Gram positive

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 bacterial strain used, with the highest levels seen in fish challenged with the most virulent (autoaggregating) strain . Immunization with a killed R . salmoninarum preparation in Freund's incomplete adjuvant significantly increased nitrate levels after challenge . Inducible nitric oxide synthase (iNOS) transcript expression was detectable in rainbow trout tissues after injection challenge with R . salmoninarum, and its induction in the gills was both quick (between 3 and 6 hr) and relatively prolonged (lasting several days) . iNOS expression in the kidney was also seen at a later stage (24 hr) but appeared to switch off relatively rapidly . Bath challenge with R . salmoninarum also induced iNOS expression in gill, and a variable expression in the gut and kidney also occurred . These results highlight the importance of the gills, not only as a point of entry of pathogens but also as a tissue capable of mounting an immune response.

Adv Perit Dial, 1998, 14, 127 - 30
Peritonitis in the extended-care facility; Troidle LK et al.; Many patients with end-stage renal disease (ESRD) require admission to an extended-care facility (ECF) . Few data are available concerning the development of peritonitis among continuous peritoneal dialysis (CPD) patients residing in an ECF . We retrospectively reviewed the charts of 77 CPD patients admitted to an ECF between November 1993 and 31 August 1997 . A total of 25 episodes of peritonitis developed among CPD patients in the ECF during this period for an overall peritonitis rate of 1 episode in 19.8 patient-months . The CPD patients developing peritonitis in the ECF were similar in age and gender to the CPD patients residing in the ECF not developing peritonitis . There were more African-Americans among the group of CPD patients residing in the ECF who developed peritonitis than among the ECF residents who did not develop peritonitis (41% vs . 23%, respectively; P < 0.05) . Patients developing peritonitis in the ECF resided in the ECF significantly longer than the remaining CPD patients not developing peritonitis in the ECF (106 vs . 77 days, respectively; P < 0.05) . The overall rate of peritonitis in the ECF was lower than that seen in the community (1 episode in 19.8 patient-months vs . 1 episode in 10.0 patient-months, respectively) . The rate of gram-positive peritonitis was lower than the rate of gram-positive peritonitis seen in the community setting (1 episode in 54.9 patient-months vs . 1 episode in 14.9 patient-months, respectively) . The rate of culture-negative peritonitis was higher among the ECF patients than among patients developing community-acquired peritonitis (1 episode in 61.9 patient-months vs . 1 episode in 106.2 patient-months, respectively) . The spectrum of organisms in the ECF was different than the spectrum noted among patients developing hospital-acquired peritonitis . Eleven of the 25 episodes of peritonitis were treated successfully at the ECF while the remaining 14 episodes of peritonitis were treated at an acute-care hospital . Continuous PD therapy was continued following 19 of the 25 episodes (76%), 1 patient transferred to hemodialysis, and 5 patients expired . We conclude that patients develop peritonitis in the ECF less frequently than in the community setting, with the spectrum of organisms different than the spectrum seen in the community and hospital settings . Seventy-six percent of the patients continue CPD therapy following an episode of peritonitis.

Bone Marrow Transplant, 1999 Dec, 24(11), 1213 - 7
Infections in patients managed at home during autologous stem cell transplantation for lymphoma and multiple myeloma; Herrmann RP et al.; A group of 51 patients with multiple myeloma, non-Hodgkin's lymphoma or Hodgkin's disease receiving high-dose chemotherapy and autologous peripheral blood stem cell rescue received chemotherapy and clinical care in the peritransplant period at home . This group was compared with 88 cases with the same diagnoses, receiving the peripheral stem cell transplant over the same time period as an inpatient in a high efficiency particulate air filtered bone marrow transplant unit . Patients were treated at home based on choice, geographic accessibility, availability of an educated care giver and a clean home environment, and comprehension of the concepts of infection and aseptic techniques . Febrile neutropenia and sepsis were not increased in the home group and no episodes of septic shock were seen in this group . Patients at home received prophylactic oral ciprofloxacin and roxithromycin during the phase when the absolute neutrophil count was < 1 x 10(9)/l . Fewer gram-negative infections, but no diminution in gram-positive infections or in the rate of fever were seen in patients at home . Empiric therapy with a third generation cephalosporin, teicoplanin and tobramycin was instituted in 31 patients who developed a fever greater than 38.5 degrees C . Of this group of 31, 18 required admission to hospital, 12 because of febrile neutropenia which persisted or was considered unsuitable for management at home due to sepsis . The remaining 13 with febrile neutropenia remained at home throughout, as did the 20 cases not developing neutropenic fever . This study demonstrates the feasibility of managing carefully selected patients in their home environment when at risk from febrile neutropenia or other septic complications following autologous peripheral stem cell support.

Biochem J, 2000 Feb 1, 345 Pt 3, 653 - 64
Androctonin, a hydrophilic disulphide-bridged non-haemolytic anti-microbial peptide: a plausible mode of action; Hetru C et al.; Androctonin is a 25-residue non-haemolytic anti-microbial peptide isolated from the scorpion Androctonus australis and contains two disulphide bridges . Androctonin is different from known native anti-microbial peptides, being a relatively hydrophilic and non-amphipathic molecule . This raises the possibility that the target of androctonin might not be the bacterial membrane, shown to be a target for most amphipathic lytic peptides . To shed light on its mode of action on bacteria and its non-haemolytic activity, we synthesized androctonin, its fluorescent derivatives and its all-D-amino acid enantiomer . The enantiomer preserved high activity, suggesting a lipid-peptide interaction between androctonin and bacterial membranes . In Gram-positive and (at higher concentrations) Gram-negative bacteria, androctonin induced an immediate perturbation of the permeability properties of the cytoplasmic membrane of the bacterial energetic state, concomitant with perturbation of the morphology of the cell envelope as revealed by electron microscopy . Androctonin binds only to negatively charged lipid vesicles and induces the leakage of markers at high concentrations and with a slow kinetics, in contrast with amphipathic alpha-helical anti-microbial peptides that bind and permeate negatively charged vesicles, and to a smaller extent also zwitterionic ones . This might explain the selective lytic activity of androctonin towards bacteria but not red blood cells . Polarized attenuated total reflection-Fourier transform infrared spectroscopy revealed that androctonin adopts a beta-sheet structure in membranes and did not affect the lipid acyl chain order, which supports a detergent-like effect . The small size of androctonin, its hydrophilic character and its physicochemical properties are favourable features for its potential application as a replacement for commercially available antibiotics to which bacteria have developed resistance.

J Pediatr Orthop, 2000 Jan-Feb, 20(1), 44 - 7
A shortened course of parenteral antibiotic therapy in the management of acute septic arthritis of the hip; Kim HK et al.; We reviewed 20 consecutive patients with a culture-proven acute septic arthritis of the hip who were treated with a shortened course of parenteral antibiotic therapy after a surgical drainage . Patients were switched over to an oral antibiotic when they showed clinical improvement . Sixteen of the 20 patients had parenteral antibiotic therapy of <10 days, whereas nine of these patients received <7 days of parenteral therapy (mean, 8.2 days) . No recurrence of infection, readmission, or osteomyelitis was observed after the discharge . At the follow-up interview (mean, 32 months), 18 patients were completely asymptomatic, and two patients had occasional hip pain with activity but no physical limitations . All 20 patients had normal hip range of motion and gait . Their latest radiographs (mean, 26 months) revealed 11 patients with normal findings, six patients with mild coxa magna, and three patients with a smaller ossific nucleus compared with the unaffected side . We conclude that a community-acquired, acute gram-positive septic arthritis of the hip can be managed safely with a surgical drainage and a shortened course of parenteral antibiotic therapy, which can be switched over to an oral therapy based on the patient's response to the therapy.

Am J Gastroenterol, 2000 Jan, 95(1), 214 - 7
Bacterial peritonitis after elective endoscopic variceal ligation: a prospective study; Lin OS et al.; OBJECTIVE: Endoscopic variceal ligation is becoming the therapy of choice for esophageal varices, replacing endoscopic variceal sclerotherapy . The latter is associated with a 5-53% incidence of port-procedural bacteremia and a 0.5-3% incidence of peritonitis, whereas the former carries a 3-6% risk of bacteremia . However, the incidence of peritonitis after variceal ligation has not been well studied . This prospective study is designed to investigate the risk of developing bacteremia and bacterial peritonitis after elective endoscopic variceal ligation . METHODS: Sixty-seven patients with esophageal varices and ascites secondary to liver cirrhosis underwent elective endoscopic variceal ligation . Before the procedure, ascitic fluid was drawn under ultrasound guidance and sent for cell counts, Gram stain, and cultures . Two to 4 days afterward, a repeat ascitic fluid sample was sent for the same studies whether or not the patient had symptoms or signs suggestive of infection . Blood cultures were drawn both immediately before and after the endoscopic ligation procedure . RESULTS: Of 67 subjects, 11 developed asymptomatic bacteremia with Gram-positive commensals . However, none of them progressed to peritonitis . Two patients who did not have bacteremia developed mild febrile peritonitis with Escherichia coli and were successfully treated with oral antibiotics . No other infectious complications were noted . CONCLUSIONS: There is a significant risk of asymptomatic bacteremia and bacterial peritonitis after elective variceal ligation . The peritonitis does not seem to be related to the bacteremia, as patients who had bacteremia did not develop peritonitis and vice versa . In addition, the involved organisms were quite different . Unlike the bacteremia, postligation peritonitis may be a consequence of severe liver cirrhosis rather than the procedure itself . The clinical significance of postligation bacteremia is doubtful . With regard to peritonitis, in our opinion the use of prophylactic antibiotics should be reserved for patients with Child's C class cirrhosis, a recent history of variceal bleeding, a past history of bacterial peritonitis, or a comorbid immunosuppressive condition.

Protein Sci, 1999 Dec, 8(12), 2580 - 8
NMR assignments, secondary structure, and global fold of calerythrin, an EF-hand calcium-binding protein from Saccharopolyspora erythraea; Aitio H et al.; Calerythrin is a 20 kDa calcium-binding protein isolated from gram-positive bacterium Saccharopolyspora erythraea . Based on amino acid sequence homology, it has been suggested that calerythrin belongs to the family of invertebrate sarcoplasmic EF-hand calcium-binding proteins (SCPs), and therefore it is expected to function as a calcium buffer . NMR spectroscopy was used to obtain structural information on the protein in solution . Backbone and side chain 1H, 13C, and 15N assignments were obtained from triple resonance experiments HNCACB, HN(CO)CACB, HNCO, CC(CO)NH, and {15N}-edited TOCSY, and HCCH-TOCSY . Secondary structure was determined by using secondary chemical shifts and characteristic NOEs . In addition, backbone N-H residual dipolar couplings were measured from a spin-state selective {1H, 15N} correlation spectrum acquired from a sample dissolved in a dilute liquid crystal . Four EF-hand motifs with characteristic helix-loop-helix patterns were observed . Three of these are typical calcium-binding EF-hands, whereas site 2 is an atypical nonbinding site . The global fold of calerythrin was assessed by dipolar couplings . Measured dipolar couplings were compared with values calculated from four crystal structures of proteins with sequence homology to calerythrin . These data allowed us to recognize an overall similarity between the folds of calerythrin and sarcoplasmic calcium-binding proteins from the sandworm Nereis diversicolor and the amphioxus Branchiostoma lanceolatum.

Arch Intern Med, 2000 Jan 10, 160(1), 113 - 5
Hepatic decompensation in patients with cirrhosis during infection with influenza A; Duchini A et al.; BACKGROUND: Patients with chronic liver disease can develop hepatic decompensation during systemic infections . Although gram-negative and gram-positive bacteria are well recognized as causes of decompensation, the effect of influenza virus infection on patients with chronic liver disease is poorly documented . METHODS: Retrospective analysis of patients with positive viral cultures who were seen at a liver transplantation clinic in a tertiary care referral center during the 1997-1998 influenza A (H3N2) epidemic in San Diego, Calif . RESULTS: Three patients with end-stage liver disease (1 with Wilson disease and 2 with alcoholic liver disease) developed hepatic decompensation and required hospitalization during infection with influenza A . Two patients had biochemical and clinical evidence of hepatic decompensation, including ascites, hepatic encephalopathy, and peripheral edema, and the third had acute hepatocellular damage, with elevated levels of aminotransferases . Viral hepatitis serologic test results, acetaminophen levels, drug and alcohol screening findings, and bacterial and fungal cultures were negative in all 3 patients . Hepatic decompensation resolved without the need for transplantation in the 2 patients with liver failure, and all patients recovered to their baseline liver function levels within 1 month of onset of acute illness . CONCLUSIONS: Influenza A infection can cause hepatic decompensation and hospitalization in patients having cirrhosis or who are awaiting liver transplantation . Effective prevention with vaccination and early recognition and treatment of influenza are strongly recommended in these individuals.

Antibiot Khimioter, 1999, 44(11), 17 - 22
{The efficacy of cefepime (Maxipime) in the treatment of abdominal sepsis in surgical patients}; Gel'fand EB et al.; The efficacy of cefepime in the treatment of 46 patients operated for general peritonitis of various genesis and severity (APACHE II not greater than 35) was studied . Cefepime was used in a dose of 2 g administered every 12 hours as slow intravenous infusions in 0.9 per cent sodium chloride solution in combination with metronidazole administered intravenously in a dose of 7.5 mg/kg body weight . The treatment course was 4 to 15 days . 45 patients were given diflucan for the prophylaxis of fungal superinfection, 3 patients were given aminoglycoside antibiotics (netilmicin or amikacin) and 2 patients were given vancomycin per os . The favourable clinical effect of the cefepime therapy was stated in 38 patients (82.6 per cent) including 4 out of 10 patients with initial APACHE II > 15 . 101 isolates of aerobic gram-negative and gram-positive microbes from 38 patients treated with cefepime in combination with metronidazole were tested to estimate the bacteriological efficacy of the therapy and it was shown that only 5.9 per cent of them was resistant . The pathogen eradication was stated in 84.2 per cent of the patients.

FEMS Microbiol Lett, 2000 Jan 15, 182(2), 197 - 205
Thiol-activated cytolysins: structure, function and role in pathogenesis; Billington SJ et al.; Members of the thiol-activated family of cytolysins are involved in the mechanism of pathogenesis of a number of Gram-positive species . While they are pore-forming toxins, their major pathogenic effects may be more subtle than simple lysis of host cells, and may include interference with immune cell function and cytokine induction . Crystal structure, electron microscopy, mutagenesis and antibody binding studies have led to the modeling of a novel mechanism of pore formation, encompassing membrane-binding, membrane insertion and oligomerization . Despite their designation as thiol-activated cytolysins, it is now clear that thiol activation is not an important property of this group of toxins.

Int J Clin Pract, 1998 Nov-Dec, 52(8), 577 - 81
Non-inpatient use of teicoplanin; Nathwani D; Teicoplanin is a glycopeptide antibiotic which was introduced into clinical use in 1988 . Its activity against most Gram-positive bacteria, including the emergent multiply resistant species, its ease of administration and safety commend its continued use well into the next millennium . The provision of health care in alternative settings such as the home or outpatient clinic has grown in importance, both in Europe and North America . One such service is the provision of parenteral antibiotics for treating a variety of infections in the non-inpatient setting . This paper aims to review and condense the current published experience of teicoplanin in this setting . Evidence suggests that teicoplanin can be administered effectively using a range of flexible, convenient and safe dosing schedules and is a highly cost-effective therapeutic option for treating in the non-inpatient setting many chronic infections e.g . osteomyelitis and endocarditis, and some acute infections.

J Ethnopharmacol, 1999 Nov 30, 67(3), 347 - 54
Screening of medicinal plants used in Lesotho for anti-bacterial and anti-inflammatory activity; Shale TL et al.; Traditional healers and herbalists from Lesotho were interviewed about plants used in traditional remedies by the Sotho . Plant roots are most often used to make water extracts . Mainly high altitude plants are used, with lowland healers obtaining most of their plant material from the highlands, either by collecting them or buying them from highland gatherers . As a result of ethnobotanical data obtained, leaves and roots of 12 plants were extracted using hexane, methanol and water, respectively and the extracts screened for anti-inflammatory activity using the cyclo-oxygenase bioassay . Six species yielded inhibitory activity above 90% . Hexane and methanol leaf and root extracts were the most active . Leaves and roots of 16 plants were extracted using hexane, methanol and water and the respective extracts screened for anti-bacterial activity using the disc-diffusion assay . Six species displayed very high anti-bacterial activity against both gram-positive and gram-negative bacteria . A number of plant extracts had medium inhibitory activity, mostly against gram-positive bacteria . The activity was mainly found in the root extracts.

Cardiovasc Pathol, 1999 Nov-Dec, 8(6), 329 - 32
Minocycline pigmentation of heart valves; Sant'Ambrogio S et al.; Minocycline, a derivative of tetracycline, is a broad spectrum antibiotic used in the treatment of gram-positive and gram-negative infections . Benitz et al . (1) were the first to report black discoloration of the thyroid gland in rats, dogs, and monkeys given minocycline . Since that time, there have been numerous reports in the literature describing minocycline related black pigmentation of the skin, thyroid gland, and other sites . We report an unusual case of minocycline induced pigmentation of the cardiac valves and coronary vessels . The pigment stained with Fontana-Masson and was reduced with bleaching . The exact nature of the pigment is unclear; however, various theories have been advocated . Ochronosis is another cause of black pigmentation of the heart valves; the clinical history should allow distinction between the two.

Biofactors, 1999, 10(2-3), 201 - 6
Thiols in formaldehyde dissimilation and detoxification; Duine JA; Glutathione is not a universal coenzyme for formaldehyde oxidation . MySH (mycothiol, 1-O-(2'-{N-acetyl-L-cysteinyl}amido-2'-deoxy-alpha-D-glucopyranosyl)-D-m yo-inositol) is GSH's counterpart as coenzyme in formaldehyde dehydrogenase from certain gram-positive bacteria . However, formaldehyde dissimilation and detoxification not only proceed via thiol-dependent but also via thiol-independent dehydrogenases . The distinct structures and enzymatic properties of MySH-dependent and GSH-dependent formaldehyde dehydrogenases could provide clues for development of selective drugs against pathogenic Mycobacteria . It is to be expected that other new types of thiol-dependent formaldehyde dehydrogenases will be discovered in the future . Indications exist that the product of thiol-dependent formaldehyde oxidation, the thiol formate ester, is not only hydrolytically converted into thiol and formate but can also be oxidatively converted in some cases by a molybdoprotein aldehyde dehydrogenase into the corresponding carbonate ester, decomposing spontaneously into CO2 and the thiol.

Int Immunol, 2000 Jan, 12(1), 113 - 7
Cellular responses to bacterial cell wall components are mediated through MyD88-dependent signaling cascades; Takeuchi O et al.; MyD88 is an adaptor molecule essential for signaling via the Toll-like receptor (TLR)/IL-1 receptor family . TLR4 is a member of the TLR family and a point mutation in the Tlr4 gene causes hyporesponsiveness to lipopolysaccharide (LPS) in C3H/HeJ mice . We have previously shown that both TLR4- and MyD88-deficient mice are hyporesponsive to LPS . In this study we examined the responsiveness of these two knockout mice to various bacterial cell wall components . Cells from TLR4-deficient mice responded to several kinds of LPS, peptidoglycan and crude cell wall preparation from Gram-positive bacteria and mycobacterial lysates . In contrast, macrophages and splenocytes from MyD88-deficient mice did not respond to any of the bacterial components we tested . These results show that MyD88 is essential for the cellular response to bacterial cell wall components.

Curr Opin Microbiol, 1999 Dec, 2(6), 582 - 7
How bacteria talk to each other: regulation of gene expression by quorum sensing; Bassler BL; Quorum sensing, or the control of gene expression in response to cell density, is used by both gram-negative and gram-positive bacteria to regulate a variety of physiological functions . In all cases, quorum sensing involves the production and detection of extracellular signalling molecules called autoinducers . While universal signalling themes exist, variations in the design of the extracellular signals, the signal detection apparatuses, and the biochemical mechanisms of signal relay have allowed quorum sensing systems to be exquisitely adapted for their varied uses . Recent studies show that quorum sensing modulates both intra- and inter-species cell-cell communication, and it plays a major role in enabling bacteria to architect complex community structures.

Antimicrob Agents Chemother, 2000 Jan, 44(1), 164 - 6
Ciprofloxacin, lomefloxacin, or levofloxacin as treatment for chronic osteomyelitis; Greenberg RN et al.; The efficacy and safety of three oral fluoroquinolones (lomefloxacin, levofloxacin, and ciprofloxacin) for the treatment of chronic osteomyelitis were analyzed . Twenty-seven patients had documented infections with quinolone-sensitive organisms and received either lomefloxacin, levofloxacin, or ciprofloxacin . Levofloxacin was effective therapy for 9 of 15 (60%) patients . Lomefloxacin was effective therapy for five of seven (71%) patients, and ciprofloxacin was effective therapy for two of five patients (40%) . Average follow-up was 11.8 months for patients who completed the course of therapy, and the average duration of therapy was 60.6 days . Gram-positive bacteria were isolated from 18 patients, and 11 patients were cured . Oral fluoroquinolones can be safe, effective therapy if they are given for a prolonged course as treatment for infections caused by susceptible gram-positive as well as gram-negative organisms and in combination with adequate surgical debridement.

Drugs, 1999, 58 Suppl 2, 6 - 10
Mode of action of fluoroquinolones; Hooper DC; The mode of action of quinolones involves interactions with both DNA gyrase, the originally recognised drug target, and topoisomerase IV, a related type II topoisomerase . In a given bacterium these 2 enzymes often differ in their relative sensitivities to many quinolones, and commonly DNA gyrase is more sensitive in gram-negative bacteria and topoisomerase IV more sensitive in gram-positive bacteria . Usually the more sensitive enzyme represents the primary drug target determined by genetic tests, but poorly understood exceptions have been documented . The formation of the ternary complex of quinolone, DNA, and either DNA gyrase or topoisomerase IV occurs through interactions in which quinolone binding appears to induce changes in both DNA and the topoisomerase that occur separately from the DNA cleavage that is the hallmark of quinolone action . X-ray crystallographic studies of a fragment of the gyrase A subunit, as well as of yeast topoisomerase IV, which has homology to the subunits of both DNA gyrase and topoisomerase IV, have revealed domains that are likely to constitute quinolone binding sites, but no topoisomerase crystal structures that include DNA and quinolone have been reported to date . Inhibition of DNA synthesis by quinolones requires the targeted topoisomerase to have DNA cleavage capability, and collisions of the replication fork with reversible quinolone-DNA-topoisomerase complexes convert them to an irreversible form . However, the molecular factors that subsequently generate DNA double-strand breaks from the irreversible complexes and that probably initiate cell death have yet to be defined.

Int J Antimicrob Agents, 1999 Oct, 13(2), 71 - 8
Cost considerations in the evaluation of new therapies for gram-positive bacteria; Nathwani D et al.; The rapid emergence of antibiotic resistance in hospitals and the community, particularly amongst Gram-positive cocci, has not been paralleled by adequate understanding of the true clinical and economic impact of infection with resistant pathogens . Furthermore, the development of novel compounds to combat this threat has been slow until recently when a number of new agents, some with a unique structure and mode of action (e.g . linezolid), are now being developed for clinical use . This paper aims to outline (1) the costs associated with infection by a resistant organism (2) the pharmacoeconomic arguments when considering a new drug and (3) the key decisions when evaluating a new drug such as linezolid, for inclusion in a formulary or policy with particular reference to the treatment of serious Gram-positive infections.

Microbiology, 1999 Nov, 145 ( Pt 11), 3305 - 15
Microbial diversity in marine sediments from Sagami Bay and Tokyo Bay, Japan, as determined by 16S rRNA gene analysis; Urakawa H et al.; 16S rDNA clone libraries were analysed to investigate the microbial diversity in marine sediments from Sagami Bay (stations SA, water depth of 1159 m, and SB, 1516 m) and Tokyo Bay (station TK, 43 m) . A total of 197 clones was examined by amplified rDNA restriction analysis (ARDRA) using three four-base-specific restriction enzymes (Hhal, Rsal and Haelll) . In SA, 57 RFLP types were detected from 77 clones . In SB, 17 RFLP types were detected from 62 clones . In TK, 21 RFLP types were detected from 58 clones . The genotypic diversity among the three sampling sites was 0.958, 0.636 and 0.821, respectively, indicating that the microbial diversity of SA was higher than at the other two stations . At SA, the most abundant RFLP type constituted 10% of all clones . The samples from SB and TK had dominant RFLP types which constituted 60% and 38% of the total clone libraries, respectively . The community structure of SA included many single-type clones, which were found only once in the clone libraries . This structure contrasted with that of the other two stations . Thirty-seven clones were selected and sequenced according to dendrograms derived from ARDRA, to cover most of the microbial diversity in the clone libraries . No clones were identical to any of the known 165 rRNA sequences or to each other . All sequences had >84.8% similarity to rDNA sequences retrieved from the DNA databases . Sequenced clones fell into five major lineages of the domain Bacteria: the gamma, delta and epsilon Proteobacteria, Gram-positive bacteria and the division Verrucomicrobia . At SA, the Verrucomicrobia and the three subclasses of the Proteobacteria were found . Most clone sequences belonged to the gamma Proteobacteria . The high-GC Gram-positive bacteria and the gamma subclass of the Proteobacteria were common at both SB and TK . Although the depths of SB and TK were very different, the community diversity inferred from ARDRA and the taxonomic position of the dominant clones were similar . All clones belonging to the highGC Gram-positive bacteria collected from both SB and TK fell into the same cluster and are regarded as members of an unknown actinomycete group . The clone compositions were different at each sampling site, and clones of the gamma Proteobacteria and high-GC Gram-positive bacteria were dominant.

Antimicrob Agents Chemother, 1999 Dec, 43(12), 3036 - 8
Novel streptomycin and spectinomycin resistance gene as a gene cassette within a class 1 integron isolated from Escherichia coli; Sandvang D; The aadA genes, encoding resistance to streptomycin and spectinomycin, have been found as gene cassettes in different gram-negative and gram-positive bacterial species . The present study has revealed the sequence of a new gene, aadA5, integrated as a gene cassette together with the trimethoprim resistance gene dfr7 in a class 1 integron . The integron was located on a plasmid and was identified in a pathogenic porcine Escherichia coli isolate.

Fogorv Sz, 1999 Oct, 92(10), 317 - 27
{Microscopic study of dental calculus in cadavers from the 18th-19th centuries}; Torok K et al.; The authors studied the dental calculus of 20 mummies with ligth microscopy, polarized ligth microscopy and scanning electron microscopy . Gram positive bacteria could be detected in all preparates, while Gram negative bacteria in 12 and fungi only in 3 dental calculus was visible . Animal food remains within five, and plant remains in all dental calculus were identified . Anorgic element and cell debris were seen in all preparates.

