In this era of bioterrorism, wherein drugs and vaccines may have to be approved for human use without clinical trials and solely on the basis of animal data, these cytokine-mediated metabolic sequelae can aid in the rational selection of drug and vaccine candidates.

Int J Syst Evol Microbiol, 2002 Nov, 52(Pt 6), 2257 - 60
Rothia amarae sp . nov., from sludge of a foul water sewer; Fan Y et al.; A Gram-positive bacterium, strain J18(T), isolated from sludge of a foul water sewer, was subjected to a polyphasic taxonomic study . Phylogenetic analysis of the bacterium based on its 16S rDNA sequence showed that it belongs to the genus Rothia and forms a distinct phyletic clade with the type strain of Rothia nasimurium . Morphological, physiological and chemotaxonomic characteristics supported the assignment of this organism to the genus Rothia and distinguished it from the type strains of all validly described Rothia species . Therefore, it is proposed that this bacterium be classified in the genus Rothia as Rothia amarae sp . nov . The type strain is strain J18(T) (= AS 4.1721(T) = JCM 11375(T)).

Am J Ophthalmol, 2003 Jan, 135(1), 89 - 91
Clinico-microbiological correlation of suture-related graft infection following penetrating keratoplasty; Sonavane A et al.; PURPOSE: To report the clinical and microbiological profiles of suture-related graft infections following penetrating keratoplasty . DESIGN: Interventional case series . METHODS: The medical and microbiology records of all patients who presented with suture-related graft infections and were seen between January 1999 and December 2001 were reviewed . RESULTS: Of 105 patients (105 eyes) who developed corneal infiltrates following penetrating keratoplasty, 37 patients (24 optical and 13 therapeutic) were identified as having suture-related graft infection . The median onset of infiltrates after penetrating keratoplasty was 87 days for 31 patients who presented within one year . Gram-stained smears of corneal scrapings were positive in 27 (73%) of 37 patients, whereas cultures were positive in 32 (only bacteria, 30; mixed bacteria and fungi, 2) . Thirty-seven isolates were obtained from 32 patients (gram-positive bacteria, 32; gram-negative bacteria, 3; fungus, 2) . All gram-positive isolates were sensitive to cefazolin, chloramphenicol, and vancomycin, whereas one of three gram-negative isolates (Pseudomanas species) was resistant to all the antibiotics tested . Of 32 patients, 30 (82%) responded satisfactorily to medical management . CONCLUSIONS: Suture-related graft infections are usually caused by bacteria that are sensitive to ciprofloxacin, cefazolin, and gentamicin, and the patients respond satisfactorily to medical therapy.

J Am Soc Mass Spectrom, 2003 Jan, 14(1), 58 - 62
GC/MS determination of fatty acid picolinyl esters by direct curie-point pyrolysis of whole bacterial cells; Kurkiewicz S et al.; A single-step method suitable for cellular fatty acid derivatization to picolinyl esters with the use of a pyrolyzer as a thermochemical micro-reactor was developed for whole bacterial cells . This reduced the preparation time from several hours to less than two minutes . In addition, the minimal bacterial mass required for analysis was reduced from several milligrams to micrograms . The profiling of cellular fatty acids of gram-positive and gram-negative bacteria was achieved using three derivatization methods: preparation of methyl esters, beta-picolinyl esters by Harvey's method and a new method based on pyrolytic derivatization to beta-picolinyl esters . It was shown that there are great similarities between profiles of bacterial fatty acids determined by the pyrolytic derivatization method and traditional preparation methods of picolinyl and methyl esters prior to GC analysis . Results obtained by application of the new technique have immense diagnostic value due to vast similarities between profiles of fatty acids derivatized to either picolinyl and methyl esters . Although the latter are referred to in the literature most often, mass spectra of picolinyl esters contain fragment ions that provide structural information about the chain branching, position of unsaturation, and other substituents.

Prostate Cancer Prostatic Dis, 1999 May, 2(3), 159 - 162
Use of prostatic massage in combination with antibiotics in the treatment of chronic prostatitis; Shoskes DA et al.; Chronic prostatitis is often refractory to antibiotics, however biopsy and molecular data indicate persistent symptoms may be due to occult infection . Combining antibiotic therapy with regular prostatic massage has been suggested as an effective therapy for some of these men . From November 1996 to December 1998, 73 men with chronic pelvic pain syndromes were treated with antibiotics and prostatic massage . Antibiotic selection was based on culture and sensitivity of prostatic fluid or empirically if cultures were negative . Prostatic massage was done 1 to 3 times per week and fluid examined for WBCs and cultured for bacteria . The average age of the group was 43.5 y (range 23-72) and average duration of symptoms 6.7 y (median 3 y, range 3 months-30 y) . Prostatic cultures were negative in 19, grew uropathogens in 2, and Gram positive bacteria in 52 patients . Overall 29 patients (40%) had complete resolution of symptoms, 14 (19%) had complete resolution followed by a recurrence, 15 (21%) had some improvement and 15 (21%) had no improvement . All positive cultures were sterilized during treatment . Combination prostatic massage and culture specific antibiotics can be an effective treatment in a proportion of men with long standing refractory chronic prostatitis.

Infect Immun, 2003 Jan, 71(1), 524 - 6
Production of macrophage inflammatory protein 3alpha (MIP-3alpha) (CCL20) and MIP-3beta (CCL19) by human peripheral blood neutrophils in response to microbial pathogens; Akahoshi T et al.; Effects of bacterial pathogens on the production of macrophage inflammatory protein 3alpha (MIP-3alpha) and MIP-3beta from human peripheral blood neutrophils were investigated . Neutrophils produced both chemokines by coincubation with either gram-positive or gram-negative bacteria . Neutrophils may initiate antigen-specific immune responses through the release of these chemokines that are capable of promoting selective recruitment of dendritic cells and T-cell subsets.

