APMIS, 1997 Feb, 105(2), 99 - 107 Ultrastructure of colony-like communities of bacteria; Tetz VV et al.; Colony-like communities are poorly studied forms of bacterial growth on agar . These communities are formed after the growth of large amounts of bacteria simultaneously plated onto a limited area of agar, while "classical" colonies are formed as a result of single bacterial cell multiplication . Colony-like communities of Gram-negative and Gram-positive bacteria differ from "classical" microbial colonies in their ultrastructural organization . Almost every cell in colony-like communities has an individual capsule-like envelope (glycocalyx) . The cells in these communities are characterized by accelerated ageing . In the course of their development both bacterial colony-like communities and "classical" colonies produce a film, the basic part of which is represented by an elementary membrane . In contrast to "classical" colonies, the thickness of the amorphous layers of this film in colony-like communities did not significantly increase after 24 h of cultivation . The formation of a three-dimensional network of cells in colony-like communities is similar to this process in "classical" colonies . The intercellular matrix of colony-like communities contains numerous membrane vesicles, and has a more amorphous structure and higher electron density than that of "classical" bacterial colonies. J Colloid Interface Sci, 1997 Feb 1, 186(1), 71 - 9 Conductivity and Dielectric Dispersion of Gram-Positive Bacterial Cells van der Wal A, Minor M, Norde W, Zehnder AJB, Lyklema J. The conductivity of bacterial cell suspensions has been studied over a wide range of ionic strengths and is interpreted in terms of their cell wall properties . The experimental data have been analyzed after improving the high kappaa double-layer theory of Fixman, by accounting for ionic mobility in the hydrodynamically stagnant layer, i.e., in the bacterial wall . Static conductivity and dielectric dispersion measurements both show that the counterions in the porous gel-like cell wall give rise to a considerable surface conductance . From a comparison of the mobile charge with the total cell wall charge it is inferred that the mobilities of the ions in the bacterial wall are of the same order but somewhat lower than those in the bulk electrolyte solution . The occurrence of surface conductance reduces the electrophoretic mobility in electrophoresis studies . If this effect is not taken into account, the zeta-potential will be underestimated, especially at low electrolyte concentrations. Appl Environ Microbiol, 1997 Feb, 63(2), 670 - 5 Biochemical and mutational analysis of a gingipain-like peptidase activity from Prevotella ruminicola B(1)4 and its role in ammonia production by ruminal bacteria; Madeira HM et al.; A chemical mutagenesis protocol was used with the ruminal bacterium Prevotella ruminicola strain B(1)4 to generate mutant strains defective in peptidase activity . Compared with the wild-type parent strain, the isolated mutants possessed 1/10 of the enzyme activity responsible for cleavage of glycine-arginine-4-methoxy-beta-naphthylamide (Gly-Arg-MNA) . A concomitant loss in activity against arginine-arginine-4-methoxy-beta-naphthylamide (Arg-Arg-MNA) was also observed . Both activities were similarly affected by various proteinase inhibitors, suggesting that the same enzyme is responsible for the Arg-Arg-MNA peptidase and Gly-Arg-MNA peptidase activities . Growth rates of wild-type and mutant strains grown in batch culture with various nitrogen sources did not differ . However, a role for the Gly-Arg-MNA peptidase activity was demonstrated in coculture experiments with gram-positive, ammonia-producing ruminal bacteria . The rate and extent of ammonia production were reduced by approximately 25% in cocultures containing the mutants when compared with that of wild-type-containing cultures . These reductions could not be accounted for simply by the decrease in ammonia production by the mutant strain alone . To our knowledge, this paper reports the first successful use of chemical mutagenesis with ruminal microorganisms. Bone Marrow Transplant, 1997 Jan, 19(2), 143 - 7 Infectious complications during neutropenia subsequent to peripheral blood stem cell transplantation; Kolbe K et al.; Type, severity and incidence of infection during the neutropenic period after peripheral blood stem cell transplantation (PBSCT) for treatment of malignant disease were studied in 66 patients treated at a single institution . Data of 34 female and 32 male patients with a median age of 43 years suffering from leukemia (12), lymphoma (35), multiple myeloma (six) or solid tumors (13) were retrospectively analyzed . All patients had received at least 2.5 x 10(6) CD34-positive cells for stem cell rescue after high-dose chemotherapy . Ninety-four percent of the patients experienced at least one febrile episode during their post-transplant course . The patients recovered quickly and defervesced after a median of 4 days . The incidence of bacteremia was 39% and gram-positive cocci