J Acquir Immune Defic Syndr, 1999 Oct 1, 22(2), 155 - 60 Bacteremia in HIV-infected patients: short-term predictors of mortality; Omenaca C et al.; To identify characteristics associated with mortality in HIV-infected patients with bacteremia, 88 bacteremic episodes in 80 HIV-infected patients were prospectively identified over a 5-month period and observed for 30 days . Demographic, clinical, laboratory, and radiologic data were collected . Mean and median age was 41 years . Most study subjects were homosexual men . Median CD4 count was 20 cells/mm3 . Gram-positive organisms predominated (65%) . The most common source of bacteremia was intravascular catheters (45%) . Overall mortality was 30% . A history of malignancy, three or more opportunistic infections, shock, low hemoglobin, source of bacteremia other than an intravascular catheter, resistance to therapy, and a second bacteremic episode during the study period, were all found to be independent predictors of mortality . In this cohort of HIV-infected patients, most of whom were severely immunosuppressed, several factors were found to be significantly and independently associated with mortality. Int J Syst Evol Microbiol, 2000 May, 50 Pt 3, 1221 - 8 Papillibacter cinnamivorans gen . nov., sp . nov., a cinnamate-transforming bacterium from a shea cake digester; Defnoun S et al.; A new, strictly anaerobic, Gram-positive, non-sporulating, mesophilic bacterium, designated strain CIN1T (T=type strain) was isolated from an anaerobic digester fed with shea cake rich in tannins and aromatic compounds . Cells of strain CIN1T were rod-shaped, had characteristically pointed ends (1.3-3.0 x 0.5-0.6 microm) and occurred singly, in pairs and sometimes in chains of up to six . The pH range for growth was 6.9-8.5 and the temperature growth range was 15-40 degrees C . Optimum growth occurred with yeast extract and cinnamate at 37 degrees C and a pH of 7.5 . The isolate transformed cinnamate by degrading the aliphatic side chain to produce acetate and benzoate rather than by aromatic ring cleavage or demethoxylation . The position of the methoxyl group appears to be important in the degradation of the aliphatic side chain of cinnamate; consequently, 3-methoxycinnamate and 4-methoxycinnamate, but not 2-methoxycinnamate, are transformed to produce acetate and methoxybenzoates, namely 3-methoxybenzoate and 4-methoxybenzoate, respectively . Crotonate is degraded to acetate and butyrate . The G+C content of the DNA is 56 mol% . Phylogenetic analysis of the 16S rRNA gene of strain CIN1T indicated that it was a member of the low-G+C-containing Gram-positive branch with a specific relationship to Sporobacter termitidis (sequence identity of 88%) . The phylogenetic results concur with the phenotypic data which reveals that the isolate is a novel bacterium and, based on these findings, strain CIN1T (= DSM 12816T = ATCC 700879T) has been designated Papillibacter cinnamivorans gen . nov., sp . nov. Int J Syst Evol Microbiol, 2000 May, 50 Pt 3, 1151 - 4 Vagococcus fessus sp . nov., isolated from a seal and a harbour porpoise; Hoyles L et al.; A polyphasic taxonomic study was performed on two strains of an unknown Gram-positive, catalase-negative, coccus-shaped bacterium isolated from a dead seal and a harbour porpoise . Comparative 16S rRNA gene sequencing demonstrated that the unknown bacterium represents a new subline within the genus Vagococcus close to, but distinct from, Vagococcus fluvialis, Vagococcus lutrae and Vagococcus salmoninarum . The unknown bacterium was readily distinguished from the three currently recognized Vagococcus species by biochemical tests and electrophoretic analysis of whole-cell proteins . Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as a new species, Vagococcus fessus . The type strain of Vagococcus fessus is CCUG 41755T. J Cardiovasc Surg (Torino), 2000 Feb, 41(1), 137 - 41 Subcarinal foregut cysts . A unique clinical problem; Zikri MA et al.; BACKGROUND: To evaluate the risks associated with a subcarinal foregut cyst in a fixed mediastinal space . METHODS: Design: Between January 1, 1986, and August 1, 1997, 8 patients who had subcarinal cysts and who underwent surgical intervention were identified . These results were analyzed to identify associated symptoms and results of surgical intervention . Mean duration of follow-up was 37.3+/-2.2 months and was 100% complete . Patients: Of the eight patients, three were men . Mean age was 45.6+/-15.6 years (range 24-66) . All patients were symptomatic . Six patient suffered respiratory distress . Four patients complained of chest pain . Preoperatively, all patients underwent routine chest radiography . Six patients underwent computed chest tomography (CT); 4 patients had magnetic resonance imagery (MRI) of the chest . Cardiac echocardiography was performed on 4 patients and esophagogastroduodenoscopy (EGD) with or without esophageal ultrasound (EUS) was done in 4 . Of 7 patients who underwent bronchoscopy, 6 patients demonstrated extrinsic airway compression . The remaining patient showed fistulous communication . Intervention: Cyst dimensions ranged from 7.33+/-1 cm (mean+/-SD) . Total resection of the cyst was accomplished in 6 patients . One patient with fistulization underwent right main bronchial sleeve resection . Histopathology revealed inflammation of the cyst in 2 cases and calcification of the cyst wall in 1 . Fluid from one cyst grew gram positive cocci . RESULTS: Length of hospital stay ranged from 10.9+/-4.4 days . There were no mortalities . Morbidity included prolonged ventilation (1), pulmonary embolism (1) and left recurrent laryngeal palsy (1) . CONCLUSIONS: Our results corroborate the need for surgical excision of subcarinal subtype cysts to prevent the development of mediastinal compression and other cyst-related complications. Eur J Clin Microbiol Infect Dis, 2000 Apr, 19(4), 243 - 7 Bacterial resistance and overgrowth due to selective decontamination of the digestive tract; Ebner W et al.; Infection of the lower airways is a major problem in ventilated patients and contributes substantially to morbidity and mortality in the intensive care unit . The selective decontamination of the digestive tract and its effect on the reduction of the gram-negative colonisation rate in patients has been studied widely . However, the findings are inconsistent . Most studies describe an increase in resistant gram-negative bacterial strains and/or an increase in the occurrence of gram-positive strains following selective decontamination of the digestive tract . In light of the unresolved questions concerning the efficacy of selective decontamination of the digestive tract, it would seem that the resultant effect of this treatment on the bacterial flora should be an important consideration when assessing the value of such treatment . To date, none of the studies available for examination have been designed to adequately assess the effect of selective decontamination of the digestive tract on the bacterial flora. Microbiology, 2000 May, 146 ( Pt 5), 1099 - 107 Microbial community changes in biological phosphate-removal systems on altering sludge phosphorus content; Liu WT et al.; Biomarkers (respiratory quinones and cellular fatty acids) and denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA genes were used to characterize the microbial community structure of lab-scale enhanced biological phosphate-removal (EBPR) systems in response to altering sludge phosphorus (P) content . All the data suggest that the microbial community structures of sludge samples with a P content between 8 and 12.3% (sludge dry weight) (i.e . good EBPR activity) were very similar, but differed from those with 2% P content (i.e . no EBPR activity) . For all samples analysed, ubiquinones Q-8 and Q-10, menaquinone MK-8(H4), and fatty acids C16:0, C16:1 omega9c and C18:1, omega11c were the major components . The dominance of Q-8, Q-10 and MK-8(H4) suggested that large numbers of organisms belonging to the beta and alpha subclasses of the Proteobacteria and the Actinobacteria from the high G+C Gram-positive bacteria, respectively, were present . DGGE analysis revealed at least 7-9 predominant DNA bands and numerous other fragments in each sample . Five major DGGE fragments from each of the 2% and 12% P-containing sludge samples, respectively, were successfully isolated and sequenced . Phylogenetic analysis of the sequences indicated that both 2% and 12% P-containing sludge samples shared three common phylotypes that were separately affiliated with a novel bacterial group from the gamma subclass of the Proteobacteria, two MK-8(H4)-containing actinobacteria previously isolated from the 2% P-containing sludge, and a Caulobacter spp . in the alpha subclass of the Proteobacteria . The phylogenetic analysis also revealed phylotypes unique to both sludge samples . Changes in sludge P content therefore had an effect on the composition and abundance of the predominant microbial populations, though specific phylotypes could not be unequivocally associated with EBPR. Appl Environ Microbiol, 2000 Jun, 66(6), 2555 - 64 Use of randomly amplified polymorphic DNA as a means of developing genus- and strain-specific Streptomyces DNA probes; Roberts MA et al.; We have analyzed 20 randomly amplified polymorphic DNA (RAPD) primers against 36 Streptomyces strains, including 17 taxonomically undefined strains, 25 nonstreptomycete actinomycetes, and 12 outgroups consisting of gram-positive and -negative species . Most of the primers were useful in identifying unique DNA polymorphisms of all strains tested . We have used RAPD techniques to develop a genus-specific probe, one not necessarily targeting the ribosomal gene, for Streptomyces, and a strain-specific probe for the biological control agent Streptomyces lydicus WYEC108 . In the course of these investigations, small-scale DNA isolations were also developed for efficiently isolating actinomycete DNA . Various modifications of isolation procedures for soil DNA were compared, and the reliability and specificity of the RAPD methodology were tested by specifically detecting the S . lydicus WYEC108 in DNA isolated from soil. Leuk Lymphoma, 2000 Jul, 38(3-4), 419 - 22 Non-Hodgkin's lymphoma presenting as anasarca: probably mediated by tumor necrosis factor alpha (TNF-alpha); Jillella AP et al.; Two patients presented with anasarca, fevers and sweats . Subsequent evaluation revealed aggressive lymphoproliferative disease . Both patients were treated with CHOP chemotherapy . One patient responded with spontaneous, vigorous diuresis and complete resolution of the edema . She relapsed two months later with recurrent edema that responded a second time to salvage chemotherapy . The second patient died of gram positive sepsis a week after diagnosis . As anasarca is an unusual presenting symptom of non-Hodgkin's lymphoma, we postulated that the malignant cells were secreting a cytokine that resulted in "vascular leakage" of fluid and development of diffuse edema . Several serum cytokine levels were tested . Both patients had elevated TNF-alpha levels, which could have been the cause of the edema; or there might be yet another unidentified mediator that was responsible for the anasarca . We report these two cases to bring to attention the unusual nature of this presentation. Infect Dis Clin North Am, 2000 Jun, 14(2), 475 - 87 Once-daily dosing of aminoglycoside antibiotics; Fisman DN et al.; Improved understanding of the pharmacodynamics and toxicity of aminoglycoside antibiotics has resulted in the study of once-daily dosing regimens . Although studies have suggested a therapeutic advantage and possibly a decrease in toxicity with once-daily administration, these effects have been modest . The cost savings associated with once-daily aminoglycoside administration, however, makes this approach appealing . Although a syndrome of fever, tachycardia, hypotension, and rigors has been associated with once-daily dosing of gentamicin, this appears to have been the result of impurities in the antibiotic from a single offshore supplier . This syndrome has not been associated with other aminoglycoside antibiotics, and the FDA has now withdrawn its recommendation that once-daily aminoglycoside use be avoided . As with any medical regimen, the decision to use once-daily dosing of aminoglycoside agents must take into account special patient characteristics and the disease state being treated . Although once-daily dosing appears effective in limited studies in children, in individuals with neutropenia, and in individuals with cystic fibrosis, its role in gram-positive coccal endocarditis and in individuals with altered volumes of distribution remains uncertain . Further data are needed to clarify the role of once-daily dosing in these situations. Int J Syst Evol Microbiol, 2000 Jan, 50 Pt 1, 171 - 8 Methanosarcina semesiae sp . nov., a dimethylsulfide-utilizing methanogen from mangrove sediment; Lyimo TJ et al.; Methanosarcina semesiae MD1T (T = type strain), a novel obligately methylotrophic methanogenic archaeon is described . Strain MD1T was isolated from an enrichment on dimethylsulfide inoculated with mangrove sediment . The cells were irregularly coccoid, non-motile, 1.4+/-0.2 microm in diameter and stained Gram-positive . The catabolic substrates used included dimethylsulfide, methanethiol, methanol and methylated amines, but not acetate, formate, H2/CO2 or a combination of these substrates . When cells grown on dimethylsulfide were transferred to trimethylamine or methanol and vice versa, a lag phase was observed . The same lag phase occurred when cells grown on trimethylamine were transferred to methanol and vice versa, indicating that for each substrate different enzymes were induced . Fastest growth occurred within a temperature range of 30-35 degrees C and a pH of 6.5-7.5 . Both Na+ and Mg2+ were required for growth, with maximum growth rates at 200-600 mM Na+ and 20-100 mM Mg2+ . The cells exhibited specific growth rates (h-1) of 0.07+/-0.02, 0.15+/-0.04 and 0.18-/+0.05 on dimethylsulfide, methanol and trimethylamine, respectively . Analysis of the 16S rRNA gene sequence showed that strain MD1T was phylogenetically closely related to members of the genus Methanosarcina, but clearly differed from all described species of this genus (94-97% sequence similarity). Int J Syst Evol Microbiol, 2000 Jan, 50 Pt 1, 127 - 34 Intrageneric relationships