J Am Coll Surg, 2002 Dec, 195(6), 759 - 67
Prophylactic antibiotics alter the bacteriology of infected necrosis in severe acute pancreatitis; Howard TJ et al.; BACKGROUND: Use of appropriate prophylactic antibiotics has been shown to decrease infectious complications and mortality rate in patients with severe acute pancreatitis, but its influence on the bacteriology of secondary pancreatic infection is poorly defined . STUDY DESIGN: Operative cultures from 61 consecutive patients with pancreatic necrosis treated during routine prophylactic antibiotic use (1993-2001) were compared with 34 consecutive patients with necrosis treated before routine antibiotic use (1977-1992) . RESULTS: The two groups of patients were similar in demographics, etiology of pancreatitis, and severity of illness . All patients in the antibiotic group received prophylactic antibiotics compared with only 38% (13 of 34) in the control group . Routine broad-spectrum prophylactic antibiotics altered the bacteriology of secondary pancreatic infection in severe acute pancreatitis from predominantly gram-negative coliforms (56% versus 26%, p = 0.005) to predominately gram-positive organisms (23% versus 52%, p = 0.009) without a significant increase in either the rate of beta-lactam resistance or fungal infections . The overall hospital stay in patients treated with prophylactic antibiotics was significantly reduced (61 +/- 24 days versus 41 +/- 28 days, p = 0.002), and there was a trend toward a decline in mortality rate in the antibiotic treatment group . CONCLUSION: Routine broad-spectrum prophylactic antibiotic use has altered the bacteriology of secondary pancreatic infection in severe acute pancreatitis from predominantly gram-negative coliforms to predominantly gram-positive organisms without altering the rate of beta-lactam resistance or fungal superinfection.

Intern Med, 2002 Nov, 41(11), 986 - 9
Pulmonary nocardiosis with bilateral diffuse granular lung shadows in a patient with subcutaneous panniculitic T-cell lymphoma; Enomoto M et al.; A 40-year-old woman undergoing prednisolone and cyclosporine therapy for subcutaneous panniculitic T-cell lymphoma complained of a cough for a few weeks . A chest X-ray revealed bilateral diffuse granular shadows . Additionally, the patient was discovered to have multiple subcutaneous abscesses . Gram-stained smears of sputum and pus from the abscess showing branched gram-positive rods led to a diagnosis of pulmonary nocardiosis with dissemination to the lungs and subcutaneous tissues . Combination therapy consisting of sulfamethoxazole/trimethoprim and panipenem/betamipron produced rapid improvement of radiographic abnormalities . It is suggested that pulmonary nocardiosis should be considered in the differential diagnosis of diffuse granular shadows on chest X-rays, especially in immunocompromised patients.

Intern Med, 2002 Nov, 41(11), 957 - 60
Infected left atrial myxoma; Uchino K et al.; A 47-year-old Japanese woman with a continuing high fever was promptly diagnosed as having infected atrial myxoma one day after admission based on transthoracic echocardiographic findings and positivity for bacteria in blood culture . The mass was removed by an urgent open heart surgery . Histopathological examination confirmed that this mass was a myxoma with gram-positive bacterial colonies . Generally, antemortem diagnosis is difficult and there is a high mortality of patients with infected myxoma; however, this patient completely recovered from the illness because of the prompt diagnosis . This is the 37th case of definite infected myxoma reported in the literature . The cause of infection of this patient might have been the acupuncture therapy she underwent for weight reduction.

J Bacteriol, 2003 Jan, 185(1), 20 - 7
Cinnamate:coenzyme A ligase from the filamentous bacterium streptomyces coelicolor A3(2); Kaneko M et al.; 4-Coumarate:coenzyme A ligase (4CL) plays a key role in phenylpropanoid metabolism, providing precursors for a large variety of important plant secondary metabolites, such as lignin, flavonoids, and phytoalexins . Although 4CLs have been believed to be specific to plants, a gene encoding a 4CL-like enzyme which shows more than 40% identity in amino acid sequence to plant 4CLs was found in the genome of the gram-positive, filamentous bacterium Streptomyces coelicolor A3(2) . The recombinant enzyme, produced in Escherichia coli with a histidine tag at its N-terminal end, showed distinct 4CL activity . The optimum pH and temperature of the reaction were pH 8.0 and 30 degrees C, respectively . The K(m) value for 4-coumarate and k(cat) were determined as 131 +/- 4 micro M and 0.202 +/- 0.007 s(-1), respectively . The K(m) value was comparable to those of plant 4CLs . The substrate specificity of this enzyme was, however, distinctly different from those of plant 4CLs . The enzyme efficiently converted cinnamate (K(m), 190 +/- 2 micro M; k(cat), 0.475 +/- 0.012 s(-1)), which is a very poor substrate for plant 4CLs . Furthermore, the enzyme showed only low activity toward caffeate and no activity toward ferulate, both of which are generally good substrates for plant 4CLs . The enzyme was therefore named ScCCL for S . coelicolor A3(2) cinnamate CoA ligase . To determine the amino acid residues providing the unique substrate specificity of ScCCL, eight ScCCL mutant enzymes having a mutation(s) at amino acid residues that probably line up along the substrate-binding pocket were generated . Mutant A294G used caffeate as a substrate more efficiently than ScCCL, and mutant A294G/A318G used ferulate, which ScCCL could not use as a substrate, suggesting that Ala(294) and Ala(318) are involved in substrate recognition . Furthermore, the catalytic activities of A294G and A294G/A318G toward cinnamate and 4-coumarate were greatly enhanced compared with those of the wild-type enzyme.