were the predominant pathogens . In contrast, severe organ infections were rare . Only 5% of the patients suffered from lung infiltrates . No invasive fungal infections were observed . No transplant-related deaths occurred in the 66 patients studied . We conclude that the severe, but shortlasting neutropenia after peripheral blood stem cell transplantation is associated with a high incidence of bacterial infection . The severity of the majority of these infections is moderate . With appropriate anti-infective therapies these infections can be managed and life-threatening infectious complications, in particular fungal infections, are rare . Empirical anti-infective regimens specifically designed for this clinical situation should be explored. Ophthalmologica, 1997, 211(5), 308 - 11 Cavernous sinus thrombosis secondary to abdominal wall abscess; Jinbayashi H et al.; An 84-year-old Japanese woman had cavernous sinus thrombosis (CST) due to abdominal wall abscess . The diagnosis was delayed because abdominal wall abscess is an unfamiliar cause of CST, but a definite diagnosis was reached with the use of magnetic resonance imaging . We started intravenous antibiotic therapy against gram-positive and gram-negative organisms and the patient responded well, although the diagnosis was delayed. Caries Res, 1997, 31(5), 336 - 48 Structural analyses of plaque and caries in relation to the morphology of the groove-fossa system on erupting mandibular third molars; Ekstrand KR et al.; Using 22 erupting human third mandibular molars, this study investigated: (1) the relationship between the morphology of the interlobal groove and the histological features of caries, (2) the ultrastructural features of the interlobal groove contents, and (3) whether the morphology of interlobal grooves influences the viability of the microorganisms . Posteruption time was classified as (1) < 3 months (7 teeth), (2) 3-12 months (8 teeth) and (3) > 12 months but < 18 months (7 teeth) . The teeth were pre- and postfixed, and buccolingual sections were prepared . The mesial/distal interlobal groove on 116 sections was classified as fissure-like or groove-like . All sections were embedded and ground, and microradiographs were made . The interlobal groove contents were analyzed on two sections from each tooth, one with a fissure-like and one with a groove-like morphology, giving a total of 44 sections . Eighteen teeth (82%) had evidence of caries . Severity of caries was associated with length of posteruption time . 'Fissures' were not more prone to caries than 'grooves' . In about 70% of the 'fissures' caries had penetrated deepest at the entrance rather than in the deeper parts . LM and TEM examinations from 1 tooth disclosed erythrocytes, enamel organ cells and pioneers of microorganisms in the interlobal grooves . All other interlobal grooves were occupied by microorganisms, mainly gram-positive coccoids and small rods . Viable microorganisms were seen at the entrance as well as at the bottom part of the 'grooves' . In 'fissures' viable microorganisms were primarily observed at the entrance, while in deeper parts the microorganisms were less viable or dead . A moderate association was observed between the viability of the microorganisms and differences in the depths of caries penetration in the fissures . Thus, the internal morphology of the interlobal grooves influenced the conditions for bacterial growth and this determined the location for caries progression within the groove-fossa system. Int Urol Nephrol, 1997, 29(3), 385 - 9 Influence of intraperitoneal vancomycin on peritoneal dialysis efficiency . A clinical study; Janicka L et al.; Peritonitis is a major complication of intermittent peritoneal dialysis (IPD); over 70% of the infections are caused by Gram-positive bacteria . Vancomycin (V) is the antibiotic of choice in the treatment of peritonitis caused by G(+) . The influence of vancomycin on peritoneal transport in IPD patients has not been described before . We have investigated the effect of intraperitoneal vancomycin on dialysis efficiency in 8 IPD patients using dialysis solutions containing either lactate or acetate . The following parameters were measured: net ultrafiltration (UF), concentration ratios (D/P) of urea, creatinine, potassium, peritoneal clearances (ml/min) of urea, creatinine, potassium, mass transfer of sodium (MTNa), sodium sieving index (SCNa) . It has been found that vancomycin significantly decreases D/P urea (p < 0.05) and creatinine (p < 0.05) . We found also a significant decrease of mean clearance of urea (p < 0.05) and creatinine (p < 0.05) . The mean clearance of potassium did not change significantly . There was no significant change in UF, MTNa, and SCNa . Our preliminary data suggest that vancomycin decreases the permeability of peritoneum for certain low molecules in IPD patients which may have a negative impact on dialysis efficiency. J Basic Microbiol, 1997, 37(3), 217 - 27 Heavy metal resistant Arthrobacter sp.