among Micromonospora species deduced from gyrB-based phylogeny and DNA relatedness; Kasai H et al.; The phylogenetic structure of genus Micromonospora within actinomycetes was examined by analysing the gyrB sequences of 15 validly described species and four subspecies . All but one of the Micromonospora strains formed a tight cluster, as had previously been demonstrated by a 16S rDNA-based phylogenetic analysis . However, the intrageneric relationships deduced from the gyrB-based phylogeny were different from those based on their 16S rDNA sequences . To examine which phylogeny would be more relevant for classifying genus Micromonospora, DNA-DNA hybridization experiments were performed . The gyrB-based classification agrees with the results of the DNA-DNA hybridization studies, indicating that this classification method is useful for analysing the phylogenetic relationships of high G+C Gram-positive bacteria at the level of the genomic species . Genus Micromonospora was reclassified into the following 14 species: Micromonospora echinospora, Micromonospora pallida, Micromonospora nigra, Micromonospora purpureochromogenes, Micromonospora aurantiaca, Micromonospora carbonacea, Micromonospora chalcea, Micromonospora chersina, Micromonospora coerulea, Micromonospora gallica, Micromonospora halophytica, Micromonospora inositola, Micromonospora olivasterospora and Micromonospora rosaria. Am J Otol, 2000 May, 21(3), 310 - 4 Octylcyanoacrylate: a new medical-grade adhesive for otologic surgery; Maw JL et al.; HYPOTHESIS: The adhesive octylcyanoacrylate is not associated with significant inner ear toxicity in a guinea pig model . BACKGROUND: Many cyanoacrylate adhesives have been investigated for use in otologic surgery, but variable ototoxicity has been reported . Octylcyanoacrylate is a medical-grade adhesive with many properties that make it ideal for use in the ear . It is free of contaminants; it forms a strong, flexible bond; and it inhibits the growth of gram-positive organisms in culture . This is the first study to assess the ototoxicity of this new adhesive . METHODS: Fourteen adult guinea pigs were used . Preoperative auditory brainstem responses (ABRs) were determined . Bilateral antrotomies were performed, and the ears were randomized to adhesive and control (saline) groups . In the adhesive ears . 0.5 or 0.1 mL of octylcyanoacrylate was instilled into the middle ear . Eight weeks later, postoperative ABRs were determined, the animals were killed, and the temporal bones were removed . Middle ear changes were noted, and the ossicular chain was assessed . Cochlear hair cell analyses were performed . Histologic assessment of the middle ear mucosa was performed . RESULTS: There was a higher incidence of conductive hearing loss in the adhesive group secondary to fixation of the ossicular chain, but there was no significant difference in bone conduction thresholds . The median postoperative bone conduction thresholds (dB peak sound pressure level) was 15.0 in the control group and 17.5 in the adhesive group, p = 0.89 . There was also no significant difference in inner hair cell counts (0.4% vs . 0.5% median hair cell loss, p = 0.72) or outer hair cell counts (3.7% vs . 3.0% median hair cell loss, p = 0.23) for the adhesive and control groups, respectively . Histopathologic analysis of the middle ear mucosa demonstrated variable mild to moderate foreign body reaction with no evidence of mucosal ulceration or necrosis . CONCLUSIONS: A large amount of octylcyanoacrylate placed in the middle ear of the guinea pig did not cause any morphologic or functional evidence of inner ear toxicity . This new adhesive is a promising tool for otologic surgery. Infect Immun, 2000 Jun, 68(6), 3581 - 6 Gram-positive bacteria are potent inducers of monocytic interleukin-12 (IL-12) while gram-negative bacteria preferentially stimulate IL-10 production; Hessle C et al.; Interleukin-10 (IL-10) and IL-12 are two cytokines secreted by monocytes/macrophages in response to bacterial products which have largely opposite effects on the immune system . IL-12 activates cytotoxicity and gamma interferon (IFN-gamma) secretion by T cells and NK cells, whereas IL-10 inhibits these functions . In the present study, the capacities of gram-positive and gram-negative bacteria to induce IL-10 and IL-12 were compared . Monocytes from blood donors were stimulated with UV-killed bacteria from each of seven gram-positive and seven gram-negative bacterial species representing both aerobic and anaerobic commensals and pathogens . Gram-positive bacteria induced much more IL-12 than did gram-negative bacteria (median, 3,500 versus 120 pg/ml at an optimal dose of 25 bacteria/cell; P < 0.001), whereas gram-negative bacteria preferentially stimulated secretion of IL-10 (650 versus 200 pg/ml; P < 0.001) . Gram-positive species also induced stronger major histocompatibility complex class II-restricted IFN-gamma production in unfractionated blood mononuclear cells than did gram-negative species (12,000 versus 3,600 pg/ml; P < 0.001) . The poor IL-12-inducing capacity of gram-negative bacteria was not remediated by addition of blocking anti-IL-10 antibodies to the cultures . No isolated bacterial component could be identified that mimicked the potent induction of IL-12 by whole gram-positive bacteria, whereas purified LPS induced IL-10 . The results suggest that gram-positive bacteria induce a cytokine pattern that promotes Th1 effector functions. J Rheumatol, 2000 May, 27(5), 1306 - 12 Nocardiosis in patients with systemic lupus erythematosus . The Singapore Lupus Study Group; Leong KP et al.; Nocardia, a gram positive variably acid-fast aerobic bacterium is an opportunistic pathogen in immunocompromised hosts . We present 5 cases of nocardiosis in patients with systemic lupus erythematosus . We emphasize the clinical features, radiologic findings, and antibiotic sensitivity . Lung involvement was the predominant manifestation; others include brain abscess, retinitis, thyroiditis, and diaphragmatic infiltration . We describe the first cases of pulmonary nocardiosis presenting as pneumothorax and the use of fine needle aspiration cytology in diagnosing nocardial thyroiditis. Nephrol Dial Transplant, 2000 May, 15(5), 680 - 3 Gel clot LAL assay in the initial management of peritoneal dialysis patients with peritonitis: a retrospective study; Hausmann MJ et al.; BACKGROUND: Indiscriminate use of broad-spectrum antibiotic treatment of peritonitis in peritoneal dialysis patients may have either unwanted side-effects or contribute to the development of antibiotic resistance . This may be avoided by improved diagnosis at presentation . The Limulus amoebocyte lysate assay is a convenient test detecting bacterial endotoxins or fungal beta glucans . This study evaluates a qualitative Limulus amoebocyte lysate test as a diagnostic tool used at presentation of a peritoneal dialysis patient with peritonitis . METHODS: One-hundred and eleven episodes of peritonitis in peritoneal dialysis patients have been analysed retrospectively . Limulus amoebocyte lysate results at presentation were compared with culture results . A Limulus amoebocyte lysate assay was performed using a commercial kit by incubating a mixture of dialysate effluent and Limulus amoebocyte lysate reagent at 37 degrees C . The development of a stable solid clot was considered positive . The specificity and sensitivity of the test were calculated . RESULTS: The specificity of the Limulus amoebocyte lysate assay was found to be 98% and the sensitivity 74% . Limulus amoebocyte lysate assay was false-negative in 13 cases of Gram-negative peritonitis (22%) . Limulus amoebocyte lysate was positive in three of seven cases of fungal peritonitis . The study included one case each with false-positive Limulus amoebocyte lysate and with culture-negative peritonitis . CONCLUSIONS: The Limulus amoebocyte lysate assay is a convenient and valuable diagnostic tool for excluding Gram-positive peritonitis in peritoneal dialysis patients . This allows more specific antibiotic treatment at presentation and may avoid the development of bacterial resistance . A negative Limulus amoebocyte lysate test is not reliable for the exclusion of Gram-negative peritonitis . In the absence of a positive culture result 48 h after presentation, accompanied by a delayed response to treatment, a positive Limulus amoebocyte lysate assay may indicate the presence of fungus . This justifies early empiric antifungal treatment before definitive culture results are made available . Routine Limulus amoebocyte lysate assay of dialysate effluent from continuous ambulatory peritoneal dialysis patients presenting with peritonitis is recommended. Neurosurgery, 2000 May, 46(5), 1149 - 53; discussion 1153-5 The value of routine cultures of the cerebrospinal fluid in patients with external ventricular drains; Hader WJ et al.; OBJECTIVE: The purpose of this study was to determine whether routine cerebrospinal fluid (CSF) bacteriological cultures in patients with external ventricular drains (EVDs) can identify infections early and prevent complications related to bacterial ventriculitis . METHODS: We retrospectively reviewed the microbiological reports and clinical data for all patients in whom an EVD was placed at a tertiary care pediatric neurosurgical center between 1984 and 1997 . EVDs were inserted in the operating room or intensive care unit, and, in most patients whose EVD remained in place for more than 2 days, daily cultures of CSF were performed . RESULTS: One hundred fifty-seven patients in whom 160 EVDs had been placed were included in the study . Forty-eight positive cultures were identified, of which the majority were determined to be contaminants . Seven infections were identified on the basis of microbiological criteria (i.e., a gram-positive stain and positive culture) and a subsequent positive culture . In all patients in whom infections developed, routine daily cultures of CSF were performed, and, in each instance, these cultures failed to identify the infections before clinical changes occurred . All seven patients with infection had fever (>38.5 degrees C) and peripheral leukocytosis (>11 x 10(3)/mm3) on the day the infection was identified, and one had a change in CSF appearance . CONCLUSION: The results of this study suggest that routine culture of CSF in children with EVDs is not necessary, and that if CSF cultures are performed for new fever (>38.5 degrees C) or peripheral leukocytosis, neurological deterioration, or a change in CSF appearance, infections will be identified in a timely fashion . In situations in which these clinical indicators might be masked, routine cultures may be valuable. Extremophiles, 2000 Apr, 4(2), 115 - 22 Molecular cloning, sequency, and expression of the heat-labile uracil-DNA glycosylase from a marine psychrophilic bacterium, strain BMTU3346; Jaeger S et al.; The gene encoding a heat-labile uracil-DNA glycosylase (UDG) from a psychrophilic, gram-positive marine strain (BMTU3346) has been cloned, sequenced, and expressed in Escherichia coli . The UDG is a cold-active enzyme with an apparent temperature optimum of 35 degrees C and a half-life of 2min at 40 degrees C . The amino acid sequence shows an identity of 39.1%-46.2% to UDGs from mesophilic bacteria . The primary structure was examined for features that could be related to the thermolability of the enzyme . The amino acid sequence of the heat-labile UDG shows 22 differences with respect to the consensus sequence derived from bacterial UDGs . Features previously recognized in cold-active enzymes such as extended surface loops or a decrease in the number of arginine residues or proline residues in loops were not observed . Because dominant features that could be related to the thermolability of the UDG from BMTU3346 cannot be identified, more subtle modifications of the conformation seem to be responsible for its thermolability. J Immunol, 2000 May 15, 164(10), 5439 - 45 Effect of CD14 blockade in rabbits with Escherichia coli pneumonia and sepsis; Frevert CW et al.; CD14, a pattern recognition receptor found on myeloid cells, is a critical component of the innate immune system that mediates local and systemic host responses to Gram-negative and Gram-positive bacterial products . Previous studies in normal animals have tested the effect of CD14 blockade on the systemic response to i.v . LPS . The goals of the study were to determine whether CD14 blockade protected against the deleterious systemic response associated with Escherichia coli pneumonia and to determine whether this strategy affected the pulmonary response to tissue infection . Rabbits were pretreated with either anti-CD14 mAb or isotype control mAb at 2.5 mg/kg . E . coli (1 x 109 CFU) was inoculated into the lungs, and the animals were observed for either 4 or 24 h . The blockade of CD14 improved the mean arterial blood pressure (p = 0.001) and decreased the i.v . fluid requirements (p = 0.01) . Although this therapy protected the vascular compartment, rabbits treated with anti-CD14 mAb had increased bacterial burdens in the bronchoalveolar lavage fluid recovered from the instilled lung (p = 0.005) and widened alveolar-arterial oxygen difference . Blockade of CD14 prevents the deleterious systemic responses that occur in sepsis; however, other measures are necessary to control bacterial proliferation at the primary site of infection. Transplantation, 2000 Apr 15, 69(7), 1511 - 4 The clinical impact of early gram-positive bacteremia and the use of vancomycin after allogeneic bone marrow transplantation; Schots R et al.; BACKGROUND: Gram-positive bacteremia (GPB) is an increasing infection after allogeneic bone marrow transplantation (BMT) . Our purpose was to identify risk factors for GPB, to evaluate its impact on early mortality and morbidity, and to compare prophylactic with empirical intravenous vancomycin . METHODS AND RESULTS: We studied 89 consecutive BMTs in adult patients . Early GPB occurred in 29% of posttransplantation episodes . T-cell depletion (odds ratio {OR}: 0.18) and vancomycin-prophylaxis (OR: 0.28) reduced the risk of GPB . Mortality at 6 weeks was not significantly different in patients with GPB (15% vs . 