J Gen Appl Microbiol, 2000 Jun, 46(3), 167 - 178
Cystite formation of Arthrobacter ureafaciens NRIC 0157(T); Tanaka N et al.; The cystite formation of Arthrobacter ureafaciens NRIC 0157(T) was studied by the use of a newly designed CT medium composed of 2.0% D-glucose, 0.28% NH(4)H(2)PO(4), 0.136% K(2)HPO(4) 0.136% KH(2)PO(4), 0.0005% MgSO(4).7H(2)O, and 0.0007% CaCl(2).2H(2)O . Cystites are drumstick-shaped and oval cells and are larger than vegetative cells . Cystites are Gram-negative, whereas vegetative cells are Gram-positive by the KOH reaction . The concentration and ratio of K(+) and Mg(2+) in CT medium mainly affected the cystite formation . Cystites are considered aberrant forms produced by nutritional imbalance.

Med Oncol, 2002, 19(3), 161 - 6
Cefepime monotherapy as an empirical initial treatment of patients with febrile neutropenia; Montalar J et al.; Currently, monotherapy is considered a valid alternative to the combination antibiotic treatments used for initial, empirical management of febrile neutropenia . The advent of new cephalosporins warrants assessment . The aim of this study was to prospectively evaluate the effectiveness of cefepime monotherapy in the treatment of cancer patients with febrile granulocytopenia (< 1000 leukocytes/muL and/or < 500 neutrophils/muL) . A prospective, multicenter, nonrandomized trial was conducted . Initial treatment consisted of iv cefepime, 2 g every 8 h . If the patient was still febrile after 72 h, amikacin, vancomycin/teicoplanin, and amphotericin B were added sequentially . Response was evaluated according to EORTC criteria . One hundred twenty episodes were analyzed in 81 males and 39 females (median age, 52 yr; range, 15-83) . The median leukocyte count at the time of diagnosis was 781 microL(-1) (range, 100-2600) and the median neutrophil count was 173 microL(-1) (range 0-500) . The median duration of neutropenia (< 1000 neutrophils/microL) was 4.8 d (range, 3-20) . Fifty-two episodes (44%) were confirmed microbiologically (42 presented as bacteremia), 31 with Gram-positive bacteria and 21 with Gram-negative bacteria, 47 (39.3%) were confirmed clinically, 16 (13.3%) were considered as probable infections, and 5 (4.2%) as doubtful infections . Protocol success was achieved in 110 episodes (91.7%), 8 (6.6%) were treatment failures, and 2 (1.7%) were not evaluable . Ninety-nine episodes (83.3%) were controlled with cefepime monotherapy, with 19 other episodes requiring additional antibiotics: amikacin in 7 (5.8%), amikacin + vancomycin/teicoplanin in 12 (10.1%) . Three patients (2,5%) died during an episode of neutropenic fever . Cefepime is effective as an initial, empirical treatment of febrile neutropenia . The early addition of amikacin and/or vancomycin resolves most of the monotherapy failures, which seem somewhat lower than with other monotherapies.

Joint Bone Spine, 2002 Oct, 69(5), 499 - 501
Septic arthritis due to Actinomyces naeslundii: report of a case; Lequerre T et al.; In a man with osteoarthritis of the knee, Actinomyces naeslundii septic arthritis developed after intra-articular injection of hyaluronate . Actinomyces is an anaerobic Gram-positive rod . The outcome was favorable after treatment with two antibiotics and arthroscopy . The nature of the organism and its location to a joint are unusual features of this case, which illustrates the need to search for a septic complication before accepting a diagnosis of inflammation related to hyaluronate injection.

Sex Transm Infect, 2002 Dec, 78(6), 413 - 5
Validation of a simplified grading of Gram stained vaginal smears for use in genitourinary medicine clinics; Ison CA et al.; OBJECTIVES: To validate a simplified grading scheme for Gram stained smears of vaginal fluid for the diagnosis of bacterial vaginosis (BV) against the accepted "gold" standard of Amsel's composite criteria . METHODS: Women attending genitourinary medicine (GUM) clinics, as part of a multicentre study, were diagnosed as having BV if three or more of the following criteria were present; homogeneous discharge, elevated vaginal pH, production of amines, and presence of "clue" cells . Women with less than three of the criteria were considered as normal . Simultaneously, smears were made of vaginal fluid and Gram stained and then assessed qualitatively as normal (grade I), intermediate (grade II), or consistent with BV (grade III) . Two new grades were used, grade 0, epithelial cells only with no bacteria, and grade IV, Gram positive cocci only . RESULTS: BV was diagnosed in 83/162 patient visits using the composite criteria, the remainder being regarded as normal . The majority of patients with BV had a smear assessed as grade III (80/83, 96%) and the majority of normal women had a smear assessed as grade I (normal, 48/79, 61%), giving a high sensitivity (97.5%), specificity (96%), and predictive value for a positive (94.1%) and negative (96%) test, kappa index = 0.91 . Smears assessed as grade II were found predominantly (12/13) among patients diagnosed as normal, with less than three of the composite criteria . Grades 0 and IV were both only found among normal women . CONCLUSION: This simplified assessment of Gram stained smears can be used as an alternative to Amsel's criteria and is more applicable for use in busy GUM clinics.