FEMS Microbiol Lett, 1999 Dec 1, 181(1), 145 - 52
A xylanase produced by the rumen anaerobic protozoan Polyplastron multivesiculatum shows close sequence similarity to family 11 xylanases from gram-positive bacteria; Devillard E et al.; We report for the first time the cloning and characterisation of a protozoal enzyme involved in plant cell wall polysaccharide degradation . A cDNA library was constructed from the ruminal protozoan Polyplastron multivesiculatum and a stable clone expressing xylanase activity was isolated . The encoded enzyme belongs to the glycoside hydrolase family 11, and phylogenetic analysis indicates a closer relationship with catalytic domains from Gram-positive bacteria than the other fibrolytic eukaryotes from the rumen, the anaerobic fungi.

FEMS Microbiol Lett, 1999 Dec 1, 181(1), 41 - 8
A small cryptic plasmid from Ruminobacter amylophilus NIAH-3 possesses functional mobilization properties; Ogata K et al.; The complete nucleotide sequence of a small cryptic plasmid designated pRAO1, from the Gram-negative ruminal bacterium Ruminobacter amylophilus NIAH-3, was determined . The plasmid is a circular DNA molecule, 2140 bp in size, with a GC content of 40% . Computer-assisted analysis identified three open reading frames (ORFs), one of which, ORF3 (347 amino acids), displayed a high degree of amino acid identity with the Mob proteins involved in conjugative mobilization and interplasmid recombination of plasmids from Gram-positive bacteria . We proved the mobilization properties of pRAO1 in the Escherichia coli system using the coresident IncW broad-host-range conjugative plasmid R388 . These data demonstrated, for the first time, the mobilization properties of small cryptic plasmids from Gram-negative inhabitants of the rumen.

Pediatr Transplant, 1999 Nov, 3(4), 322 - 7
Utility of liver biopsy culture in pediatric liver transplant recipients; Cohen MS et al.; The purpose of our study was to determine the utility of the practice of culturing percutaneous liver biopsy specimens obtained from pediatric LT recipients for evaluation of fever and/or elevated serum aminotransferases . Accordingly, a retrospective analysis was done of the 101 liver biopsies obtained during an eight-year period which had been submitted for bacterial, fungal and/or viral culture (out of a total of 174 biopsies in 38 patients) . The purpose of the analysis was to ask three questions . (1) What organisms were cultured? (2) Were there clinical profiles that were characteristic of the type of organism? (3) Was the practice cost-effective? The analysis indicated that 34/86 biopsy cultures were positive for bacteria, 4/75 for fungus and 2/81 for virus . Clinical and laboratory data for children with cultures positive for enteric flora (n = 9) were compared to those with cultures positive for Gram-positive organisms (n = 17), laboratory contaminants (n = 8), and those with negative cultures (n = 52) . Children with biopsies positive for enteric flora had a 'cholestatic profile': mean direct bilirubin 7 mg/dl, ALT 78 IU/l, direct bilirubin/ALT 0.09, in comparison to children with biopsies positive for Gram-positive flora . These children had a 'hepatocellular profile': mean direct bilirubin 4 mg/dl, ALT 332 IU/l, direct bilirubin/ALT 0.01 (p = 0.04 versus the enteric flora values) and a high percentage of polymorphonuclear leukocytes (mean 69% versus 38% for those with negative cultures, p = 0.001.) The charge for performing each bacterial culture was $28 (total $28 x 86 = $2408: $268 per enteric flora-positive biopsy; $93 per biopsy positive for either enteric flora or Gram-positive flora) . The charge for each fungal culture was also $28 (total $28 x 75 = $2100: $525 per positive culture), while the cost for each viral culture was $140 (total $140 x 81 = $11,340: $5670 per positive culture) . Thus, discounting the eight cultures positive for laboratory contaminants, a total of $15,848 was spent for 32 positive cultures . Given the high cost of liver transplantation, this information suggests that discretion should be used in submission of liver biopsy samples for culture in pediatric transplant patients . We recommend that when liver biopsies are performed for evaluation of elevated serum aminotransferases and/or fever, culture of biopsy specimens for bacteria should be considered in children with a 'cholestatic profile': direct bilirubin > or = 7 mg/dl and direct bilirubin/ALT > 0.08, or a 'hepatocellular profile': direct bilirubin < or = 4 mg/dl and direct bilirubin/ALT < 0.05, together with polymorphonuclear leukocytes > 70% . Following these guidelines might provide valuable information pertinent to patient management (especially since Gram-negative organisms can sometimes be cultured from the liver and not from blood) while reducing costs . Fungal cultures should be restricted to critically ill children . However, our data suggest that the practice of obtaining fungal and viral cultures of the liver in most pediatric transplant patients has an unacceptably high cost/benefit ratio, particularly since recovery of the organism from the peripheral blood is likely.

Kidney Int Suppl, 1999 Nov, 72, S15 - 9
Use of adsorptive mechanisms in continuous renal replacement therapies in the critically ill; Tetta C et al.; The pathophysiology of sepsis is becoming a more complicated scenario . In sepsis, endotoxin or other gram-positive derived products induce a complex and dynamic cellular response giving rise to several mediators known to be relevant in the pathogenesis of septic shock, such as specific mediators . substances responsible for up- or down-regulation of cytokine receptors and cytokine antagonists, inactivators of nuclear factor-kappaB or signal transduction pathways, and precursor molecules . In this article, we delve into some new concepts stemming from the use of sorbents in continuous plasma filtration . The rationale is based on the assumption that the nonspecific removal of several mediators of the inflammatory cascade and cytokine network may improve outcome in a rabbit model of septic shock and hemodynamics in a pilot clinical study . The importance of looking for innovative treatments specifically targeted for the special needs of the critically ill patients rather than using concepts and technology applied to the treatment of chronic renal failure is underlined.

Chest, 1999 Nov, 116(5), 1354 - 9
Persistent preload defect in severe sepsis despite fluid loading: A longitudinal echocardiographic study in patients with septic shock; Jardin F et al.; STUDY OBJECTIVE: To investigate the rate of recovery from septic shock in patients with suspected left ventricular (LV) preload deficiency and LV systolic dysfunction . DESIGN: A monitoring period was defined by the need for inotropic/vasopressor support, and LV function was assessed daily during this period by bedside two-dimensional echocardiography (2D-ECHO) . SETTING: University hospital ICU . PATIENTS: During a 5-year period, 90 patients with an episode of septic shock (60% with gram-positive bacteria as the causative agent) were consecutively enrolled in the study (mean age, 55 +/- 18 years) . Standard volume resuscitation combined with inotropic/vasopressor support was used to maintain systolic arterial pressure > 90 mm Hg . All patients received mechanical ventilation because of associated respiratory failure . The average duration of hemodynamic support was 4.4 +/- 1.6 days . Thirty-four patients were weaned from hemodynamic support during the monitoring period and ultimately recovered (group I) . Twenty-eight patients died from refractory circulatory failure during the monitoring period, and 28 died later from ARDS or multiple organ dysfunction syndrome, leading to a 62% overall mortality rate (group II) . METHODS: Daily bedside LV volumes and ejection fraction (LVEF) were recorded using 2D-ECHO . Data obtained at the start (day 1 and day 2) and end of the monitoring period (day n) were compared . RESULTS: LV end-diastolic volume was within the normal range of our laboratory values in all patients, but was initially smaller in group II than in group I, and remained so despite fluid loading . LVEF was significantly depressed in all patients, resulting in severe reduction in LV stroke volume (LVSV), which was initially more marked in group I . In group I patients, LVEF significantly improved during the monitoring period, resulting in an increase in LVSV . CONCLUSION: 2D-ECHO changes during hemodynamic support in 90 septic patients confirmed defective LV preload with a propensity to worsen despite fluid loading in nonsurvivors (62% in the present study) . Our results are also in agreement with previous studies reporting depressed LV systolic function at the initial phase of septic shock . Since LV dysfunction was more marked in patients who recovered, we suggest that the exact significance of this finding should be reevaluated.

Int J Syst Bacteriol, 1999 Oct, 49 Pt 4, 1717 - 24
Ornithinicoccus hortensis gen . nov., sp . nov., a soil actinomycete which contains L-ornithine; Groth I et al.; Two Gram-positive coccoid, non-motile bacteria with L-ornithine as diagnostic diamino acid of the peptidoglycan and an interpeptide bridge of L-Orn<--Gly(1,2)<--D-Glu were isolated from a sample of garden soil . The major menaquinone is MK-8(H4) . 13-methyl and 12-methyl tetradecanoic acids are the predominant fatty acids . The polar lipids are phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylserine and two unknown phospholipids . Mycolic acids are absent . The DNA base composition is 72 mol% G + C . Recent comparative 16S rDNA studies revealed that strains HKI 0125T and HKI 0131 represent a novel lineage adjacent to the family Intrasporangiaceae of the order Actinomycetales but distinct from the previously described genera of this family . On the basis of the genotypic, chemotaxonomic, morphological and physiological characteristics of these two isolates it is proposed to classify HKI 0125T and HKI 0131 in a new genus and species for which the name Ornithinicoccus hortensis gen . nov., sp . nov . is proposed . The type strain is HKI 0125T (= DSM 12335T).

Int J Syst Bacteriol, 1999 Oct, 49 Pt 4, 1523 - 6
Characterization of a Gemella-like organism from the oral cavity of a dog: description of Gemella palaticanis sp . nov; Collins MD et al.; A hitherto unknown Gram-positive, catalase-negative, facultatively anaerobic coccus isolated from a vesicle on the gum of a dog was characterized by phenotypic and molecular taxonomic methods . Comparative 16S rRNA gene sequencing studies demonstrated that the isolate represents a new subline within the genus Gemella . The unknown bacterium was readily distinguished from all currently described members of this genus, Gemella haemolysans, Gemella bergeri, Gemella morbillorum and Gemella sanguinis, by biochemical tests and electrophoretic analysis of whole-cell proteins . On the basis of phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Gemella palaticanis sp . nov . The type strain of Gemella palaticanis is CCUG 39489T.

Int J Syst Bacteriol, 1999 Oct, 49 Pt 4, 1439 - 42
Dolosicoccus paucivorans gen . nov., sp . nov., isolated from human blood; Collins MD et al.; Phenotypic and phylogenetic studies were performed on a hitherto undescribed Gram-positive, catalase-negative, chain-forming coccus isolated from human blood . Comparative 16S rRNA gene sequencing studies demonstrated that the unknown organism constitutes a new phylogenetic line, close to, but distinct from, Facklamia and Globicatella . The unknown bacterium was readily distinguished from currently recognized Facklamia species and Globicatella sanguinis by biochemical tests and electrophoretic analysis of whole-cell proteins . On the basis of phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Dolosicoccus paucivorans gen . nov., sp . nov . The type strain of Dolosicoccus paucivorans is CCUG 39307T.

Int J Syst Bacteriol, 1999 Oct, 49 Pt 4, 1429 - 32
Helcococcus ovis sp . nov., a gram-positive organism from sheep; Collins MD et al.; Two strains of a hitherto undescribed Gram-positive, catalase-negative, facultatively anaerobic coccus isolated from sheep were characterized by phenotypic and molecular taxonomic methods . Comparative 16S rRNA gene sequencing studies demonstrated that the unknown strains were genealogically highly related and constitute a new line close to, but distinct from, Helcococcus kunzii . The unknown bacterium was readily distinguished from H . kunzii 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 Helcococcus ovis sp . nov . The type strain of Helcococcus ovis is CCUG 37441T.

Int J Syst Bacteriol, 1999 Oct, 49 Pt 4, 1381 - 5
Characterization of a novel gram-positive, catalase-negative coccus from horses: description of Eremococcus coleocola gen . nov., sp . nov; Collins MD et al.; Two strains of an unknown Gram-positive, catalase-negative, facultatively anaerobic coccus originating from the reproductive tract of horses were characterized by phenotypic and molecular taxonomic methods . Comparative 16S rRNA gene sequencing studies demonstrated that the two strains constitute a new subline within the lactic-acid group of bacteria, close to, but distinct from, Abiotrophia defectiva, Globicatella sanguinis and close relatives . The unknown bacterium was readily distinguished from other described Gram-positive, catalase-negative cocci by biochemical tests and electrophoretic analysis of whole-cell proteins . On the basis of phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Eremococcus coleocola gen . nov., sp . nov . The type strain of Eremococcus coleocola is CCUG 38207T.

Nature, 1999 Oct 21, 401(6755), 811 - 5
The Toll-like receptor 2 is recruited to macrophage phagosomes and discriminates between pathogens; Underhill DM et al.; Macrophages orchestrate innate immunity by phagocytosing pathogens and coordinating inflammatory responses . Effective defence requires the host to discriminate between different pathogens . The specificity of innate immune recognition in Drosophila is mediated by the Toll family of receptors; Toll mediates anti-fungal responses, whereas 18-wheeler mediates anti-bacterial defence . A large number of Toll homologues have been identified in mammals, and Toll-like receptor 4 is critical in responses to Gram-negative bacteria . Here we show that Toll-like receptor 2 is recruited specifically to macrophage phagosomes containing yeast, and that a point mutation in the receptor abrogates inflammatory responses to yeast and Gram-positive bacteria, but not to Gram-negative bacteria . Thus, during the phagocytosis of pathogens, two classes of innate immune receptors cooperate to mediate host defence: phagocytic receptors, such as the mannose receptor, signal particle internalization, and the Toll-like receptors sample the contents of the vacuole and trigger an inflammatory response appropriate to defence against the specific organism.

Annu Rev Microbiol, 1999, 53, 217 - 44
DNA uptake in bacteria; Dubnau D; Natural competence is widespread among bacterial species . The mechanism of DNA uptake in both gram-positive and gram-negative bacteria is reviewed . The transformation pathways are discussed, with attention to the fate of donor DNA as it is processed by the competent cell . The proteins involved in mediating various steps in these pathways are described, and models for the transformation mechanisms are presented . Uptake of DNA across the inner membrane is probably similar in gram-positive and gram-negative bacteria, and at least some of the required proteins are orthologs . The initial transformation steps differ, as expected, from the presence of an outer membrane only in the gram-negative organisms . The similarity of certain essential competence proteins to those required for the assembly of type-4 pili and for type-2 protein secretion is discussed . Finally several hypotheses for the biological role of transformation are presented and evaluated.

Cytometry, 1999 Dec 1, 37(4), 320 - 6
Photothermal studies of modulating effect of photoactivated chlorin on interaction of blood cells with bacteria; Lapotko D et al.; BACKGROUND: Bactericidal application of photosensitizers (PS) is a new and promising area . Up to now the action of PS against bacteria was studied without regard to immunocompetent cells, doses of drugs and radiation being used usually are not safe for such cells and therefore there is still no efficient model for PS application . METHODS: Action of chlorin, a plant derived photosensitizer, on a system of interacting cells and microbes was studied . Cell monitoring was done with a photothermal microscopy method . The two bacteria used were gram-positive (S . aureus) and gram-negative (E . coli) in a mixture of basic blood cells and neutrophils . The latter were used to model phagocytosis as fast cell response reaction . RESULTS: The strongest bactericidal effect was found when the cells and photoactivated chlorin act together . Selective bactericidal regime of PS application which does not affect cell viability was demonstrated . CONCLUSIONS: The results obtained suggest that this photosensitizer may enhance cell natural defense capabilities .

Pediatrics . 1999 Nov;104(5):e63.
Dexamethasone therapy increases infection in very low birth weight infants; Stoll BJ et al.; BACKGROUND: Infection is a major complication of preterm infants, resulting in increased morbidity and mortality . We recently reported the results of a multicenter trial of dexamethasone initiated at 14 or 28 days in very low birth weight (VLBW) infants who were at risk for chronic lung disease; the results showed an increase in nosocomial bacteremia in the group receiving dexamethasone . This study is an in-depth analysis of bacteremia/sepsis and meningitis among infants enrolled in the trial . METHODS: Data on cultures performed and antibiotic therapy were collected prospectively . Infections were classified as definite or possible/clinical . RESULTS: A total of 371 infants were enrolled in the trial . There were no baseline differences in risk factors for infection . For the first 14 days of study, infants received either dexamethasone (group I, 182) or placebo (group II, 189) . During this period, infants in group I were significantly more likely than those in group II to have a positive blood culture result (48% vs 30%) and definite bacteremia/sepsis/meningitis (22% vs 14%) . Over the 6-week study period, 47% of those cultured had at least one positive blood culture result (53% in group I vs 41% in group II) and 25% of the infants had at least one episode of definite bacteremia/sepsis/meningitis (29% in group I vs 21% in group II) . Among infants with definite infections, 46.8% were attributable to Gram-positive organisms, 26.6% to Gram-negative organisms and 26.6% to fungi . The factors present at randomization were evaluated for their association with infection . Group I assignment and H(2) blocker therapy (before study entry) were associated with increased risk of definite infection, whereas cesarean section delivery and increasing birth weight were associated with decreased risk . CONCLUSIONS: Infants who received a 14-day course of dexamethasone initiated at 2 weeks of age were more likely to develop a bloodstream or cerebrospinal fluid infection while on dexamethasone therapy than were those who received placebo . Physicians must consider this increased risk of infection when deciding whether to treat VLBW infants with dexamethasone.

J Bacteriol, 1999 Nov, 181(21), 6634 - 41
The Oenococcus oeni clpX homologue is a heat shock gene preferentially expressed in exponential growth phase; Jobin MP et al.; Using degenerated primers from conserved regions of previously studied clpX gene products, we cloned the clpX gene of the malolactic bacterium Oenococcus oeni . The clpX gene was sequenced, and the deduced protein of 413 amino acids (predicted molecular mass of 45,650 Da) was highly similar to previously analyzed clpX gene products from other organisms . An open reading frame located upstream of the clpX gene was identified as the tig gene by similarity of its predicted product to other bacterial trigger factors . ClpX was purified by using a maltose binding protein fusion system and was shown to possess an ATPase activity . Northern analyses indicated the presence of two independent 1.6-kb monocistronic clpX and tig mRNAs and also showed an increase in clpX mRNA amount after a temperature shift from 30 to 42 degrees C . The clpX transcript is abundant in the early exponential growth phase and progressively declines to undetectable levels in the stationary phase . Thus, unlike hsp18, the gene encoding one of the major small heat shock proteins of Oenococcus oeni, clpX expression is related to the exponential growth phase and requires de novo protein synthesis . Primer extension analysis identified the 5' end of clpX mRNA which is located 408 nucleotides upstream of a putative AUA start codon . The putative transcription start site allowed identification of a predicted promoter sequence with a high similarity to the consensus sequence found in the housekeeping gene promoter of gram-positive bacteria as well as Escherichia coli.

Virology, 1999 Oct 25, 263(2), 427 - 35
Similarly organized lysogeny modules in temperate Siphoviridae from low GC content gram-positive bacteria; Lucchini S et al.; Temperate Siphoviridae from an evolutionarily related branch of low GC content gram-positive bacteria share a common genetic organization of lysogeny-related genes and the predicted proteins are linked by many sequence similarities . Their compact lysogeny modules {integrase/1-2 orfs (phage exclusion? and metalloproteinase motif proteins)/cI-like repressor/cro-like repressor/antirepressor (optional)} differ clearly from that of lambda-like and L5-like viruses, the two currently established genera of temperate Siphoviridae, while they resemble those of the P2-like genus of Myoviridae . In all known temperate Siphoviridae from low GC content gram-positive bacteria the lysogeny module is flanked by the lysis module and the DNA replication module . This modular organization is again distinct from that of the known genera of temperate Siphoviridae . On the basis of comparative sequence analysis we propose a new genus of Siphoviridae: "Sfi21-like" phages . With a larger database of phage sequences it might be possible to establish a genomics-based phage taxonomy and to retrace the evolutionary history of selected phage modules or individual phage genes . The antirepressor of Sfi21-like phages has an unusual widespread distribution since proteins with high aa similarity (40%) were found not only in phages from gram-negative bacteria, but also in insect viruses .

Clin Infect Dis, 1999 Sep, 29(3), 503 - 7
Vancomycin as part of initial empirical antibiotic therapy for febrile neutropenia in patients with cancer: pros and cons; Feld R; Gram-positive organisms predominate as the bacterial pathogens identified in episodes of febrile neutropenia . This has led to increased use of antibiotics with efficacy against gram-positive organisms (often vancomycin) as part of empirical antibiotic regimens for treating febrile neutropenia . Among 101 children randomized to receive amikacin, ticarcillin, and vancomycin or ticarcillin/clavulanate and amikacin along with vancomycin placebo, treatment success in those treated with vancomycin was higher (85% vs . 62%) . In 1990, the European Organization for Research and Treatment of Cancer-National Cancer Institute of Canada Clinical Trials Group compared amikacin and ceftazidime with and without vancomycin and concluded that there was no need to include vancomycin in initial empirical antibiotic therapy . Results from another study and a retrospective review of a large clinical trial also support the previous conclusion . In 1999, most experts in the field recommend vancomycin not be part of the initial empirical therapy regimen for treating febrile neutropenia in patients with cancer.

Intern Med, 1999 Oct, 38(10), 833 - 6
Peritonitis due to Mycobacterium fortuitum infection following gastric cancer surgery; Kasamatsu Y et al.; Mycobacterium fortuitum is a well-documented cause of nosocomial infection . However, no studies have reported peritonitis with M . fortuitum as a postoperative complication . We describe a case of peritonitis with M . fortuitum biovariant peregrinum following gastric cancer surgery . Gram-positive bacterial infection coexisted . Although the source of the infection was unclear, the patient was successfully treated with drainage tube exchange and combination therapy consisting of sparfloxacin, clarithromycin, and imipenem/cilastatin sodium . Thus for postoperative infectious pathogens, not only bacteria but also nontuberculous mycobacteria should be considered.

FEMS Microbiol Ecol, 1999 Nov 1, 30(3), 237 - 251
Analysis of bacterial communities in heavy metal-contaminated soils at different levels of resolution; Sandaa R et al.; The impact of heavy metal contamination on soil bacterial communities was studied in soils amended for many years with sewage sludge contaminated with heavy metals to varying extents . At the broad level of resolution, DNA reassociation analysis indicated a dramatic decrease in bacterial diversity from 16{ omitted}000 bacterial genomes (g soil {wet wt})(-1) in the non-contaminated soil to 6400 bacterial genomes (g soil {wet wt})(-1) in soil with low metal amendments and only 2000 bacterial genomes (g soil {wet wt})(-1) in soil with high metal amendments . No differences between bacterial communities of these soils, however, were displayed in the %G+C profiles analysed by thermal denaturation . At a coarse level of characterisation, in situ hybridisation analysing larger phylogenetic groups of bacteria revealed a general decrease in the percentage of cells detected with probes ARCH915, BET42a, GAM42a, SRB385, CF319a, LGCb and HGC69a with increasing metal amendment . Only cells detected with probe ALF1b increased significantly from 3.1+/-0.8% of the cells detected by the domain-specific probe EUB338 in the non-contaminated soil to 6.5+/-1.3% in soil with high metal amendments . These shifts in populations of larger phylogenetic groups were largely confirmed by dot blot analysis of 16S and 23S rDNA clone libraries from bacteria in soil with low metal and high metal amendments, respectively . For a fine-level characterisation, 72 clones of 16S rDNA libraries were identified by comparative sequence analysis . A few sequences could not be assigned to the major taxa described . Most of the sequences were assigned to the Gram-positive bacteria with a high DNA G+C content (45%) and the alpha-subdivision of Proteobacteria (24%) . However, only minor differences were seen between bacterial communities from the low and high metal soils . In the soil with high metal amendment, more sequences clustered to the alpha-subdivision of Proteobacteria, while in the low metal soil, more sequences clustered to the Gram-positive bacteria with a high DNA G+C content.

FEMS Microbiol Rev, 1999 Oct, 23(5), 615 - 27
Bacterial respiration of arsenic and selenium; Stolz JF et al.; Oxyanions of arsenic and selenium can be used in microbial anaerobic respiration as terminal electron acceptors . The detection of arsenate and selenate respiring bacteria in numerous pristine and contaminated environments and their rapid appearance in enrichment culture suggest that they are widespread and metabolically active in nature . Although the bacterial species that have been isolated and characterized are still few in number, they are scattered throughout the bacterial domain and include Gram-positive bacteria, beta, gamma and epsilon Proteobacteria and the sole member of a deeply branching lineage of the bacteria, Chrysiogenes arsenatus . The oxidation of a number of organic substrates (i.e . acetate, lactate, pyruvate, glycerol, ethanol) or hydrogen can be coupled to the reduction of arsenate and selenate, but the actual donor used varies from species to species . Both periplasmic and membrane-associated arsenate and selenate reductases have been characterized . Although the number of subunits and molecular masses differs, they all contain molybdenum . The extent of the environmental impact on the transformation and mobilization of arsenic and selenium by microbial dissimilatory processes is only now being fully appreciated.

DNA Seq, 1998, 9(2), 79 - 88
The Sip(Sli) gene of Streptomyces lividans TK24 specifies an unusual signal peptidase with a putative C-terminal transmembrane anchor; Schacht S et al.; Type I signal peptidases (SPases) are a widespread family of enzymes which remove signal peptides from proteins translocated across cellular membranes . Here, we report the first isolation of a gene coding for type I signal peptidase of Streptomyces, denoted Sip(Sli) . The sip(sli) gene specifies a protein of 291 amino acids . Thus Sip(Sli) is much larger (approximately 100 amino acids) than other known SPases of Gram-positive bacteria and resembles SPases of Gram-negative bacteria, showing the highest degree of similarity to an SPase of the cyanobacterium Phormidium laminosum . Sip(Sli) contains conserved serine and lysine residues, which are believed to be required for the catalytic activity . Similar to other known SPases from Gram-positive bacteria, Sip(Sli) seems to have only one N-terminal transmembrane anchor . In addition, Sip(Sli) seems to contain a second transmembrane anchor at the C-terminus, which is an unusual feature for type I signal peptidases.

DNA Seq, 1998, 9(2), 71 - 7
A new signal peptidase gene from Streptomyces lividans TK21; Parro V et al.; Using synthetic oligonucleotides derived from known signal peptidase genes and a multicopy plasmid as a vector, a signal peptidase gene (sipZ) from Streptomyces lividansTK21 has been cloned . The primary structure of the gene has been determined and the amino acid composition of the SipZ protein inferred . SipZ is 258 aa long and showed homology to other type I signal peptidases, containing like them an N-terminal transmembrane anchor . Alignment of SipZ with other known SPases allowed the identification of a conserved sequence of amino acids specific for Gram-positive bacteria.

Carbohydr Res, 1999 Jun 30, 319(1-4), 112 - 23
A novel trisaccharide glycolipid biosurfactant containing trehalose bears ester-linked hexanoate, succinate, and acyloxyacyl moieties: NMR and MS characterization of the underivatized structure; Esch SW et al.; A Gram-positive actinomycete growing on n-hexadecane secreted a family of anionic glycolipid surfactant homologs . The major homolog, with a molecular weight of 1210.6347, had the formula C58H98O26 . Following mild alkaline saponification, 1H and 13C NMR spectroscopy were used to characterize the non-reducing trisaccharide backbone: beta-Glcp-(1-->3)-alpha-Glcp-(1<-->1)-alpha-Glcp ('laminaratrehalose') . Hexanoate, succinate, 3-hydroxyoctanoate, and 3-hydroxydecanoate were found in 3:1:1:1 molar ratio using GC-EIMS analysis of fatty acid methyl esters (FAME) prepared by transesterification . We found that the beta-hydroxy acids bore secondary hexanoate chains in 3-O-ester linkage, giving acyloxyacyl anions of appropriate m/z in FABMS and FABMS/MS spectra . COSY, HETCOR, HMBC, and HMQC NMR experiments established the acylation pattern: succinate at C-2 of the terminal alpha-glucopyranose ring; hexanoate at C-3" of the beta-glucopyranose ring; 3-hexanoyloxyoctanoate and 3-hexanoyloxydecanoate at the 2'- and 4-positions . In FABMS spectra, the homologs flanked the molecular ion by +/- 14 and +/- 28 amu, suggesting heterogeneity in acyl chain length.