9.5%, P = 0.669) . GPB was associated with the development of major complications, the use of amphotericin B, and prolonged neutropenia . Vancomycin prophylaxis led to an increased risk of early renal dysfunction (OR: 18.7) . CONCLUSION: GPB contributes to overall morbidity during the early post-BMT episode but has no impact on mortality . Vancomycin prophylaxis is effective to reduce GPB but has a negative effect on renal function. Diagn Microbiol Infect Dis, 2000 May, 37(1), 5 - 9 Evaluation of the BacT/Alert microbial detection system with FAN aerobic and FAN anaerobic bottles for culturing normally sterile body fluids other than blood; Simor AE et al.; We evaluated the BacT/Alert Microbial Detection System (Organon Teknika Corporation, Durham, NC, USA) by using FAN bottles compared to conventional culture methods for the recovery of microorganisms from normally sterile body fluids other than blood and dialysates . Clinically significant pathogens were isolated from 116 (11%) of 1, 099 consecutive specimens (80 from both conventional media and FAN bottles; 23 from FAN bottles only; 13 from conventional media only) . Gram-positive cocci were more likely to be recovered from FAN bottles than from conventional media (p = 0.04) . Contaminants were also more likely to have grown in FAN bottles (3%) than on conventional media (1%) (p = 0.04) . The mean time to detection of significant pathogens was 20.9 h using FAN bottles as compared to 30 . 9 h using conventional media (p = 0.0001) . These results indicate that the BacT/Alert Microbial Detection System using FAN blood culture bottles improves the yield of clinically significant Gram-positive isolates from normally sterile body fluids with a reduced time to detection. Kidney Int, 2000 May, 57(5), 2151 - 5 Bacteremia associated with tunneled dialysis catheters: comparison of two treatment strategies; Tanriover B et al.; BACKGROUND: Tunneled dialysis catheters are often used for temporary vascular access in hemodialysis patients, but are complicated by frequent systemic infections . The treatment of bacteremia associated with infected tunneled catheters requires both antibiotic therapy and catheter replacement . We compared the outcomes of two treatment strategies for catheter-associated bacteremia: exchange of the existing catheter with a new one over a guidewire versus catheter removal with delayed replacement . METHODS: We retrospectively analyzed the outcomes of all cases of tunneled dialysis catheter-associated bacteremia during a two-year period . The infection-free survival time of the subsequent catheter was evaluated in two groups of patients: group A (31 catheters), exchange of the existing infected catheter with a new catheter over a guidewire, and group B (38 catheters), removal of the infected catheter followed by delayed catheter replacement 3 to 10 days later . Patients in both groups received three weeks of systemic antibiotic therapy . Cox proportional hazard models were used to evaluate the factors predictive of infection-free survival time of the replacement catheter . RESULTS: On univariate proportional hazard regression analysis, the infection-free survival time of the replacement catheter was similar for groups A and B (P = 0.72), whereas the hazard of infection was significantly greater for patients with hypoalbuminemia (serum albumin < 3.5 g/dL), as compared with patients with a normal serum albumin (hazard ratio 2.81, 95% CI, 1 . 21, 6.53, P = 0.016) . The infection-free survival time was not affected by patient age, sex, diabetic status, or type of organism (gram-positive coccus vs . gram-negative rod) . CONCLUSIONS: The infection-free survival time associated with the subsequent catheter is similar for the two treatment strategies . However, exchanging the catheter for a new one over a guidewire minimizes the number of separate procedures required by the patient . Hypoalbuminemia is the major risk factor for recurrent bacteremia in the replacement catheter. Microb Ecol, 2000 Jan, 39(1), 92 - 99 Physiological Status and Community Composition of Microbial Mats of the Ebro Delta, Spain, by Signature Lipid Biomarkers; Navarrete A et al.; Physiological status of microbial mats of the Ebro Delta (Tarragona, Spain) based on the extraction of lipids considered "signature lipid biomarkers" (SLB) from the cell membranes and walls of microorganisms has been analyzed . Data from a day-night cycle show significant differences in viable cells countings (PLFA cells counts) ranging from 1.5 x 10(10) to 5.0 x 10(10) cells g(-1) of sediment . Minimum values were observed at 18:00 and 6:00, when physicochemical conditions change drastically . The diversity of the microbial community was assessed by GC/MS analysis of phospholipid fatty acids (PLFA) . The ratio of PLFA, representative of Gram-negative bacteria, comprises 47.8% of the total PLFA of the microbial mat community . The remaining PLFA was representative of Gram-positive (10.0%), anaerobic (5.7%), and eukaryotic microorganisms (5.7%), and other common lipids . Two different approaches were used as a comparative study to assess the physiological status of the microbial mats . Two parameters (cyclopropane fatty acids/omega7c monoenoic fatty acids, and measurement of the trans/cis monoenoic PLFA ratio) showed a minimum at midnight, suggesting the highest microbial activity . Higher values were observed at 18:00 and 6:00, coinciding with lower PLFA cell counts . </hea J Clin Microbiol, 2000 May, 38(5), 1753 - 7 Detection of and discrimination between gram-positive and gram-negative bacteria in intraocular samples by using nested PCR; Carroll NM et al.; A nested PCR protocol has been developed for the detection of and discrimination between 14 species of gram-positive and -negative bacteria in samples of ocular fluids . First-round PCR with pan-bacterial oligonucleotide primers, based on conserved sequences of the 16S ribosomal gene, was followed by a gram-negative-organism-specific PCR, which resulted in a single 985-bp amplification product, and a multiplex PCR which resulted in two PCR products: a 1,025 bp amplicon (all bacteria) and a 355 bp amplicon (gram-positive bacteria only) . All products were detected by gel electrophoresis . The sensitivity of the assay was between 10 fg and 1 pg of bacterial DNA, depending on the species tested, equivalent to between 24 and 4 live bacteria spiked in water . The identification was complete in 3.5 h . The molecular techniques were subsequently applied to four samples of intraocular fluid, (three vitreous and one aqueous) from three patients with clinical signs of bacterial endophthalmitis (test samples) and two samples of vitreous from a patient with chronic intraocular inflammation (control samples) . In all culture-positive samples (two of three vitreous and one of one aqueous), a complete concordance was observed between molecular methods and culture results . PCR correctly identified the gram stain classification of the organisms . The bacterial etiology was also identified in a culture-negative patient with clinical history and signs highly suggestive of bacterial endophthalmitis . Furthermore, control samples from a patient with chronic intraocular inflammation remained PCR negative . In summary, this protocol has demonstrated potential as a rapid diagnostic test in confirming the diagnosis of infection and also determining the Gram status of bacteria with high specificity and sensitivity. Appl Environ Microbiol, 2000 May, 66(5), 2208 - 10 Evaluation of a fluorescent lectin-based staining technique for some acidophilic mining bacteria; Fife DJ et al.; A fluorescence-labeled wheat germ agglutinin staining technique (R . K . Sizemore et al., Appl . Environ . Microbiol . 56:2245-2247, 1990) was modified and found to be effective for staining gram-positive, acidophilic mining bacteria . Bacteria identified by others as being gram positive through 16S rRNA sequence analyses, yet clustering near the divergence of that group, stained weakly . Gram-negative bacteria did not stain . Background staining of environmental samples was negligible, and pyrite and soil particles in the samples did not interfere with the staining procedure. Appl Environ Microbiol, 2000 May, 66(5), 1826 - 33 The chromosomal arsenic resistance genes of Thiobacillus ferrooxidans have an unusual arrangement and confer increased arsenic and antimony resistance to Escherichia coli; Butcher BG et al.; The chromosomal arsenic resistance genes of the acidophilic, chemolithoautotrophic, biomining bacterium Thiobacillus ferrooxidans were cloned and sequenced . Homologues of four arsenic resistance genes, arsB, arsC, arsH, and a putative arsR gene, were identified . The T . ferrooxidans arsB (arsenite export) and arsC (arsenate reductase) gene products were functional when they were cloned in an Escherichia coli ars deletion mutant and conferred increased resistance to arsenite, arsenate, and antimony . Therefore, despite the fact that the ars genes originated from an obligately acidophilic bacterium, they were functional in E . coli . Although T . ferrooxidans is gram negative, its ArsC was more closely related to the ArsC molecules of gram-positive bacteria . Furthermore, a functional trxA (thioredoxin) gene was required for ArsC-mediated arsenate resistance in E . coli; this finding confirmed the gram-positive ArsC-like status of this resistance and indicated that the division of ArsC molecules based on Gram staining results is artificial . Although arsH was expressed in an E . coli-derived in vitro transcription-translation system, ArsH was not required for and did not enhance arsenic resistance in E . coli . The T . ferrooxidans ars genes were arranged in an unusual manner, and the putative arsR and arsC genes and the arsBH genes were translated in opposite directions . This divergent orientation was conserved in the four T . ferrooxidans strains investigated. Intensive Crit Care Nurs, 1999 Aug, 15(4), 239 - 41 New antibiotics for gram positive infections: linezolid and combination quinapristin/dalfopristin; MacConnachie AM; Considerable research over the past decade has thrown up two novel antibiotic preparations which are effective in 'difficult' Gram positive infections . Their imminent arrival is welcomed at a time when the emergence of resistance to last line drugs is rapidly spreading . Their careful use is, however, crucial if long-term value is to be preserved. Microbiology, 2000 Apr, 146 ( Pt 4), 903 - 10 An acyl-coenzyme A carboxylase encoding gene associated with jadomycin biosynthesis in Streptomyces venezuelae ISP5230; Han L et al.; Analysis of a region of chromosomal DNA lying between jadR1 and jadI in the gene cluster for jadomycin biosynthesis in Streptomyces venezuelae ISP5230 detected an ORF encoding 584 amino acids similar in sequence to the biotin carboxylase (BC) and biotin carboxyl carrier protein (BCCP) components of acyl-coenzyme A carboxylases . Multiple sequence alignments of the deduced Jad protein with acyl-coenzyme A carboxylases from various sources located the BC and BCCP components in the N- and C-terminal regions, respectively, of the deduced polypeptides . The organization and amino acid sequence of the deduced polypeptide most closely resembled those in other Gram-positive bacteria broadly classified as actinomycetes . Disrupting the gene, designated jadJ, severely reduced but did not eliminate jadomycin production . The disruption had no effect on growth or morphology of the organism, implying that the product of jadJ is not essential for fatty acid biosynthesis . It is concluded that jadJ supplies malonyl-coenzyme A for biosynthesis of the polyketide intermediate that is eventually processed to form the antibiotic jadomycin B. Arch Pathol Lab Med, 2000 May, 124(5), 712 - 6 Pyogenic spondylodiskitis: a radiologic/pathologic and culture correlation study; Lucio E et al.; BACKGROUND: Intervertebral disk tissue is resistant to hematogenous infection because of its avascularity . However, spondylodiskitis is being diagnosed with increasing frequency because of advancement in magnetic resonance imaging technology . There is a dearth of information regarding the bacteriology, histomorphologic features, and radiopathologic correlation of spondylodiskitis . DESIGN: The study population consisted of 20 patients diagnosed as having spondylodiskitis by magnetic resonance imaging with and without gadolinium 67 enhancement and bone scans with technetium Tc 99m or gallium citrate Ga 67 . Twenty-seven biopsy and debridement specimens were obtained from these patients . The specimens were cultured for microorganisms and also processed for histopathologic testing . Tissue sections were examined with hematoxylin-eosin and stains for infectious agents (Gomori's methenamine-silver, Gram, and Ziehl-Neelsen stains) . RESULTS: Where intervertebral disk tissue was present (23 of 27 cases), the morphologic changes included vascularization (with or without granulation tissue), myxoid degeneration, and necrosis . Chronic osteomyelitis was present in all 27 specimens and was associated with acute osteomyelitis in 7 cases (25%) . Twenty-one of 27 cases had positive culture results (mostly pyogenic bacteria), but special stains revealed microorganisms in sections of the disk in only 4 cases (3 cases with gram-positive cocci and 1 with yeast consistent with Blastomyces) . Florid acute inflammation was present in all the 4 cases . CONCLUSION: Histopathologic features of acute spondylodiskitis include vascular proliferation, myxoid degeneration, and necrosis of the disk tissue with adjacent chronic osteomyelitis . Acute inflammation is variable and when florid is usually associated with identifiable organisms on histologic examination . At biopsy, tissue should be submitted for culture, since culture has a high sensitivity and specificity for detecting the etiologic organism. Proc Natl Acad Sci U S A, 2000 Apr 25, 97(9), 4850 - 5 Development of an extrachromosomal cloning vector system for use in Borrelia burgdorferi; Sartakova M et al.; Molecular genetic analysis of Borrelia burgdorferi, the cause of Lyme disease, has been hampered by the absence of any means of efficient generation, identification, and complementation of chromosomal and plasmid null gene mutants . The similarity of borrelial G + C content to that of Gram-positive organisms suggested that a wide-host-range plasmid active in Gram-positive bacteria might also be recognized by borrelial DNA replication machinery . One such plasmid, pGK12, is able to propagate in both Gram-positive and Gram-negative bacteria and carries erythromycin and chloramphenicol