Curr Opin Chem Biol, 2002 Dec, 6(6), 786 - 93
Substrate analogues to study cell-wall biosynthesis and its inhibition; Lazar K et al.; It has been known for more than 30 years that Lipid II is an intermediate in peptidoglycan synthesis . Recently, it has become apparent that it is also an important target of numerous antibiotics, including the glycopeptides, the lantibiotics and ramoplanin . It is also utilized by sortases in the construction of Gram-positive cell walls . Recent progress has been made in the synthesis of peptidoglycan intermediates that can be used to study enzymes which make peptidoglycan . These intermediates also enable studies to probe the mechanism of action of a variety of substrate-binding antibiotics.

Curr Top Microbiol Immunol, 2002, 270, 121 - 44
TLR2: cellular sensor for microbial and endogenous molecular patterns; Kirschning CJ et al.; Toll-like receptor (TLR) 2 is a member of the vertebrate protein family of TLRs that has been studied in substantial detail over the last years . The extracellular domain of the type I receptor molecule TLR2 contains 18 to 20 leucine rich repeat (LRR) and LRR like motives . The intracellular domain of TLR2 contains a Toll/IL-1 receptor/resistance protein typical TIR domain . After the first implication of TLR4 in immunity thereinafter followed by the discovery of the lipopolysaccharide signal transducer function of TLR4, TLR2 was the first of ten mammalian TLRs proven to be directly involved in recognition of pathogen associated molecular patterns (PAMPs) . Among the TLR2 specific agonists are microbial products representing broad groups of species such as Gram-positive and Gram-negative bacteria, as well as mycobacteria, spirochetes, and mycoplasm . PAMP induced phagosomal localization of TLR2 and TLR2 dependent apoptosis have been shown . Complex formation with other molecules involved in pattern recognition such as CD14, MD2, TLR1, and TLR6 has been implicated for TLR2 . Surprisingly even proteinaceous host material such as heat shock protein (HSP) 60 has been demonstrated to activate cells through TLR2 . Thus, TLR2 may be a sensor and inductor of specific defense processes, including oxidative stress and cellular necrosis initially spurred by microbial compounds . Here we summarize the current knowledge on the structure and function of TLR2, which is far from being complete . Detailed understanding of the biology of TLR2 will probably contribute to the characterization of a number of infectious diseases and potentially help in the development of novel intervention strategies.

Appl Microbiol Biotechnol, 2002 Dec, 60(4), 428 - 36 Epub 2002 Nov 06.
Gene cloning, sequencing, and characterization of a family 9 endoglucanase (CelA) with an unusual pattern of activity from the thermoacidophile Alicyclobacillus acidocaldarius ATCC27009; Eckert K et al.; A gene encoding a beta-1,4-glucanase (CelA) belonging to subfamily E1 of family 9 of glycoside hydrolases was cloned and sequenced from the gram-positive thermoacidophile Alicyclobacillus acidocaldarius strain ATCC27009 . The translated protein contains an immunoglobulin-like domain but lacks a cellulose-binding domain . The enzyme, when overproduced in Escherichia coli and purified, displayed a temperature optimum of 70 degrees C and a pH optimum of 5.5 . CelA contained one zinc and two calcium atoms . Calcium and zinc are likely to be important for temperature stability . The enzyme was most active against substrates containing beta-1,4-linked glucans (lichenan and carboxy methyl cellulose), but also exhibited activity against oat spelt xylan . A striking pattern of hydrolysis on p-nitrophenyl-glycosides was observed, with highest activity on the cellobioside derivative, some on the cellotetraoside derivative, and none on the glucoside and cellotrioside derivatives . Unmodified cellooligosaccharides were also hydrolyzed by CelA . No signal peptide for transport across the cytoplasmic membrane was detected . This, together with the substrate specificity displayed, near neutral pH optimum and irreversible inactivation at low pH, suggests a role for CelA as a cytoplasmic enzyme for the degradation of imported oligosaccharides.

Appl Microbiol Biotechnol, 2002 Dec, 60(4), 377 - 80 Epub 2002 Oct 24.
Impact of the first Streptomyces genome sequence on the discovery and production of bioactive substances; Donadio S et al.; An important addition to the field of bacterial genomics is the recent publication of the complete genome sequence of Streptomyces coelicolor . This strain has been for some decades the model organism for streptomycetes and other filamentous actinomycetes, Gram-positive bacteria highly valuable for their ability to produce thousands of bioactive metabolites, many of which have found important applications in medicine and agriculture . We discuss here the impacts that the S . coelicolor genome sequence is likely to have on the production of bioactive metabolites by current industrial strains, on the possible development of future superhost(s) for the production of valuable drugs, and on the search for new bioactive substances from microbial sources.