Ann Biol Clin (Paris), 1999 Sep-Oct, 57(5), 545 - 54
{Nocardia infections: clinical and biological aspects}; Laurent F et al.; The nocardiosis is an infection caused by a bacterial pathogen agent, Nocardia, belonging to the Actinomycetales order . They are Gram-positive, strictly aerobic bacteria . Members of the genus Nocardia are ubiquitous . They are frequently isolated from soil, water, air dusts . The mode of contamination occurs by inhalation or by cutaneous or ocular traumatic lesion . Clinically, nocardiosis is essentially characterized by pulmonary diseases . Others secondary localizations are described, such as in the central nervous system . Nocardia can be responsible for important cutaneous, subcutaneous and lymphocutaneous manifestations . In the same way, some extrapulmonary diseases and spread nocardiosis are more rarely observed . Several factors seem to favour the development of Nocardia . The immunocompromised patients, particularly those with organ transplant and the patients treated with immunosuppressor treatments, offer strong predispositions to this opportunistic disease . The nocardiosis is nevertheless observed in healthy persons . In front of polymorphic and specific-less clinical manifestations, large phenotypic heterogeneity, and resistance profiles to specific antibiotics, a correct diagnosis for Nocardia species is necessary to apply an adequate treatment . The techniques of identification based on the chemotaxonomic analysis and the susceptibility to different inhibitors are efficient for the identification of genus and species . However, because of the slow growth rate of Nocardia, the reading of these tests can require several weeks of incubation . With the intention of the rapid identification of genus and species, the molecular techniques (PCR-RFLP) seem to be efficient . The technique of RAPD allows an efficient molecular typing, which will give a better knowledge concerning transmission, ecological niches and epidemic reservoirs.

Eur J Clin Microbiol Infect Dis, 1999 Aug, 18(8), 539 - 44
Prognostic factors influencing mortality in cancer patients with neutropenia and bacteremia; Gonzalez-Barca E et al.; The purpose of this study was to identify risk factors for mortality in neutropenic patients with cancer and bacteremia . A consecutive sample of 438 neutropenic patients (granulocyte count <0.5 x 10(9)/l) with cancer and bacteremia was studied to identify the clinical characteristics associated with mortality at the onset of bacteremia . The mean age of the subjects was 48 years (range, 15-87 years) . Most cases of bacteremia (77%) were hospital-acquired and occurred in patients with acute leukemia (48%) . Gram-positive organisms caused 233 (53%) episodes of bacteremia, gram-negative organisms caused 151 (34%) episodes, and 48 (11%) episodes were polymicrobial . The overall mortality within 30 days of the onset of bacteremia was 24.4% . The variables found to be independently associated with increased mortality using logistic regression techniques were as follows: shock at the onset of bacteremia (OR, 10; 95% CI, 4.2-23.8), pneumonia (OR,4.4; 95% CI, 1.9-10), uncontrolled cancer (OR,4.3; 95% CI, 1.5-12.7), and absence of prophylaxis with norfloxacin (OR,2.4; 95% CI, 1.3-4.5) . The prognostic factors ascertained in this study may help to identify those patients at higher risk of death . Medical intervention addressing some of these factors may improve the outcome of bacteremia in neutropenic patients with cancer.

J Nat Prod, 1999 Sep, 62(9), 1295 - 7
New bromopyrrole alkaloids from the Indopacific sponge Agelas nakamurai; Eder C et al.; Two new dimeric bromopyrrole alkaloids, nakamuric acid (1) and its corresponding methyl ester (2), have been isolated from the Indopacific sponge Agelas nakamurai along with the known metabolites sceptrin (3), debromosceptrin (4), and ageliferin (5) . Their structures were identified by analysis of spectral data . All compounds inhibited the growth of several Gram-positive and Gram-negative bacteria in the agar plate diffusion assay.

J Antimicrob Chemother, 1999 Sep, 44(3), 367 - 76
An open, randomized, multicentre study comparing the use of low-dose ceftazidime or cefotaxime, both in combination with netilmicin, in febrile neutropenic patients . German Multicentre Study Group; Hoffken G et al.; To reduce drug acquisition costs, the clinical and bacteriological efficacy of low-dose ceftazidime i.v . (1 g tid) was compared with cefotaxime i.v . (2 g tid) . Both regimens were combined with netilmicin i.v . (2 mg/kg bodyweight tid), in an open, randomized, multicentre trial in febrile neutropenic patients . The addition of antibiotics for gram-positive coverage was part of the protocol; alteration in the antibiotics for gram-negative cover or premature discontinuation of the study antibiotics were judged as failure . One hundred and eighty six patients were randomized by nine German centres, the patients matched for age, underlying diseases and duration of neutropenia (median duration 14 days) in both treatment arms . Infections were documented microbiologically in 29% of the patients, clinically in 16% and suspected (fever of unknown origin) in 102/186 patients (55%) . The 82 pathogens isolated were predominantly gram-positive bacteria . In an intent-to-treat analysis, the overall response rate without modification at the final evaluation was 58% in the ceftazidime group and 34% in the cefotaxime group (P < 0.01) . The success rates with modification were 84% and 64%, respectively . The failure rate in a highly immunosuppressed subgroup of the patients (bone marrow transplant recipients) was higher for cefotaxime (53%) than for the ceftazidime arm (14%) (P < 0.001) . Response rates were significantly higher in the ceftazidime group for patients with microbiologically documented and possible infections . No major bacterial superinfections occurred in the low-dose treatment arm . The tolerability was good for both regimens . Low-dose ceftazidime combined with netilmicin proved to be superior to recommended doses of cefotaxime/netilmicin in febrile neutropenic patients.

J Immunol, 1999 Oct 15, 163(8), 4095 - 9
Cutting edge: bacterial DNA and LPS act in synergy in inducing nitric oxide production in RAW 264.7 macrophages; Gao JJ et al.; LPS is well recognized for its potent capacity to activate mouse macrophages to produce NO, an important inflammatory mediator in innate host defense . We demonstrate here that, although inducing little NO alone, DNA from both Gram-negative and Gram-positive bacteria synergizes with subthreshold concentrations of LPS (0.3 ng/ml) to induce NO in cultures of RAW 264.7 macrophages . The effects of the DNA are mimicked by synthetic CpG-containing oligodeoxynucleotides but not by non-CpG-containing oligodeoxynucleotides . This synergistic activity is not inhibited by neutralizing Abs against IFN . Preincubation of macrophages with DNA for 8-24 h suppresses subsequent synergistic macrophage responses to DNA/LPS, whereas prolonged pretreatment with LPS enhances synergy . RT-PCR analysis indicates that the mRNA levels of the inducible NO synthase gene are also coordinately suppressed or induced . These findings indicate that temporally controlled, synergistic interactions exist between microbial DNA and LPS in the induction of macrophage NO via enhanced inducible NO synthase gene expression.

Curr Opin Biotechnol, 1999 Oct, 10(5), 498 - 504
Genetics and engineering of microbial exopolysaccharides for food: approaches for the production of existing and novel polysaccharides; van Kranenburg R et al.; Microbial exopolysaccharides (EPSs) are used in the food industry for their unique properties as viscosifiers, stabilisers, emulsifiers or gelling agents . In recent years, significant progress in the understanding of the genetics and biochemistry of microbial EPS synthesis by both Gram-negative and Gram-positive bacteria has been made . Biosynthesis pathways have been elucidated, and several of the genes involved have been characterised . This knowledge can now be applied to start EPS engineering or to improve EPS production.

Perit Dial Int, 1998 Jul-Aug, 18(4), 371 - 5
A randomized prospective comparison of oral levofloxacin plus intraperitoneal (IP) vancomycin and IP netromycin plus IP vancomycin as primary treatment of peritonitis complicating CAPD; Cheng IK et al.; OBJECTIVE: To compare the therapeutic efficacy of daily oral levofloxacin plus intermittent intraperitoneal (IP) vancomycin (group 1) versus daily IP netromycin and intermittent IP vancomycin (group 2) in the primary treatment of peritonitis complicating continuous ambulatory peritoneal dialysis (CAPD) . DESIGN: A randomized multicenter prospective open-label comparative clinical study . SETTING: University and Hospital Authority hospitals in Hong Kong . PATIENTS: All CAPD patients who developed bacterial or culture-negative peritonitis beyond 28 days of a previous episode and without evidence of septicemia, associated tunnel infection, or known sensitivity to trial medications were accepted into the clinical trial . RESULTS: A total of 101 patients entered the trial . The primary cure rate was 74.5% for group 1 and 73.6% for group 2 . Baseline culture results appeared to influence the clinical outcome: the primary cure rate for culture-negative, gram-positive, and gram-negative episodes was 83.3%, 78.6%, and 42.9% for group 1 and 69.1%, 76.9%, and 71.3% for group 2, respectively . The primary cure rate also varied considerably among individual centers and was particularly noticeable in group 1 . In the latter group, it correlated closely with in vitro levofloxacin resistance which in turn correlated closely with previous exposure to fluoroquinolones . CONCLUSION: Oral levofloxacin in combination with intermittent IP vancomycin has comparable efficacy to IP netromycin combined with intermittent IP vancomycin as primary treatment in CAPD peritonitis, but is simpler and more cost-effective to administer . It may be recommended as primary therapy in centers with relatively low exposure and, therefore, low background resistance to fluoroquinolones.

Eur J Cancer, 1999 May, 35(5), 770 - 4
Bloodstream infections in children with cancer: a multicentre surveillance study of the Italian Association of Paediatric Haematology and Oncology . Supportive Therapy Group-Infectious Diseases Section; Viscoli C et al.; A one-year prospective, multicentre surveillance study on aetiology, main clinical features and outcome of bloodstream infections in children with cancer was conducted in 18 paediatric haematology centres belonging to the Italian Association for Paediatric Haematology and Oncology . A total of 191 bloodstream infections were reported during the study period . Of them, 123 (64%) occurred in neutropenic and 68 (36%) in non-neutropenic patients . Gram-positive cocci caused 45% (85/191) of the episodes, gram-negative rods 41% (78/191), and fungi 9% (18/191) . The remaining 5% (10/191) of the episodes were poly-microbial infections . A total of 204 pathogens were isolated (46% gram-positive cocci; 44% gram-negative rods; and 10% fungi) . The aetiologic distribution was similar among neutropenic and non-neutropenic patients . A correlation between the infection and the presence of an indwelling central venous catheter was found in 20% (23/114) of the episodes among neutropenic patients and in 55% (23/62) among non-neutropenic patients . Gram-negative micro-organisms were isolated in an unusually high proportion of catheter-related infections (48%) . The overall mortality rate from any cause within 30 days from the first positive blood culture was 11%, and was higher among patients who were neutropenic at the onset of the infection than among those who were not neutropenic (15 versus 4%, P = 0.03) . In addition, the mortality was significantly higher in recipients of bone marrow transplantation than in patients with acute leukaemia or solid tumour (21, 11 and 6%, respectively) and was also higher in fungaemias and poly-microbial infections (22 and 30%) than in single gram-positive and gram-negative bacteraemias (11 and 6%).

Comb Chem High Throughput Screen, 1998 Jun, 1(2), 89 - 99
A simple, inexpensive apparatus for performance of preparative scale solution phase multiple parallel synthesis of drug analogs . II . Biological evaluation of a retrospective library of quinolone antiinfective agents; Frank KE et al.; A series of pure fluoroquinolone antiinfective agents was prepared by multiple parallel synthesis using a simple new apparatus . These compounds were evaluated biologically against Gram-positive and Gram-negative microorganisms and against a BCG strain transfected with luciferase in a fluorescence-based antitubercular assay . Activity against relatively fast growing, acid-fast Mycobacterium smegmatis was determined in part by agar-dilution streak assays . Data obtained against Escherichia coli-derived DNA gyrase does not correlate well with whole cell assays against E . coli . These compounds were assayed by a convenient glass-fiber filter binding method modified for high throughput screening . In these analogs, the results with a N-1 cyclopropyl substituent were often inferior to those obtained with a N-1 2',4'-difluorophenyl substituent . None of the new compounds prepared was superior in its antimycobacterial potency to ciprofloxacin or temafloxacin.

Leuk Lymphoma, 1999 Aug, 34(5-6), 585 - 9
Intensified prophylaxis of febrile neutropenia with ofloxacin plus rifampin during severe short-duration neutropenia in patients with lymphoma; Munoz L et al.; To analyse the impact of intensified prophylaxis with ofloxacin plus rifampin (O+R) in neutropenic patients we used this combination in 40 consecutive cycles of ifosfamide, cytarabine, prednisolone and etoposide (IAPVP-16) . This salvage chemotherapy regimen for lymphoma usually produces four to six days of severe neutropenia without significant extrahematologic toxicities . We compared the infectious morbidity during neutropenia under O+R with 58 consecutives cycles using either norfloxacin or no prophylaxis (control group) . Fifty-three percent of control group patients and 20% of the O+R group developed febrile neutropenia that required hospital admission (p<0.001, 95% CI for the difference between both proportions of 16% to 51%) . Bacteremia was documented in two patients in the O+R group and six in the control group (p=0.08) . Gram-positive cocci (GPC) accounted for all six bacteremias in the control group, while both cases in O+R group were due to a quinolone-resistant gram-negative bacteria (GNB) (p<0.01 for GPC) . Five patients (13%) who received O+R and 23 (40%) in control group developed fever of unknown origin, p<0.001, while the total duration of hospitalization due to febril neutropenia was 42 days and 158 days, respectively (p<0.001) . In conclusion, intensified prophylaxis with O+R appears to reduce the rate of febrile neutropenia and GPC bacteremia in patients with short and severe neutropenia, which translates into a reduction in the need for hospitalization.

Mar Biotechnol (NY), 1999 Jul, 1(4), 391 - 400
Microbial Diversity in Sediments Collected from the Deepest Cold-Seep Area, the Japan Trench; Li L et al.; : The Japan Trench land slope at a depth of 6,400 m is the deepest cold-seep environment with Calyptogena communities . Sediment samples from inside and beside the Calyptogena communities were collected, and the microbial diversity in the sediment samples was studied by molecular phylogenetic techniques . From DNA extracted directly from the sediment samples, 16S rDNAs were amplified by the polymerase chain reaction method . The sequences of the amplified 16S rDNAs selected by restriction fragment length polymorphism analysis were determined and compared with sequences in DNA databases . The results showed that 33 different bacterial 16S rDNA sequences from the two samples analyzed fell into similar phylogenetic categories, the alpha-, gamma-, delta-, and varepsilon-subdivisions of Proteobacteria, Cytophaga, and gram-positive bacteria; some of the 16S rDNA sequences were common to both samples . delta- and varepsilon-Proteobacteria-related sequences were abundant in both sediments . These sequences are mostly related to sulfate-reducing or sulfur-reducing bacteria and epibionts, respectively . Eight different archaeal 16S rDNA sequences were cloned from the sediments . The majority of the archaeal 16S rDNA sequences clustered in Crenarchaeota and showed high similarities to marine group I archaeal rDNA . A Methanococcoides burtonii-related sequence obtained from the sediment clustered in the Euryarchaeota indicating that M . burtonii-related strains in the area of Calyptogena communities may contribute to production of methane in this environment . From these results, we propose a possible model of sulfur circulation within the microbial community and that of Calyptogena clams in the cold-seep environment.

Int J Food Microbiol, 1999 Sep 15, 50(1-2), 65 - 91
Microbial stress response in minimal processing; Abee T et al.; "Bacteria have evolved adaptive networks to face the challenges of changing environments and to survive under conditions of stress . Therefore, the efficiencies of inactivation and preservation methods need to be assessed, especially with regard to the enormous potential of food pathogens to adapt to a wide variety of stress conditions . All adaptive responses, whether to changing nutrients or to various stresses encountered in minimal processing, involve a series of genetic switches that control the metabolic changes taking place . A common regulatory mechanism involves the modification of sigma (sigma) factors whose primary role is to bind to core RNA polymerase conferring promoter specificity directing expression of specialty regulons involved in heat-shock response, the chemotactic response, sporulation, and general stress response . Examples of the latter regulon in Gram-positive bacteria (the sigmaB regulon) and in Gram-negative bacteria (the RpoS regulon) will be discussed in more detail . Cellular adaptive mechanisms to starvation, cold shock, heat shock, (weak) acids, high osmolarity and high hydrostatic pressure will be described and their significance in food preservation and safety will be discussed."

Support Care Cancer, 1999 Sep, 7(5), 336 - 42
Ceftazidime in combination with glycopeptide antibiotic is an effective first-line therapy for patients undergoing high-dose therapy with autologous peripheral blood stem cell support; Egerer G et al.; It was the objective of this study to evaluate the efficacy and toxicity of an empirical antibiotic therapy consisting in ceftazidime and a glycopeptide antibiotic . All patients enrolled in the study had hematological malignancies and underwent high-dose therapy with peripheral blood stem cell (PBSC) support . In this retrospective study, 183 of 207 patients who had received a PBSC-supported high-dose therapy were evaluable . Any patients who had fever higher than 38.5 degrees C received ceftazidime in combination with vancomycin (105 patients) or teicoplanin (69 patients) . In 80 of 174 patients with fever (45%) the fever resolved within 72 h as a result of the treatment with ceftazidime and the glycopeptide antibiotic . In nonresponding patients, the changes included the replacement of ceftazidime by imipenem/cilastin (94 patients) and the addition of erythromycin (12 patients) or metronidazole (3 patients) . Amphotericin B was administered in 29 patients . Following hematological reconstitution, the fever and clinical signs, including radiographic findings, resolved in 20 primarily nonresponding patients . In blood cultures, a significantly higher incidence of gram-positive than of gram-negative bacteria was observed (26 vs 7) . The toxicity of the first-line antibiotic therapy was limited to allergic skin reactions in 12 patients . Ceftazidime in combination with a glycopeptide antibiotic provides an effective and safe first-line therapy for patients with neutropenic fever following PBSC-supported high-dose therapy.

J Bacteriol, 1999 Sep, 181(18), 5750 - 7
Distribution of tetrahydromethanopterin-dependent enzymes in methylotrophic bacteria and phylogeny of methenyl tetrahydromethanopterin cyclohydrolases; Vorholt JA et al.; The methylotrophic proteobacterium Methylobacterium extorquens AM1 possesses tetrahydromethanopterin (H(4)MPT)-dependent enzymes, which are otherwise specific to methanogenic and sulfate-reducing archaea and which have been suggested to be involved in formaldehyde oxidation to CO(2) in M . extorquens AM1 . The distribution of H(4)MPT-dependent enzyme activities in cell extracts of methylotrophic bacteria from 13 different genera are reported . H(4)MPT-dependent activities were detected in all of the methylotrophic and methanotrophic proteobacteria tested that assimilate formaldehyde by the serine or ribulose monophosphate pathway . H(4)MPT-dependent activities were also found in autotrophic Xanthobacter strains . However, no H(4)MPT-dependent enzyme activities could be detected in other autotrophic alpha-proteobacteria or in gram-positive methylotrophic bacteria . Genes encoding methenyl H(4)MPT cyclohydrolase (mch genes) were cloned and sequenced from several proteobacteria . Bacterial and archaeal Mch sequences have roughly 35% amino acid identity and form distinct groups in phylogenetic analysis.

J Antibiot (Tokyo), 1999 May, 52(5), 491 - 500
AS-924, a novel bifunctional prodrug of ceftizoxime; Kasai M et al.; To improve the oral absorption of ceftizoxime (CZX), 7beta-{(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido}- 3-cephem-4- carboxylic acid, we synthesized and evaluated a novel series of bifunctional prodrugs, in which L-alanine was introduced into the aminothiazole-oxime moiety at the C-7 position of the various lipophilic esters of CZX . Among these prodrugs, pivaloyloxymethyl 7beta-{(Z)-2-(2-(S)-alanylaminothiazol-4-yl)-2-methoxyiminoa cetamido}-3-cephem-4-carboxylate hydrochloride (ceftizoxime alapivoxil, AS-924) was well absorbed after oral administration in experimental animals and showed potent therapeutic effects in mice infected with gram-positive and gram-negative bacteria.

Nature, 1999 Aug 26, 400(6747), 897 - 9
A new pathway for polyketide synthesis in microorganisms; Funa N et al.; Chalcone synthases, which biosynthesize chalcones (the starting materials for many flavonoids), have been believed to be specific to plants . However, the rppA gene from the Gram-positive, soil-living filamentous bacterium Streptomyces griseus encodes a 372-amino-acid protein that shows significant similarity to chalcone synthases . Several rppA-like genes are known, but their functions and catalytic properties have not been described . Here we show that a homodimer of RppA catalyses polyketide synthesis: it selects malonyl-coenzyme-A as the starter, carries out four successive extensions and releases the resulting pentaketide to cyclize to 1,3,6,8-tetrahydroxynaphthalene (THN) . Site-directed mutagenesis revealed that, as in other chalcone synthases, a cysteine residue is essential for enzyme activity . Disruption of the chromosomal rppA gene in S . griseus abolished melanin production in hyphae, resulting in 'albino' mycelium . THN was readily oxidized to form 2,5,7-trihydroxy-1,4-naphthoquinone (flaviolin), which then randomly polymerized to form various coloured compounds . THN formed by RppA appears to be an intermediate in the biosynthetic pathways for not only melanins but also various secondary metabolites containing a naphthoquinone ring . Therefore, RppA is a chalcone-synthase-related synthase that synthesizes polyketides and is found in the Streptomyces and other bacteria.

Clin Infect Dis, 1999 Aug, 29(2), 414 - 9
Assessment of measuring circulating levels of interleukin-6, interleukin-8, C-reactive protein, soluble Fc gamma receptor type III, and mannose-binding protein in febrile children with cancer and neutropenia; Lehrnbecher T et al.; Circulating levels of interleukin (IL)-6, IL-8, soluble Fc gamma receptor type III (sFc gammaRIII), mannose-binding protein (MBP), and C-reactive protein (CrP) were assessed among febrile children with cancer and neutropenia . Levels of IL-6, IL-8, sFc gammaRIII, MBP, and CrP were measured in serum from 56 pediatric cancer patients at the time of admission for 121 episodes of febrile neutropenia (88 febrile episodes without identifiable source, 5 clinically documented infections, 20 episodes of bacteremia due to gram-positive and 5 due to gram-negative organisms, and 3 fungal infections) . IL-6 and IL-8 levels were higher in patients with either bacteremia due to gram-negative organisms or fungal infections than in patients with febrile episodes without an identifiable source (P < .00001 for each) . IL-6 and IL-8 levels were higher in children with bacteremia due to gram-negative organisms than in those with bacteremia due to gram-positive organisms (P = .0011 and P = .0003, respectively) . The measured levels of CrP, MBP, and sFc gammaRIII were not useful for identifying the type of infection . These preliminary results show the potential usefulness of IL-6 and IL-8 as early indicators for life-threatening infections in febrile cancer patients with neutropenia.

Clin Infect Dis, 1999 Aug, 29(2), 346 - 51
Morbidity associated with long-term use of totally implantable ports in patients with AIDS; Domingo P et al.; To determine the morbidity associated with long-term use of a totally implantable central venous access device (Port-A-Cath {PAC}) in patients with AIDS, we studied 68 consecutive patients with AIDS requiring 79 such devices for long-term use, inserted over a period of 5 years . The total number of PAC-days was 20,159 . At least one PAC-related complication occurred with 40 of 79 PACs (50.6% {95% confidence interval (CI): 39.6%-61.6%}), and 16 devices (20.2% {95% CI, 11.4%-29.0%}) had to be removed because of complications . Device-related infection occurred with 33 of 79 PACs (41.7% {95 CI, 30.8%-52.6%}) . The predominant infection occurring with PACs was chamber infection, with an incidence of 0.16 per 100 PAC-days . The predominant organisms isolated from patients with chamber infections but also from those with device-related bacteremia were gram-positive cocci (79.4%) . The presence of neutropenia (odds ratio {OR} = 9.72; 95% CI, 3.0-31.3; P < .001) and a CD4 cell count lower than 0.025 x 10(9)/L (OR = 6.14; 95% CI, 1.9-19.2; P = .002) were independent predictors of infection . The antibiotic lock technique was associated with decreased device loss when compared with isolated systemic antibiotic therapy (OR = 0.05; 95% CI, 0.0-0.59; P = .008) . This technique may be useful to treat PAC infection in patients with AIDS, for whom the risk of PAC-related complications is very high.

Crit Care Med, 1999 Aug, 27(8), 1608 - 16
Clinical gram-positive sepsis: does it fundamentally differ from gram-negative bacterial sepsis?
Opal SM, Cohen J.
OBJECTIVE: To review the basic differences between gram-positive and gram-negative sepsis and to assess the effect of these differences on current and future therapeutic strategies for sepsis . DESIGN: Literature review of the past 30 yrs of laboratory and clinical reports that analyze the microbial aspects of sepsis and the immunologic response to systemic infection . RESULTS: The increasing prevalence of sepsis from gram-positive bacterial pathogens necessitates reevaluation of many of the basic assumptions about the molecular pathogenesis of septic shock . It has been assumed that the initiation of the systemic inflammatory response with activation of the proinflammatory cytokine networks and other mediators results in a similar pathophysiologic process, regardless of the causative microbic pathogen . Yet, there is increasing experimental evidence that fundamental differences exist in the host response to gram-positive bacterial pathogens compared with the host response to gram-negative organisms . Systemic immune activation during sepsis may promote the clearance of the microbic pathogen; however, generalized inflammation also contributes to the pathogenesis of septic shock . The balance between these beneficial and deleterious effects may differ between gram-positive and gram-negative pathogens . CONCLUSIONS: Results of antimediator therapies in clinical trials in septic shock are inconclusive but suggest that the response may differ, depending on the type of microbic pathogen . The immune-mediated pathophysiologic mechanisms that underlie gram-positive sepsis and the potential interactions between the infecting microorganism and efficacy of anticytokine therapies require further investigation . Treatment strategies that explain the causative organism may be necessary for optimal use of immunoadjuvants in the future.