resistance markers . pGK12 propagated extrachromosomally in B . burgdorferi B31 after electroporation but conferred only erythromycin resistance . pGK12 was used to express enhanced green fluorescent protein in B31 under the control of the flaB promoter . Escherichia coli transformed with pGK12 DNA extracted from B31 expressing only erythromycin resistance developed both erythromycin and chloramphenicol resistance, and plasmid DNA isolated from these transformed E . coli had a restriction pattern similar to the original pGK12 . Our data indicate that the replicons of pGK12 can provide the basis to continue developing efficient genetic systems for B . burgdorferi together with the erythromycin resistance and reporter egfp genes. J Assoc Physicians India, 1999 Aug, 47(8), 779 - 83 Pulmonary infections after kidney transplantation; Jha R et al.; OBJECTIVE: To retrospectively analyse the epidemiology, aetiology, temporal profile and outcome of lung infection following kidney transplantation . METHODS: Out of 142 consecutive renal transplant (RT) recipients who underwent live donor transplantation from June, 1990 to May, 1998, 43 (33%) had serious infection requiring hospitalisation of which 27 were pulmonary . All such pneumonia were included for retrospective analysis . All had a minimum follow up of six months (if alive) and were on triple drug immunosuppression . All had detailed and appropriate investigations for definitive diagnosis . RESULTS: The aetiological agents were Gram negative bacterial infection (2), Gram positive bacterial infection (1), nocardia (2), tuberculosis (10), aspergillosis (2), mixed bacterial and fungal infection (4), Pneumocystis (2) and unconfirmed (4) . Four patients had pneumonia because of probable nosocomial exposure . Radiologically lobar/segmental pneumonia was observed in five, nodular lesion six, reticulonodular lesion eight, patchy consolidation five and pleural effusion three . Nodular pneumonias were due to aspergillosis or nocardiosis . Four patients developed secondary cavitation . Pulmonary infections were significantly associated with leucopenia (8/27) (p < 0.01) but not with renal dysfunction (creat > 2 mg%), diabetes, old age or additional immunosuppression (p > 0.05) . There were 11 deaths . Mortality was related to failure to reach diagnosis (3) and delayed institution of therapy (6 patients) . Pneumonia within first six months had a higher mortality (9/16) compared to late pneumonia (2/11) . Immunomodulating virus (CMV 4, HEP B 2) was present in six patients of whom four succumbed . CONCLUSION: Pulmonary infection is a common and serious post-transplant infection requiring hospitalisation, is associated with high mortality . Patients with leucopenia are predisposed to these infections . Prophylaxis for Pneumocystis, Nocardia and tuberculosis needs strong consideration to reduce mortality of such infection . Nosocomial exposure risk needs careful consideration in outbreaks of opportunistic infection. J Leukoc Biol, 2000 Apr, 67(4), 508 - 14 The interleukin-1 receptor/Toll-like receptor superfamily: signal generators for pro-inflammatory interleukins and microbial products; Bowie A et al.; The interleukin-1 (IL-1) receptor/Toll-like receptor (TLR) superfamily is a recently defined and expanding group of receptors that participate in host responses to injury and infection . The superfamily is defined by the Toll/IL-1 receptor (TIR) domain, which occurs in the cytosolic region of family members, and is further subdivided into two groups based on homology to either the Type I IL-1 receptor or Drosophila Toll receptor extracellular domain . The former group includes the receptor for the important Th1 cytokine IL-18, and T1/ST2, which may have a role in Th2 cell function . The latter group includes six mammalian TLRs, including TLR2 and TLR4, that largely mediate the host response to gram-positive and gram-negative bacteria, respectively . Whether bacterial products are actual ligands for TLRs, or whether they generate ligands via as yet unidentified pattern recognition receptors, has yet to be determined . Signaling pathways activated via the TIR domain trigger the activation of downstream kinases, and transcription factors such as NF-kappaB, and involve the adaptor protein MyD88, which itself contains a TIR domain. FEMS Immunol Med Microbiol, 2000 May, 28(1), 49 - 53 Detection of peptidoglycan in human plasma using the silkworm larvae plasma test; Kobayashi T et al.; Silkworm larvae plasma (SLP) reagent, which is prepared from the body fluid of the silkworm, reacts with peptidoglycan (PG), a fragment of both the Gram-positive and Gram-negative bacterial cell wall, as well as with beta-glucan, a component of fungi . We developed a quantitative method for the detection of PG in human plasma from cases with bacterial infection using the SLP reagent . Tested in this way, the SLP method showed 86.2% sensitivity, 90.6% specificity, 89.3% positive predictive value, and 88.5% efficiency . The SLP method provides a valuable tool for the diagnosis of systemic infection using patients' blood. Cancer, 2000 Apr 15, 88(8), 1964 - 9 Substituting dexamethasone for prednisone complicates remission induction in children with acute lymphoblastic leukemia; Hurwitz CA et al.; BACKGROUND: The authors report the occurrence of fatal or near-fatal sepsis in 16 of 38 children with newly diagnosed acute lymphoblastic leukemia (ALL) treated with a new induction regimen that differed from its predecessor by the substitution of dexamethasone for prednisone . METHODS: The frequency of septic deaths among 38 children who received multiagent remission induction therapy, including dexamethasone (6 mg/m(2)) daily for 28 days (pilot protocol 91-01P), was compared with the frequency of septic deaths among children previously treated (protocol 87-01) and subsequently treated (protocol 91-01) in consecutive Dana-Farber Cancer Institute (DFCI) ALL trials with induction therapy that included 21 and 28 days of prednisone (40 mg/m(2)), respectively . Except for dexamethasone in protocol 91-01P, the remission induction agents used were identical in substance to those used in protocol 87-01 . Protocol 91-01, the successor 91-01P, was also similar, with the exception of the deletion of a single dose of L-asparaginase . RESULTS: Sixteen of the 38 children (42%) treated on the DFCI 91-01P had documented gram positive or gram negative sepsis (17 episodes) during remission induction, including 4 toxic deaths (11%) . In contrast, there were 4 induction deaths among 369 children (1%) treated on protocol 87-01 (P = 0.0035) and 1 induction death among 377 children (<1%) treated on protocol 91-01 (P = 0.0003) . CONCLUSIONS: Substitution of dexamethasone for prednisone or methylprednisolone in an otherwise intensive conventional induction regimen for previously untreated children with ALL resulted in an alarmingly high incidence of septic episodes and toxic deaths . Awareness of this complication, considering that the substitution has no apparent benefit in the efficacy of remission induction, argues against its routine use in intensive induction regimens for children with ALL . Mol Microbiol, 2000 Apr, 36(1), 44 - 54 The carbon metabolism-controlled Synechocystis gap2 gene harbours a conserved enhancer element and a Gram-positive-like -16 promoter box retained in some chloroplast genes; Figge RM et al.; The two glyceraldehyde-3-phosphate dehydrogenase-encoding genes (gap) of Synechocystis were shown to be expressed as monocistronic transcripts . Whereas gap1 expression is slow and weak, gap2 gene induction is rapid and strong . Transcription of the gap2 gene was shown to depend on functional photosynthetic electron transport and on active carbon metabolism . The basal promoter of gap2 (P, -45 to +34, relative to the transcription start site) is controlled by three cis-acting elements designated A (-443 to -45), B (+34 to +50, in the untranslated leader region) and C (+50 to +167, in the coding region) that, together, promote a 100-fold stimulation of P activity . Element B was found to behave as a transcriptional enhancer, in that it was active regardless of its position, orientation and distance relative to P . All three cis-acting stimulatory elements exhibit a common 5'-agaTYAACg-3' nucleotide motif that appears to be conserved in cyanobacteria and may be the target for a transcriptional enhancer . We also report that gap2 transcription depends on a Gram-positive-like -16 promoter box (5'-TRTG-3') that was obviously conserved throughout the evolution of chloroplasts . This is the first report on the occurrence of a -16 promoter element in photoautotrophic organisms. Int J Syst Evol Microbiol, 2000 Mar, 50 Pt 2, 529 - 36 Marmoricola aurantiacus gen . nov., sp . nov., a coccoid member of the family Nocardioidaceae isolated from a marble statue; Urzi C et al.; A Gram-positive, aerobic bacterium with coccoid cells occurring singly, in pairs and in clusters was isolated from the surface of a marble statue . The peptidoglycan contain LL-diaminopimelic acid as diagnostic diamino acid and a single glycine residue as interpeptide bridge (type A3 gamma) . The major menaquinone is MK-8(H4) . The cellular fatty acid pattern consists of straight chain saturated and monounsaturated components and 10-methyl octadecanoic (tuberculostearic) acid as the only branched chain fatty acid . Phosphatidylinositol, phosphatidylglycerol and diphosphatidylglycerol occur as characteristic polar lipids . The DNA G + C composition is 72 mol% . According to its phylogenetic position and 16S rDNA signature nucleotides, the organism is a member of the family Nocardioidaceae . The combination of chemotaxonomic characteristics is unique within this family and supports the description of a new genus and new species, Marmoricola aurantiacus . The type strain is strain BC 361T (= DSM 12652T). Microbes Infect, 2000 Mar, 2(3), 251 - 5 Toll-like receptors; Muzio M et al.; Toll-like receptors (TLRs) are a growing family of molecules involved in innate immunity . Accumulating evidence suggests that TLR molecules are involved in signalling receptor complexes which recognise components of Gram-positive and Gram-negative bacteria and mycobacteria . Differential expression and regulation as well as distinct though overlapping ligand recognition patterns may underlie the existence of a vast TLR family . Apparent structural and functional redundancy may render certain outputs of the TLR family robust. J Antimicrob Chemother, 2000 Apr, 45(4), 533 - 5 Endothelial cell compatibility of trovafloxacin and levofloxacin for intravenous use; Armbruster C et al.; Levofloxacin and trovafloxacin have excellent activity against a variety of Gram-positive and Gram-negative organisms resistant to the established agents . One local side-effect closely related to the use of parenteral fluoroquinolones is phlebitis . To evaluate the effect of trovafloxacin and levofloxacin on endothelial cell viability, intracellular levels of adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), guanosine 5'-triphosphate (GTP) and guanosine 5'-diphosphate (GDP) levels were measured using high-performance liquid chromatography . Trovafloxacin at concentrations of 2 and 1 mg/mL reduced the intracellular ATP content from 12.5 +/- 1.7 to 1.9 +/- 0.3 nmol/10(6) cells and 9.3 +/- 0.8 nmol/10(6) cells, respectively, within 60 min . In addition, ADP, GTP and GDP levels were extensively depleted . Levofloxacin at concentrations of 5 and 2.5 mg/mL led to a significant ATP decline from 12.5 +/- 1.7 to 2.3 +/- 0.2 nmol/10(6) cells and 10.3 +/- 0.9 nmol/10(6) cells, respectively, within 60 min . These data indicate that infusions of high doses of trovafloxacin or levofloxacin are not compatible with maintenance of endothelial cell function . Commercial preparations have to be diluted and should be administered into large veins. Lett Appl Microbiol, 2000 Mar, 30(3), 178 - 82 A simple HPLC method for analysing diaminopimelic acid diastereomers in cell walls of Gram-positive bacteria; McKerrow J et al.; A simple and sensitive method for separating and detecting the LL, DD and meso diastereomers of the dibasic amino acid diaminopimelic acid (DAP) in the peptidoglycan of Gram-positive bacteria is described . This method is based on reverse phase HPLC separation of chiral derivatives of DAP followed by fluorescence detection of the o-phthaldehyde derivatives . Its application to the analyses of cell walls of several Gram-positive bacteria is described, where 10 mg or less of dry cells is required. Clin Ther, 2000 Feb, 22(2), 154 - 66 Cefaclor revisited; Meyers BR; OBJECTIVE: This paper describes the rationale for choosing cefaclor for the management of respiratory tract infections . BACKGROUND: Since 1979, cefaclor has established a record of efficacy in the management of respiratory tract infections . Factors contributing to the efficacy and tolerability of this drug include its molecular stability, activity against the most prevalent gram-positive and gram-negative respiratory tract pathogens, rapid absorption, >90% bioavailability, and good penetration into respiratory mucosa . After 2 decades of widespread use, this agent remains clinically effective in patients with respiratory tract infections, making it competitive with other cephalosporins and with macrolides and fluoroquinolones, including many newer agents used for respiratory tract infections . Cefaclor extended-release tablets, the newest formulation, retain the positive efficacy and tolerability attributes of immediate-release cefaclor, varying mainly in the rate of dissolution . The approved indications for extended-release cefaclor include bacterial bronchitis, pharyngitis, and skin infections . METHODS: A MEDLINE search showed that the few adverse effects related to therapy with cefaclor are usually minor and transient and that drug-drug interactions involving cefaclor are rare . CONCLUSIONS: Multiple clinical trials have shown that extended-release cefaclor in 375-mg and 500-mg doses BID demonstrates tolerability and efficacy comparable to those of immediate-release cefaclor 250 mg TID . Extended-release cefaclor is indicated for BID dosing, which should encourage greater compliance. Nat Struct Biol, 2000 Apr, 7(4), 287 - 91 The structure of the ferric siderophore binding protein FhuD complexed with gallichrome; Clarke TE et al.; Siderophore binding proteins play a key role in