J Med Microbiol, 2002 Dec, 51(12), 1097 - 101
Development of a PCR assay specific for Peptostreptococcus anaerobius; Riggio MP et al.; Peptostreptococcus anaerobius is a gram-positive anaerobic coccus that is widely distributed in the normal human flora . The organism has also been implicated as a causative agent of several systemic infections, including endocarditis and infections of the genitourinary and gastrointestinal tracts . Its role in oral disease is less well defined, although it has been implicated in periodontal disease, gingivitis and root canal infections . Identification of P . anaerobius in clinical samples is currently reliant upon traditional culture and biochemical methods . The aim of this study was to develop a novel PCR assay for the detection of P . anaerobius and to attempt detection of this organism in oral samples . PCR primers specific for P . anaerobius DNA were developed by alignment of bacterial 16S ribosomal RNA gene sequences and selection of sequences specific at their 3' ends for P . anaerobius . When used in a PCR assay, positivity for P . anaerobius DNA was indicated by the amplification of a 943-bp product . The primers were shown to be specific for P . anaerobius DNA, as no PCR products were obtained when genomic DNA from a wide range of other Peptostreptococcus species and other oral bacteria were used as templates . The PCR assay was then applied to the detection of P . anaerobius DNA in subgingival plaque samples from adult periodontitis patients and pus aspirates from subjects with acute dento-alveolar abscesses . All of 60 subgingival plaque samples from 16 patients were negative for P . anaerobius DNA . None of the 43 pus samples analysed contained P . anaerobius DNA . These results suggest that P . anaerobius is not a major pathogen in adult periodontitis and dento-alveolar abscesses . The PCR assay is a more rapid, sensitive and specific alternative to culture-based methods for identification of P . anaerobius in clinical samples.

Burns, 2002 Dec, 28(8), 746 - 51
Septicaemia after burn injury: a comparative study; Bang RL et al.; Seventy-nine (8.4%) patients during June 1992-May 1996 (Group-1) and 68 (7.2%) patients from June 1996 to May 2000 (Group-2) who developed septicaemia at the burns unit of Al-Babtain Centre for Plastic Surgery and Burns, Kuwait, were retrospectively studied and compared . The mean age of 26 years, male predominance, flame burns as main aetiology and mean burn percentage of >or=40% was observed in both the groups . Both groups revealed extensive flame burn, inhalation injury, intubation and difficult resuscitation as the risk factors . The proportion of satisfactory resuscitation increased significantly (P<0.001) in Group-2 . The septicaemia commonly occurred within 2 weeks postburn but the number of episodes during 5 days postburn was less in Group-2 . The surface wound was found to be the likely source of entry of the organisms into the blood stream in both the groups . The gram positive organisms were dominant aetiologic factor in both groups but an increase frequency of Acnetobacter was found in Group-2 . The proportion of MRSE and Pseudomonas septicaemia was significantly higher (P<0.01) in the Group-1 . The rate of survivors, in both the groups was higher among operated patients but it was significantly higher (P<0.001) in the Group-1 . A mortality rate 20.6% in Group-2 decreased against Group-1, which can be attributed to better resuscitation, nutritional care, early detection of septicaemia, appropriate antibiotics and early wound excision and skin grafting . MOF was the cause of death of 60.9% in Group-1 and 85.7% in Group-2 . There was no role of prophylactic antibiotic in burn patients in the incidence of septicaemia and mortality.

Med Mycol, 2002 Oct, 40(5), 525 - 7
Mycetoma due to Pseudallescheria boydii and co-isolation of Nocardia abscessus in a patient injured in road accident; Horre R et al.; We report the case of a patient who developed a mycetoma after experiencing a road accident . From surgical biopsies Pseudallescheria boydii was isolated . Subsequently, after the infection had been treated with itraconazole, a Gram-positive bacterium, identified as the newly described species Nocardia abscessus, was cultured from wound fluids.

Mutat Res, 2002 Dec 29, 510(1-2), 9 - 22
poliota-dependent lesion bypass in vitro; Vaisman A et al.; Based upon phylogenetic relationships, the broad Y-family of DNA polymerases can be divided into various subfamilies consisting of UmuC (polV)-like; DinB (polIV/polkappa)-like; Rev1-like, Rad30A (poleta)-like and Rad30B (poliota)-like polymerases . The polIV/polkappa-like polymerases are most ubiquitous, having been identified in bacteria, archaea and eukaryotes . In contrast, the polV-like polymerases appear restricted to bacteria (both Gram positive and Gram negative) . Rev1 and poleta-like polymerases are found exclusively in eukaryotes, and to date, poliota-like polymerases have only been identified in higher eukaryotes . In general, the in vitro properties of polymerases characterized within each sub-family are quite similar . An exception to this rule occurs with the poliota-like polymerases, where the enzymatic properties of Drosophila melanogaster poliota are more similar to that of Saccharomyces cerevisiae and human poleta than to the related human poliota . For example, like poleta, Drosophila poliota can bypass a cis-syn thymine-thymine dimer both accurately and efficiently, while human poliota bypasses the same lesion inefficiently and with low-fidelity . Even in cases where human poliota can efficiently insert a base opposite a lesion (such as a synthetic abasic site, the 3'T of a 6-4-thymine-thymine pyrimidine-pyrimidone photoproduct or opposite benzo{a}pyrene diol epoxide deoxyadenosine adducts), further extension is often limited . Thus, although poliota most likely arose from a genetic duplication of poleta millions of years ago as eukaryotes evolved, it would appear that poliota from humans (and possibly all mammals) has been further subjected to evolutionary pressures that have "tailored" its enzymatic properties away from lesion bypass and towards other function(s) specific for higher eukaryotes . The identification of such functions and the role that mammalian poliota plays in lesion bypass in vivo, should hopefully be forthcoming with the construction of human cell lines deleted for poliota and the identification of mice deficient in poliota.