Insect Mol Biol, 1999 Aug, 8(3), 311 - 8
Immunity proteins from mosquito cell lines include three defensin A isoforms from Aedes aegypti and a defensin D from Aedes albopictus; Gao Y et al.; An Aedes aegypti mosquito cell line, Aag-2, exhibits a response to immune stimulation that is qualitatively similar to that of C7-10 cultured cells from the related mosquito, Aedes albopictus . Using SDS polyacrylamide gels, we found that a small peptide was preferentially induced by the treatment of growing cells with heat-killed, Gram-positive bacteria . By an analogy with other studies, this small peptide was postulated to be a member of the defensin family of insect immunity peptides . A differential display was used to obtain partial polymerase chain reaction products corresponding to mRNAs that were preferentially expressed in induced cells . One of these products, which contained the partial sequence of a defensin gene, was used to screen cDNA libraries from Ae . aegypti and Ae . albopictus cells . From Ae . aegypti cells, we found two previously described isoforms (A1 and A4) of mosquito defensin A, as well as a new isoform which we defined as A5 . From Ae . albopictus cells, we found a new mature mosquito defensin, named D, which contains proline and isoleucine as the final amino acids . In both Ae . aegypti and Ae . albopictus cell lines, the expression of defensin mRNA was visible on Northern blots as early as 3 h after exposure to heat-killed bacteria, and defensin mRNA abundance was maximal at 12-36 h after induction.

Br J Haematol, 1999 Sep, 106(3), 756 - 61
Defective functional activity and accelerated apoptosis in neutrophils from children with cancer are differentially corrected by granulocyte and granulocyte-macrophage colony stimulating factors in vitro; Lejeune M et al.; We have previously shown that polymorphonuclear leucocytes (PMN) harvested from children with cancer and exposed to chemotherapy exhibit defective bactericidal activities against both Gram-positive and Gram-negative microorganisms as well as accelerated apoptosis . In this study, PMN from children with cancer were evaluated to compare in vitro the corrective effects of the two myeloid colony stimulating factors G-CSF and GM-CSF on these defective pathways . Both G-CSF and GM-CSF were able to increase the defective bactericidal activities against S . aureus and E . coli . However, GM-CSF was consistently superior to G-CSF in correcting PMN microbicidal activity; this correction was incomplete since it did not reach the level observed in normal PMN exposed to GM-CSF . The accelerated apoptosis of PMN was not affected by G-CSF . In contrast, GM-CSF significantly prolonged the survival of the PMN although it did not reach the level of survival observed with normal PMN exposed to GM-CSF . These observations were consistent with other studies indicating that in PMN, microbicidal activities and apoptosis are differentially sensitive to the myeloid growth factors G-CSF and GM-CSF.

Infect Control Hosp Epidemiol, 1999 Aug, 20(8), 557 - 60
Vancomycin in Oregon: who's using it and why; Cieslak PR et al.; OBJECTIVE: To determine the proportion of vancomycin orders that are appropriate according to national guidelines and to identify targets for educational messages . DESIGN: Population-based study of vancomycin use in Oregon during a 3-week period . Survey of pharmacists, prospective flagging of vancomycin orders, and data abstraction from patient charts using standardized forms . SETTING: Nonpsychiatric hospitals in Oregon . RESULTS: Four (6%) of the 66 Oregon hospitals had pharmacy restrictions on initial vancomycin orders . Sixty-four (97%) of the hospitals participated in the study of indications for use; 293 vancomycin orders were reported; 3.8 courses were initiated per 1,000 patient-days . Indications for use were determined for 266 (91%); of these, 159 (60%) were deemed appropriate . Of uses for prophylaxis, empirical treatment of suspected gram-positive infection, and treatment of documented gram-positive infection, 57%, 56%, and 65%, respectively, were appropriate . Of hospitals with <250, 251-475, and >475 licensed beds, 65%, 58%, and 57% of vancomycin orders were appropriate . No single medical specialty accounted for >16% of inappropriate vancomycin use . CONCLUSIONS: Vancomycin was used inappropriately by physicians of many different specialties, in hospitals of all sizes, and in sundry clinical situations . The problem of inappropriate vancomycin use does not lend itself to solution by educational strategies targeted at specific subgroups; restrictions by hospital pharmacies may be required.

Am Fam Physician, 1999 Aug, 60(2), 510 - 4
Endogenous endophthalmitis: case report and brief review; Romero CF et al.; Endogenous endophthalmitis is a potentially blinding ocular infection resulting from hematogenous spread from a remote primary source . The condition is relatively rare but may become more common as the number of chronically debilitated patients and the use of invasive procedures increase . Many etiologic organisms (gram-positive, gram-negative and fungal) have been reported to cause endogenous endophthalmitis . Risk factors are well defined and include most reasons for immune suppression . A high clinical suspicion is needed for early diagnosis and treatment . Early intravenous antibiotic therapy remains the cornerstone of treatment . The roles of intravitreal antibiotics and vitrectomy are evolving and may become more widely accepted as therapeutic modalities . The authors report a case of endogenous endophthalmitis and provide a brief review of the literature.

Bull Acad Natl Med, 1999, 183(5), 973 - 82; discussion 983
{Is it possible to reduce the incidence of aminoglycoside-induced nephrotoxicity?}; Fillastre JP; The incidence of nephrotoxicity due to aminoglycosides should be sharply reduced . The indications for prescribing these antibiotics should be limited to infectious disorders induced by aerobic Gram-negative bacteria and by some Gram-positive bacteria requiring treatment in specialized hospital units using an association of aminoglycosides and another antibiotic . Daily doses should not exceed those indicated by the manufacturer, and the length of treatment should be as short as possible, with a relay to other antibiotics that are not or are less nephrotoxic . The possibilities for reducing the incidence of nephrotoxicity are few . It is not possible to prevent the antibiotic from entering the renal tubular cell or from producing deleterious effects therein . However, by using short-term intravenous infusion as the administration route, prolonged contact between the antibiotic and its receptors on the brush borders of the proximal tubular cells can be avoided, particularly since the process of cellular absorption is saturable . Essentially, doses should be adapted according to the age and the glomerular filtration of the patient, since renal function usually decreases with age . Volemic and hydroelectrolytic disorders favour nephrotoxicity and should be corrected . Associations with other nephrotoxic drugs should either be avoided or used with increased caution . The same is true in special situations such as endotoxaemia, severe renal parenchymatous infections and cholestasis . In any case, given the well-known insidious onset of nephropathy, aminoglycoside treatment always requires laboratory follow-up consisting of repeated testing of creatinemia during the two weeks of treatment.

J Bacteriol, 1999 Sep, 181(17), 5414 - 8
The frequency of conjugative transposition of Tn916 is not determined by the frequency of excision; Marra D et al.; Excision and formation of a covalently closed circular transposon molecule are required for conjugative transposition of Tn916 but are not the only factors that limit the frequency of conjugative transposition from one host to another . We found that in gram-positive bacteria, an increase in the frequency of excision and circularization of Tn916 caused by expression of integrase (Int) and excisionase (Xis) from a xylose-inducible promoter does not lead to an increase in the frequency of conjugative transposition . We also found that the concentration of Int and Xis in the recipient cell does not limit the frequency of conjugative transposition and that increased excision does not result in increased expression of transfer functions required to mobilize a plasmid containing the Tn916 origin of transfer . We conclude that in gram-positive hosts in which the Tn916 functions Int and Xis are overexpressed, the frequency of conjugative transposition is limited by the availability of transfer functions.

Int J Antimicrob Agents, 1999 Aug, 12(3), 253 - 6
Clinical and microbiological efficacy of single dose cefuroxime prophylaxis for dental surgical procedures; Wahlmann U et al.; In a prospective controlled randomised trial of 59 patients the effect of a single dose of cefuroxime before multiple tooth extractions on the clinical findings and occurrence of bacteraemia was studied . In one group 1.5 g cefuroxime was administered intravenously 10 min before start of surgery . A total of 118 blood samples were collected after 10 and 40 min . The cefuroxime group had a significantly lower rate of bacteraemia (33%) compared with the control group (86%) . The predominant bacteria were gram-positive cocci . Fifty four different strains of bacteria were isolated, of which most were susceptible to cefuroxime . However seven strains which were susceptible to cefuroxime in vitro, were isolated from blood cultures of the treated group . Serum concentrations of cefuroxime ranged from 52.3 to 141.6 mg/l at 10 min and 32.6 to 91.8 mg/l at 40 min with a mean of 78.6 and 55.1 mg/l respectively . The only other factor which had a significant influence on the rate of bacteraemia was the number of extracted teeth . Oral hygiene and periodontal status did not significantly affect bacteraemia . There were no wound infections in any of the groups . During these dental interventions, sufficient plasma levels of antibiotic prevented bacteraemia . Selected patients might benefit from single-dose-prophylaxis with cefuroxime in preparation for dental surgery.

Childs Nerv Syst, 1999 Jul, 15(6-7), 309 - 17
Neurosurgical management of brain abscesses in children; Ciurea AV et al.; The authors review the management of brain abscesses (BAbs) in 59 pediatric cases . The major surgical procedure used to treat them was repeated puncture and aspiration (51 cases, or 86.44%), excision (8 cases - 13.56%) being of secondary importance . The main etiology was metastatic lesions (24 cases - 40.68%), those due to cyanotic congenital heart disease being the most frequent . Single lesions dominated (41 cases - 69.49%) . Gram-positive cocci were the main bacteria involved (31 cases, 52.5%) . CT scan represented the main tool in the diagnosis and follow-up . Puncture and aspiration in BAbs led to a significant decrease in mortality (7 cases - 11.86%), and the incidence of seizures and neurological deficits was also reduced . Mortality was significantly correlated with the following factors: consciousness status, multiple BAbs location and hematogenous dissemination . There were recurrences in 11 cases (18.64%), all observed after aspiration procedures . The current concepts for complex management of BAbs, as reflected in recent literature data, are reviewed, with particular emphasis on the use of real-time CT or ultrasound-guided operative techniques.

Korean J Intern Med, 1999 Jul, 14(2), 15 - 9
Imipenem-cilastatin versus sulbactam-cefoperazone plus amikacin in the initial treatment of febrile neutropenic cancer patients; Ozyilkan O et al.; The treatment of infectious complications in cancer patients has evolved as a consequence of the developments in the chemotherapy of cancer patients . In this prospective, randomized study, we compared imipenem-cilastatin and sulbactam-cefoperazone with amikacin in the empiric therapy of febrile neutropenic (< 1000/mm3) patients with liquids and solid tumours . Of 30 evaluable episodes, 15 were treated with imipenem-cilastatin and 15 were treated with sulbactam-cefoperazone plus amikacin . 73% of episodes were culture-positive: gram-positive pathogens accounted for 62% of the isolates . Bacteremia was the most frequent site of infection . The initial clinical response rate for both regimens was 60% (p > 0.05) . No major adverse effects occurred . This study demonstrated that imipenem-cilastatin monotherapy and combination therapy of sulbactam-cefoperazone plus amikacin were equally effective empiric therapy for febrile granulocytopenic cancer patients.

Ann Hematol, 1999 Aug, 78(8), 385 - 7
Disseminated nocardiosis as a complication of Evans' syndrome; Urbaniak-Kujda D et al.; Nocardiosis is an opportunistic infection caused by gram-positive, weakly acid-fast filamentous aerobic organisms . Three species cause infection in man: N . asteroides, N . brasiliensis, and N . caviae, the first one being the most common . With increased use of immunosuppressive therapy for various autoimmune diseases, opportunistic infection by Nocardia has increasingly been reported . N . asteroides infections manifest in various ways; the lungs, skin, and brain are the organs most frequently involved . We describe a patient with Evans' syndrome, a disease requiring long-term immunosuppression, who acquired systemic nocardiosis . The infection was primarily pulmonary, misdiagnosed as tuberculosis, with subsequent hematogenous dissemination to the skin and central nervous system . The diagnosis of cerebral involvement was difficult to prove, as the patient presented with stroke-like episodes . After a positive blood culture was obtained, antibiotic therapy was introduced . The patient's condition deteriorated and the brain with infiltration of the meninges, lungs, skin, and kidneys . Nocardia is an important but often overlooked opportunistic infectious agent in immunocompromised hosts, causing diagnostic and therapeutic problems . As the mortality of cerebral nocardiosis is greater than 80%, early diagnosis and appropriate therapy are crucial.

Clin Infect Dis, 1999 Jun, 28(6), 1212 - 5
Herpes simplex peritonitis: case report; Yakulis R et al.; It has been previously reported that the most common cause of peritonitis in patients undergoing chronic ambulatory peritoneal dialysis (CAPD) is infection by a single gram-positive bacterial species . Polymicrobial bacterial infections are identified that may be secondary to bowel perforation . In 20% of cases bacterial cultures are negative . Although cultures may be negative when infection is due to a fastidious organism, when antibiotic therapy has been administered, and in cases of chemical peritonitis, a viral etiology should also be considered . We report the first documented case of herpes simplex peritonitis, which involved a 60-year-old female undergoing CAPD . Viral peritonitis may be an important form of peritonitis that has been previously unrecognized and should be considered in the differential diagnosis.

Acta Otolaryngol, 1999, 119(4), 446 - 52
Cytokine and chemokine induction using cell wall component and toxin derived from gram-positive bacteria in the rat middle ear; Kita H et al.; Lipoteichoic acid (LTA) is one of the components present within the cell wall layer of most gram-positive bacteria . It plays an important role in the initiation and progression of bacterial infection . In this study, we performed a rat middle ear and nasal perfusion with LTA purified from two different gram-positive bacterial species . Using ELISA and RT-PCR, the production and mRNA expression of rat chemokine, GRO/CINC-1, proinflammatory cytokine, TNF-alpha, in rat middle ear lavage and mucosa were investigated . GRO/CINC-1 in middle ear lavage was produced by stimulation of LTA in a time-dependent fashion; however, TNF-alpha production into the lavage was not detectable using ELISA assay . The mRNA expressions of GRO/CINC-1 and TNF-alpha in the middle ear mucosa were both induced after LTA and exotoxin exposures . The expression of IL-10 mRNA was also induced after 6 h of LTA and exotoxin exposures . The profile of the production of GRO/CINC-1 and TNF-alpha in rat nasal lavage was similar to that in the middle ear; however, the mRNA expressions of GRO/CINC-1, TNF-alpha and IL-10 in the nasal mucosa were different from those in the middle ear mucosa . These results suggest that the cell wall component and exotoxin of gram-positive bacteria can induce several cytokines in vivo and play an important role in the initiation of the inflammatory cascade in the middle ear.

Ther Drug Monit, 1999 Aug, 21(4), 379 - 88
Some international approaches to aminoglycoside monitoring in the extended dosing interval era; Morris RG et al.; Aminoglycosides have rightly remained a cost-effective anti-microbial strategy for the treatment of gram-positive infections for some 25 years . However, in recent years there has been a review of the traditional thrice-daily administration regimen in favor of an extended dosing interval strategy that takes into account the individual patient's renal function . The general recommendations that have been provided to date have been adopted in various ways internationally . These approaches were a matter of discussion for the Clinical Pharmacokinetics Committee of the International Association of Therapeutic Drug Monitoring and Clinical Toxicology at its congress (Vancouver, Canada; November 1997), and will again be a workshop issue at the Cairns (Australia) congress of the Association (September 1999) . The present report provides examples of how these practices have been applied at a group of centers from Canada (2 centers), The Netherlands, Egypt, and Australia . These reports demonstrate a variety of approaches and highlight the need for further research for assessing clinical outcomes from different dosing strategies.

J Bacteriol, 1999 Aug, 181(16), 5090 - 3
Moraxella (Branhamella) catarrhalis BRO beta-lactamase: a lipoprotein of gram-positive origin?
Bootsma HJ, Aerts PC, Posthuma G, Harmsen T, Verhoef J, van Dijk H, Mooi FR.
In the past 20 years, BRO beta-lactamase-producing Moraxella catarrhalis strains have emerged . We show that BRO is expressed as a 33-kDa lipoprotein associated with the inner leaflet of the outer membrane . To our knowledge, this is the first description of a lipidated beta-lactamase in a gram-negative species.

Lancet, 1999 Jul 31, 354(9176), 386 - 9
Detection of Alloiococcus otitis in mixed bacterial populations from middle-ear effusions of patients with otitis media; Beswick AJ et al.; BACKGROUND: Otitis media is a potentially serious disorder, since there is a risk of permanent hearing loss . Culture methods are not useful in characterisation of populations of bacteria in the middle ear . We have used a PCR-based method that does not depend on prior knowledge of the bacteria identified by culture . METHODS: Middle-ear effusion fluid was obtained from 12 patients with chronic otitis media with effusion . Total DNA was extracted from the samples, and the hypervariable regions of bacterial 16S rRNA genes were amplified by means of broad-range PCR primers . Individual PCR products were segregated by cloning to allow analysis of mixed bacterial populations . FINDINGS: Many bacterial species were detected by PCR, whereas with culture-based approaches, no bacterial growth was detected for ten of the 12 patients . The gram-positive bacterium Alloiococcus otitis (A . otitidis) was detected by 16S rDNA amplification in six of the twelve samples, but not by culture techniques . Interpretation The method may have general usefulness in characterising bacterial populations at the site of infection and may indicate, from small sample numbers, organisms that are candidates for further investigation.

Insect Biochem Mol Biol, 1999 Jul, 29(7), 585 - 97
Immulectin, an inducible C-type lectin from an insect, Manduca sexta, stimulates activation of plasma prophenol oxidase; Yu XQ et al.; Immulectin, a C-type lectin from the tobacco hornworm, Manduca sexta, was cloned from a larval fat body cDNA library . The immulectin cDNA encodes a 309 residue polypeptide . Immulectin synthesis was induced by injection of killed gram-positive or gram-negative bacteria or yeast . After injection of bacteria, immulectin mRNA appeared in fat body and immulectin protein was detected in hemolymph . Immulectin contains two carbohydrate recognition domains . The carboxyl-terminal carbohydrate recognition domain is most similar (36% identity) to a lipopolysaccharide-binding protein from the American cockroach, Periplaneta americana . It also shares 26-35% identity to carbohydrate recognition domains of various mammalian C-type lectins . Two immulectin isoforms were identified in the hemolymph of bacteria-injected larvae . Recombinant immulectin agglutinated gram-positive and gram-negative bacteria and yeast . Addition of recombinant immulectin to M . sexta plasma stimulated activation of phenol oxidase . A combination of immulectin with lipopolysaccharide from E . coli activated phenol oxidase more rapidly and to a higher level than immulectin alone, whereas lipopolysaccharide by itself had little effect on phenol oxidase activation . Immulectin synthesized in response to bacterial or fungal infection may help to trigger protective responses in M . sexta in a manner similar to mannose-binding protein, a C-type lectin that functions in the mammalian innate immune system.

Medicina (B Aires), 1999, 59 Suppl 1, 17 - 22
{Pharmacokinetics of trovafloxacin: its clinical significance}; Clara LO et al.; Trovafloxacin, once administrated by oral route, is rapidly absorbed and reaches its maximum seric concentration in about one hour . Alatrofloxacin is the prodrug, that is rapidly hydrolyzed to its original state when administered intravenously . Its bioavailability is equivalent when administered either oral or intravenously . It is a fluoroquinolone of extensive in vitro spectrum, with a higher activity for Gram positive coccus, anaerobic and atypical pneumonia-producing bacteria . The average half-life is around eleven hours, with an enhanced protein bound that allows to administrate it only once a day . The renal excretion is under 8%; it doesn't require any doses adjustment in renal insufficiency . Trovafloxacin reaches higher tissue and intracelular concentrations than classic fluoroquinolones . All these pharmacokinetic and pharmacodynamic characteristics allow to consider trovafloxacin as an interesting drug for the treatment of mixed infections or those resistant to the first line drugs.

Br J Pharmacol, 1999 Jun, 127(4), 853 - 62
E5531, a synthetic non-toxic lipid A derivative blocks the immunobiological activities of lipopolysaccharide; Kawata T et al.; 1 . The major pathological responses to Gram-negative bacterial sepsis are triggered by endotoxin or lipopolysaccharide . As endotoxin is shed from the bacterial outer membrane, it induces immunological responses that lead to release of a variety of cytokines and other cellular mediators . As part of a program aimed at developing a therapeutic agent for septic shock, we have developed E5531, a novel synthetic lipopolysaccharide antagonist . 2 . As measured by release by tumour necrosis factor-alpha, human monocytes or whole blood can be activated by lipopolysaccharide, lipid A, and lipoteichoic acid (from Gram-positive bacteria) . E5531 potently antagonizes activation by all these agents while itself being devoid of agonistic activity . 3 . The inhibitory activity of E5531 was dependent on time of addition . When 10 nM E5531 was added simultaneously with lipopolysaccharide or 1 - 3 h before addition of lipopolysaccharide, production of tumour necrosis factor-alpha was inhibited by more than 98% . The addition of E5531 1 h after lipopolysaccharide reduced the efficacy of E5531 by 47% . 4 . Antagonistic activity of E5531 was specific for lipopolysaccharide as it was ineffective at inhibiting interferon-gamma mediated NO release of RAW 264.7 cells, phorbor 12-myristate 13-acetate stimulated superoxide anion production in human neutrophils, concanavalin A stimulated mitogenic activity in murine thymocytes and tumor necrosis factor-alpha induced E-selectin expression in human umbilical vein endothelial cells . 5 . E5531 as well as MY4, an anti-CD14 antibody, inhibited radiolabelled lipopolysaccharide binding in human monocytes . 6 . These results support our contention that E5531 is a potent antagonist of lipopolysaccharide-induced release of tumour necrosis factor-alpha and other cellular mediators and may be an effective therapeutic agent for human septic shock due to Gram-negative bacteria.

EMBO J, 1999 Aug 2, 18(15), 4292 - 8
RsrA, an anti-sigma factor regulated by redox change; Kang JG et al.; SigR (sigma(R)) is a sigma factor responsible for inducing the thioredoxin system in response to oxidative stress in the antibiotic-producing, Gram-positive bacterium Streptomyces coelicolor A3(2) . Here we identify a redox-sensitive, sigma(R)-specific anti-sigma factor, RsrA, which binds sigma(R) and inhibits sigma(R)-directed transcription in vitro only under reducing conditions . Exposure to H(2)O(2) or to the thiol-specific oxidant diamide caused the dissociation of the sigma(R)-RsrA complex, thereby allowing sigma(R)-dependent transcription . This correlated with intramolecular disulfide bond formation in RsrA . Thioredoxin was able to reduce oxidized RsrA, suggesting that sigma(R), RsrA and the thioredoxin system comprise a novel feedback homeostasis loop that senses and responds to changes in the intracellular thiol-disulfide redox balance.

Int J Syst Bacteriol, 1999 Jul, 49 Pt 3, 1251 - 4
Vagococcus lutrae sp . nov., isolated from the common otter (Lutra lutra); Lawson PA et al.; Phenotypic and phylogenetic studies were performed on an unknown Gram-positive catalase-negative coccus isolated from a common otter (Lutra lutra) . 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 and Vagococcus salmoninarum . The unknown bacterium was readily distinguished from the two 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 lutrae, the type strain of which is CCUG 39187T.

Int J Syst Bacteriol, 1999 Jul, 49 Pt 3, 1247 - 50
Facklamia tabacinasalis sp . nov., from powdered tobacco; Collins MD et al.; An unknown Gram-positive, catalase-negative, facultatively anaerobic, coccus-shaped organism originating as a contaminant of snuff was characterized by phenotypic and molecular taxonomic methods . Comparative 16S rRNA gene sequencing studies demonstrated that the bacterium represents a new subline within the genus Facklamia . The unknown bacterium was readily distinguished from Facklamis hominis and Facklamia ignava by biochemical tests and electrophoretic analysis of whole-cell proteins . On the basis of phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Facklamia tabacinasalis sp . nov., the type strain of which is CCUG 30090T.

Int J Syst Bacteriol, 1999 Jul, 49 Pt 3, 1217 - 20
Arthrobacter rhombi sp . nov., isolated from Greenland halibut (Reinhardtius hippoglossoides); Osorio CR et al.; Two strains of a hitherto undescribed Gram-positive coryneform bacterium isolated from Greenland halibut (Reinhardtius hippoglossoides) were characterized by phenotypic and molecular taxonomic methods . Comparative 16S rRNA gene sequencing studies demonstrated that the unknown strains constitute a new line within the genus Arthrobacter . The nearest relatives of the bacterium from fish were members of the Arthrobacter nicotianael Arthrobacter sulfureus group . The unknown bacterium was readily distinguished from these species by phenotypic methods . Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Arthrobacter rhombi sp . nov . The type strain of Arthrobacter rhombi is CCUG 38813T.

Support Care Cancer, 1999 Jul, 7(4), 253 - 9
Analysis of early infectious complications in pediatric patients undergoing bone marrow transplantation; Busca A et al.; The purpose of the present study was to analyze the characteristics of infectious complications occurring during the first 100 days after bone marrow transplantation (BMT) in a cohort of 123 pediatric patients with hematological malignancies (n = 73), solid tumors (n = 32) and nonmalignant disorders (n = 18) . Fifty-eight patients received allogeneic grafts, and 65 patients an autologous transplant . Fever developed in 107 (87%) children; 82% of infectious complications occurred during the neutropenic period . Documented infection developed in 33 (31%) patients, while 74 (69%) patients had possible infection (i.e . fever of unknown origin) . The incidence of bacteremia was 21%, and gram-positive cocci were the predominant pathogens; non-bacteremic microbiologically documented infection developed in 6% of patients; clinically evident infection developed in 4% of subjects . The incidence of primary febrile episodes was not significantly different between autologous and allogeneic BMT (86% vs 88%); nor did the median number of days to the onset of fever (5 days in both groups) or the median duration of fever (5 days in both groups) differ . In contrast, the frequency of secondary febrile episodes was significantly higher (P = 0.0001) in allogeneic BMT recipients (40%) than in autologous recipients (15%) . The mortality rate due to infections was 2/36 (5%) for matched sibling donor BMT, and 1/13 (8%) for matched unrelated donor BMT . No deaths occurred in the 65 patients who were autografted . Invasive fungal infections accounted for 2 of the 3 infectious deaths . In conclusion, the majority of children undergoing BMT experienced at least one infectious episode; allogeneic BMT recipients were at high risk of developing secondary febrile episodes, but the overall mortality rate due to infection in the first 100 days after transplantation was low.

Vet Pathol, 1999 Jul, 36(4), 345 - 7
Nocardia nova causing pulmonary nocardiosis of black crakes (Limnocorax flavirostra); Bacciarini LN et al.; Natural nocardial infection has been reported in many different species including mammals and fish, but reports in birds remain uncommon . Eight juvenile Black Crakes (Limnocoraxflavirostra) died unexpectedly at the Basle Zoo . Necropsy revealed disseminated white, firm nodules, 1-3 mm in diameter, throughout the lung parenchyma . Histologically, the lungs contained multiple, often confluent granulomas with central necrosis . Delicate, gram-positive, 0.5- to 1.0-microm-wide, branching, occasionally beaded, filamentous organisms were visible in necrotic centers . These organisms were acid fast when stained with Fite-Faraco . No histologic lesions were seen in other organs . Nocardia nova was isolated from liver, spleen, kidney, and lung . Granulomatous and necrotizing nocardial pneumonia with agonal septicemia was diagnosed, suggesting an aerogenous infection . To our knowledge, this is the first reported epizootic outbreak of nocardiosis in birds, which is additionally unusual because it was caused by N . nova.