the uptake of iron in many gram-positive and gram-negative bacteria . FhuD is a soluble periplasmic binding protein that transports ferrichrome and other hydroxamate siderophores . The crystal structure of FhuD from Escherichia coli in complex with the ferrichrome homolog gallichrome has been determined at 1.9 inverted question mark resolution, the first structure of a periplasmic binding protein involved in the uptake of siderophores . Gallichrome is held in a shallow pocket lined with aromatic groups; Arg and Tyr side chains interact directly with the hydroxamate moieties of the siderophore . FhuD possesses a novel fold, suggesting that its mechanisms of ligand binding and release are different from other structurally characterized periplasmic ligand binding proteins. Biol Blood Marrow Transplant, 2000, 6(2), 109 - 14 Early outcomes after allogeneic stem cell transplantation for leukemia and myelodysplasia without protective isolation: a 10-year experience; Russell JA et al.; Although it is common practice to use some form of isolation to protect allogeneic stem cell transplant patients from infection, the necessity for these practices in all environments has not been demonstrated . The current study evaluated patterns of infection and 100-day transplant-related mortality in 288 patients with myelodysplasia and leukemia transplanted without isolation . Patients were allowed out of hospital at any time within constraints of the medication schedule . Fever, foci of infection, and positive cultures within 28 days and death within 100 days because of the transplant procedure were recorded . Fever occurred in 57% of patients, and 10% had a clinical or radiographic focus of infection . Most infections were apparently endogenous; blood cultures from 24% of recipients grew organisms, 87% of which were gram-positive bacteria . Four patients (1%) died with aspergillus infection in circumstances indicating that isolation would not have been helpful . Twenty percent of patients remained without evidence of infection throughout . Transplant-related mortality at 100 days was 1% for 108 patients with early leukemia receiving transplants from matched siblings . For patients at higher risk, by virtue of donor and/or disease status, mortality was 21% . These figures compare favorably with those reported to the International Bone Marrow Transplant Registry, the majority of patients having been subjected to some form of isolation . We conclude that allogeneic stem cell transplantation can be safely performed in some environments without confining patients continuously to the hospital. J Appl Microbiol, 2000 Feb, 88(2), 248 - 59 Spore-forming, Desulfosporosinus-like sulphate-reducing bacteria from a shallow aquifer contaminated with gasoline; Robertson WJ et al.; Previous studies on the geochemistry of a shallow unconfined aquifer contaminated with hydrocarbons suggested that the degradation of some hydrocarbons was linked to bacterial sulphate reduction . There was attenuation of naphthalene, 1,3,5-trimethylbenzene (TMB), toluene, p-xylene and ethylbenzene in the groundwater with concomitant loss of sulphate . Here, the recovery of eight strains of sulphate-reducing bacteria (SRB) from the contaminated site is reported . All were straight or curved rod-shaped cells which formed endospores . Amplification and sequencing of the 16S rDNA indicated that the strains were all sulphate reducers of the Gram-positive line of descent, and were most closely related to Desulfosporosinus (previously Desulfotomaculum) orientis DSM 8344 (97-98.9% sequence similarity) . The strains clustered in three phylogenetic groups based on 16S rRNA sequences . Whole cell fatty acid compositions were similar to those of D . orientis DSM 8344, and were consistent with previous studies of fatty acids in soil and groundwater from the site . Microcosms containing groundwater from this aquifer indicated a role for sulphate reduction in the degradation of {ring-UL-14C}toluene, but not for the degradation of {UL-14C}benzene which could also be degraded by the microcosms . Adding one of the strains that was isolated from the groundwater (strain T2) to sulphate-enriched microcosms increased the rate of toluene degradation four- to 10-fold but had no effect on the rate of benzene degradation . The addition of molybdate, an inhibitor of sulphate reduction, to the groundwater samples decreased the rate of toluene mineralization . There was no evidence to support the mineralization of {UL-14C}benzene, {ring-UL-14C}toluene or unlabelled m-xylene, p-xylene, ethylbenzene, TMB or naphthalene by any of the strains in pure culture . Growth of all the strains was completely inhibited by 100 micromol l-1 TMB. J Bacteriol, 2000 Apr, 182(8), 2163 - 9 The net charge of the first 18 residues of the mature sequence affects protein translocation across the cytoplasmic membrane of gram-negative bacteria; Kajava AV et al.; This statistical study shows that in proteins of gram-negative bacteria exported by the Sec-dependent pathway, the first 14 to 18 residues of the mature sequences have the highest deviation between the observed and expected net charge distributions . Moreover, almost all sequences have either neutral or negative net charge in this region . This rule is restricted to gram-negative bacteria, since neither eukaryotic nor gram-positive bacterial exported proteins have this charge bias . Subsequent experiments performed with a series of Escherichia coli alkaline phosphatase mutants confirmed that this charge bias is associated with protein translocation across the cytoplasmic membrane . Two consecutive basic residues inhibit translocation effectively when placed within the first 14 residues of the mature protein but not when placed in positions 19 and 20 . The sensitivity to arginine partially reappeared again 30 residues away from the signal sequence . These data provide new insight into the mechanism of protein export in gram-negative bacteria and lead to practical recommendations for successful secretion of hybrid proteins. Bioseparation, 1999, 8(1-5), 99 - 109 Cell/adsorbent interactions in expanded bed adsorption of proteins; Feuser J et al.; Expanded bed adsorption (EBA) is an integrated technology for the primary recovery of proteins from unclarified feedstock . A method is presented which allows a qualitative and quantitative understanding of the main mechanisms governing the interaction of biomass with fluidized resins . A pulse response technique was used to determine the adsorption of various cell types (yeast, Gram positive and Gram negative bacteria, mammalian cells and yeast homogenate) to a range of commercially available matrices for EBA . Cells and cell debris were found to interact with the ligands of agarose based resins mainly by electrostatic forces . From the adsorbents investigated the anion exchange matrix showed the most severe interactions, while cation exchange and affinity adsorbents appeared to be less affected . Within the range of biologic systems under study E . coli cells had the lowest tendency of binding to all matrices while hybridoma cells attached to all the adsorbents except the protein A affinity matrix . The method presented may be employed for screening of suitable biomass/adsorbent combinations, which yield a robust and reliable initial capture step by expanded bed adsorption from unclarified feedstock. Nucleic Acids Res . 2000 Apr 15;28(8):E37. Amplifiable DNA from gram-negative and gram-positive bacteria by a low strength pulsed electric field method; Vitzthum F et al.; An efficient electric field-based procedure for cell disruption and DNA isolation is described . Isoosmotic suspensions of Gram-negative and Gram-positive bacteria were treated with pulsed electric fields of <60 V/cm . Pulses had an exponential decay waveform with a time constant of 3.4 micros . DNA yield was linearly dependent on time or pulse number, with several thousand pulses needed . Electrochemical side-effects and electrophoresis were minimal . The lysates contained non-fragmented DNA which was readily amplifiable by PCR . As the method was not limited to samples of high specific resistance, it should be applicable to physiological fluids and be useful for genomic and DNA diagnostic applications. Eur J Obstet Gynecol Reprod Biol, 2000 Apr, 89(2), 197 - 200 Treatment strategy for pelvic actinomycosis: case report and review of the literature; Hamid D et al.; BACKGROUND: Pelvic actinomycosis is a chronic granulomatous suppurative disease caused by an anaerobic Gram positive organism Actinomyces israelii usually associated with intra-uterine devices . Pelvic actinomycosis can mimick pelvic or intra-abdominal malignancy leading to mutilating surgical exeresis . RESULTS: We present a pelvic actinomycosis secondary to long-standing intra-uterine device use in a 50-year old European woman treated by intravenous antibiotic therapy, and then by a total abdominal hysterectomy and bilateral salpingoophorectomy to free the pelvis from abscess . We point out the difficulty in diagnosis, and the importance of high-dose intravenous antibiotic therapy to reduce the very high risk for nearby pelvic structure injuries, reported in the literature, leading to post-operative morbidity. Infect Immun, 2000 Apr, 68(4), 2156 - 60 Upregulation of monocyte urokinase plasminogen activator receptor during human endotoxemia; Dekkers PE et al.; The receptor for urokinase-type plasminogen activator (uPAR) (CD87) plays an important role in leukocyte adhesion and migration . To assess the effect of endotoxin on cellular uPAR, uPAR expression was determined on leukocytes by fluorescence-activated cell sorter analysis in seven healthy subjects following intravenous injection of endotoxin (lot G; 4 ng/kg) . Endotoxin induced a transient increase in uPAR expression on monocytes, reaching a 92% +/- 46% increase over baseline expression after 6 h (P < 0.05) . Endotoxin did not influence uPAR expression on granulocytes, while uPAR remained undetectable on lymphocytes . Endotoxin also increased soluble uPAR levels in plasma (P < 0.05) . Stimulation of human whole blood with endotoxin or gram-positive stimuli in vitro also resulted in an upregulation of monocyte uPAR expression . Although tumor necrosis factor alpha (TNF) upregulated monocyte uPAR expression, anti-TNF did not influence the endotoxin-induced increase in monocyte uPAR expression . These data suggest that infectious stimuli may influence monocyte function in vivo by enhancing the expression of uPAR. Antimicrob Agents Chemother, 2000 Apr, 44(4), 916 - 9 Pharmacokinetics of an everninomicin (SCH 27899) in mice, rats, rabbits, and cynomolgus monkeys following intravenous administration; Lin C et al.; The pharmacokinetics of SCH 27899, a novel oligosaccharide compound of the everninomicin class with excellent activity against gram-positive strains, was studied with mice, rats, rabbits, and cynomolgus monkeys following intravenous administration as SCH 27899-N-methylglucamine-hydroxypropyl beta-cyclodextrin . Concentrations of SCH 27899 in mouse serum, rat plasma, and rabbit serum were determined by a high-pressure liquid chromatography method on a poly(styrene-divinyl benzene) column, and those in monkey plasma were determined by a paired-ion chromatographic method . Plasma and serum concentrations of SCH 27899 exhibited a biexponential decline in all species following intravenous administration . The half-lives at beta phase were 3.0 to 7.9 h in mice, rats, and rabbits and 24 h in cynomolgus monkeys . There was a linear relationship between the area under the curve extrapolated to infinity {AUC(I)} in mice and dose . Rabbits also exhibited dose proportionality in AUC(I) . However, in rats, increasing the dose from 3 to 60 mg/kg of body weight resulted in a 49-fold increase in AUC(I) . When the species was changed from mouse to rat, rabbit, or cynomolgus monkey, AUC(I) increased, whereas clearance (CL) decreased . It was concluded that the pharmacokinetics of SCH 27899 in animals varied with species; CL was the highest in mice and rats, followed by rabbits and cynomolgus monkeys. Arch Immunol Ther Exp (Warsz), 2000, 48(1), 31 - 7 Effective phage therapy is associated with normalization of cytokine production by blood cell cultures; Weber-Dabrowska B et al.; The aim of this study was to investigate the effect of phagotherapy on tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) serum levels and the ability of blood cells to produce these cytokines in culture . Fifty one patients with long-term, suppurative infections of various tissues and organs were enrolled . The ability of cells to secrete cytokines was tested using whole blood cell cultures, unstimulated or stimulated with lipopolysaccharide (LPS) from E . coli . In addition, cytokine serum levels were determined . Measurement of cytokine activity was performed using bioassays . We showed that TNF-alpha, but not IL-6 serum levels, were regulated upon division of patients into categories exhibiting initial: low, moderate and high cytokine levels . The low spontaneous production of IL-6 by blood cell cultures was elevated significantly on day 21 of phage therapy, whereas high release of this cytokine was inhibited . No such correlation was observed with LPS-induced IL-6 production in cell cultures when cells from low-, moderately- or highly-reactive patients were studied . Phage therapy modified TNF release according to the initial ability to produce that cytokine: it reduced TNF production in high responders and increased it in low responders . Patients infected only with Gram-positive bacteria demonstrated analogous changes in the spontaneous and LPS-induced TNF-alpha production as in the whole studied group . A similar kind of regulation was observed in TNF-alpha and LPS-induced production, i.e . low production was significantly elevated, high strongly inhibited, and moderate only slightly affected . In summary, we demonstrated for the first time that effective phage therapy can normalize TNF-alpha serum levels and the production of TNF-alpha and IL-6 by blood cell cultures. J Infect Dis, 2000 Mar, 181(3), 1055 - 61 Endotoxin down-regulates monocyte and granulocyte interleukin-6 receptors without influencing gp130 expression in humans; Dekkers PE et al.; Interleukin (IL)-6 is important for host defense against various pathogens . The IL-6 receptor (IL-6R) complex consists of a ligand-binding component (IL-6R) and a signal-transducing component (gp130) . In a study designed to obtain insight into the regulation of this receptor complex during inflammation, 8 healthy subjects received an intravenous injection of lipopolysaccharide (LPS; 4 ng/kg), and receptor expression was determined on blood leukocytes by use of fluorescence-activated cell cytometry . LPS induced a transient decrease in monocyte and granulocyte IL-6R expression but did not influence gp130 . The plasma concentrations of soluble IL-6R and soluble gp130 did not change after LPS administration . Expression of the receptor for leukemia inhibitory factor, a member of the IL-6R family, remained unaltered after LPS injection . In whole blood in vitro, LPS and gram-positive stimuli and proinflammatory cytokines were capable of down-modulating the IL-6R . Monocytes and granulocytes may down-regulate IL-6R at their surface upon their first interaction with bacterial antigens. Scand J Infect Dis, 2000, 32(1), 63 - 7 Effects of anti-neoplastic agents on the recovery of bacteria and yeasts in an automated blood culture system; Kinnunen U et al.; The effects of 7 anti-neoplastic agents on the recovery of 5 aerobic gram-positive cocci, gram-negative rods and yeasts were studied in 2 different automated blood culture systems using an experimental model . In the absence of anti-neoplastic agents, the growth of gram-positive cocci was detected significantly earlier in standard than in FAN aerobic bottles . In the presence of 100 microM doxorubicin, however, the growth of gram-positive cocci was totally inhibited in standard culture conditions, while in FAN bottles the agent has no inhibitory effect . Etoposide at a concentration of 100 micromol/l also significantly delayed the growth of cocci in standard conditions . Neither the culture bottles nor the anti-neoplastic agents tested had any effect on the growth of gram-negative rods and yeasts . The results suggest that the anti-neoplastic agents present in blood might disturb the growth of gram-positive cocci in blood culture . This should be considered when validating blood culture systems and evaluating blood cultures of chemotherapy-receiving febrile patients. J Bacteriol, 2000 Apr, 182(7), 2001 - 9 Pyruvate kinase of the hyperthermophilic crenarchaeote Thermoproteus tenax: physiological role and phylogenetic aspects; Schramm A et al.; Pyruvate kinase (PK; EC 2.7.1.40) of Thermoproteus tenax was purified to homogeneity, and its coding gene was cloned and expressed in Escherichia coli . It represents a homomeric tetramer with a molecular mass of 49 kDa per subunit . PK exhibits positive binding cooperativity with respect to phosphoenolpyruvate and metal ions such as Mg(2+) and Mn(2+) . Heterotropic effects, as commonly found for PKs from bacterial and eucaryal sources, could not be detected . The enzyme does not depend on K(+) ions . Heterotrophically grown cells exhibit specific activity of PK four times higher than autotrophically grown cells . Since the mRNA level of the PK coding gene is also accordingly higher in heterotrophic cells, we conclude that the PK activity is adjusted to growth conditions mainly on the transcript level . The enzymic properties of the PK and the regulation of its expression are discussed with respect to the physiological framework given by the T . tenax-specific variant of the Embden-Meyerhof-Parnas pathway . T . tenax PK shows moderate overall sequence similarity (25 to 40% identity) to its bacterial and eucaryal pendants . Phylogenetic analyses of the known PK sequences result in a dichotomic tree topology that divides the enzymes into two major PK clusters, probably diverged by an early gene duplication event . The phylogenetic divergence is paralleled by a striking phenotypic differentiation of PKs: PKs of cluster I, which occur in eucaryal cytoplasm, some gamma proteobacteria, and low-GC gram-positive bacteria, are only active in the presence of fructose-1,6-bisphosphate or other phosphorylated sugars, whereas PKs of cluster II, found in various bacterial phyla, plastids, and in Archaea, show activity without effectors but are commonly regulated by the energy charge of the cell. FEMS Microbiol Lett, 2000 Mar 15, 184(2), 265 - 71 Design and uses of Bdellovibrio 16S rRNA-targeted oligonucleotides; Jurkevitch E et al.; An 18-mer oligonucleotide almost exclusively targeting Bdellovibrio spp . was designed based on available 16S rRNA sequence data . The specificity of this oligonucleotide used as a PCR primer in combination with a Bacteria domain-targeted primer as well as used as a probe in rRNA dot blot hybridizations was experimentally confirmed using a variety of alpha-, beta-, gamma-, delta-Proteobacteria and Gram-positive bacteria . Similarly, combinations of the Bacteria primer with oligonucleotides targeting the 16S rRNA gene of Bdellovibrio bacteriovorus and Bdellovibrio stolpii were shown to be species specific by PCR . Positive amplification products were obtained from an irrigation water sample in which a low level of bdellovibrios was detected by plating as well as from soil detached from potato tubers, using the Bdellovibrio spp.-Bacteria and the B . bacteriovorus-Bacteria primer pairs. J Biol Chem, 2000 Mar 17, 275(11), 7505 - 14 A beta1,3-glucan recognition protein from an insect, Manduca sexta, agglutinates microorganisms and activates the phenoloxidase cascade; Ma C et al.; Pattern recognition proteins function in innate immune responses by binding to molecules on the surface of invading pathogens and initiating host defense reactions . We report the purification and molecular cloning of a cDNA for a 53-kDa beta1,3-glucan-recognition protein from the tobacco hornworm, Manduca sexta . This protein is constitutively expressed in fat body and secreted into hemolymph . The protein contains a region with sequence similarity to several glucanases, but it lacks glucanase activity . It binds to the surface of and agglutinates yeast, as well as gram-negative and gram-positive bacteria . Beta1,3-glucan-recognition protein in the presence of laminarin, a soluble glucan, stimulated activation of prophenoloxidase in plasma, whereas laminarin alone did not . These results suggest that beta1,3-glucan-recognition protein serves as a pattern recognition molecule for beta1,3-glucan on the surface of fungal cell walls . After binding to beta1,3-glucan, the protein may interact with a serine protease, leading to the activation of the prophenoloxidase cascade, a pathway in insects for defense against microbial infection. Med Klin (Munich), 2000 Feb 15, 95(2), 69 - 74 {Effects of restrictions on use of vancomycin in a German university hospital}; Gluck T et al.; BACKGROUND: Recently, increasing antibiotic resistance has been observed among gram-positive bacteria . However, only few isolates were found to be resistant against glycopeptides . Therefore, internationally accepted guidelines recommend a restricted use of vancomycin and other glycopeptide antibiotics in order to prevent the development of resistance against these clinically important antibiotics . In many countries, the hospital pharmacies play a key role in control and reinforcement of antibiotic formulary restrictions . In Germany, however, the hospital pharmacies usually do not take over such control functions, and most wards keep a stock of regularly used drugs including antibiotics, which makes reinforcement of restrictions difficult . METHODS: In an attempt to achieve a restriction of vancomycin use, the pharmacy of our university hospital was advised to deliver vancomycin to the wards only on request with a special order form signed by an attending, individually for every patient who should receive vancomycin . The efficacy of this restriction measure was evaluated in 3-month periods before and after the restriction became effective . RESULTS: Hospitalwide, this led to a 20.1% reduction of i.v . vancomycin and an 85.7% reduction of oral vancomycin use per 1000 patient days . If the hematology/oncology units were not considered, the reduction of i.v . vancomycin use was 41.8%, and the total use after the restriction 24.2 g per 1000 patient days . Microbiology results which justified the use of vancomycin decreased by 8.3% (10.9% hematology/oncology units not considered) between the 2 observation periods . Assuming a 7-day mean course of i.v . vancomycin therapy, the empirical use of i.v . vancomycin decreased from 39.9% to 8% after the restriction had been instituted . CONCLUSION: Allowing only experienced physicians (attendings) to decide on the use of vancomycin therapy, proved in our experience to be an effective measure to reduce unnecessary vancomycin use. Crit Care Med, 2000 Feb, 28(2), 451 - 7 Coagulation system and platelets are fully activated in uncomplicated sepsis; Mavrommatis AC et al.; OBJECTIVE: To test the hypothesis that the coagulation system and platelets are activated in sepsis, the uncomplicated and usually earliest stage of the septic process, and to compare the findings detected in sepsis with those found in severe sepsis and septic shock . DESIGN: Prospective study comparing patients with sepsis, severe sepsis, and septic shock, and healthy volunteers . SETTING: General intensive care unit in a tertiary university hospital . PATIENTS: Seventy-four consecutive septic patients (45 with sepsis, 15 with severe sepsis, and 14 with septic shock) . Fourteen healthy volunteers served as control subjects . INTERVENTION: None . MEASUREMENTS AND MAIN RESULTS: After blood sampling, molecular activation markers of coagulation (prothrombin fragments 1 and 2, fibrinopeptide A, thrombin-antithrombin complexes, and monomers of fibrin) and of platelets (beta-thromboglobulin and platelet factor 4), several coagulation factors, global tests of coagulation (prothrombin time and activated partial thromboplastin time), and platelet count (PTL) were measured . In sepsis, prothrombin fragments 1 and 2, fibrinopeptide A, thrombin-antithrombin complexes, and monomers of fibrin were increased to 2.52+/-0.21 nmol/L, 20.88+/-2.52 ng/mL, 33.8+/-2.9 microg/L, and 69% positive, respectively, compared with control subjects (0.86+/-063 nmol/L, 1.14+/-0.15 ng/mL, 16.07+/-1.01 microg/L, and 0%, respectively) . Beta-Thromboglobulin and the beta-thromboglobulin-to-platelet factor 4 ratio were also increased to 107.87+/-11.87 IU/mL and 8.86+/-1.06, compared with controls (18.36 +/-2.99 IU/mL and 2.67+/-0.52, respectively) . With the exception of a decrease in factor XII and an increase in fibrinogen, coagulation factors, global coagulation tests, and PTL were not changed in sepsis . In severe sepsis and mainly in septic shock, coagulation factors were markedly decreased, global coagulation tests were prolonged, and PTL was reduced . All changes were independent of the causative infectious pathogen . CONCLUSION: Coagulation system and platelets are strongly activated in sepsis . In this stage, only factor XII is decreased . In contrast, in severe sepsis and mainly in septic shock, most of the coagulation factors are depleted, PTL is decreased, and global coagulation tests are prolonged, indicating exhaustion of hemostasis . Finally, Gram-positive, Gram-negative, and other microorganisms produce identical impairment of coagulation. J Hosp Infect, 2000 Mar, 44(3), 206 - 13 A one-year prospective study of nosocomial bacteraemia in ICU and non-ICU patients and its impact on patient outcome; Garrouste-Orgeas M et al.; A one-year, prospective, two-observational cohort study was performed to evaluate the incidence and outcome in hospitalized patients (ICU and non-ICU) of nosocomial bacteraemia, and to assess its prognostic value in the ICU group . A group of 18 098 hospitalized patients and a group of 291 consecutive ICU patients were followed . Prognostic factors were determined using single and multivariable analyses . 109 (90 non-ICU and 19 ICU) patients developed 118 nosocomial bacteraemic episodes . The incidence of nosocomial bacteraemia was 6.0 per 1000 admissions (95% confidence interval (CI): 5-7%) and 65 per 1000 admissions in ICU patients (95% CI: 4.5-8.5%) . Gram-positive and Gram-negative bacteria were 63/133 (47%) and 70/133 (53%) of the isolated micro-organisms respectively . Crude mortality rates were 41/109 (38%) with adverse outcome associated with mechanical ventilation (OR: 3.6; 95% CI: 1.4-9.2, P =0.01), neutropenia (OR: 7.7; 95% CI: 0.8-73.1;P =0.07) while gastro-intestinal surgery was associated with an improved outcome (OR: 0.4; 95% CI: 0.16-0.96;P =0.04) . Of the 291 ICU patients, 19 acquired 22 episodes of nosocomial bacteraemia, and 18 were referred from the wards with documented nosocomial bacteraemia . Of these 37 bacteraemic patients, 17 (46%) died . When adjusting for predictors of death (SAPS II>/=40, cardiac and neurological failure), nosocomial bacteraemia markedly influence the outcome in ICU patients (OR: 3.4; 95% CI: 1.3-8.7;P =0.010) . This study suggests that the outcome of nosocomial bacteraemia in hospitalized patients is poor in ventilated and neutropenic patients and that nosocomial bacteraemia per se influenced outcome in ICU patients . Curr Microbiol, 2000 May, 40(5), 327 - 32 Isolation and identification of ruminal methanogens from grazing cattle; Jarvis GN et al.; To obtain information on the diversity of ruminal methanogens in grazing animals, three ruminal methanogens from grazing cattle were characterized and identified . Two of the isolates were rod-shaped, with one staining Gram-positive and being non-motile (BRM9), and the other (BRM16) staining Gram-negative and being motile . These isolates grew only on H(2)/CO(2) and formate, and optimally at 38 degrees C and pH 6.5-7.0 . The third isolate (CM1) was non-motile, pseudosarcina-shaped, and grew on H(2)/CO(2), acetate, and methyl-containing compounds, with optimal growth at 40 degrees C and pH 6.5 . DNA was prepared from the three isolates, and their 16S rRNA genes were sequenced . Phenotypic data and comparisons of nearly complete 16S rDNA sequences showed that BRM9, BRM16, and CM1 are strains of Methanobacterium formicicum, Methanomicrobium mobile, and Methanosarcina barkeri respectively . To the best of our knowledge, this is the first information on ruminal methanogens in cattle maintained under grazing management. Cent Afr J Med, 1999 Aug, 45(8), 217 - 20 Not everything acid-fast is Mycobacterium tuberculosis--a case of Nocardia; Bhagat K et al.; We report a