Front Biosci, 2003 Jan 01, 8, d20 - 31
The T box and S box transcription termination control systems; Grundy FJ et al.; The T box and S box transcription termination control systems are widely used for control of gene expression in Gram-positive bacteria, but are rare in Gram-negative organisms . Both of these systems can be recognized in genomic data because of high conservation of primary sequence and structural elements . The T box system regulates a variety of amino acid-related genes, while the S box system is dedicated to genes involved in methionine metabolism . While both systems involve gene regulation at the level of premature termination of transcription, the molecular mechanisms employed are very different . In the T box system, expression is induced by stabilization of an antiterminator structure in the leader by interaction with the cognate uncharged tRNA; this prevents formation of the competing terminator helix, allowing synthesis of the full-length mRNA . Disruption of conserved leader features results in loss of readthrough . In the S box system, the antiterminator form of the leader is the more stable form . A competing anti-antiterminator must be stabilized by an unknown factor during growth in methionine to prevent formation of the antiterminator, thereby allowing formation of the terminator helix . Disruption of conserved leader elements results in constitutive expression.

Mol Microbiol, 2002 Dec, 46(5), 1295 - 304
Ribosomal and non-ribosomal resistance to oxazolidinones: species-specific idiosyncrasy of ribosomal alterations; Sander P et al.; A derivative of Mycobacterium smegmatis, which carries only one functional rRNA (rrn) operon, was used to isolate mutants resistant to the ribosome-targeted antibiotic linezolid . Isolation and characterization of linezolid-resistant clones revealed two classes of mutants . Ribosomes from class I mutants are resistant to oxazolidinones in an in vitro peptidyl transferase assay, indicating that resistance maps to the ribosome component . In contrast, ribosomes from class II mutants show wild-type susceptibility to a linezolid derivative in vitro, pointing to a non-ribosomal mechanism of resistance . Introduction of a wild-type ribosomal RNA operon into linezolid-resistant strains restored linezolid sensitivity in class I mutants, indicating that resistance (i) maps to the rRNA and (ii) is recessive . Sequencing of the entire rrn operon identified a single nucleotide alteration in 23S rRNA of class I mutant strains, 2447G --> T (Escherichia coli numbering) . Introduction of mutant rrl2447T into M . smegmatis rrn- resulted in a linezolid-resistant phenotype, demonstrating a cause-effect relationship of the 2447G --> T alteration . The 2447G --> T mutation, which renders M . smegmatis linezolid resistant, confers lethality in E . coli . This finding is strong evidence of structural and pos-sibly functional differences between the ribosomes of Gram-positive and Gram-negative bacteria . In agreement with the results of the in vitro assay, class II mutants show a wild-type sequence of the complete rRNA operon . The lack of cross-resistance of the class II mutants to other antibiotics suggests a resistance mechanism other than activation of a broad-spectrum multidrug transporter.

Wound Repair Regen, 2002 Nov-Dec, 10(6), 346 - 53
Potential role of anaerobic cocci in impaired human wound healing; Wall IB et al.; Although more than 80% of infected and 70% of noninfected leg ulcers have been shown to harbor anaerobic organisms, their role in mediating impaired wound healing in the skin is frequently overlooked . There is now increasing evidence that the gram-positive anaerobic cocci play a role (both directly and indirectly) in mediating impaired wound healing in vivo . This article discusses the mechanisms by which these microorganisms may interfere with the inflammation, repair, and remodeling phases of the wound healing process.

Infect Control Hosp Epidemiol, 2002 Nov, 23(11), 683 - 8
Effectiveness of pharmacy policies designed to limit inappropriate vancomycin use: a population-based assessment; Thomas AR et al.; OBJECTIVE: In Oregon in 1994, a population-based study of 66 nonpsychiatric hospitals indicated that 40% of vancomycin orders were inappropriate according to Centers for Disease Control and Prevention guidelines . We repeated the study to determine whether vancomycin use had been affected by pharmacy policies implemented following the 1994 study . METHODS: We surveyed pharmacists in nonpsychiatric hospitals in Oregon regarding vancomycin use policies in their hospitals . Using pharmacy records, we identified and abstracted the charts of all patients in Oregon hospitals receiving vancomycin during a 3-week period to determine appropriate use of vancomycin . RESULTS: Thirteen (20%) of 64 hospitals had implemented a vancomycin restriction policy since 1994; none ofthe remaining hospitals in the study had a policy . In 1999, hospitals with vancomycin restriction policies had substantially decreased rates of inappropriate vancomycin use compared with hospitals without such policies (1.0 vs 1.8 orders per 1,000 patient-days; P = .01) . Compared with 1994 baseline rates of inappropriate use, hospitals that adopted policies experienced a decrease (from 1.5 orders per 1,000 patient-days in 1994 to 1.0 in 1999; P= .13), whereas hospitals without policies experienced a statistically significant increase (from 0.9 orders per 1,000 patient-days in 1994 to 1.8 in 1999; P= .001) . Restriction policies were most effective at reducing rates of inappropriate use for treatment of confirmed gram-positive infections and prophylaxis . CONCLUSION: Vancomycin restriction policies were associated with a decrease in inappropriate therapeutic and prophylactic vancomycin use, but had no effect on inappropriate empiric use . Hospitals considering limits regarding inappropriate use should consider implementation of institution-based vancomycin restriction policies as part of an overall strategy.