Int J Biol Macromol, 1999 Jun-Jul, 25(1-3), 3 - 19
Biochemical and genetic analysis of PHA synthases and other proteins required for PHA synthesis; Rehm BH et al.; Polyhydroxyalkanoic acids (PHA) represent a complex class of storage polyesters that are synthesized by a wide range of different gram-positive and gram-negative bacteria as well as by some Archaea and that are deposited as insoluble cytoplasmic inclusions . PHA synthases, which are the key enzymes for PHA biosynthesis, have been characterized in much detail . At present 42 PHA synthase structural genes from 38 different bacteria have been cloned, and from 30 genes the nucleotide sequences were obtained . The strategies successfully employed to clone these genes and the current knowledge on the organization of the PHA synthase genes and other genes encoding proteins related to PHA metabolism will be compiled . In addition, the primary structures of the 30 PHA synthases were aligned and analyzed with respect to highly conserved amino acids and biochemical features . The direction, in which research should proceed, in order to increase our knowledge on biosynthesis of PHAs and to utilize this knowledge for the development of technically and economically feasible processes for the production of these polyesters will be outlined.

Immunopharmacology, 1999 May, 42(1-3), 61 - 74
Therapeutic intervention with complement and beta-glucan in cancer; Ross GD et al.; Complement (C) has two major effector systems available for host defense . The membrane attack complex (MAC) generated from components C5-C9 can form membrane-penetrating lesions that lead to cell death by causing a rapid loss of cytoplasmic components . The MAC is only effective against pathogens with outer phospholipid membranes, and cannot kill gram-positive bacteria or yeast whose membranes are protected by cell walls . The most important effector mechanism of C is the opsonization of microbial pathogens with the serum protein C3 that leads to their high avidity attachment to the C3-receptors of phagocytic cells . Pathogens that activate complement are first coated with the C3b fragment of C3, which is rapidly proteolyzed into the iC3b fragment by serum factor I . These iC3b fragments serve to promote the high avidity attachment of the 'iC3b-opsonized' pathogens to the iC3b-receptors (CR3, CD11b/CD18) of phagocytic cells and natural killer (NK) cells, stimulating phagocytosis and/or cytotoxic degranulation . Host cells, including neoplastic tumor cells, have been endowed with natural mechanisms for self-protection against both the MAC and the cytotoxic activation of CR3 . This review discusses a novel type of immunotherapy for cancer that uses soluble yeast beta-glucan to override the normal resistance of iC3b-opsonized tumor cells to the cytotoxic activation of phagocyte and NK cell CR3, allowing this important effector mechanism of the C system to function against tumor cells in the same way that it normally functions against bacteria and yeast . Moreover, the cytotoxic activation of beta-glucan-primed NK cell CR3 by iC3b-opsonized tumors is shown to be accompanied by a tumor-localized secretion of the cytokines TNFalpha, IFNalpha, IFNgamma, and IL-6.

Intensive Care Med, 1999 May, 25(5), 469 - 74
Bacterial sepsis-induced rhabdomyolysis; Betrosian A et al.; OBJECTIVE: To describe the syndrome of rhabdomyolysis during bacterial sepsis with regard to incidence, blood bacteriology and complications and to examine the association between hyperosmolal state and rhabdomyolysis, evaluating the relationship between plasma osmolality (Posm) and serum creatine phosphokinase (CPK) levels . DESIGN: Prospective study including all patients admitted to the intensive care unit (ICU) for sepsis with positive blood culture and rhabdomyolysis over a 3-year period . SETTING: Seven-bed medical/surgical ICU of a teaching hospital . PATIENTS: 35 patients (group 1) with bacterial sepsis-induced rhabdomyolysis (15 males, 20 females; mean age 71+/-13 years) and 122 (group 2) bacteraemic septic patients without rhabdomyolysis (49 males, 73 females; mean age 68+/-15) were studied . Patients with rhabdomyolysis were divided into gram(+) and gram(-) subgroups according to the blood culture growth . RESULTS: From 491 patients recorded, 35 fulfilled the inclusion criteria for bacterial sepsis-induced rhabdomyolysis (7.1%) . Gram-positive bacteria predominated in group 1 (69%), while gram-negative predominated (60%) in group 2 . There was a correlation between CPK and Posm levels in the rhabdomyolysis Group (r = 0.52, R2 = 0.27, p = 0.003) . There was a stronger correlation between these two variables (r = 0.67, R2 = 0.45, p = 0.001) in the gram(+).subgroup . Acute renal failure (68.5%) and electrolyte disorders such as hyperkalaemia (34%) and hypocalcaemia (48.5%) were the major complications in the rhabdomyolysis group . Sixteen (45.7%) patients in group 1 and 49 (40%) in group 2 died during their stay in the ICU from sepsis and multiple organ failure . Rhabdomyolysis was not considered a contributing factor to their death, as none of our patients died during or immediately after the syndrome . CONCLUSION: Bacterial sepsis-induced rhabdomyolysis results from certain types of microorganisms, mainly gram-positive and to a lesser extent gram-negative . Hyperosmolality is a predisposing mechanism for rhabdomyolysis during bacteraemic sepsis from any type of bacterial microorganism.

J Biol Chem, 1999 Jul 16, 274(29), 20287 - 92
Purification and molecular characterization of ortho-chlorophenol reductive dehalogenase, a key enzyme of halorespiration in Desulfitobacterium dehalogenans; van de Pas BA et al.; ortho-Chlorophenol reductive dehalogenase of the halorespiring Gram-positive Desulfitobacterium dehalogenans was purified 90-fold to apparent homogeneity . The purified dehalogenase catalyzed the reductive removal of a halogen atom from the ortho position of 3-chloro-4-hydroxyphenylacetate, 2-chlorophenol, 2,3-dichlorophenol, 2,4-dichlorophenol, 2,6-dichlorophenol, pentachlorophenol, and 2-bromo-4-chlorophenol with reduced methyl viologen as electron donor . The dechlorination of 3-chloro-4-hydroxyphenylacetate was catalyzed by the enzyme at a Vmax of 28 units/mg protein and a Km of 20 microM . The pH and temperature optimum were 8.2 and 52 degrees C, respectively . EPR analysis indicated one {4Fe-4S} cluster (midpoint redox potential (Em) = -440 mV), one {3Fe-4S} cluster (Em = +70 mV), and one cobalamin per 48-kDa monomer . The Co(I)/Co(II) transition had an Em of -370 mV . Via a reversed genetic approach based on the N-terminal sequence, the corresponding gene was isolated from a D . dehalogenans genomic library, cloned, and sequenced . This revealed the presence of two closely linked genes: (i) cprA, encoding the o-chlorophenol reductive dehalogenase, which contains a twin-arginine type signal sequence that is processed in the purified enzyme; (ii) cprB, coding for an integral membrane protein that could act as a membrane anchor of the dehalogenase . This first biochemical and molecular characterization of a chlorophenol reductive dehalogenase has revealed structural resemblance with haloalkene reductive dehalogenases.

Avian Dis, 1999 Apr-Jun, 43(2), 338 - 41
Erysipelas in a free-ranging Hawaiian crow (Corvus hawaiiensis); Work TM et al.; We describe a case of erysipelas in a free-ranging endangered Hawaiian crow . The partially scavenged carcass exhibited gross emaciation and petechial hemorrhages in both lungs . Microscopy revealed multiple necrotic foci associated with gram-positive rods in the liver and adrenal, diffuse acute proximal tubular necrosis of kidney, diffuse necrosis and inflammation of proventricular mucosa associated with gram-positive rods, and multiple intravascular aggregates of gram-positive rods associated with thrombi . Culture of the kidney revealed the bacterium to be Erysipelothrix rhusiopathiae . The implications of this finding to free-ranging crows remain unclear.

Biochemistry (Mosc), 1999 Jun, 64(6), 625 - 30
Purification and characterization of maleylacetate reductase from Nocardioides simplex 3E utilizing phenoxyalcanoic herbicides 2,4-D and 2,4,5-T; Travkin VM et al.; Maleylacetate reductase was isolated and purified from the Gram-positive strain Nocardioides simplex 3E which is able to utilize the phenoxyalcanoic herbicides 2,4-D and 2,4,5-T . Cells were grown on 2,4-D as the sole carbon source . The enzyme was purified by 380-fold with 3.0% yield . The purified maleylacetate reductase is a homodimer with subunit molecular mass of 37 kD . The enzyme required NADH as a cofactor; the Km for maleylacetate is 25 microM; Vmax (with NADH as cofactor) and kcat are 185 U/mg and 6845 min-1, respectively . The enzyme is very unstable; its pH and temperature optima are at 7.0-7.1 and 50 degrees C, respectively.

Syst Appl Microbiol, 1999 May, 22(2), 269 - 79
Sulfate-reducing bacteria in rice field soil and on rice roots; Wind T et al.; Rice plants that were grown in flooded rice soil microcosms were examined for their ability to exhibit sulfate reducing activity . Washed excised rice roots showed sulfate reduction potential when incubated in anaerobic medium indicating the presence of sulfate-reducing bacteria . Rice plants, that were incubated in a double-chamber (phylloshpere and rhizosphere separated), showed potential sulfate reduction rates in the anoxic rhizosphere compartment . These rates decreased when oxygen was allowed to penetrate through the aerenchyma system of the plants into the anoxic root compartment, indicating that sulfate reducers on the roots were partially inhibited by oxygen or that sulfate was regenerated by oxidation of reduced S-compounds . The potential activity of sulfate reducers on rice roots was consistent with MPN enumerations showing that H2-utilizing sulfate-reducing bacteria were present in high numbers on the rhizoplane (4.1 x 10(7) g-1 root fresh weight) and in the adjacent rhizosperic soil (2.5 x 10(7) g-1 soil dry weight) . Acetate-oxidizing sulfate reducers, on the other hand, showed highest numbers in the unplanted bulk soil (1.9 x 10(6) g-1 soil dry weight) . Two sulfate reducing bacteria were isolated from the highest dilutions of the MPN series and were characterized physiologically and phylogenetically . Strain F1-7b which was isolated from the rhizoplane with H2 as electron donor was related to subgroup II of the family Desulfovibrionaceae . Strain EZ-2C2, isolated from the rhizoplane on acetate, grouped together with Desulforhabdus sp . and Syntrophobacter wolinii . Other strains of sulfate-reducing bacteria originated from bulk soil of rice soil microcosms and were isolated using different electron donors . From these isolates, strains R-AcA1, R-IbutA1, R-PimA1 and R-AcetonA170 were Gram-positive bacteria which were affiliated with the genus Desulfotomaculum . The other isolates were members of subgroup II of the Desulfovibrionaceae (R-SucA1 and R-LacA1), were related to Desulforhabdus sp . (strain BKA11), Desulfobulbus (R-PropA1), or culstered between Desulfobotulus sapovorans and Desulfosarcina variabilis (R-ButA1 and R-CaprA1).

Syst Appl Microbiol, 1999 May, 22(2), 174 - 8
Organization of the chromosomal region containing the genes lexA and topA in Thermotoga neapolitana . Primary structure of LexA reveals phylogenetic relevance; Zverlov VV et al.; The chromosomal region of Thermotoga neapolitana surrounding the gene lexA (4283 bp) was sequenced . In addition to the topoisomerase gene top2A it contained five open reading frames . A part of the cloned region showed high sequence homology with a previously published sequence of Th . maritima and indicated an identical arrangement of genes in both microorganisms . Structural analysis of the LexA protein showed significant, but relatively low overall homology with LexA proteins of other bacteria, especially in the DNA binding region . However, key amino acids for processing and secondary structure elements like the helix-turn-helix motif are well conserved . Sequence alignment analysis of the whole protein and the DNA-binding sites of all known LexA sequences uncovers groups of similarity reminding the phylogenetic tree of the Bacteria . A consensus sequence with the SOS- or Cheo-box upstream of the lexA gene of Th . maritima and Th . neapolitana was absent . Together with the phylogenetic distance of the Thermotogales from other bacteria this suggests the presence of a new operator target sequence specific for the Thermotogales, in analogy to the SOS-box for the gamma-group Proteobacteria and the Cheo-box for low- and high-GC Gram-positive bacteria.

Appl Environ Microbiol, 1999 Jul, 65(7), 3192 - 204
Phylogenetic analysis of particle-attached and free-living bacterial communities in the Columbia river, its estuary, and the adjacent coastal ocean; Crump BC et al.; The Columbia River estuary is a dynamic system in which estuarine turbidity maxima trap and extend the residence time of particles and particle-attached bacteria over those of the water and free-living bacteria . Particle-attached bacteria dominate bacterial activity in the estuary and are an important part of the estuarine food web . PCR-amplified 16S rRNA genes from particle-attached and free-living bacteria in the Columbia River, its estuary, and the adjacent coastal ocean were cloned, and 239 partial sequences were determined . A wide diversity was observed at the species level within at least six different bacterial phyla, including most subphyla of the class Proteobacteria . In the estuary, most particle-attached bacterial clones (75%) were related to members of the genus Cytophaga or of the alpha, gamma, or delta subclass of the class Proteobacteria . These same clones, however, were rare in or absent from either the particle-attached or the free-living bacterial communities of the river and the coastal ocean . In contrast, about half (48%) of the free-living estuarine bacterial clones were similar to clones from the river or the coastal ocean . These free-living bacteria were related to groups of cosmopolitan freshwater bacteria (beta-proteobacteria, gram-positive bacteria, and Verrucomicrobium spp.) and groups of marine organisms (gram-positive bacteria and alpha-proteobacteria {SAR11 and Rhodobacter spp.}) . These results suggest that rapidly growing particle-attached bacteria develop into a uniquely adapted estuarine community and that free-living estuarine bacteria are similar to members of the river and the coastal ocean microbial communities . The high degree of diversity in the estuary is the result of the mixing of bacterial communities from the river, estuary, and coastal ocean.

Hindustan Antibiot Bull, 1997 Feb-Nov, 39(1-4), 50 - 5
A new variety of Chainia olivacea from marine sediment off Gulf of Mannar; Ellaiah P et al.; A new variety of Chainia olivacea was isolated from marine sediment off Gulf of Mannar . The morphological, cultural, physiological and biochemical characters were studied, compared to known species and identified as a new variety of Chainia olivacea . Antibiotic activity of the strain was tested against both Gram positive and Gram negative bacteria as well as fungi and yeasts . Sodium chloride tolerance was also tested.

Eur J Clin Microbiol Infect Dis, 1999 Apr, 18(4), 283 - 5
Procalcitonin concentrations in patients with neutropenic fever; Ruokonen E et al.; To assess the usefulness of markers of inflammation in distinguishing bacterial infection from severe systemic nonbacterial inflammation, concentrations of procalcitonin, neopterin, endotoxin, tumor necrosis factor, and interleukin-6 were measured in 28 neutropenic patients at the onset of fever and twice thereafter at 4 h intervals . Infection was found in 11 patients, and 17 patients had fever of undetermined origin . The procalcitonin concentration increased rapidly in patients with infection: the response was detectable within 8 h of the onset of fever . Procalcitonin is a specific but not a sensitive marker of infection in patients with neutropenic fever . Its poor sensitivity was related to an absent or delayed response in patients with gram-positive infections . Considerable overlap between infected and noninfected patients was found in levels of endotoxin, tumor necrosis factor, and interleukin-6.

J Antimicrob Chemother, 1999 May, 43(5), 683 - 8
Pharmacodynamics of trovafloxacin in experimental pneumococcal meningitis: basis for dosage selection in children with meningitis; McCoig CC et al.; Trovafloxacin is a recently approved fluoroquinolone with excellent activity against gram-positive and gram-negative organisms that offers a potential alternative for treatment of beta-lactam-resistant pneumococcal meningitis . Using the rabbit meningitis model, we sought to characterize the pharmacodynamic properties of trovafloxacin in the cerebrospinal fluid (CSF) . Animals were given single doses of trovafloxacin of 10, 15, 20 or 30 mg/kg; 1 h after Infusion mean CSF concentrations were 0.59+/-0.18, 0.74+/-0.14, 1.12+/-0.12 and 1.07+/-0.35 mg/L, respectively . The bacterial killing rate Increased with increasing dosages of trovafloxacin, indicating that its activity is concentration dependent . All three pharmacodynamic Indices (area under the concentration curve (AUC)/MBC, peak concentration (Cmax)/MBC, and time above MBC (T > MBC)) correlated with bacterial killing; however, AUC/MBC correlated best (r = 0.71) . In a second experiment we found comparable bacterial killing with multiple doses of trovafloxacin given either every serum half-life or every two serum half-lives . In both experiments bacterial regrowth occurred when the concentration of trovafloxacin in CSF fell below the MBC . These data have been used in formulating an appropriate regimen for trovafloxacin treatment of bacterial meningitis in children.

J Antimicrob Chemother, 1999 Jan, 43(1), 141 - 3
Study to assess the reliability of a disc diffusion method for determining the sensitivity of gram-positive pathogens to dalfopristin/quinupristin; Andrews JM et al.; A standardized method of disc testing the sensitivity of gram-positive pathogens to dalfopristin/quinupristin was developed, and then 'field tested' in ten centres in the UK . For a 15 microg disc, zone diameter breakpoints of 20 mm and 15 mm are suggested when organisms are tested on Iso-Sensitest agar and Iso-Sensitest agar supplemented with 5% whole horse blood, respectively.

Microbiology, 1999 May, 145 ( Pt 5), 1245 - 51
Expression of the Oenococcus oeni trxA gene is induced by hydrogen peroxide and heat shock; Jobin MP et al.; Sequencing of the DNA region located upstream of the alpha-acetolactate synthase and decarboxylase (alsS-alsD) cluster of Oenococcus oeni allowed identification of an ORF, named trxA . This encodes a protein of 104 amino acids very similar to known thioredoxins . The protein encoded by the cloned fragment was able to complement Escherichia coli strains lacking a functional thioredoxin . Considering the results of protein sequence comparisons and complementation experiments, it was concluded that the trxA gene encodes a functional thioredoxin . Studies of trxA expression showed that the abundance of trxA mRNA was similar during all growth stages . A significant increase in trxA mRNA levels was observed in the presence of hydrogen peroxide in the medium or after heat shock . A single transcriptional start site was determined with total RNA isolated from cells subjected or not subjected to oxidative stress or heat shock . In each case the same promoter region was identified and shown to have a high similarity to the consensus promoter sequence of Gram-positive bacteria, as well as to that of E . coli and the previously mapped promoters from O . oeni.

Microb Pathog, 1999 Jul, 27(1), 43 - 53
Differential induction of NO synthesis by gram-positive and gram-negative bacteria and their components in bovine monocyte-derived macrophages; Jungi TW et al.; The ability of Gram-positive and Gram-negative bacteria to promote the induction of NO synthesis in bovine monocyte-derived macrophages (MDM) was tested . Heat-killed Gram-negative organisms induced NO synthesis at low concentrations (optimum 0.2 to 2 microg/ml wet weight), regardless of the strain, and the response was only moderately enhanced by co-administration of recombinant bovine interferon-gamma (rboIFN-gamma) . The activity was largely, but not exclusively, due to lipopolysaccharide (LPS), since it was largely abrogated by co-incubation with polymyxin-B . Diphosphoryl-lipid-A and rough-strain LPS were two orders of magnitude more active than monophosphoryl-lipid A, but two orders of magnitude less active than smooth-strain LPS, suggesting that O side chains contribute to increasing the affinity of LPS or to act as a costimulus . Gram-positive bacteria as single stimuli were four orders of magnitude less potent in inducing NO synthesis than Gram-negative organisms, but upon costimulation with rboIFN-gamma, some of them were excellent inducers of NO synthesis . A similar rboIFN-gamma-enhanced NO synthesis induction was also observed for zymosan, muramyl dipeptide, lipoteichoic acid and lipoarabinomannan, although to a lesser extent than for the whole heat-inactivated prototypic organisms . Thus, bovine macrophages exposed to rboIFN-gamma have mechanisms by which they universally react to bacterial compounds distinct from LPS by induction of NO synthesis .

Arch Microbiol, 1999 May-Jun, 171(6), 355 - 63
Biochemical and phylogenetic analyses of psychrophilic isolates belonging to the Arthrobacter subgroup and description of Arthrobacter psychrolactophilus, sp . nov; Loveland-Curtze J et al.; During our work on psychrophilic microorganisms we obtained a large collection of new isolates . In order to identify six of these, we examined their growth properties, cell wall compositions, and their 16S rRNA gene sequences . The results showed that all of the isolates are gram-positive, aerobic, contain lysine in their cell walls, and belong to the high mol% G+C Arthrobacter subgroup . Phylogenetic analysis of the 16S rRNA genes grouped five isolates obtained from a small geographical region into a monophyletic clade . Isolate B7 had a 16S rRNA sequence that was 94.3% similar to that of Arthrobacter polychromogenes and 94.4% similar to that of Arthrobacter oxydans . Primary characteristics that distinguish isolate B7 from the Arthrobacter type strain (Arthrobacter globiformis) and A . polychromogenes include lack of growth at 37 degrees C, growth at 0-5 degrees C, the ability to use lactose as a sole carbon source, and the absence of blue pigments . Because of these differences, isolate B7 was chosen as a type strain representing a new Arthrobacter species, Arthrobacter psychrolactophilus . The sixth isolate, LV7, differed from the other five because it did not have the rod/ coccus morphological cycle and was most closely related to Arthrobacter agilis.

Ann Pharmacother, 1999 May, 33(5), 600 - 6
Once-daily aminoglycosides in the treatment of gram-positive endocarditis; Tam VH et al.; OBJECTIVE: To examine the role of once-daily aminoglycosides (ODA) in the treatment of gram-positive endocarditis . DATA SOURCES: A MEDLINE search was conducted from January 1984 to August 1998, and a Current Contents search was performed from September 1998 to December 1998, using endocarditis or aminoglycoside as key words . In addition, relevant articles were cross-referenced to screen for additional information . DATA EXTRACTION: Data published in English regarding the use of aminoglycosides in endocarditis are cited . Emphasis was placed on animal and human studies, but in vitro studies and review articles are also included . DATA SYNTHESIS: Endocarditis and the pharmacology of aminoglycosides are briefly reviewed . ODA is an alternative to conventional dosing in the treatment of endocarditis . Extensive work in endocarditis has been done recently in animals and humans to add to our understanding . Limited clinical data exist to support the theoretical advantages of increased efficacy, reduced toxicity, and potential cost savings versus traditional synergistic aminoglycoside dosing . Optimal monitoring of ODA remains undefined . CONCLUSIONS: Routine use of ODA for the treatment of endocarditis is not yet advocated . Promising supporting evidence and speculation of success of ODA in gram-positive endocarditis justify well-designed trials to further define its role in therapy.

J Clin Microbiol, 1999 Jul, 37(7), 2255 - 61
Development of amplified 16S ribosomal DNA restriction analysis for identification of Actinomyces species and comparison with pyrolysis-mass spectrometry and conventional biochemical tests; Hall V et al.; Identification of Actinomyces spp . by conventional phenotypic methods is notoriously difficult and unreliable . Recently, the application of chemotaxonomic and molecular methods has clarified the taxonomy of the group and has led to the recognition of several new species . A practical and discriminatory identification method is now needed for routine identification of clinical isolates . Amplified 16S ribosomal DNA restriction analysis (ARDRA) was applied to reference strains (n = 27) and clinical isolates (n = 36) of Actinomyces spp . and other gram-positive rods . Clinical strains were identified initially to the species level by conventional biochemical tests . However, given the low degree of confidence in conventional methods, the findings obtained by ARDRA were also compared with those obtained by pyrolysis-mass spectrometry . The ARDRA profiles generated by the combination of HaeIII and HpaII endonuclease digestion differentiated all reference strains to the species or subspecies level . The profiles correlated well with the findings obtained by pyrolysis-mass spectrometry and by conventional tests and enabled the identification of 31 of 36 clinical isolates to the species level . ARDRA was shown to be a simple, rapid, cost-effective, and highly discriminatory method for routine identification of Actinomyces spp . of clinical origin.

J Biol Chem, 1999 Jun 18, 274(25), 17406 - 9
Peptidoglycan- and lipoteichoic acid-induced cell activation is mediated by toll-like receptor 2; Schwandner R et al.; The life-threatening complications of sepsis in humans are elicited by infection with Gram-negative as well as Gram-positive bacteria . Recently, lipopolysaccharide (LPS), a major biologically active agent of Gram-negative bacteria, was shown to mediate cellular activation by a member of the human Toll-like receptor family, Toll-like receptor (TLR) 2 . Here we investigate the mechanism of cellular activation by soluble peptidoglycan (sPGN) and lipoteichoic acid (LTA), main stimulatory components of Gram-positive bacteria . Like LPS, sPGN and LTA bind to the glycosylphosphatidylinositol-anchored membrane protein CD14 and induce activation of the transcription factor NF-kappaB in host cells like macrophages . We show that whole Gram-positive bacteria, sPGN and LTA induce the activation of NF-kappaB in HEK293 cells expressing TLR2 but not in cells expressing TLR1 or TLR4 . The sPGN- and LTA-induced NF-kappaB activation was not inhibited by polymyxin B, an antibiotic that binds and neutralizes LPS . Coexpression together with membrane CD14 enhances sPGN signal transmission through TLR2 . In contrast to LPS signaling, activation of TLR2 by sPGN and LTA does not require serum . These findings identify TLR2 as a signal transducer for sPGN and LTA in addition to LPS.

Crit Care Med, 1999 May, 27(5), 937 - 40
The use of polymerase chain reaction to detect septicemia in critically ill patients; Cursons RT et al.; OBJECTIVE: To describe the use of bacterial DNA amplification of conserved bacterial 16S ribosomal DNA nucleotide sequences by polymerase chain reaction (PCR) to detect the presence of septicemia in critically ill septic patients . DESIGN: Case series of blood samples from septic patients comparing the PCR results with conventional blood culture results . SETTING: A general intensive care unit in a tertiary referral hospital . PATIENTS: Two sets of samples (n = 101 and n = 55) from patients diagnosed as clinically septic and requiring blood cultures . They were classified by internationally accepted criteria into systemic inflammatory response syndrome, severe sepsis, and septic shock groups . INTERVENTIONS: Blood samples taken in a sterile fashion concurrently for blood culture, and PCR of the bacterial 16S ribosomal RNA gene in leukocytes and plasma . Two different DNA extraction techniques for PCR were tried sequentially . MEASUREMENTS AND MAIN RESULTS: Blood culture and PCR positivity were measured in relation to the clinical classification of severity of sepsis . Using the initial extraction method (n = 101), ten patients were positive by both PCR and blood culture, eight patients were PCR positive and blood culture negative, and seven patients were blood culture positive and PCR negative . From the clinical criteria, PCR detected at least six true positives that had been missed on blood culture and missed four true Gram-positive bacteremias . When the initial code was broken, this deficiency was rectified using the improved extraction technique (n = 55), in which ten patients were positive by PCR and blood culture, 29 patients were PCR positive and blood culture negative, and two patients were PCR negative and PCR positive . CONCLUSIONS: We conclude that the use of PCR (for the 16S ribosomal DNA in the plasma) was significantly more sensitive than the use of conventional blood culturing techniques for the detection of bacteremia in seriously ill patients . This could prove to be a valuable adjunct to conventional blood cultures.