case of a 47 year old woman who presented with a history of motor convulsions and a three month history of an increasingly painful and progressively enlarging mass on the right side of her back . Neurological examination revealed generalised wasting and a right sided hemiplegia . A biopsy of the mass was taken for microbiology which reported growing branching gram positive rods after three days of incubation . A mycological diagnosis of Nocardia asteroides was made . An MRI scan revealed extensive infiltration of the fungal mass into extending from the base of the skull to fifth cervical vertebra. J Heart Valve Dis, 2000 Jan, 9(1), 88 - 95; discussion 95-6 Structural changes in porcine bioprosthetic valves of a left ventricular assist system in human patients; Moczar M et al.; BACKGROUND AND AIM OF THE STUDY: Porcine, specially manufactured bioprosthetic valves regulate blood flow from the left ventricle to pump sac (inflow valve) and from the pump to the aorta (outflow valve) in a wearable, electrically powered left ventricular support system (LVAS, Novacor) . The increased need for long-term circulatory assistance requires information on the evolution of these valves when exposed to specific hemodynamic conditions and inflammatory reactions in the device . The study aim was to examine structural changes in valves from explanted LVASs . METHODS: Thirteen patients (11 males, two females; mean age 42 years (range: 17-64 years) were supported for a mean of 285 days (range: 37-1,293 days) with LVAS . Histologic sections from explanted inflow and outflow valves were studied immunohistochemically using peroxidase-labeled antibodies and avidin-biotinylated peroxidase complex for detection . RESULTS: In the macroscopically normal inflow valves (11/13), the outflow surface (facing the pump) was covered with a discontinuous deposit of fibrin, macrophages and granulocyte elastase . Fibrinogen, IgG, complement proteins C1q and C3 had infiltrated the extracellular matrix (ECM) between 37 and 1,293 days . The crevices were enlarged during circulatory support, and fibrinogen/fibrin insudations were detected in the spongiosa . The collagen layers in the fibrosa were disrupted after 293 days, and eroded on the inflow surface in the ventricularis after 1,293 days . In a deteriorated valve from a patient with endocarditis, Gram-positive bacteria and metalloproteinases were concentrated in the ECM . In the macroscopically normal (11/13) outflow valves, fibrin and complement proteins had penetrated the ECM from the inflow side (facing the pump), while macrophages and granulocytes were localized mainly on the outflow surface . IgG and complement proteins were detected on and beneath the cusp surface up to 200 days and covered the disrupted ECM as implant time progressed . CONCLUSIONS: Structural changes appear to progress more rapidly in the inflow than in the outflow of bioprosthetic valves . This difference indicates that the effects of biological factors are modulated by mechanical stress. Biochim Biophys Acta, 2000 Mar 10, 1469(1), 43 - 61 Topology and transport of membrane lipids in bacteria; Huijbregts RP et al.; The last two decades have witnessed a break-through in identifying and understanding the functions of both the proteins and lipids of bacterial membranes . This development was parallelled by increasing insights into the biogenesis, topology, transport and sorting of membrane proteins . However, progress in research on the membrane distribution and transport of lipids in bacteria has been slow in that period . The development of novel biochemical in vitro approaches and recent genetic studies have increased our understanding of these subjects . The aim of this review is to present an overview of the current knowledge of the distribution and transport of lipids in both Gram-positive and Gram-negative bacteria . Special attention is paid to recently obtained results, which are expected to inspire further research to finally unravel these poorly understood phenomena. J Bacteriol, 2000 Mar, 182(6), 1680 - 92 Biochemical and physical properties of the Methanococcus jannaschii 20S proteasome and PAN, a homolog of the ATPase (Rpt) subunits of the eucaryal 26S proteasome; Wilson HL et al.; The 20S proteasome is a self-compartmentalized protease which degrades unfolded polypeptides and has been purified from eucaryotes, gram-positive actinomycetes, and archaea . Energy-dependent complexes, such as the 19S cap of the eucaryal 26S proteasome, are assumed to be responsible for the recognition and/or unfolding of substrate proteins which are then translocated into the central chamber of the 20S proteasome and hydrolyzed to polypeptide products of 3 to 30 residues . All archaeal genomes which have been sequenced are predicted to encode proteins with up to approximately 50% identity to the six ATPase subunits of the 19S cap . In this study, one of these archaeal homologs which has been named PAN for proteasome-activating nucleotidase was characterized from the hyperthermophile Methanococcus jannaschii . In addition, the M . jannaschii 20S proteasome was purified as a 700-kDa complex by in vitro assembly of the alpha and beta subunits and has an unusually high rate of peptide and unfolded-polypeptide hydrolysis at 100 degrees C . The 550-kDa PAN complex was required for CTP- or ATP-dependent degradation of beta-casein by archaeal 20S proteasomes . A 500-kDa complex of PAN(Delta1-73), which has a deletion of residues 1 to 73 of the deduced protein and disrupts the predicted N-terminal coiled-coil, also facilitated this energy-dependent proteolysis . However, this deletion increased the types of nucleotides hydrolyzed to include not only ATP and CTP but also ITP, GTP, TTP, and UTP . The temperature optimum for nucleotide (ATP) hydrolysis was reduced from 80 degrees C for the full-length protein to 65 degrees C for PAN(Delta1-73) . Both PAN protein complexes were stable in the absence of ATP and were inhibited by N-ethylmaleimide and p-chloromercuriphenyl-sulfonic acid . Kinetic analysis reveals that the PAN protein has a relatively high V(max) for ATP and CTP hydrolysis of 3.5 and 5.8 micromol of P(i) per min per mg of protein as well as a relatively low affinity for CTP and ATP with K(m) values of 307 and 497 microM compared to other proteins of the AAA family . Based on electron micrographs, PAN and PAN(Delta1-73) apparently associate with the ends of the 20S proteasome cylinder . These results suggest that the M . jannaschii as well as related archaeal 20S proteasomes require a nucleotidase complex such as PAN to mediate the energy-dependent hydrolysis of folded-substrate proteins and that the N-terminal 73 amino acid residues of PAN are not absolutely required for this reaction. Gene, 2000 Feb 22, 244(1-2), 97 - 107 Description and characterization of IS994, a putative IS3 family insertion sequence from the salmon pathogen, Renibacterium salmoninarum; Rhodes LD et al.; Renibacterium salmoninarum, a slowly growing, Gram-positive bacterium, is responsible for bacterial kidney disease in salmonid fishes world-wide . To date, no mobile genetic elements have been reported for this pathogen . Here, we describe the first insertion sequence (IS) identified from R . salmoninarum . This element, IS994, has a significant predicted amino acid sequence homology (64.8 and 71.9%) to the two open reading frames encoding the transposase of IS6110 of Mycobacterium tuberculosis . Protein parsimony and protein distance matrix analyses show that IS994 is a member of group IS51 of the IS3 family . From a conservative estimate, there are at least 17 chromosomal insertions of IS994 or closely related elements . Sequence analysis of seven of these loci reveals single nucleotide polymorphisms throughout the element (including the terminal inverted repeats), a 15bp insertion in three of the seven loci, and an absence of flanking direct repeats or conserved insertion site . Restriction fragment length polymorphism analysis of XbaI-digested chromosomal DNA shows variations among European and North American isolates, indicating that IS994 may be a useful molecular marker for epizootiological studies. Infect Immun, 2000 Mar, 68(3), 1600 - 7 Bacterial induction of beta interferon in mice is a function of the lipopolysaccharide component; Sing A et al.; We investigated the reason for the inability of lipopolysaccharide (LPS)-resistant (Lps-defective {Lps(d)}) C57BL/10ScCr mice to produce beta interferon (IFN-beta) when stimulated with bacteria . For this purpose, the IFN-beta and other macrophage cytokine responses induced by LPS and several killed gram-negative and gram-positive bacteria in LPS-sensitive (Lps-normal {Lps(n)}; C57BL/10ScSn and BALB/c) and Lps(d) (C57BL/10ScCr and BALB/c/l) mice in vitro and in vivo were investigated on the mRNA and protein levels . In addition, double-stranded RNA (dsRNA) was used as a nonbacterial stimulus . LPS and all gram-negative bacteria employed induced IFN-beta in the Lps(n) mice but not in the Lps(d) mice . All gram-positive bacteria tested failed to induce significant amounts of IFN-beta in all four of the mouse strains used . As expected, all other cytokines tested (tumor necrosis factor alpha, interleukin 1alpha {IL-1alpha}, IL-6, and IL-10) were differentially induced by gram-negative and gram-positive bacteria . Stimulation with dsRNA induced IFN-beta and all other cytokines mentioned above in all mouse strains, regardless of their LPS sensitivities . The results suggest strongly that LPS is the only bacterial component capable of inducing IFN-beta in significant amounts that are readily detectable under the conditions used in this study . Consequently, in mice, IFN-beta is inducible only by gram-negative bacteria, but not in C57BL/10ScCr or other LPS-resistant mice. J Endod, 1999 Sep, 25(9), 605 - 8 Evaluation of microbial infiltration in restored cavities--an alternative method; Moreira Junior G et al.; This work evaluated the efficacy of an improved method used to determine the frequency of bacterial infiltration and bacterial population levels and morphotypes in cavities restored with adhesive composites in conventional mice . By using the alternative methodology suggested in this work, bacteria from microleakage were recovered and identified in cavities subjected to restoration procedures that used acid etching of the dentin and dentin adhesives used with light-curing resin . The methodology presented herein seems to be more effective than the one normally used to investigate the presence of bacteria, which uses acid demineralization of dental structures for the histological processing of tissues . The results suggest that the methodology presented in this work made it possible to recover and identify Gram-negative and Gram-positive bacteria from microleakage . Frequencies of microleakage and bacterial population levels in restored cavities using two different adhesive systems were not statistically different (p < 0.05). Indian J Exp Biol, 1999 Sep, 37(9), 936 - 8 Studies on anti-ulcer properties of Cissampelos mucronata leaf extract; Akah PA et al.; The methanolic extract of the leaves of C . mucronata was screened for anti-ulcer properties using animal models . On isolated guinea pig ileum the extract inhibited contractions evoked by acetylcholine, histamine and serotonin . The extract remarkably decreased the propulsive movement of gastrointestinal content . The extract exhibited significant anti-ulcer activity protecting rats from indomethacin, histamine and stress-induced ulcers . It inhibited the growth of both Gram-positive and Gram-negative microorganisms . The oral LD50 value of the extract in mice was estimated to be 8.5 +/- 0.35 g/kg . The results revealed that the plant C . mucronata has potential medicinal value as an anti-ulcer agent. Adv Perit Dial, 1999, 15, 205 - 8 Simultaneous catheter removal and replacement in peritoneal dialysis infections: update and current recommendations; Swartz RD et al.; Problematic peritoneal dialysis infection is a major cause of catheter loss and interruption of peritoneal dialysis (PD) therapy . In selected instances, problematic infection can be successfully treated by removing and replacing the catheter while continuing with PD . Accumulated experience has helped to define the circumstances under which a removal/replacement procedure is likely to be safe and under which complications are likely to arise . It appears that simultaneous removal and replacement can be expected to succeed when problematic infection is associated with tunnel infection, with recurring peritonitis repetitively culturing the same organism but clearing between episodes, and with gram-positive organisms . Success is less likely in the presence of ongoing inflammation, of active infection, of gram-negative or fungal organisms, or of any evidence of intra-abdominal adhesions . We review the literature on which these criteria are based and conclude with updated recommendations. Adv Perit Dial, 1999, 15, 193 - 6 Comparing peritonitis in continuous ambulatory peritoneal dialysis patients versus automated peritoneal dialysis patients; Locatelli AJ et al.; The purpose of our study was to compare the incidence of peritonitis between continuous ambulatory peritoneal dialysis (CAPD) treatment (Group I) and automated peritoneal dialysis (APD) treatment (Group II) taking into account the same population . We compared 20 patients with a follow-up of 215 patient-months on CAPD and 252 patient-months on APD . Demographic data, diagnosis, peritoneal equilibration test (PET) results, adequacy, and peritonitis rate were analyzed . Diagnoses included glomerulopathy 35%, autosomal dominant polycystic kidney disease (ADPKD) 20%, Type II diabetes 10%, systemic lupus erythematosus 5%, interstitial nephritis 5%, nephrolitiasis 5%, and unknown 20% . PET results showed that the group consisted of 30% high transporters, 45% high-average transporters, and 25% low-average transporters . Kt/V for Group I was 1.3 +/- 0.3, and for Group II, 1.83 +/- 0.48 . Creatinine clearance for Group I was 43.64 +/- 7.31 L/week/1.73 m2, and for Group II, 52.42 +/- 13.47 L/week/1.73 m2 . Group I presented a peritonitis rate of 8.3 episodes/patient-month, and Group II presented a rate of 18.9 episodes/patient-month . Gram-positive organisms were responsible for 49.8% of episodes of peritonitis in Group I (S . aureus 26.6%, S . epidermidis 16.6%, others 10%) and 