Appl Environ Microbiol, 2002 Dec, 68(12), 6094 - 105
Impact of protozoan grazing on bacterial community structure in soil microcosms; Ronn R et al.; The influence of grazing by a mixed assemblage of soil protozoa (seven flagellates and one amoeba) on bacterial community structure was studied in soil microcosms amended with a particulate resource (sterile wheat roots) or a soluble resource (a solution of various organic compounds) . Sterilized soil was reinoculated with mixed soil bacteria (obtained by filtering and dilution) or with bacteria and protozoa . Denaturing gradient gel electrophoresis (DGGE) of PCR amplifications of 16S rRNA gene fragments, as well as community level physiological profiling (Biolog plates), suggested that the mixed protozoan community had significant effects on the bacterial community structure . Excising and sequencing of bands from the DGGE gels indicated that high-G+C gram-positive bacteria closely related to Arthrobacter spp . were favored by grazing, whereas the excised bands that decreased in intensity were related to gram-negative bacteria . The percentages of intensity found in bands related to high G+C gram positives increased from 4.5 and 12.6% in the ungrazed microcosms amended with roots and nutrient solution, respectively, to 19.3 and 32.9% in the grazed microcosms . Protozoa reduced the average bacterial cell size in microcosms amended with nutrient solution but not in the treatment amended with roots . Hence, size-selective feeding may explain some but not all of the changes in bacterial community structure . Five different protozoan isolates (Acanthamoeba sp., two species of Cercomonas, Thaumatomonas sp., and Spumella sp.) had different effects on the bacterial communities . This suggests that the composition of protozoan communities is important for the effect of protozoan grazing on bacterial communities.

J Invest Dermatol, 2002 Nov, 119(5), 1090 - 5
Cathelicidin anti-microbial peptide expression in sweat, an innate defense system for the skin; Murakami M et al.; The eccrine gland is one of the major cutaneous appendages and secretes sweat . Its principal function is thermoregulation during exposure to a hot environment or physical exercise . In addition to this function, we show that LL-37, a member of cathelicidin family of anti-microbial peptides, is expressed in sweat . LL-37 protein and mRNA was seen in the eccrine structures of normal human skin by immunohistochemistry and in situ hybridization . LL-37 was localized to both the eccrine gland and sweat ductal epithelial cells, whereas dermcidin, a previously described natural antibiotic in sweat, was expressed only in the gland itself . The anti-microbial activity of LL-37 and dermcidin against various bacteria in the sweat ionic environment was demonstrated by solution colony forming assay using synthetic peptides, and in sweat obtained from normal volunteers . These results indicate that cathelicidin is secreted in human sweat, has potent anti-microbial activity against both gram-positive and gram-negative bacteria, and can, after processing from the preproform, provide a barrier for protection against infection . Thus, sweat represents a unique mode of delivery for potent innate immune effector molecules in the absence of inflammation.

J Dermatol Sci, 2002 Dec, 30(3), 185 - 94
Expression of functional Toll-like receptor 2 on human epidermal keratinocytes; Kawai K et al.; Epidermal keratinocytes secrete cytokines, chemokines, and anti-microbial peptides in response to various microbial pathogens and their components including lipopolysaccharide (LPS) . To identify the receptor(s) involved in the anti-microbial responses of epidermal keratinocytes, we analyzed expression of CD14, Toll-like receptor 2 (TLR2), and TLR4 on cultured normal human epidermal keratinocytes (NHEK) . Although CD14 and TLR2 mRNA were expressed in cultured NHEK, only TLR2 was detected on the cell surface . Cultured NHEK did not express TLR4 mRNA or protein . Commercial LPS preparations could stimulate epidermal keratinocytes to produce beta-defensin-2 and IL-8, and the LPS response was inhibited with mAb specific for TLR2, but not for CD14 or TLR4 . Repurified LPS and lipid A did not stimulate epidermal keratinocytes, whereas peptidoglycan (PGN) from Gram-positive bacteria and yeast cell wall particle induced beta-defensin-2 and IL-8 production . Thus, cultured NHEK express functional TLR2, but not CD14 or TLR4, and the "LPS" response of epidermal keratinocytes shown in the previous studies might be mediated by TLR2-dependent recognition of non-LPS bacterial components contaminating in commercial LPS preparations . In the normal human skin, however, epidermal keratinocytes expressed both TLR2 and TLR4 . Because TLR4 was induced in epidermal keratinocytes by in vitro stimulation with PGN from Gram-positive bacteria, constitutive expression of TLR4 on epidermal keratinocytes in vivo might also be induced by continuous recognition of the resident skin flora containing Gram-positive bacteria through TLR2.

Clin Orthop, 2002 Nov, (404), 125 - 31
Results of direct exchange or debridement of the infected total knee arthroplasty; Silva M et al.; In this literature review, 30 reports provided outcome data on 37 direct exchange arthroplasties, 530 open debridements, and 23 arthroscopic debridements . The average followup was approximately 4 years, but the range was broad (range, 0.02-14 years) . Infection was controlled in 33 of the 37 infected total knee arthroplasties (89.2%) treated by direct exchange arthroplasty, in only 173 of the 530 infected total knee arthroplasties (32.6%) treated by open debridement and retention of the prosthetic components, and in 12 of the 23 infected total knee arthroplasties (52.2%) treated by arthroscopic debridement . There was wide variability in associated antibiotic therapy . Factors associated with successful direct exchange included infections by gram-positive organisms, absence of sinus formation, use of antibiotic-impregnated bone cement for the new prosthesis, and 12 weeks of antibiotic therapy . Direct exchange arthroplasty failed in four of 37 knees; two were in patients with rheumatoid arthritis who were taking corticosteroids . Factors associated with successful debridements included those done within 4 months of the index procedure, or in patients with less than 4 weeks of symptoms, antibiotic sensitive gram-positive organisms, well-fixed components with no radiologic evidence of osteitis, and in young healthy patients . Factors associated with the failed debridements included postoperative drainage for more than 2 weeks, sinus tracts present at the time of the debridement, a hinged prosthesis, and an immunocompromised host . Direct exchange can be successful with a sensitive organism in a healthy host with prolonged antibiotic therapy . Debridement can be successful in early infections in a healthy host.