Protein Eng, 1999 May, 12(5), 371 - 3
Proteolytic cleavage of gram-positive beta recombinase is required for crystallization; Orth P et al.; Beta recombinase, a DNA resolvase-invertase, catalyzes in the presence of a chromatin-associated protein such as Hbsu, DNA resolution or DNA inversion on supercoiled substrates containing two directly or inversely oriented target (six) sites . Single crystals of the beta recombinase from plasmid pSM19035 were obtained using the vapor diffusion technique with ammonium phosphate as the precipitating agent . The crystals diffracted X-rays to a maximum resolution of 2.5A . Due to proteolytic degradation during the crystallization experiment, the crystals contain only the N-terminal catalytic domain of beta recombinase corresponding to about 60% of the molecular mass of the initially assayed native protein . The proteolytic removal of the C-terminal DNA-binding domain demonstrated that protein modification can be essential to provide material suitable for X-ray analysis.

Life Sci, 1999, 64(19), 1733 - 8
Significantly improved oral uptake of amikacin in FVB mice in the presence of CRL-1605 copolymer; Jagannath C et al.; Amikacin is an aminoglycoside which is used in the treatment of infection from gram negative bacteria . Amikacin is also used synergistically with penicillin against gram positive cocci . Amikacin cannot be delivered orally probably due to efflux of drug by P-glycoprotein pump in the brush border of intestine . We studied the possibility of delivering amikacin orally in mice using a copolymer (CRL-1605) as a vehicle . This copolymer inhibits P-glycoprotein pump . Two different doses of amikacin were used (500 mg/kg and 100 mg/kg) . The concentration of polymer used was 132 mg/kg . The liquid formulation was fed to mice by gavage and serum amikacin concentrations were estimated after one hour and two hours using fluorescence polarization immunoassay . We observed a two fold increase in serum amikacin concentration when amikacin was orally delivered in the presence of CRL-1605 compared to controls (amikacin alone) . We conclude that gastrointestinal absorption of amikacin is significantly increased in the presence of CRL-1605 in mice.

J Nat Prod, 1999 May, 62(5), 750 - 1
A new antibiotic nortriterpene quinone methide from Maytenus catingarum; Alvarenga NL et al.; 15alpha-Hydroxy-21-keto-pristimerine (1), a new nortriterpene quinone methide was isolated from the root bark of Maytenus catingarum along with other well-known related compounds, including pristimerine (2), tingenone, and 20alpha-hydroxy-tingenone . The structure of 1 was determined by means of 1H and 13C NMR spectroscopy, including homonuclear and heteronuclear correlations . Compound 1 showed antibiotic activity against Gram-positive bacteria.

J Vet Med Sci, 1999 Apr, 61(4), 421 - 3
Nasal nocardiosis in a calf; Takahashi K et al.; Nasal nocardiosis was found in a female Japanese Black calf, 11 months of age . Macroscopically, the posterior half of the left nasal passage was completely obstructed by yellowish brown caseous substance and the mucosa was irregularly thickened . In the brain, a few soft brown foci were present in the olfactory bulb and frontal lobe . Microscopically, there were closely packed granulomas in the nasal cavity and brain . The lesions were characterized by a center of cellular debris surrounded by epithelioid macrophages, neutrophils, lymphocytes and multinucleated giant cells of the Langhans type . Special stains revealed the presence of a large number of filamentous branching gram-positive, partially acid-fast organisms in these epithelioid cells and giant cells, and in cellular debris.

Infect Dis Clin North Am, 1999 Jun, 13(2), 397 - 412
Gram-positive sepsis . Mechanisms and differences from gram-negative sepsis; Sriskandan S et al.; This article has reviewed the mechanisms by which gram-positive bacteria lead to septic shock, with regard to bacterial structure and toxicology and the host responses elicited both in animal models and in the clinical setting . Gram-positive organisms are better suited to invade host tissues and elicit, in general, a brisker phagocytic response than gram-negative organisms . The lack of endotoxin in the outer cell wall is compensated for by the presence of exposed peptidoglycan and a range of other toxic secreted products . It appears that cell wall components of gram-positive bacteria may signal via the same receptor as gram-negative endotoxin, although the type of signal and coreceptor may differ . Both animal and clinical data suggest that, unlike endotoxin-mediated shock, gram-positive infection produces a modest TNF response only and does not respond well to anti-TNF therapies . This leads one to conclude that the mechanisms leading to shock in gram-positive infection may be multifactorial and perhaps more difficult to treat . A thorough review of gram-positive mechanisms of sepsis is hampered by a lack of basic research in this field . Understanding of gram-negative bacterial structure and the regulation of virulence genes is at an advanced stage, yet the molecular tools to analyse virulence factors in the gram-positive genome have only recently become available . There is a paucity of good animal models of gram-positive infection and a lack of microbiologic data from some of the major trials in sepsis that might have given greater insight into the mechanisms leading to shock in various infections.

Respirology, 1999 Mar, 4(1), 31 - 5
Pulmonary actinomycosis in Korea; Baik JJ et al.; Pulmonary actinomycosis is a chronic pulmonary infection caused by Actinomyces, a Gram-positive, microaerophilic bacterium . Pulmonary involvement, other than cervicofacially or abdominopelvically, is uncommon and often leads to a misdiagnosis of pulmonary tuberculosis or lung cancer . In order to investigate the clinical, radiological, diagnostic and therapeutic characteristics of pulmonary actinomycosis, we reviewed a total of 25 cases reported in Korea . Thirteen were diagnosed at our hospital between 1985 and 1997 and 12 were reported in Korean publications . The condition occurred most frequently in middle-aged males, the most common symptom being haemoptysis, followed by cough and sputum . The main radiological features were peripherally located mass or nodule and consolidation, with central low attenuation . Complications such as empyema, sinus fistula or mediastinitis did not occur . Diagnosis was confirmed by percutaneous needle aspiration (n = 8), bronchoscopic biopsy (n = 3) or thoracotomy (n = 13) . Eleven of 25 cases were treated medically and in nine others, surgical resection was followed by treatment with antibiotics . In conclusion, when a middle-aged male patient presents with haemoptysis and cough, together with radiologic findings of a peripheral mass or nodule with/without central low attenuation, pulmonary actinomycosis should be suspected.

Chest, 1999 May, 115(5), 1312 - 5
Empyema complicating successful lung transplantation; Nunley DR et al.; OBJECTIVE: To assess the prevalence and etiology of empyema complicating successful lung transplantation . DESIGN: Retrospective review . SETTING: University medical center transplant service . PATIENTS: All recipients (n = 392) of single-lung, double-lung, and heart-lung transplantation between May 1984 and April 1997 . RESULTS: Of the 392 transplant recipients, empyema was documented in 14 patients (3.6%) at a mean time (+/- SD) of 46 days after transplantation (range, 14 to 167 days) . Of these 14 recipients with empyema, 4 recipients (28.6%) died of infectious complications related to empyema . Empyema was seen secondary to Gram-positive, Gram-negative, and saprophytic organisms; however, there was no predominance of a particular organism recovered from the empyemic fluid (chi2 = 0.53; p = 0.75) . The development of empyema was not related to whether the transplant was performed secondary to a septic or nonseptic lung disorder (chi2 = 1.06; p = 0.67), nor was it related to the type of transplant procedure performed (ie, single-lung, double-lung, or heart-lung allografts; chi2 = 4.39; p = 0.30) . CONCLUSION: Empyema, a relatively uncommon complication of lung transplantation, is not related to the type of allograft received or to whether the recipient had a septic or a nonseptic lung disorder . If empyema does occur, the mortality associated with this infection is substantial.

J Fr Ophtalmol, 1999 Mar, 22(2), 213 - 4
{Intraocular contamination during cataract surgery according to surgical technique and type of implant}; Feys J et al.; PURPOSE: To determine wether the cataract extraction method and intraocular lens material affect bacterial contamination of the eye during surgery . METHODS: This retrospective study evaluated microbial contamination of the anterior chamber fluid in three group of patients . Group 1: extracapsular extraction and PMMA lens (354 cases) . Group 2: phacoemulsification and PMMA lens (474 cases) . Group 3: phacoemulsification and silicone lens with PMMA haptic (264 cases) . RESULTS: Microbial contamination rate was in group 1: 5.65%, in group 2: 4.75%, and in group 3: 4.54% . The difference was not statistically significant (chi: 0.321) . All contaminants were Gram positive . CONCLUSION: Bacterial contamination rate of the anterior chamber is similar during extracapsular extraction and phacoemulsification, and is not modified when using PMMA or silicone lens.

Curr Opin Microbiol, 1999 Apr, 2(2), 153 - 8
Regulation of the heat-shock response; Yura T et al.; Current models of both heat induction and the chaperone-mediated feedback control of the sigma32 regulon in Escherichia coli have been further substantiated, and the extent of conservation among Gram-negative bacteria has been assessed . Analyses of the 'CIRCE' and other regulons or operons in Gram-positive and Gram-negative bacteria have provided new insights into their significance and regulatory mechanisms.

J Bacteriol, 1999 May, 181(10), 3039 - 50
Identification and transcriptional control of the genes encoding the Caulobacter crescentus ClpXP protease; Osteras M et al.; The region of the Caulobacter crescentus chromosome harboring the genes for the ClpXP protease was isolated and characterized . Comparison of the deduced amino acid sequences of the C . crescentus ClpP and ClpX proteins with those of their homologues from several gram-positive and gram-negative bacteria revealed stronger conservation for the ATPase regulatory subunit (ClpX) than for the peptidase subunit (ClpP) . The C . crescentus clpX gene was shown by complementation analysis to be functional in Escherichia coli . However, clpX from E . coli was not able to substitute for the essential nature of the clpX gene in C . crescentus . The clpP and clpX genes are separated on the C . crescentus chromosome by an open reading frame pointing in the opposite direction from the clp genes, and transcription of clpP and clpX was found to be uncoupled . clpP is transcribed as a monocistronic unit with a promoter (PP1) located immediately upstream of the 5' end of the gene and a terminator structure following its 3' end . PP1 is under heat shock control and is induced upon entry of the cells into the stationary phase . At least three promoters for clpX (PX1, PX2, and PX3) were mapped in the clpP-clpX intergenic region . In contrast to PP1, the clpX promoters were found to be downregulated after heat shock but were also subject to growth phase control . In addition, the clpP and clpX promoters showed different activity patterns during the cell cycle . Together, these results demonstrate that the genes coding for the peptidase and the regulatory subunits of the ClpXP protease are under independent transcriptional control in C . crescentus . Determination of the numbers of ClpP and ClpX molecules per cell suggested that ClpX is the limiting component compared with ClpP.

Clin Ther, 1999 Mar, 21(3), 513 - 22
Absolute bioavailability of moxifloxacin; Ballow C et al.; Moxifloxacin (BAY 12-8039) is an investigational 8-methoxy-fluoroquinolone with broad-spectrum gram-positive and gram-negative activity . To determine the absolute bioavailability of moxifloxacin, this open-label, randomized, crossover study compared the pharmacokinetic characteristics of a single 100-mg dose administered either orally or intravenously as a 60-minute infusion in 10 healthy male volunteers (mean age {+/- SD}, 29.3+/-7.1 years; mean weight {+/- SD}, 77.7+/-8.7 kg) . Geometric mean values for oral/IV moxifloxacin were as follows: peak serum concentration, 1.15/1.34 mg/L, and area under the concentration-time curve over 48 hours, 9.86/10.89 mg x h/L . The geometric mean absolute bioavailability of oral moxifloxacin was 91.8% . Mean renal clearance was approximately 2.3 L/h after administration of both the single oral and IV formulations, which suggests lack of active tubular secretion of moxifloxacin . Both the oral and IV formulations were well tolerated, with 5 reported possible or probable drug-related adverse events; they included headache, nausea, and localized urticaria . In summary, a single oral dose of moxifloxacin was extensively absorbed in healthy young men . Further studies are necessary in actual patients to confirm the viability of IV to oral conversion at the same dose of moxifloxacin.

Int J Syst Bacteriol, 1999 Apr, 49 Pt 2, 635 - 8
Facklamia sourekii sp . nov., isolated from human sources; Collins MD et al.; Two strains of a Gram-positive catalase-negative, facultatively anaerobic coccus originating from human sources were characterized by phenotypic and molecular taxonomic methods . The strains were found to be identical to each other based on 16S rRNA gene sequencing and constitute a new subline within the genus Facklamia . The unknown bacterium was readily distinguished from Facklamis hominis and Facklamia ignava 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 Facklamia sourekii sp . nov., the type strain of which is CCUG 28783AT.

J Biol Chem, 1999 May 14, 274(20), 14012 - 20
Bacterial peptidoglycan induces CD14-dependent activation of transcription factors CREB/ATF and AP-1; Gupta D et al.; Peptidoglycan (PGN), the major cell wall component of Gram-positive bacteria, induces secretion of cytokines in macrophages through CD14, the pattern recognition receptor that binds lipopolysaccharide and other microbial products . To begin to elucidate the mechanisms that regulate the transcription of cytokine genes, we wanted to determine which transcription factors are activated by PGN in mouse RAW264.7 and human THP-1 macrophage cells . Our results demonstrated that: (i) PGN induced phosphorylation of the transcription factors ATF-1 and CREB; (ii) ATF-1 and CREB bound DNA as a dimer and induced transcriptional activation of a CRE reporter plasmid, which was inhibited by dominant negative CREB and ATF-1; (iii) PGN induced phosphorylation of c-Jun, protein synthesis of JunB and c-Fos, and transcriptional activation of the AP-1 reporter plasmid, which was inhibited by dominant negative c-Fos; and (iv) PGN-induced activation of CREB/ATF and AP-1 was mediated through CD14 . This is the first study to demonstrate activation of CREB/ATF and AP-1 transcription factors by PGN or by any other component of Gram-positive bacteria.

Ter Arkh, 1999, 71(3), 35 - 8
{Clinical, bactericidal and pharmacological evaluation of the effects of netromycin}; Sokolova VI et al.; AIM: To analyze clinical, bactericidal effectiveness and pharmacokinetics of wide-spectrum antibiotic netromycin (NM) . MATERIALS AND METHODS: The trial entered 29 patients: 21 with bronchopulmonary diseases (pneumonia and chronic bronchitis), 5 with exacerbation of chronic pyelonephritis, 2 with infectious endocarditis and 1 with peritonsillar abscess . RESULTS: Microbiologically, most of the agents (80.36%) showed sensitivity to NM . Pharmacologically, NM persisted long in blood serum and sputum irrespective of the administration mode . Positive clinical dynamics after NM treatment was achieved in all the patients but one who had a peritonsillar abscess . CONCLUSION: NM is highly active against both gram-positive and gram-negative flora . Side effects are minimal.

J Infect Dis, 1999 Jun, 179(6), 1459 - 68
Bacterial peptidoglycan polysaccharides in sterile human spleen induce proinflammatory cytokine production by human blood cells; Schrijver IA et al.; Peptidoglycan (PG) is the major component of the cell wall of gram-positive bacteria . In vitro, PG isolated from conventional bacterial cultures can induce secretion of proinflammatory cytokines by human monocytes, indicating that PG may be involved in immune responses against infections by gram-positive bacteria . To investigate the biologic activity of PG in human tissues, an improved method was developed to isolate significant amounts of PG from sterile human spleen tissue . Biochemical analysis demonstrated that PG isolated from human spleen is largely intact . Human whole blood cell cultures were able to produce the proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1 and -6 after stimulation with PG isolated from human spleen . Cytokine induction was not sensitive to inhibition by polymyxin B, in contrast to lipopolysaccharide . Collectively, the data show that intact PG in sterile human tissue is biologically active and may induce local proinflammatory cytokine production.

J Immunoassay, 1999 Feb-May, 20(1-2), 79 - 89
Differentiation between endotoxin and non-endotoxin pyrogens in human albumin solutions using an ex vivo whole blood culture assay; Pool EJ et al.; Purified E.coli endotoxin, Gram negative bacteria and Gram positive bacteria induce IL-6 secretion by whole blood cultures (WBC's) . Polymyxin B at concentrations greater than 2 U/ml completely inhibits IL-6 secretion caused by 10 EU/ml of endotoxin . Polymyxin B has no effect on IL-6 secretion by WBC's in the absence of endotoxin . The inhibition of endotoxin induced IL-6 secretion is Polymyxin B concentration dependent at concentrations less than 1 U/ml . IL-6 induction caused by E.coli is only partially inactivated by 8 U/ml Polymyxin B . Polymyxin B has no effect on IL-6 secretion caused by B.subtilis . Two pyrogenic batches of human serum albumin (HSA), as tested by the rabbit assay for pyrogens, were also investigated . Polymyxin B at 4 U/ml inhibits less than 40 % of IL-6 secretion caused by these pyrogenic HSA batches . All the endotoxin activity in HSA samples spiked with purified endotoxin is inhibited by Polymyxin B indicating that HSA does not protect endotoxin against Polymyxin B inhibition . These results indicate that the pyrogenicity of these HSA batches are caused by Polymyxin B inhibitable and non-inhibitable fractions . This study shows that pyrogenic substances other than endotoxin can contaminate batches of pharmaceutical products and that results obtained using the Limulus Amoebocyte Lysate (LAL) assay does not necessarily indicate the pyrogenic status of pharmaceutical products . The WBC assay for pyrogens, having a broader sensitivity range than the LAL assay, is a better indicator of the pyrogenic status of pharmaceutical products.

Bone Marrow Transplant, 1999 Mar, 23(6), 599 - 605
Neutropenic infections in 100 patients with non-Hodgkin's lymphoma or Hodgkin's disease treated with high-dose BEAM chemotherapy and peripheral blood progenitor cell transplant: out-patient treatment is a viable option; Seropian S et al.; A retrospective analysis was performed on 100 patients with non-Hodgkin's lymphoma (NHL, n = 75) or Hodgkin's disease (HD, n = 25) who underwent peripheral blood progenitor cell transplant (PBPCT) following high-dose chemotherapy (HDCT) with BCNU, etoposide, cytarabine and melphalan (BEAM) between March 1994 and June 1997 . Following PBPCT and until engraftment all patients received oral ciprofloxacin and fluconazole, patients with positive Herpes simplex virus serology received acyclovir and 91 patients received filgrastim . The median days of neutropenia and days to an absolute neutrophil count (ANC) >500/mm3 were 6 and 9, respectively . Febrile neutropenia occurred in 68 patients . Gram-positive bacteremia occurred in 14 patients . No gram-negative infections, invasive fungal infections, intensive care visits or deaths occurred during the period of neutropenia or in the first 30 days following transplant . In multivariate logistic regression the risk of development of any infection was associated only with the duration of neutropenia (P = 0.02) and the risk of bacteremia was associated only with the number of CD34+ cells infused (P = 0.046) . Among 49 patients treated in the outpatient setting, 14 (28%) were never admitted . High-dose chemotherapy with BEAM supported by PBPCT, prophylactic antibiotics and filgrastim resulted in a low incidence of infections and no acute mortality . WBC engraftment occurred rapidly allowing for a predictable course during which lengthy hospital stays and amphotericin therapy could be avoided.

J Vet Pharmacol Ther, 1999 Feb, 22(1), 35 - 40
Comparison of plasma pharmacokinetics and bioavailability of ceftiofur sodium and ceftiofur hydrochloride in pigs after a single intramuscular injection; Brown SA et al.; Ceftiofur sodium, a broad-spectrum cephalosporin, is active against gram-positive and gram-negative pathogens of veterinary importance . Two studies were designed to compare the intramuscular bioavailability of the current sodium salt and the new hydrochloride salt in pigs at doses of either 3 mg or 5 mg ceftiofur equivalents (CE)/kg body weight . Twenty-six healthy young pigs were selected for these two-period, two-treatment crossover studies, 12 for the 3 mg/kg study and 14 for the 5 mg/kg study . Each animal received one intramuscular (i.m.) injection of ceftiofur sodium and one i.m . injection of ceftiofur hydrochloride with a 14-day washout period between the two treatments . Blood samples were collected serially for up to 96 h postinjection . Plasma samples were then analysed using a validated assay that measures ceftiofur and all desfuroylceftiofur-related metabolites by high-performance liquid chromatography . In the 3 mg/kg dosage study, average maximum plasma concentration (C(max)) after administration of ceftiofur sodium was 15.8+/-3.40 microg/mL at 0.4-4 h after injection . After administration of ceftiofur hydrochloride, the C(max) was 11.8+/-1.67 microg/mL at 1-4 h after injection . Concentrations of ceftiofur and metabolites 72 h after the injection were 0.392+/-0.162 microg/mL for ceftiofur hydrochloride and 0.270+/-0.118 microg/mL for ceftiofur sodium . The mean area under the curve (AUC), from time 0 to the limit of quantitation (AUC(O-LOQ)) after ceftiofur hydrochloride administration, was 216+/-28.0 microg x h/mL, compared to 169+/-45.4 microg x h/mL after ceftiofur sodium administration . The calculated time during which plasma concentrations remained above 0.02 microg/mL (t(>0.2)) was 85.3+/-10.6 h for ceftiofur sodium and 77.2+/-10.7 h for ceftiofur hydrochloride . In the 5 mg/kg dosage study, C(max) after administration of ceftiofur sodium was 28.3+/-4.45 microg/mL at 0.33-2 h after injection . After administration of ceftiofur hydrochloride, the C(max) was 29.7+/-6.72 microg/mL at 0.66-2 h after injection . Concentrations of ceftiofur and metabolites 96 h after the injection were 0.274+/-0.0550 microg/mL for ceftiofur hydrochloride and 0.224+/-0.0350 microg/mL for ceftiofur sodium . The mean AUC(O-LOQ) after ceftiofur hydrochloride administration was 382+/-89.8 microg x h/mL compared to 302+/-54.4 microg x h/mL after ceftiofur sodium administration . The t(>0.2) was 78.9+/-9.65 h for ceftiofur sodium and 94.2+/-8.64 h for ceftiofur hydrochloride . Based on the similarity of the pharmacokinetic parameters of the sodium and hydrochloride formulations of ceftiofur, similar therapeutic efficacy can be inferred for the two products.

Gene, 1999 Feb 18, 227(2), 125 - 35
Isolation and characterization of the naphthocyclinone gene cluster from Streptomyces arenae DSM 40737 and heterologous expression of the polyketide synthase genes; Brunker P et al.; Streptomyces arenae produces the aromatic polyketide naphthocyclinone, which exhibits activity against Gram-positive bacteria . A cosmid clone containing the putative naphthocyclinone gene cluster was isolated from a genomic library of S . arenae by hybridization with a conserved region from the actinorhodin PKS of S . coelicolor . Sequence analysis of a 5.5-kb DNA fragment, which hybridizes with the actI probe, revealed three open reading frames coding for the minimal polyketide synthase . A strong sequence similarity was found to several previously described ketosynthases, chain length factors and acyl carrier proteins from other polyketide gene clusters . An additional open reading frame downstream of the PKS genes of S . arenae showed 53% identity to act VII probably encoding an aromatase . Another open reading frame was identified in a region of 1.436 bp upstream of the PKS genes, which, however, had no similarity to known genes in the database . Approximately 8 kb upstream of the PKS genes, a DNA fragment was identified that hybridizes to an actVII--actIV specific probe coding for a cyclase and a putative regulatory protein, respectively . Disruption of the proposed naphthocyclinone gene cluster by insertion of a thiostrepton resistance gene completely abolished production of naphthocyclinones in the mutant strain, showing that indeed the naphthocyclinone gene cluster had been isolated . Heterologous expression of the minimal PKS genes in S . coelicolor CH999 in the presence of the act ketoreductase led to the production of mutactin and dehydromutactin, indicating that the S . arenae polyketide synthase forms a C-16 backbone that is subsequently dimerized to build naphthocyclinone . The functions of the proposed cyclase and aromatase were examined by coexpression with genes from different polyketide core producers.

Med Microbiol Immunol (Berl), 1999 Mar, 187(3), 173 - 81
Layered murein revisited: a fundamentally new concept of bacterial cell wall structure, biogenesis and function; Dmitriev BA et al.; The classical concept of the architecture of microbial murein assumes cross-linked glycan chains to be arranged in horizontal layers outside of the plasma membrane . It necessitates elaborate hypotheses to explain processes such as the biosynthesis, growth and division of the bacterial cell wall and provides no explanation for transenvelope macromolecular transport . Moreover, this model is difficult to reconcile with a number of basic chemical and electron microscopical data . According to a fundamentally distinct concept which is presented here, glycan strands in the microbial wall run perpendicular to the plasma membrane, each strand being cross-linked by peptide bridges with four other strands . This arrangement allows the formation of a structured matrix pierced with ordered ionophoric channels potentially harboring either lipoprotein or teichoic (lipoteichoic) acid molecules in Gram-negative and Gram-positive bacteria, respectively . New wall structures are synthesized in toto emerging from the cytoplasmic membrane as a condensed gel-like network below the old wall without being covalently attached to it, expanding due to inherent elasticity as the old wall is lyzed . This model reflects published genetic and biochemical data and offers a simple explanation for peptidoglycan biogenesis . As the biosynthesis is terminated by enzymic cleavage of all glycan strands, murein is irreversibly released from the membrane . The murein detachment prepares the membrane for de novo assembly of both the new wall synthesis machinery and the multicomponent factory for protein, DNA and phospholipid transfer . Being assembled in parallel, both new murein and the traffic complexes grow from the membrane together . This concept eliminates the necessity for the traffic complexes to penetrate intact murein . In the process of simultaneous assembly, the expanding murein functions as a lifting platform driven by the force of turgor pressure, transporting macromolecules through the perisplasmic space.

Science, 1999 Apr 16, 284(5413), 507 - 11
Vancomycin derivatives that inhibit peptidoglycan biosynthesis without binding D-Ala-D-Ala; Ge M et al.; Vancomycin is an important drug for the treatment of Gram-positive bacterial infections . Resistance to vancomycin has begun to appear, posing a serious public health threat . Vancomycin analogs containing modified carbohydrates are very active against resistant microorganisms . Results presented here show that these carbohydrate derivatives operate by a different mechanism than vancomycin; moreover, peptide binding is not required for activity . It is proposed that carbohydrate-modified vancomycin compounds are effective against resistant bacteria because they interact directly with bacterial proteins involved in the transglycosylation step of cell wall biosynthesis . These results suggest new strategies for designing glycopeptide antibiotics that overcome bacterial resistance.

Jpn J Antibiot, 1999 Jan, 52(1), 57 - 67
Comparative studies on in vitro activities of kasugamycin and clinically-used aminoglycoside antibiotics; Tamamura T et al.; Kasugamycin, a unique aminoglycoside antibiotic, has been used for many years solely for crop protections . In general, aminoglycoside antibiotics possess broad and strong inhibitory activity against both Gram-positive and -negative bacteria, however, kasugamycin merely exhibits limited activity against phytopathogenic microbes such as Pyricularia oryzae and certain strains of pseudomonads . Recently, in human and animal chemotherapy, it has been seriously concerned about the emergence of multiply resistant bacteria by consumption of a large amount of antibiotics not only for therapy but also for growth-promotion of farm animals . It was believed that kasugamycin, the agricultural antibiotic, does not undergo any cross-resistance with other clinically important aminoglycoside antibiotics because of its weak or almost no activity against common pathogenic microbes except for some phytopathogenic fungi and pseudomonads . However, no confirmative study on this fact has been published so far . In this study, we compared activity of kasugamycin with those of twelve clinically used aminoglycoside antibiotics against susceptible standard strains and well-characterized aminoglycoside-resistant strains as well as clinically isolated strains, in order to show the least potential of kasugamycin to create cross-resistant human pathogens against clinically important aminoglycoside antibiotics.