83% of peritonitis episodes in Group II (S . epidermidis 46.6%, S . aureus 20%) . Gram-negative organisms were responsible for 16.5% of episodes of peritonitis in Group I . No gram-negative peritonitis was seen in Group II . APD patients developed two cases of candida peritonitis . Our preliminary results show that Group II exhibited a decrease in peritonitis rate while achieving better adequacy . In CAPD and APD peritonitis, gram-positive organisms predominated . In APD, we observed an increase in S . epidermidis incidence . No gram-negative organisms were observed in APD . It seems that APD is a safer treatment owing to the lower peritonitis incidence. FEMS Microbiol Lett, 2000 Feb 15, 183(2), 201 - 7 Hyaluronidases of Gram-positive bacteria; Hynes WL et al.; Bacterial hyaluronidases, enzymes capable of breaking down hyaluronate, are produced by a number of pathogenic Gram-positive bacteria that initiate infections at the skin or mucosal surfaces . Since reports of the hyaluronidases first appeared, there have been numerous suggestions as to the role of the enzyme in the disease process . Unlike some of the other more well studied virulence factors, much of the information on the role of hyaluronidase is speculative, with little or no data to substantiate proposed roles . Over the last 5 years, a number of these enzymes from Gram-positive organisms have been cloned, and the nucleotide sequence determined . Phylogenetic analysis, using the deduced amino acid sequences of the Gram-positive hyaluronidases, suggests a relatedness among some of the enzymes . Molecular advances may lead to a more thorough understanding of the role of hyaluronidases in bacterial physiology and pathogenesis. Zentralbl Chir, 1999, 124 Suppl 4, 13 - 8 {The influence of taurolidine on physiological and pathological blood coagulation and implications for its use}; Reinmuller J; Taurolidine is an anti-infective agent with an unusually broad spectrum of effectivity against gram-positive and gram-negative bacteria, anaerobic organisms, and fungi . The effective principle is explained by the decomposition of the substance and the transfer of methylol groups to specific molecular structures of the cell walls of microbes . The acceptors are amino groups of the amino sugars and the lysine residues of glycoproteids . More recent investigations have shown that the effect of taurolidine is not limited to microorganisms, but can be detected in cells of the macrorganism as well . Here the influence of taurolidine on different blood clotting factors is described . The results can be explained by a transfer of methylol groups to residues of arginine and histidine in the active region, in analogy to the transfer to lysine residues in microorganisms . It is therefore to be expected that taurolidine will influence other vital systems of the macroorganism, dependently of concentration, as long as their biological function is connected to residues of arginine or histidine . Examples are the complement system and the fibrinolysis system . The implications of these observations have to do with new indications in connection with clotting phenomena in extracorporal circulatory systems or with thrombolysis, as well as with known indications in cases of shock and sepsis. Biotechnol Appl Biochem, 2000 Feb, 31 ( Pt 1), 15 - 20 Affinity adsorbents for the vancomycin group of antibiotics; Yan H et al.; The vancomycin group of antibiotics kill Gram-positive bacteria by binding to nascent bacterial cell-wall peptidoglycan bearing the C-terminal sequence-D-Ala-D-Ala . In this paper, affinity adsorbents for the vancomycin group of antibiotics were prepared by immobilizing the peptidoglycan analogues -D-Ala-D-Ala, -succinyl-D-Ala and -succinyl-Gly on to crosslinked poly(N, N-dimethylacrylamide) . The adsorption capacities of the three adsorbents for demethylvancomycin were 0.59, 0.35 and 0.29 mmol/g, respectively . The adsorption capacity of the adsorbent with-D-Ala-D-Ala for vancomycin was 0.53 mmol/g . In contrast, the adsorbent bearing -succinyl-L-Ala hardly adsorbed demethylvancomycin . Aqueous sodium carbonate (0.4 M)/acetonitrile (7/3, v/v) completely desorbed demethylvancomycin adsorbed on the adsorbents. Eur J Pediatr, 2000 Mar, 159(3), 193 - 7 Glycopeptide prescribing in a tertiary referral paediatric hospital and applicability of hospital infection control practices advisory committee (HICPAC) guidelines to children; Nourse C et al.; This study was undertaken to investigate the frequency of, indications for and appropriateness of glycopeptide prescription in a paediatric tertiary referral hospital and to assess the usefulness of the Hospital Infection Control Practices Advisory Committee (HICPAC) guidelines . A prospective audit of all systemic glycopeptide prescriptions over a 2-month period was undertaken . Clinical and microbiological data were recorded . Of 2810 hospital admissions, systemic IV glycopeptides were prescribed on 57 occasions to 50 patients, 30 (52.6%) for vancomycin and 27 (47.4%) for teicoplanin . Prescriptions were for 34 males and 23 females aged from 2 weeks to 11 years (mean 15 months; median 9 months) . Median hospital stay was 50 days . Glycopeptides were given to the following patient groups: cardiology 7 (12%), prophylaxis for cardiac surgery 11 (19%), post-cardiac surgery 1 (1.8%), oncology 14 (24.6%), post-gastrointestinal tract surgical 8 (14%), general surgical 9 (15 . 8%) and medical 7 (12.3%) . Twenty three children (41.8%) had central lines in situ . Reason for use of glycopeptide was therapeutic in 7 (12.3%), empiric in 38 (66.7%), and as prophylaxis in 12 (21.1%) . Eight (14%) prescriptions met strict HICPAC criteria, but a further 22 (39%) prescriptions were considered appropriate in this high-risk population . Glycopeptides were chosen appropriately for cardiac surgery prophylaxis in a further 10 (18%) but timing and duration of use in this group was inappropriate . Of all prescriptions, use was empiric in 38 (76%) and appropriate cultures were obtained at the time of commencement in only 13 (34%) of these . Glycopeptides were not used for routine surgical prophylaxis or for first-line empiric treatment of febrile neutropenia . CONCLUSIONS: The strict implementation of HICPAC guidelines may not always be appropriate for children at risk of beta-lactam resistant gram-positive infections . Hospital guidelines need to be tailored to the patient population and microbial susceptibility patterns of each institution . Appropriate cultures should be obtained at the time glycopeptide treatment is begun so that duration of exposure can be limited. J Antimicrob Chemother, 2000 Feb, 45(2), 213 - 6 Penetration of cefpirome into the anterior chamber of the human eye after intravenous application; Egger SF et al.; The penetration of intravenously administered cefpirome into the anterior chamber of the non-inflamed human eye was investigated in this study . A total of 42 patients, all hospitalized for cataract extraction, received a dosage of 1 g or 2 g of cefpirome by iv infusion 1, 2 or 6 h preoperatively . An aqueous humour sample was collected immediately after paracentesis and a blood specimen was simultaneously obtained from each patient . All samples were analysed for cefpirome concentrations using high-pressure liquid chromatography . Mean aqueous humour levels of cefpirome in patients receiving a dosage of 1 g were 1.33 mg/L (1 h), 1.67 mg/L (2 h) and 1.29 mg/L (6 h after application), respectively . When patients received a dosage of 2 g cefpirome the resulting mean aqueous humour concentrations were 1.60 mg/L (1 h), 2.27 mg/L (2 h) and 2.39 mg/L (6 h after application), respectively . A statistically significant difference in aqueous humour concentrations between patients receiving 1 g or 2 g of cefpirome could not be proven . In conclusion, cefpirome penetrates well into the anterior chamber of the non-inflamed human eye in concentrations that are therapeutic for many gram-positive and gram-negative organisms, frequently responsible for anterior segment eye infections. J Antimicrob Chemother, 2000 Feb, 45(2), 159 - 65 Accumulation of rifampicin by Mycobacterium aurum, Mycobacterium smegmatis and Mycobacterium tuberculosis; Piddock LJ et al.; The characteristics of the accumulation of 2 mg/L {(14)C}rifampicin by wild-type strains of Mycobacterium aurum (A(+)), Mycobacterium smegmatis (mc(2)155) and Mycobacterium tuberculosis (H37Rv) were determined . After 10 min exposure M . aurum had accumulated 220 ng rifampicin/mg cells, M . smegmatis had accumulated 120 ng rifampicin/mg cells and M . tuberculosis had accumulated 154 ng rifampicin/mg cells . A steady-state concentration (SSC) of rifampicin was accumulated rapidly by M . aurum and M . tuberculosis within minutes of drug exposure, unlike M . smegmatis, which accumulated rifampicin more slowly . With an increase in the concentration of rifampicin from 0.12 mg/L to 2 mg/L there was an increase in the concentration of rifampicin accumulated by M . tuberculosis, with no detectable loss of viability over the 20 min of the accumulation experiment . With an increase in temperature there was also an increase in the concentration of rifampicin accumulated by M . tuberculosis; between 15 and 30 degrees C the increase was linear . For all three species sub-inhibitory concentrations of ethambutol increased the concentration of rifampicin accumulated . However, both growth and accumulation of rifampicin were lower in the presence of 0.05% Tween 80 . Accumulation of rifampicin by M . smegmatis was unaffected by the presence of the proton motive force inhibitor, 2,4-dinitrophenol (1 mM), whether added before or after the addition of rifampicin to the mycobacterial culture . For all three species, the Gram-positive bacterial efflux inhibitor reserpine (20 mg/L) slightly increased the SSC of rifampicin, but the increase was not statistically significant . Addition of glucose to energize a putative efflux pump had little effect on the accumulation of rifampicin in the presence or absence of reserpine for M . tuberculosis; however, for M . aurum and M . smegmatis the reserpine effect was abolished by the addition of glucose . These data suggest that rifampicin may be removed from wild-type mycobacteria by efflux, but that the pump(s) is expressed at low level. J Antibiot (Tokyo), 1999 Nov, 52(11), 998 - 1006 Tylopeptins A and B, new antibiotic peptides from Tylopilus neofelleus; Lee SJ et al.; Two new peptides, tylopeptins A and B, were isolated from the methanol extract of the fruiting body of the mushroom, Tylopilus neofelleus . These peptides were identified as peptaibols possessing an acetylated N-terminal residue, fourteen amino acids, and leucinol as the C-terminal amino alcohol . Sequential determination and complete 1H and 13C resonance assignments were based on positive ion FAB mass spectroscopy and two dimensional NMR techniques . These peptides were subsequently shown to be active against some gram-positive bacteria, but inactive against pathogenic fungi and gram-negative bacteria. Kansenshogaku Zasshi, 1999 Dec, 73(12), 1222 - 6 {A clinical evaluation of the new laboratory method that diagnoses bacterial infection, using silkworm larvae plasma}; Hiyoshi M et al.; Based upon the phenomenon that the peptidoglycan, a common component of Gram positive and negative bacteria, reacts specifically with silkworm larvae plasma (SLP), a new laboratory method named "SLP test" was developed to measure the reaction products in plasma quantitatively as SLP . This SLP test seems to be able to diagnose both Gram positive and negative bacterial infection . So we evaluated its usefulness in diagnosing clinical infectious diseases . This study included 14 patients with result to positive bacterial blood culture, 22 patients with bacterial local infection, 7 patients without any evidence of bacterial infection, and 19 healthy volunteers . It seemed that the cut-off value of this SLP test should be set at 0.6 ng/ml . The sensitivity and specificity of this SLP test were 57.1%, 100%, respectively . A significant difference was not detected statistically between SLP values of patients with Gram positive and Gram negative bacterial infectious diseases . So the SLP test did not appeared specific to either Gram positive or Gram negative bacteria . This test may become a new method diagnosing bacterial infectious disease. Bone Marrow Transplant, 2000 Jan, 25(1), 59 - 65 Fever and neutropenia in pediatric hematopoietic stem cell transplant patients; Mullen CA et al.; The objective of this study was to identify patterns of fever and neutropenia in pediatric patients undergoing initial hospitalization for hematopoietic stem cell transplantation . A retrospective review of 75 HSCTs over a 4-year period at a single institution was performed, of which 68% were allogeneic and 32% were autologous . Stem cell sources included bone marrow (29%), PBSC (52%) and umbilical cord blood (16%) . Fever occurred in 74 (98%) of the episodes . Unexplained fever (FUO) occurred in 43% . Bacteremia without an anatomic focus occurred in 29%, while CVC associated infections occurred in 17% . In 49% of transplants at least one blood culture was positive . The incidence of bacteremia was higher in allogeneic HSCTs (58%) than in autologous transplants (29%) . Gram-positive bacteria accounted for 71% of the isolates . Lower rates of bacteremia were observed in patients receiving oral fluoroquinolone prophylaxis . The median duration of fever was 12.5 days and time to engraftment 14 days . Regression analysis demonstrated that duration of fever was strongly associated with time to engraftment, and that time to engraftment was associated with source of cells and number of CD34+ cells/kg administered . Recipients of autologous PBSC had the shortest durations of fever and time to engraftment, while recipients of allogeneic umbilical cord blood had the longest . Bone Marrow Transplantation (2000) 25, 59-65. Immunology, 2000 Jan, 99(1), 153 - 61 The gills are an important site of iNOS expression in rainbow trout Oncorhynchus mykiss after challenge with the gram-positive pathogen Renibacterium salmoninarum; Campos-Perez JJ et al.; Following injection challenge of rainbow trout with the Gram-positive pathogen Renibacterium salmoninarum, serum nitrate levels increased indicative of NO production . The timing and amount of nitrate produced varied with the virulence of the ba