Microbiology, 2002 Nov, 148(Pt 11), 3583 - 97
A new regulatory DNA motif of the gamma subclass Proteobacteria: identification of the LexA protein binding site of the plant pathogen Xylella fastidiosa; Campoy S et al.; Escherichia coli LexA protein is the repressor of a gene network whose members are directly involved in the repair of damaged DNA and in the survival of bacterial cells until DNA lesions have been eliminated . The lexA gene is widely present in bacteria, although the sequences of only three LexA-binding sites are known: Gram-positive, alpha Proteobacteria and some members of gamma Proteobacteria represented by E . coli . Taking advantage of the fact that the genome sequence of the plant-pathogenic bacterium Xylella fastidiosa has been determined, its lexA gene has been cloned and overexpressed in E . coli to purify its product . After demonstration that X . fastidiosa lexA and recA genes are co-transcribed, gel mobility shift assays and directed mutagenesis experiments using the promoter of the lexA-recA transcriptional unit demonstrated that the X . fastidiosa LexA protein specifically binds the imperfect palindrome TTAGN(6)TACTA . This is the first LexA binding sequence identified in the gamma Proteobacteria differing from the E . coli-like LexA box . Although a computational search has revealed the presence of TTAGN(6)TACTA-like motifs upstream of X . fastidiosa genes other than lexA, X . fastidiosa LexA only binds the promoter of one of them, XF2313, encoding a putative DNA-modification methylase . Moreover, X . fastidiosa LexA protein does not bind any of the other genes whose homologues are regulated by the LexA repressor in E . coli (uvrA, uvrB, ssb, ruvAB, ftsK, dinG, recN and ybfE) . RT-PCR quantitative analysis has also demonstrated that lexA-recA and XF2313 genes, as well as the X . fastidiosa genes which are homologues to those of E . coli belonging to the LexA regulon, with the exception of ssb, are DNA damage-inducible in X . fastidiosa.

J Bacteriol, 2002 Dec, 184(23), 6437 - 47
Impact of phosphorylation of specific residues in the tyrosine autokinase, Wzc, on its activity in assembly of group 1 capsules in Escherichia coli; Paiment A et al.; Wzc(CPS) is a tyrosine autokinase essential for the assembly of a high-molecular-weight (HMW) group 1 capsular polysaccharide (CPS) in Escherichia coli . Homologues of Wzc participate in the formation of CPS and exopolysaccharides in a variety of gram-positive and gram-negative bacteria . Phosphorylation of tyrosine residues in the Wzc(CPS) C terminus is essential for HMW CPS assembly . Overexpression of Wzb(CPS) (phosphatase) in a wild-type background caused a 3.7-fold decrease in the amount of cell-associated K30 CPS produced, confirming the importance of Wzc(CPS) phosphorylation for capsule assembly . In this study, the tyrosine-rich region was dissected in an attempt to identify residues critical for Wzc(CPS) phosphorylation and/or capsule expression . Site-directed mutagenesis demonstrated that no single tyrosine residue in this region is sufficient for detectable phosphorylation of Wzc(CPS) in vivo or for HMW CPS expression . Furthermore, no single tyrosine residue is essential for phosphorylation or capsule assembly, since removal of any one tyrosine residue has no detectable effect . Altering combinations of tyrosine residues (from two to five) led to Wzc(CPS) derivatives that were still competent for phosphorylation but that could not support assembly of HMW CPS, showing that phosphorylation of Wzc per se is not an accurate measure of its ability to function in capsule assembly . One interpretation of these data is that the overall level of phosphorylation in this region, rather than the precise combination of residues accessible to phosphorylation, is important for the activity of Wzc(CPS) . Tyrosine 569, a residue shown to modulate the in vitro phosphorylation of Wzc(CA) from E . coli K-12, was also mutated . The derivative with this mutation still functioned in capsule assembly . Quantitation of K30(CPS) from this mutant revealed no difference in the amount of polymer produced . Finally, dithiobis(succinimidylpropionate) cross-linking was used to confirm that Wzc(CPS) forms complexes in vivo, independent of the phosphorylation state of the protein.

Biochimie, 2002 May-Jun, 84(5-6), 545 - 57
Mode of action of modified and unmodified bacteriocins from Gram-positive bacteria; Hechard Y et al.; The antibiotic activity of bacteriocins from Gram-positive bacteria, whether they are modified (class I bacteriocins, lantibiotics) or unmodified (class II), is based on interaction with the bacterial membrane . However, recent work has demonstrated that for many bacteriocins, generalised membrane disruption models as elaborated for amphiphilic peptides (e.g . tyriodal pore or carpet model) cannot adequately describe the bactericidal action . Rather, specific targets seem to be involved in pore formation and other activities . For the nisin and epidermin family of lantibiotics, the membrane-bound cell wall precursor lipid II has recently been identified as target . The duramycin family of lantibiotics binds specifically to phosphoethanolamine which results in inhibition of phospholipase A2 and various other cellular functions . Most of the class II bacteriocins dissipate the proton motive force (PMF) of the target cell, via pore formation . The subclass IIa bacteriocin activity likely depends on a mannose permease of the phosphotransferase system (PTS) as specific target . The subclass IIb bacteriocins (two-component) also induce dissipation of the PMF by forming cation- or anion-specific pores; specific targets have not yet been identified . Finally, the subclass IIc comprises miscellaneous peptides with various modes of action such as membrane permeabilization, specific inhibition of septum formation and pheromone activityPublication Types:

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