Bioorg Med Chem Lett, 1999 Mar 8, 9(5), 673 - 8
Synthesis and activity of 2-(sulfonamido)methyl-carbapenems: discovery of a novel, anti-MRSA 1,8-naphthosultam pharmacophore; Wilkening RR et al.; A series of 1beta-methyl carbapenems substituted at the 2-position with lipophilic, acyclic and cyclic (sulfonamido)methyl groups was prepared and evaluated for activity against resistant gram-positive bacteria . From these studies, the 1,8-naphthosultamyl group emerged as a novel, PBP2a-binding, anti-MRSA pharmacophore worthy of further exploration.

Arch Microbiol, 1999 Feb, 171(3), 173 - 82
The cell wall porin of the gram-positive bacterium Nocardia asteroides forms cation-selective channels that exhibit asymmetric voltage dependence; Riess FG et al.; Detergent-solubilized cell wall extracts of the gram-positive, strictly aerobic bacterium Nocardia asteroides contain channel-forming activity as judged from reconstitution experiments using lipid bilayer membranes . The cell wall porin was identified as a protein with an apparent molecular mass of about 84 kDa based on SDS-PAGE . The porin was purified to homogeneity using preparative SDS-PAGE . The 84-kDa protein was no longer observed after heating in SDS buffer . The presumed dissociation products were not observed on SDS-polyacrylamide gels . The cell wall porin increased the specific conductance of artificial lipid bilayer membranes from phosphatidylcholine/phosphatidylserine mixtures by the formation of cation-selective channels, which had an average single-channel conductance of 3.0 nS in 1 M KCl . The single-channel conductance was only moderately dependent on the bulk aqueous KCl concentration, which indicated negative point charge effects on the channel properties . The analysis of the concentration dependence of the single-channel conductance using the effect of negative charges on channel conductance suggested that the diameter of the cell wall channel is about 1.4 nm . Asymmetric addition of the cell wall porin to lipid bilayer membranes resulted in an asymmetric voltage dependence . The cell wall channel switched into substates, when the cis side of the membrane, the side of the addition of the protein, had negative polarity . Positive potentials at the cis side had no influence on the conductance of the cell wall channel.

Arch Microbiol, 1999 Feb, 171(3), 159 - 65
Cell wall of Thermoanaerobacterium thermosulfurigenes EM1: isolation of its components and attachment of the xylanase XynA; Brechtel E et al.; Thermoanaerobacterium thermosulfurigenes EM1 has a gram-positive type cell wall completely covered by a surface layer (S-layer) with hexagonal lattice symmetry . The components of the cell envelope were isolated, and the S-layer protein was purified and characterized . S-layer monomers assembled in vitro into sheets with the same hexagonal symmetry as in vivo . Monosaccharide analysis revealed that the S-layer is associated with fucose, rhamnose, mannosamine, glucosamine, galactose, and glucose . The N-terminal 31 amino acid residues of the S-layer protein showed significant similarity to SLH (S-layer homology) domains found in S-layer proteins of different bacteria and in the exocellular enzymes pullulanase, polygalacturonate hydrolase, and xylanase of T . thermosulfurigenes EM1 . The xylanase from T . thermosulfurigenes EM1 was copurified with the S-layer protein during isolation of cell wall components . Since SLH domains of some structural proteins have been shown to anchor these proteins noncovalently to the cell envelope, we propose a common anchoring mechanism for the S-layer protein and exocellular enzymes via their SLH domains in the peptidoglycan-containing layer of T . thermosulfurigenes EM1.

J Bacteriol, 1999 Apr, 181(8), 2485 - 91
Cloning of fibA, encoding an immunogenic subunit of the fibril-like surface structure of Peptostreptococcus micros; Kremer BH et al.; Although we are currently unaware of its biological function, the fibril-like surface structure is a prominent characteristic of the rough (Rg) genotype of the gram-positive periodontal pathogen Peptostreptococcus micros . The smooth (Sm) type of this species as well as the smooth variant of the Rg type (RgSm) lack these structures on their surface . A fibril-specific serum, as determined by immunogold electron microscopy, was obtained through adsorption of a rabbit anti-Rg type serum with excess bacteria of the RgSm type . This serum recognized a 42-kDa protein, which was subjected to N-terminal sequencing . Both clones of a lambdaTriplEx expression library that were selected by immunoscreening with the fibril-specific serum contained an open reading frame, designated fibA, encoding a 393-amino-acid protein (FibA) . The 15-residue N-terminal amino acid sequence of the 42-kDa antigen was present at positions 39 to 53 in FibA; from this we conclude that the mature FibA protein contains 355 amino acids, resulting in a predicted molecular mass of 41,368 Da . The putative 38-residue signal sequence of FibA strongly resembles other gram-positive secretion signal sequences . The C termini of FibA and two open reading frames directly upstream and downstream of fibA exhibited significant sequence homology to the C termini of a group of secreted and surface-located proteins of other gram-positive cocci that are all presumably involved in anchoring of the protein to carbohydrate structures . We conclude that FibA is a secreted and surface-located protein and as such is part of the fibril-like structures.

Can J Hosp Pharm, 1994 Apr, 47(2), 59 - 64
A vancomycin drug use evaluation and economic analysis in a cancer treatment centre; Dranitsaris G et al.; Princess Margaret Hospital is a 140-bed university affiliated cancer treatment centre . Vancomycin was the only formulary agent available for the treatment of methicillin-resistant gram-positive organisms . The high cost and potential toxicity of this drug warranted a closer examination of its use . The purpose of this study was to evaluate vancomycin use and to determine the economic impact when it was used contrary to newly developed hospital guidelines . A sample of 100 vancomycin orders was randomly selected from all prescriptions filled in 1992 . The indication, dose, and duration of therapy for each order were compared against the hospital guidelines . The cost savings associated with altering the sample of prescriptions to meet hospital guidelines were then determined . Nine percent of the prescriptions were for nonapproved indications . The actual dose used did not meet criteria in 32% of cases and the length of therapy was beyond the approved duration in 45% of the orders . If the cases had been altered to meet the guidelines then a total savings of $13,581 would have been realized . The projected savings for the entire year (1992) would have been $100,907 . The critical problem areas in vancomycin prescribing were the duration of therapy and dose . The results have provided the impetus to initiate a hospital wide prospective Drug Utilization Evaluation (DUE) study to optimize vancomycin prescribing . The program costs would be easily covered by the expected savings.

Appl Environ Microbiol, 1999 Apr, 65(4), 1731 - 7
Wide distribution and diversity of members of the bacterial kingdom Acidobacterium in the environment; Barns SM et al.; To assess the distribution and diversity of members of the recently identified bacterial kingdom Acidobacterium, members of this kingdom present in 43 environmental samples were surveyed by PCR amplification . A primer designed to amplify rRNA gene sequences (ribosomal DNAs {rDNAs}) from most known members of the kingdom was used to interrogate bulk DNA extracted from the samples . Positive PCR results were obtained with all temperate soil and sediment samples tested, as well as some hot spring samples, indicating that members of this kingdom are very widespread in terrestrial environments . PCR primers specific for four phylogenetic subgroups within the kingdom were used in similar surveys . All four subgroups were detected in most neutral soils and some sediments, while only two of the groups were seen in most low-pH environments . The combined use of these primers allowed identification of a novel lineage within the kingdom in a hot spring environment . Phylogenetic analysis of rDNA sequences from our survey and the literature outlines at least six major subgroups within the kingdom . Taken together, these data suggest that members of the Acidobacterium kingdom are as genetically and metabolically diverse, environmentally widespread and perhaps as ecologically important as the well-known Proteobacteria and gram-positive bacterial kingdoms.

Infect Control Hosp Epidemiol, 1999 Mar, 20(3), 187 - 91
Epidemiology of device-associated infections related to a long-term implantable vascular access device; Sotir MJ et al.; OBJECTIVE: To examine risk factors for, and determine the incidence of, device-associated infections among patients with an implantable vascular access device . SETTING: Grady Health System, including a 1,000-bed, inner-city, public, teaching hospital and human immunodeficiency virus (HIV), oncology, and sickle cell clinics in Atlanta, Georgia . PATIENTS: 123 consecutive patients who received a PAS-Port implantable venous access device between January 1 and June 30, 1995 . DESIGN: Retrospective cohort study with follow-up > or = 1 year following device implantation . RESULTS: Underlying illnesses included HIV infection in 66 patients (median CD4 count, 24.4 cells/mm3), malignancy in 51, and sickle cell disease in 6 . Mean age of patients was 43.7 years, 50% were male, and 74% were black . Thirty-one (25%) of 123 patients developed a primary or device-associated bloodstream infection (BSI), and 3 of the 31 patients experienced two separate episodes of infection . The overall rate of infection was 1.23 primary BSIs per 1,000 device days . Patients with cancer had a lower rate of infection than those with HIV infection, but the difference was not statistically significant (0.96 vs 1.50 BSIs/1,000 device days; relative risk, 0.58; 95% confidence interval, 0.27-1.26) . Subgroup analysis of patients with different malignancies indicated that infection rates differed according to type of cancer, and there was a trend for heterogeneity across the different cancer strata (P=.06) . Gram-positive pathogens accounted for 60% of the pathogens recovered . Six (19%) of 31 patients who developed an infection did so within the first 14 days after implantation . In 11 (32%) of the 34 BSIs, the port required removal; two patient deaths were attributed to device-associated bacteremias (0.072 deaths/1,000 device days) . CONCLUSIONS: Approximately one fourth of patients who had a vascular access device implanted developed a primary BSI, but the overall infection rate (per 1,000 device days) was relatively low, even among those with HIV infection . Primary BSI rates in patients with vascular access devices appeared to differ according to the specific underlying illness.

Hautarzt, 1999 Feb, 50(2), 91 - 7
{The role of bacterial superantigens in pathophysiology of the skin}; Boehncke WH et al.; In contrast to conventional antigens, bacterial superantigens activate a large percentage of an organism's total T-cell repertoire . This has clinical implications particularly in the case of infections with Gram-positive bacteria . Unravelling the molecular mechanisms of superantigen-mediated T-cell activation has yielded new insights in the pathogenesis of a number of diseases, in the field of dermatology namely atopic dermatitis and psoriasis . Moreover, therapeutic applications of superantigens have begun to emerge.

J Commun Dis, 1998 Sep, 30(3), 167 - 70
Evaluation of efficacy of Dettol-H in hospital use; Kapoor H et al.; A study was conducted at Safdarjang Hospital, New Delhi, to elucidate the bactericidal effect of a new disinfectant having benzalkonium chloride (40%) + disodium edetate (1.5%) and commercially available as Dettol-H, in comparison with that of a disinfectant already in-use in this hospital and having chlorhexidine gluconate (7.5%) + cetrimide (15%) + isopropyl alcohol (commercially available as Alpilon) . The modified Capacity test of Kelsey-Sykes and In-use test of Kelsey-Maurer were carried out using control strains and random gram positive and gram negative bacterial isolates from clinical specimens . The two disinfectant solutions were analysed at three dilutions viz . weak, recommended and strong . From the study it could be inferred that both Dettol-H and Alpilon were equally effective in recommended and strong dilutions against the organisms tested . Weak dilutions were not effective in both and hence Dettol-H and Alpilon are effective only if used in prescribed concentrations.

Bioorg Med Chem Lett, 1999 Feb 8, 9(3), 375 - 80
Synthesis and structure-activity relationships of a series of novel thiazoles as inhibitors of aminoacyl-tRNA synthetases; Yu XY et al.; A series of novel aminoacyl adenylate mimics has been prepared and evaluated for their inhibitory activity against aminoacyl-tRNA synthetases . Several of these thiazole derivatives displayed potent and selective enzyme activity against both Gram-positive and Gram-negative bacteria.

Plasmid, 1999 Mar, 41(2), 128 - 34
Nucleotide sequence analysis of pRS1, a cryptic plasmid from Oenococcus oeni; Alegre MT et al.; A new cryptic plasmid, pRS1, from an Oenococcus oeni strain isolated from Spanish wines is reported . Nucleotide sequence analysis (2523 bp) revealed the presence of three major open reading frames (ORFs) whose nucleotide sequence and encoded proteins exhibit high homology with those of pOg32, a previously described plasmid of O . oeni . Common features in other plasmids from O . oeni (i.e., pLo13 and pOg32) have been found in pRS1 . They have three major ORFs in the same strand; the putative encoded proteins by two of these ORFs exhibit homology with the replication (Rep) and the recombination (Pre) proteins, respectively, of the pT181 plasmid family and related gram-positive bacteria plasmids; these plasmids contain the DNA sequences required for plasmid replication by the rolling circle mechanism and for recombination (i.e., double-strand origin, DSO; single-strand origin, SSO; recombination-specific sites, RSA and RSB); and finally, all these plasmids have a third ORF of unknown function . These features suggest that pRS1 could constitute together with pLo13 and pOg32 a family of small cryptic plasmids of O . oeni .

Eur J Med Res, 1999 Mar 26, 4(3), 126 - 30
Effects of topical honey on post-operative wound infections due to gram positive and gram negative bacteria following caesarean sections and hysterectomies; Al-Waili NS et al.; The possible therapeutic effect of topical crude undiluted honey in the treatment of severe acute postoperative wound infections was studied . Fifty patients having postoperative wound infections following caesarean sections or total abdominal hysterectomies with gram positive or gram negative bacterial infections were allocated in two groups . Twenty-six patients (group A) were treated with 12 hourly application of crude honey and 24 patients (group B) were treated with local antiseptics: spirit (70% Ethanol) and povidone-iodine . Both groups received systemic antibiotics according to culture and sensitivity . Results showed that eradication of bacterial infections was obtained after 6 +/- 1.9 days (mean +/- SD) in group A and after 14.8 +/- 4.2 days in group B (p <0.05) . Period for antibiotics use was 6.88 +/- 1.7 days in-group A and 15.45 +/- 4 . 37 in-group B (p <0.05) . Complete wound healing was evident after 10 . 73 +/- 2.5 days in group A and after 22.04 +/- 7.33 in group B (p <0 . 05) . Size of postoperative scar was 3.62 +/- 1.4 mm after using topical honey and was 8.62 +/- 3.8 mm after local antiseptics (p <0 . 05) . The mean hospital stay was 9.36 +/- 1.8 days in group A and 19 . 91 +/- 7.35 days in group B (p <0.05) . After using honey, 22/26 patients (84.4%) showed complete wound healing without wound disruption or need for re-suturing and only 4 patients showed mild dehiscence . In group B, 12/24 patients (50%) showed complete wound healing and 12 patients showed wound dehiscence, six of them needed re-suturing under general anesthesia . We concluded that topical application of crude undiluted honey could (1) faster eradication of bacterial infections, (2) reduce period of antibiotic use and hospital stay, (3) accelerate wound healing, (4) prevent wound dehiscence and need for re-suturing and (5) result in minimal scar formation.

Infect Immun, 1999 Apr, 67(4), 1646 - 51
Immunological characterization of a protective antigen of Erysipelothrix rhusiopathiae: identification of the region responsible for protective immunity; Shimoji Y et al.; The gene encoding a protective protein antigen of the gram-positive bacterium Erysipelothrix rhusiopathiae, an important veterinary pathogen responsible for erysipelas in swine and a variety of diseases in animals, was cloned and sequenced . The gene encodes a polypeptide of 597 amino acids plus a putative signal sequence of 29 amino acids, resulting in a mature protein with a molecular mass of 69,017 Da . Sequence analysis of the gene product revealed a C-terminal region composed of nine tandem repeats of 20 amino acids and a total sequence that is nearly identical to that of the 64-kDa cell surface protein (SpaA) of the bacterium . Because of this similarity, the protein was designated SpaA.1 . In this study, we examined whether the SpaA.1 protein could induce protective antibodies and whether we could identify the region involved in protective immunity . Both the mature SpaA.1 protein and its C-terminal repeat region, but not the N-terminal segment, were expressed in Escherichia coli and purified as a histidine-tagged fusion recombinant protein . Rabbit antiserum raised against the mature SpaA.1 protein passively protected mice from lethal challenge with a virulent homologous strain, Fujisawa-SmR, suggesting that protection is mediated by humoral antibodies . To determine which domain of the SpaA.1 protein is responsible for the observed protection, mice were actively immunized with either the mature SpaA . 1 protein or the C-terminal repeat region and then challenged with Fujisawa-SmR . The result showed that mice immunized with the mature SpaA.1 protein, but not the C-terminal repeat region, were protected, suggesting that the protection-eliciting epitope(s) is located within the N-terminal two-thirds of the SpaA.1 molecule . This was confirmed by passive immunization experiments in which the protective activity of rabbit antiserum, raised against mature SpaA . 1 protein, was not abolished by absorption with the purified recombinant C-terminal repeat region . In addition, antibodies specific for the C-terminal repeat region were unable to protect mice from lethal challenge . These results show that the N-terminal two-thirds of the SpaA.1 molecule may constitute a good vaccine candidate against erysipelas.

J Dent, 1999 Mar, 27(3), 243 - 6
Patient discomfort and cross-infection control in bitewing examination with a storage phosphor plate and a CCD-based sensor; Wenzel A et al.; OBJECTIVES: The aim was to compare a CCD-based sensor and a storage phosphor plate with respect to patient discomfort and the efficacy of a simple cross-infection control procedure in connection with a posterior bitewing examination . METHODS: 130 patients accepted to have one posterior bitewing of the left and right side taken with two digital radiography systems, the Digora phosphor plate and the Trophy RVG XL, CCD-based sensor system . The patients assessed their feeling of discomfort after the examination on a 100-mm . Visual Analogue Scale (VAS) . Microbiological samples were taken from the RVG sensor and cord and from the Digora envelope, plate and scanner during examination of 14 patients . The samples were plated and incubated anaerobically, and the colony-forming units counted . RESULTS: Median VAS score for discomfort was 20 min for Digora and 32 min for the RVG sensor (P < 0.001) . Median total counts of cultivable bacteria were low (< 20), the majority being catalase-positive, Gram-positive cocci and Gram-positive rods, presumably skin bacteria . Only the samples taken from the enveloped Digora plate and the rubber tube coated RVG sensor immediately after exposure yielded large numbers of oral bacteria . CONCLUSION: The phosphor plate was less unpleasant than the CCD sensor . Cross-contamination posed a minor problem for both systems when a simple, standard hygiene procedure was followed.

FEMS Microbiol Lett, 1999 Mar 1, 172(1), 53 - 60
Stimulation of bacterial growth by heat-stable, norepinephrine-induced autoinducers; Freestone PP et al.; The ability of norepinephrine to increase the growth of Escherichia coli in a serum-based medium has previously been shown to be due to the production of an autoinducer of growth during early log phase . Seventeen Gram-negative and 6 Gram-positive clinical isolates were examined for a similar ability to respond to norepinephrine, and to synthesise autoinducer . The majority of Gram-negative strains both produced and responded to heat-stable norepinephrine-induced autoinducers of growth . Most of these autoinducers showed a high degree of cross-species activity, suggesting the existence of a novel family of Gram-negative bacterial signalling molecules . In contrast, although a number of Gram-positive strains were able to respond to norepinephrine, the majority failed to produce autoinducers in the presence of norepinephrine.

J Clin Microbiol, 1999 Apr, 37(4), 1161 - 4
Facklamia languida sp . nov., isolated from human clinical specimens; Lawson PA et al.; Three strains of a gram-positive catalase-negative, facultatively anaerobic coccus-shaped organism originating from human clinical samples were characterized by phenotypic and molecular taxonomic methods . Sequencing of genes encoding 16S rRNA showed that the strains are phylogenetically closely related (99.9 to 100% sequence similarity) and represent a new subline within the genus Facklamia . The unknown bacterium was readily distinguished from all currently described species of the genus Facklamia (viz., Facklamia hominis, Facklamia ignava, and Facklamia sourekii) 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 Facklamia languida sp . nov . The type strain of F . languida is CCUG 37842.

Ann Vasc Surg, 1999 Mar, 13(2), 188 - 90
Mycotic aneurysms of the tibioperoneal arteries; McKee MA et al.; This case report describes a pediatric patient with mycotic aneurysms of both posterior tibial arteries and the right peroneal artery associated with an episode of endocarditis . To our knowledge, this case represents the first reported occurrence of multiple mycotic aneurysms of the tibioperoneal arteries . It is also unique in that the pathogen was Brucella canis instead of the usual gram-positive pathogens associated with intravenous drug injection . Vascular reconstruction can be accomplished; however, management of this complex problem should be individualized.

Curr Opin Microbiol, 1998 Feb, 1(1), 56 - 65
Superantigens of gram-positive bacteria: structure-function analyses and their implications for biological activity; Kotb M; Just as we thought that we know everything about superantigens, new molecular and structural studies indicate that we have only just begun to unravel the secrets of these fascinating molecules . Recent structure-function analysis of superantigens from Gram-positive bacteria, with emphasis on their interaction with major histocompatibility complex molecules, could help us decipher the role of superantigens in disease, identify host factors that potentiate their effects and design drugs that specifically block their activity.

Curr Opin Microbiol, 1998 Feb, 1(1), 49 - 55
Molecular basis of host-pathogen interaction in septic shock; Heumann D et al.; Specific mechanisms of recognition of microbial products have been developed by host cells . Among these mechanisms, recognition of lipopolysaccharide of Gram-negative bacteria by CD14, a glycoprotein expressed at the surface of myelomonocytic cells, plays a major role . There is increasing evidence that CD14 also serves as a receptor for other microbial products including peptidoglycan of Gram-positive bacteria . A common theme is that CD14 represents a key molecule in innate immunity . Recognition of microbial products by host cells leads to cell activation and production of a large array of mediators that are necessary for the development of controlled inflammatory processes . When the activation process is out of control, such as in septic shock, these mediators can be detrimental to the host.

Eur J Clin Microbiol Infect Dis, 1998 Dec, 17(12), 849 - 52
Comparative evaluation of a commercial system for identification of gram-positive cocci; von Baum H et al.; The performance of a new commercial system for the identification of different groups of gram-positive cocci {BBL Crystal Gram-Positive (GP) Identification System; Becton Dickinson Microbiology Systems, Germany} was evaluated in comparison with two currently used commercial systems, the API Staph and the API Strep (bioMerieux Diagnostic, Germany) . A total of 191 strains from seven different gram-positive genera comprising 32 different species were tested . For the BBL Crystal GP system, the correct identification rate without additional tests was 89.5% at the species level and 97.9% at the genus level . The findings suggest that the newly introduced BBL Crystal GP ID system provides an accurate method for the identification of gram-positive cocci, with an overall rate of correct species identification of about 90%, similar to that of the established API systems . Its major advantage is the extended spectrum of taxa included in a single test panel in contrast to the two different API test kits . Furthermore, the simplicity of use and the safe and rapid handling in a closed system conveniently accommodate existing laboratory workflow.

Ann Pathol, 1998 Dec, 18(6), 466 - 72
{Placental infection in Rwanda: comparison an HIV infected population and a control population}; Hofman V et al.; We report an histological study from term placentas of 286 HIV positive women born in Rwanda . We observed chorioamnionitis without any pathogen in 15% of the cases, cocci Gram positive infection in 12 observations and malaria infection in 75% of placentas . We noted 71 cases of active malaria infection with Plasmodium falciparum trophozoites in the erythrocytes of the intervillous spaces, and 135 cases of chronic infection with malaria pigment without any parasite . An ultrastructural study performed in 8 cases of active malaria infection showed characteristic features of trophozoites and schizontes, and malaria pigment . No viral particle were seen . We did not observe any significative difference concerning the incidence of chorioamnionitis and of malaria infection in 275 HIV negative placentas . In the literature as well as in the present study, the main lesions observed in the placentas of AIDS patients were chorioamnionitis . Opportunistic infections and neoplasias of the placenta are exceptional . Detection of HIV proteins by immunochemistry or in situ hybridization is possible, but the HIV could not be identified in the trophoblast by electron microscopy . Mechanisms of the materno-fetal transmission for HIV are currently unknown.

Radiol Med (Torino), 1998 Nov, 96(5), 434 - 8
{Magnetic resonance findings in chronic osteomyelitis fistula}; Capparelli G et al.; INTRODUCTION: Osteomyelitis is a common inflammatory process caused by an infection which is usually from gram-positive germs . Acute, subacute and chronic forms can be distinguished both clinically and radiographically, each of them presenting different patterns but which are not always easy to recognize . Besides clinical-laboratory data, imaging methods are also useful to make the diagnosis; in chronic forms, MRI has higher sensitivity in recognizing the active foci and the presence of sinus tracts than the other techniques . MATERIAL AND METHODS: We examined nine patients with suspected chronic osteomyelitis with a sinus tract, all of them submitted to radiographic and MR examinations . MRI was performed with a .5 T magnet using a surface coil for joint studies; T1-weighted SE (TR 300-500, TE 20), T2-weighted GE (TR 500-700, TE 20, FA 35 degrees) and FIR sequences with fat suppression (TR 3000, TE 20, TI 100) were performed on the axial, sagittal and coronal planes . Three patients underwent CT too . RESULTS: Radiography showed the osteostructural changes as osteolytic and osteosclerotic areas . CT depicted not only the changes seen radiographically, but also increased bone marrow density . MRI demonstrated active foci of bone marrow appearing as low-intensity areas in T1, with increased intensity in T2 GE and fat-suppressed FIR images; it also depicted the sinus tracts as areas of decreased signal on T1 and of increased signal on FIR images . CONCLUSIONS: Based on our experience and in agreement with the literature data, we believe that MRI is more useful to diagnose the active foci of bone marrow and the sinus tracts in chronic osteomyelitis, especially with T1-weighted SE and fat-suppressed FIR sequences.

Methods, 1998 Dec, 16(4), 396 - 406
Identification of genes involved in innate responsiveness to bacterial products by differential display; Jin F et al.; To explore gene regulation by bacterial lipopolysaccharide (LPS), we compared mRNA profiles of macrophage cell lines from two strains of mice congenic for a locus markedly affecting their ability to respond to LPS . Differential display detected four differentially expressed transcripts . One transcript encoded the mouse homolog of human secretory leukocyte protease inhibitor (SLPI), which was expressed by LPS-hyporesponsive macrophage cells (Lps(d)) but not by LPS-normoresponsive cells (Lps(n)) . Among five macrophage cell lines, secretion of SLPI was inversely correlated with ability to produce nitric oxide (NO) and tumor necrosis factor alpha in response to LPS . Stable transfection of LPS-responsive macrophages with SLPI suppressed LPS-induced responses . Interferon-gamma (IFN-gamma), which corrects the defective LPS response in Lps(d) macrophages, suppressed the LPS-induced expression of SLPI and restored LPS response to SLPI-overexpressing macrophages . Besides its role as a LPS response inhibitor, mouse SLPI is also a lipoteichoic acid response inhibitor . The expression of SLPI was strongly enhanced by interleukin-10 and -6 . SLPI may be an important antiinflammatory molecule in host defense against gram-negative and gram-positive bacteria.

Int J Syst Bacteriol, 1999 Jan, 49 Pt 1, 97 - 101
Ignavigranum ruoffiae sp . nov., isolated from human clinical specimens; Collins MD et al.; Two strains of a hitherto undescribed Gram-positive catalase-negative, facultatively anaerobic coccus isolated from human sources were characterized by phenotypic and molecular taxonomic methods . Comparative 16S rRNA gene sequencing studies demonstrated the unknown strains were genealogically identical, and constitute a new line close to, but distinct from, the genera Facklamia and Globicatella . The unknown bacterium was readily distinguished from Facklamia species and Globicatella sanguinus 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 Ignavigranum ruoffiae gen . nov., sp . nov . The type strain of Ignavigranum ruoffiae is CCUG 37658T.

Mol Microbiol, 1999 Jan, 31(2), 609 - 21
Regulation of excision of the conjugative transposon Tn916; Marra D et al.; Excision from the donor DNA molecule is the first step in conjugative transposition of Tn916 and is followed by circularization of the transposon and its transfer to a new host . We have demonstrated that, in Gram-positive hosts, the Xis protein, as well as the site-specific recombinase Int, is required for the excision of Tn916 . Using assays for closure of the excised covalently closed transposon and for repair of the donor DNA molecule, we found that neither protein alone is rate limiting for excision, but overexpression of Int and Xis together results in increased excision . After excision, the frequency of Tn916 circle formation was found to be the same as the frequency of repair of the donor DNA molecule . This suggests that a single reaction results in the closure of both molecules . We have also identified two transcripts that encode Int, one of which also encodes Xis and one of which does not, suggesting that there are steps in conjugative transposition of Tn916 that require Int without Xis.

Mol Microbiol, 1999 Jan, 31(2), 521 - 32
The ClpB ATPase of Streptomyces albus G belongs to the HspR heat shock regulon; Grandvalet C et al.; The clpB gene of Streptomyces albus was cloned by polymerase chain reaction (PCR) using degenerate oligonucleotides . Transcriptional analysis showed that the clpB gene was heat induced . Primer extension identified a transcription start site preceded by typical vegetative -10 and -35 hexamer sequences . The Streptomyces HspR repressor is known to bind to three inverted repeat motifs (IR1, IR2, IR3) upstream from the S . coelicolor dnaK operon . We identified an inverted repeat motif identical to IR3 upstream from the S . albus clpB gene . DNA-binding experiments showed that HspR regulates clpB transcription by interacting directly with this motif . Streptomyces albus is the first Gram-positive organism for which the co-regulation of DnaK and ClpB has been described . Such co-regulation suggests that there is a physiological relationship between these two proteins in this bacterium . Genes similar to hspR were also identified in Mycobacterium leprae, M . tuberculosis and in bacteria unrelated to the actinomycetales order, such as Helicobacter pylori and Aquifex aeolicus . HspR binding sites were found in these bacteria upstream from various heat shock genes, suggesting that these genes are regulated by HspR . The HspR binding site, here called HAIR (HspR associated inverted repeat), has the consensus sequence CTTGAGT N7 ACTCAAG.

Ann Surg, 1999 Feb, 229(2), 246 - 54
HLA-DR expression and soluble HLA-DR levels in septic patients after trauma; Ditschkowski M et al.; OBJECTIVE: To determine if cellular and soluble HLA-DR molecules may be relevant in severely injured patients for the development of gram-positive or gram-negative sepsis . SUMMARY BACKGROUND DATA: HLA-DR molecules play a central role in the specific immune response to infection . The reduced HLA-DR expression on monocytes is considered to correlate with infectious complications and the development of sepsis . Data on the role of HLA-DR expression on T cells and soluble HLA-DR molecules are rare . METHODS: HLA-DR expression on monocytes and T cells was measured by flow cytometry . Plasma levels of soluble HLA-DR were studied by enzyme-linked immunosorbent assay . RESULTS: HLA-DR expression on circulating T cells, calculated as mean fluorescence intensity in channels, was reduced at day 1 after admission in 20 patients with subsequent severe sepsis compared with 46 patients without sepsis . The septic patients immediately after trauma had significantly lower soluble HLA-DR plasma levels than the nonseptic patients . At day 2 after admission, HLA-DR expression on monocytes was significantly lower in the severe sepsis group than in the patients without sepsis, and lasted until day 14 after injury . CONCLUSIONS: In severely injured patients, decreased levels of cellular and soluble HLA-DR appear as early indicators of an immune deviation associated with the development of severe sepsis . Moreover, immune alterations of different cell types may promote distinct kinds of septicemia.

FEMS Microbiol Lett, 1999 Feb 1, 171(1), 1 - 9
Biocontrol of plant disease: a (gram-) positive perspective; Emmert EA et al.; Biological control offers an environmentally friendly alternative to the use of pesticides for controlling plant diseases . Unfortunately, growers continue to use chemical control over biological agents, and lack of knowledge often contributes to the downfall of a biocontrol agent . Knowledge of the biological environment in which the agent will be used and of how to produce a stable formulation are both critical to successful biocontrol . Certain Gram-positive bacteria have a natural formulation advantage over their Gram-negative counterparts: the spore . Although the Gram-positive bacteria have not been as well represented in the biocontrol literature, their spore-forming abilities and historical industrial uses bode well for biocontrol success . Here we describe several systems utilizing Gram-positive biocontrol agents that have been researched in depth and provide models for the future of biocontrol.

Mol Gen Mikrobiol Virusol, 1998, (4), 12 - 8
{Repression of catabolites in gram-positive and gram-negative bacteria}; Gershanovich VN; Analyzes the mechanism of catabolite repression of grampositive and gramnegative bacteria . The role of cyclic adenosine monophosphate and CRP protein, forming a complex, is shown . Contribution of ATP kinase to manifestation of the catabolic repression phenomenon in grampositive bacteria is discussed.

Minerva Urol Nefrol, 1998 Dec, 50(4), 225 - 31
Simultaneous detection of HPV and other sexually transmitted agents in chronic urethritis; Chiarini F et al.; BACKGROUND: Many pathogens may be responsible of Non Gonococcal Urethritis (NGU) with the possible occurrence of symptomatic and asymptomatic mixed viral and bacterial infections . In particular, genital papillomaviruses (HPVs) have been searched since they are linked to both benign and malignant lesions of the penis and urethra and the presence of a potential male carried state has received limited scrutiny while the screening of sexually active females has received substantial attention . METHODS: In male patients affected by chronic NGU, the presence of DNA of Chlamydia trachomatis, herpes simplex virus (HSV) type 1 and 2 and human papillomaviruses by PCR and the occurrence of Gram positive and Gram negative micro-organisms, of Mycoplasma hominis and Ureaplasma urealyticum, by conventional cultural methods have been investigated . RESULTS: Results obtained indicated a high percentage of mixed infections, up to 36% . Genital HPV DNA was detected in 31% of specimens positive for two or more agents, and HSV DNA was detected in 10% of studied population . CONCLUSIONS: The concomitant presence of different infectious agents could determine latent, sub-clinical or chronic infections with periodic reactivation . In particular results suggest that HPV and HSV may stimulate cytokine production which can up regulate the expression of other infectious agents and may be responsible for latent chlamydial infections characterised by the persistence of this micro-organism in an altered form, viable but in a culture negative state . Therefore an increased awareness of mixed infections is relevant to define the management and treatment of chronic urethritis.

Oral Dis, 1998 Sep, 4(3), 194 - 9
An immunohistochemical study of p53 and PCNA in inflammatory papillary hyperplasia of the palate: a dilemma of interpretation; Kaplan I et al.; OBJECTIVE: Inflammatory papillary hyperplasia of the palate (IPHP) or the granular type of denture stomatitis, is a non-neoplastic lesion characterized histologically by a significant epithelial hyperplasia and inflammatory infiltrate usually caused by trauma and Candida infection . p53 and proliferative cell nuclear antigen (PCNA) are cell-cycle regulators, that when overexpressed, are considered by many investigators as markers of malignant transformation . The objective of this study was to investigate the immunodetection of p53 and PCNA in IPHP, and to correlate these results with the degree of epithelial hyperplasia and inflammatory infiltrate . MATERIALS AND METHODS: In 12 cases diagnosed clinically as IPHP, Candida was cultured from the denture base and the palatal mucosa . Lesions were biopsied and stained with H&E for histomorphometric analysis of the epithelial width and inflammatory infiltrate . PAS and Gram stains were used for screening of Candida . Sections were immunostained with DO-7 for p53 and PC-10 for PCNA . Fifteen palatal biopsies obtained from autopsies of edentulous subjects with normal palatal mucosa served as controls . RESULTS: All cultures of swabs from both the palatal mucosa and denture base were positive for Candida . Candidal hyphae could not be identified in PAS stained sections . Small foci of Gram-positive organisms were found in two cases of IPHP . Epithelial width and inflammation were significantly higher in IPHP than in controls (P < 0.001) . A three-fold increase in positively stained cells for p53 and a two-fold increase in positively stained cells for PCNA were seen in IPHP compared with controls (P < 0.001) . CONCLUSIONS: Although a significant increase in the immunodetection of p53 and PCNA may indicate a malignant potential, IPHP has never been reported to undergo malignant transformation nor is it associated with cytologic signs of dysplasia . The increase in the epithelial width and inflammation degree is probably associated with the colonization of the denture bases with Candida organisms . The increased detection of p53 and PCNA can be a secondary effect of cytokines originating from both the inflammatory cells and the keratinocytes . Thus, immunodetection of p53 and PCNA by current immunohistochemical methods on archival tissues is neither specific nor sensitive enough to be used as indicators for malignant potential in the absence of cytological dysplastic changes or genetic proof of mutated cell cycle genes.

Otolaryngol Head Neck Surg, 1999 Feb, 120(2), 279 - 82
Microbiology of sinusitis in patients undergoing bone marrow transplantation; Imamura R et al.; As a result of increasing use of bone marrow transplantation and new cytotoxic chemotherapy, more patients have become susceptible to sinus disease caused by unusual organisms . Sinusitis caused by fungi and gram-negative bacteria can be difficult to treat, may lead to severe complications, and should be managed promptly in the bone marrow transplant patient . Here we present the results of 41 cultures of the paranasal sinuses obtained from 18 bone marrow transplant patients in whom sinusitis developed . The most common agents were gram-negative bacteria (56.7%), followed by gram-positive bacteria (26.7%) and fungi (16.6%) . In 13 samples the cultures were negative . Nasal cultures were performed ipsilateral to the sinus drained in 28 cases . Concordance was obtained in only 5 (17.8%) samples . The antibiogram of the isolated agents from the maxillary sinuses in this series revealed that the most efficient antibiotics were those that covered gram-negative bacteria . Treatment was usually prolonged in these patients, and different antibiotics were necessary to clear infections from the sinuses . In conclusion, treating sinusitis in bone marrow transplant patients may be challenging . Considerations about the microbiology and antibiogram susceptibilities of this specific population should be kept in mind when dealing with such cases.

J Lipid Res, 1999 Feb, 40(2), 245 - 52
Lipoproteins inhibit macrophage activation by lipoteichoic acid; Grunfeld C et al.; Regulation of lipid metabolism during infection is thought to be part of host defense, as lipoproteins neutralize endotoxin (LPS) and viruses . Gram-positive infections also induce disturbances in lipid metabolism . Therefore, we investigated whether lipoproteins could inhibit the toxic effects of lipoteichoic acid (LTA), a fragment of gram-positive bacteria . LTA activated RAW264.7 macrophage cells, stimulating production of tumor necrosis factor (TNF) in a dose-dependent matter, but produced less TNF than that seen after LPS activation . High density (HDL) or low density lipoprotein (LDL) alone inhibited the ability of LPS to stimulate TNF production, but had little effect on the activation by LTA . When a maximally effective dose of LTA was mixed with lipoproteins and 10% lipoprotein-depleted plasma (LPDP), the ability of LTA to stimulate macrophage production of TNF was inhibited . HDL, LDL, and the synthetic particle, Soyacal, when mixed with LPDP, were able to inhibit the ability of LTA to activate macrophages . Lipopolysaccharide-binding protein (LBP) substituted for LPDP in catalyzing lipoprotein neutralization of LTA by HDL . Antibody to LBP inhibited the ability of LPDP to induce LTA neutralization by HDL.Thus, lipoproteins can prevent macrophage activation by fragments from both gram-positive and gram-negative microorganisms.-Grunfeld, C., M . Marshall, J . K . Shigenaga, A . H . Moser, P . Tobias, and K . R . Feingold . Lipoproteins inhibit macrophage activation by lipoteichoic acid.

Syst Appl Microbiol, 1998 Dec, 21(4), 579 - 87
Analysis of broad-scale differences in microbial community composition of two pristine forest soils; Chatzinotas A et al.; Broad-scale differences in soil microbial community composition were analyzed in two contrasting soils using DNA reassociation and % G + C profiles for analysis on the community-level, and filter- and whole cell hybridization techniques for a coarse-level characterization of larger phylogenetic groups of bacteria . Reassociation analysis of DNA from bacterial fractions extracted from the organic soil Seim and the mineral soil Hau revealed similar complexity of the communities with 5700 and 4900 different bacterial genomes (g soil {dry wt})-1, respectively . Thermal denaturation studies showed wide % G + C distributions in DNA from bacteria of both soils . Differences in the median % G + C with 55 to 61% for the bacterial community in soil Seim and 61 to 66% for that in soil Hau indicated a higher proportion of bacteria with a high DNA G + C content in soil Hau . In situ hybridization with fluorescent (Cy3-labeled) probes targeting larger phylogenetic groups showed minor differences between both soils, and between direct detection of bacteria in dispersed soil slurries and in bacterial fractions extracted from soils through about 90% of the total bacteria were lost during extraction . In dispersed slurries of both soils, only probes ALF1b, SRB385, and PLA46 hybridized to cells accounting for more than 1% of the DAPI-stained cells, while numbers obtained after hybridization with probes ARCH915, BET42a, GAM42a, HGC69a, and CF319a were below the detection limit set at < 1% . These results were confirmed by in situ hybridization with horseradish peroxidase (HRP)-labeled probes and subsequent Cy3-tyramide signal amplification . In contrast, dot blot hybridization with probe HGC69a indicated significant amounts of Gram-positive bacteria with a high DNA G + C content in both soils . These could subsequently be visualized in non-dispersed soil slurries by in situ hybridization with HRP-labeled probe HGC69a and Cy3-tyramide signal amplification . Filamentous Gram-positive bacteria with a high DNA G + C content, likely actinomycetes, which are present in soil Hau in significant numbers are obviously destroyed by procedures used for soil dispersion.

J Clin Periodontol, 1999 Jan, 26(1), 19 - 25
Microbiological features and crevicular fluid aspartate aminotransferase enzyme activity in early onset periodontitis patients; Kuru B et al.; Gingival crevicular fluid (GCF) reflects the immune and inflammatory reactions and the specific host-microbe interactions that lead to periodontal diseases . Aspartate aminotransferase enzyme (AST) is one of the components of GCF that is released as a result of cell death . In this study, periodontal sites (4 sites/patient) with a probing depth of > or =5 mm in early onset periodontitis (EOP) patients were first examined for the AST levels in GCF by the Periogard periodontal tissue monitor.To be eligible for the study, each of the patients had at least 1 AST positive site with clinical inflammatory changes (AST+, CIC+) and 1 AST negative site with no or minimum clinical inflammatory changes (AST-, CIC-) . In 15 EOP patients who met the entry criteria, 30 AST+, CIC+ sites (1st group) and 19 AST-, CIC- sites (2nd group) were evaluated for microbiological variables . Certain microbial species, including Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Prevotella intermedia were detected more frequently (p<0.001, p<0.001 and p<0.05, respectively) in the 1st group, while gram-positive facultative organisms such as Actinomyces species were found more often (p<0.001) in the 2nd group . Parallel to the AST levels, the 2nd group had a lower number of total bacteria and proportion of obligate anaerobic and capnophilic micro-organisms than the first group (p<0.05 and p<0.05, respectively) . Within the scope of this study, AST activity and microbiological data were found in agreement in the examined groups . These findings are encouraging and indicate the need for further studies to evaluate the ability of the AST test to differentiate the microbial flora of progressing sites and those that are inflamed, but not progressing.

Rapid Commun Mass Spectrom, 1999, 13(1), 39 - 49
Liquid chromatography/microspray mass spectrometry for bacterial investigations; Krishnamurthy T et al.; Cellular proteins (biomarkers) specific to any individual microorganism, determined by the direct mass spectral analysis of the corresponding intact cellular suspension, can be applied for the rapid and specific identification of the organisms present in unknown samples . The components of the bacterial suspensions, after a rapid separation over a C18 reversed-phase microcapillary column, were directly subjected to on-line electrospray ionization followed by analysis using an ion trap tandem mass spectrometer . This approach is equally effective for gram-positive as well as gram-negative bacteria but has a distinct advantage over our earlier reported method involving matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) . During electrospray ionitation mass spectrometry (ESI-MS), liquid samples can be directly analyzed and there is the potential for developing tandem mass spectral methods for more specific identification of the individual organisms present in crude bacterial mixtures . The total analysis time leading to unambiguous bacterial identification in samples was less than 10 minutes and the results were quite reproducible . Miniaturization of the instrumentation along with total automation of this simple process could have immense impact on field operations . Routine, rapid, cost-effective field monitoring of environmental samples, agricultural products, samples from food processing, industrial sites and health institutions for suspected bacterial contamination could be a reality in the near future . Potential utility in biological, medical, bioprocessing, pharmaceutical, and other industrial research is also enormous.

Ann N Y Acad Sci, 1998 Sep 29, 856, 116 - 31
Tolerance to pyrogens; Zeisberger E et al.; In humans or experimental animals, the repeated confrontation with lipopolysaccharides (LPS) from gram-negative bacteria, but not with muramyl dipeptide (MDP) from gram-positive bacteria, leads to attenuation of almost all pathophysiologic effects mediated by proinflammatory cytokines . Our experiments in guinea pigs and rats demonstrate that attenuation of the febrile response during the development of LPS tolerance is associated with a reduced production of cytokines rather than a decrease in responsiveness to cytokines . Cross-tolerance experiments demonstrate that different stimuli influencing LPS-induced tumor necrosis factor (TNF) release and nitric oxide (NO) synthesis can modify the development of tolerance . On the other hand, the lack of cross-tolerance between LPS and MDP indicates that MDP can activate the cytokine cascade and induce the febrile response in animals tolerant to LPS . This may indicate distinct receptors and signal pathways for LPS and MDP, leading to activation of the cytokine cascade . LPS tolerance has also been demonstrated in ex vivo and in vitro studies . In cultures of monocytes, diminished synthesis of TNF and NO reported after LPS restimulation could be prevented and reversed by interferon and granulocyte-macrophage colony-stimulating factor . These findings add an additional hypothesis in tolerance development.

Int J Antimicrob Agents, 1998 Nov, 10(4), 263 - 70
Imipenem versus targeted therapy in cancer patients; Aoun M et al.; In many instances, broad-spectrum antibiotics are initiated empirically in febrile cancer patients and continued for the whole duration of therapy . An alternative is to narrow the spectrum whenever the offending pathogen is identified . This study is aimed at comparing these two options . Non-neutropenic cancer patients with severe infections received empiric imipenem . After 72 h, those with microbiologically documented infection were randomized either to continue imipenem or to receive a targeted therapy . After 72 h of imipenem 76.1% were improved . After randomization, a higher efficacy was observed with imipenem (88.5 vs . 72.1%: P = 0.025) . Bacterial and fungal superinfections were comparable . Costs were lower for targeted therapy in gram-positive infection and higher in gram-negative infection.

Clin Rheumatol, 1998, 17(6), 538 - 9
Spondylitis caused by Peptostreptococcus; Rousseau MC et al.; Peptostreptococcus is an anaerobic gram-positive coccus that is encountered in dental and sinus infections, pelvic infections in women, osteomyelitis, arthritis, skin and soft-tissues infections, conjunctivitis and bacteraemia {1,2} . We report here, what is to our knowledge, the first cases of spondylitis with discitis caused by Peptostreptococcus.

J Am Soc Nephrol, 1999 Jan, 10(1), 136 - 45
Intermittent versus continuous intraperitoneal glycopeptide/ceftazidime treatment in children with peritoneal dialysis-associated peritonitis . The Mid-European Pediatric Peritoneal Dialysis Study Group (MEPPS); Schaefer F et al.; Intermittent intraperitoneal antibiotic administration appears as a practical and economical therapeutic concept in continuous peritoneal dialysis (CPD)-related peritonitis, but the equivalence of this principle with standard continuous treatment awaits confirmation by prospective, randomized clinical trials . This study evaluates the efficacy, safety, and clinical acceptance of an initial combination treatment including a glycopeptide (vancomycin or teicoplanin) and ceftazidime, each applied either intermittently or continuously, in a cohort of pediatric patients with CPD-related peritonitis . Patients randomized for continuous treatment received an intraperitoneal loading dose of glycopeptide and ceftazidime followed by maintenance doses added to each dialysate bag . In the intermittent treatment groups, the glycopeptide was administered in two loading doses 7 d apart, and ceftazidime during one dialysis cycle per day . Initial treatment response was evaluated after 60 h by the change in a Disease Severity Score and by the clinical decision to continue initial treatment . Of 152 patients observed for a total of 234 patient years, 90 patients developed 195 episodes of peritonitis (including 27 relapses within 4 wk after end of treatment) . Dialysate cultures were positive in 83% of the episodes . In gram-positive peritonitis (79% of culture-positive cases), the primary success (overall 95%) and relapse rates (21%) were not different between continuous and intermittent, or between vancomycin and teicoplanin treatment . Oversensitivity reactions occurred in three and ototoxicity in one vancomycin-treated patient, whereas no such side effects were observed with teicoplanin . Residual renal function declined during peritonitis episodes regardless of treatment modality . In gram-negative peritonitis (18% of cases), intermittent ceftazidime treatment was less successful than continuous treatment according to clinical judgment (3 of 11 versus 10 of 14, P < 0.05), but not when rated by Disease Severity Score (8 of 11 versus 12 of 14) . In conclusion, intermittent and continuous intraperitoneal treatment of CPD-related peritonitis with glycopeptides and ceftazidime is equally efficacious and safe when measured by objective clinical criteria . This contrasts with a strong tendency of clinicians to move from intermittent to continuous treatment in severe peritonitis.

Gene, 1999 Jan 8, 226(1), 83 - 93
Gene organization in a central DNA fragment of Oenococcus oeni bacteriophage fOg44 encoding lytic, integrative and non-essential functions; Parreira R et al.; The nucleotide sequence of a DNA fragment previously shown to contain the attachment site (attP) of Oenococcus oeni phage fOg44 ( . Arch . Virol . 143, 523-536) has been determined . Sequence analysis indicated that this 6226bp EcoRI fragment harbours an integrase gene, in the vicinity of a direct repeat rich region defining attP, as well as genes encoding a muramidase-related lysin (Lys) and a holin polypeptide (Hol) . Transcriptional studies suggested that lys and hol are mainly co-expressed, late in the lytic cycle, from a promotor located upstream of lys . Between the lytic cassette and the phage integration elements three additional open reading frames were found: orf217 and orf252 of unknown function and orf72, the putative product of which bears 32% identity with acidic excisionases from other Gram positive phages . We have established that the first two orfs, as well as the predicted promotor of orf72, are included in a 2143-bp DNA segment missing from the genome of the deletion mutant fOg44Delta2 . Although lysogens of fOg44 and fOg44Delta2 exhibited similar properties, each phage produced two distinguishable types of lysogenic strains, differing in inducibility and immunity to other oenophages.

J Clin Microbiol, 1999 Feb, 37(2), 464 - 6
Gram type-specific broad-range PCR amplification for rapid detection of 62 pathogenic bacteria; Klausegger A et al.; Broad-range PCR has proven to be useful for the detection of bacteria . A set of broad-range PCR primers directed against conserved regions in the 16S rRNA gene was designed to specifically amplify either gram-positive or gram-negative bacteria . The gram type-specific broad-range PCR correctly classified all 62 pathogenic species tested.

Chem Biol, 1999 Jan, 6(1), 11 - 8
Aminoglycoside antibiotic phosphotransferases are also serine protein kinases; Daigle DM et al.; BACKGROUND: Bacterial resistance to aminoglycoside antibiotics occurs primarily through the expression of modifying enzymes that covalently alter the drugs by O-phosphorylation, O-adenylation or N-acetylation . Aminoglycoside phosphotransferases (APHs) catalyze the ATP-dependent phosphorylation of these antibiotics . Two particular enzymes in this class, APH(3')-IIIa and AAC(6')-APH(2"), are produced in gram-positive cocci and have been shown to phosphorylate aminoglycosides on their 3' and 2" hydroxyl groups, respectively . The three-dimensional structure of APH (3')-IIIa is strikingly similar to those of eukaryotic protein kinases (EPKs), and the observation, reported previously, that APH(3')-IIIa and AAC(6')-APH(2") are effectively inhibited by EPK inhibitors suggested the possibility that these aminoglycoside kinases might phosphorylate EPK substrates . RESULTS: Our data demonstrate unequivocally that APHs can phosphorylate several EPK substrates and that this phosphorylation occurs exclusively on serine residues . Phosphorylation of Ser/Thr protein kinase substrates by APHs was considerably slower than phosphorylation of aminoglycosides under identical assay conditions, which is consistent with the primary biological roles of the enzymes . CONCLUSIONS: These results demonstrate a functional relationship between aminoglycoside and protein kinases, expanding on our previous observations of similarities in protein structure, enzyme mechanism and sensitivity to inhibitors, and suggest an evolutionary link between APHs and EPKs.

J Mol Biol, 1998 Dec 18, 284(5), 1323 - 39
Gene structures and properties of enzymes of the plasmid-encoded nicotine catabolism of Arthrobacter nicotinovorans; Schenk S et al.; Arthrobacter nicotinovorans is a Gram-positive aerobic soil bacterium able to grow on nicotine as its sole source of carbon and nitrogen . The initial steps of nicotine catabolism are catalyzed by nicotine dehydrogenase, the l- and d-specific 6-hydroxynicotine oxidases, and ketone dehydrogenase . The genes encoding these enzymes reside on a 160 kb plasmid, pAO1 . The cccDNA of this plasmid was isolated in high purity and reasonable yield . It served as template material for the construction of a lambda-phage DNA library of the plasmid . The genes coding for 6-hydroxy-l-nicotine oxidase and for the subunits of the heterotrimeric ketone dehydrogenase were identified, subcloned and sequenced . The 6-hlno gene was identified as a 1278 bp open reading frame; its regulatory elements were also recognized . The derived primary structure of the monomer of apo-6-hydroxy-l-nicotine oxidase (46,264.5 Da) agrees with the data obtained by partial amino acid sequencing . 6-Hydroxy-l-nicotine oxidase and 6-hydroxy-d-nicotine oxidase were expressed in Escherichia coli and obtained in a state of high purity and crystallized.Ketone dehydrogenase (KDH) was found to be a heterotrimer with subunits of molecular mass 89,021.71, 26,778.65 and 17,638.88 . The genes of KDH-A and KDH-B are juxtaposed; the A of the stop codon of KDH-A is used in the start codon of KDH-B, eliciting a frame shift . KDH-C is separated from KDH-A by 281 bp .

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