--a tool for studying conjugational plasmid transfer between gram-negative and gram-positive bacteria; Margesin R et al.; The role of two heavy metal-resistant strains of the Gram-positive genus Arthrobacter sp . as a tool in studying conjugational plasmid transfer between Gram-positive and Gram-negative bacteria is described . The high nickel resistance and the cobalt resistance of Arthrobacter sp . strain RM1/6 could be transferred to Arthrobacter sp . strain WS14 . IncQ plasmids (pKT240, pKT240::czc, pML10) could be mobilized from E . coli into Arthrobacter spp . strains; antibiotic (Km, Ap, Tc) and heavy metal (Co) resistance genes were expressed in the recipient stains . IncQ plasmid pKT240 could be mobilized between Arthrobacter spp . strains . IncP plasmid RP4::Tn4371 was transferred from A . eutrophus to Arthrobacter sp., RP4-mediated antibiotic resistance to Km was expressed in the recipient strain. Folia Microbiol (Praha), 1997, 42(3), 171 - 8 Regulation of carbon metabolism in gram-positive bacteria by protein phosphorylation; Deutscher J et al.; The main function of the bacterial phosphotransferase system is to transport and to phosphorylate mono- and disaccharides as well as sugar alcohols . However, the phosphotransferase system is also involved in regulation of carbon metabolism . In Gram-positive bacteria, it is implicated in carbon catabolite repression and regulation of expression of catabolic genes by controlling either catabolic enzyme activities, transcriptional activators or antiterminators . All these different regulations follow a protein phosphorylation mechanism. Exp Clin Endocrinol Diabetes, 1997, 105(3), 182 - 6 Adrenal functions in patients with sepsis; Aygen B et al.; The basal cortisol level and cortisol response to ACTH stimulation test were assessed in patients with sepsis, the results being compared to a control group of 30 healthy persons . The study group included 49 patients with sepsis and 30 healthy subjects as a control group . The mean age in the study group was 42.6 +/- 18.7 years and 41.4 +/- 12.1 years in the control group . Fifteen of the 49 (30.6%) patients had hospital-acquired and 34 (69.4%) patients community-acquired sepsis . Etiological agent was isolated in 35 (71.4%) patients (57.1% gram negative bacteria and 34.3% gram positive bacteria, plus 8.6% polymicrobial) . Fourteen of 49 (28.6%) patients died . Mean basal cortisol level was 597.1 +/- 304.6 nmol/l (range 217.8-1667.9) in the study group and 460.2 +/- 180.8 nmol/l (range 253.6-988.9) in the control group . Mean basal cortisol level in the study group was significantly higher than that of the control group (p < 0.05) . Mean basal cortisol level was found to be 725.5 +/- 448.9 nmol/l in the patients who died and 545.8 +/- 210.9 nmol/l in the patients who recovered . The difference between the two groups was found to be significant (p < 0.05) . ACTH stimulation test was performed in 43 of the patients and 30 healthy subjects . Cortisol response was significantly lower (mean 277.7 +/- 216.9 nmol/l) in the patients than that detected in the control group (mean 519.6 +/- 279.2) (p < 0.001) . Mean cortisol response in the patients who died was 227.2 +/- 224.5 nmol/l and 302.1 +/- 212.7 nmol/l in the patients who recovered (p > 0.05) . Adrenocortical insufficiency was detected in 16.3% of the patients and 42.9% of these patients died . In conclusion, sepsis is characterized by high basal cortisol level which may show a poor prognosis and a blunted cortisol response to ACTH stimulation . A small percentage of patients with sepsis may develop adrenocortical insufficiency. Ciba Found Symp, 1997, 207, 206 - 18; discussion 219-22 Genetic mobility and distribution of tetracycline resistance determinants; Roberts MC; Since 1953, tetracycline-resistant bacteria have been found increasingly in humans, animals, food and the environment . Tetracycline resistance is normally due to the acquisition of new genes and is primarily due to either energy-dependent efflux of tetracycline or protection of the ribosomes from its action . Gram-negative efflux genes are frequently associated with conjugative plasmids, whereas Gram-positive efflux genes are often found on small mobilizable plasmids or in the chromosome . The ribosomal protection genes are generally associated with conjugative transposons which have a preference for the chromosome . Recently, tetracycline resistance genes have been found in the genera Mycobacterium, Nocardia, Streptomyces and Treponema . The Tet M determinant codes for a ribosomal protection protein which can be found in Gram-positive, Gram-negative, cell-wall-free, aerobic, anaerobic, pathogenic, opportunistic and normal flora species . This promiscuous nature may be correlated with its location on a conjugative transposon and its ability to cross most biochemical and physical barriers found in bacteria . The Tet B efflux determinant is unlike other efflux gene products because it confers resistance to tetracycline, doxycycline and minocycline and has the widest host range of all Gram-negative efflux determinants . We have hypothesized that mobility and the environment of the bacteria may help influence the ultimate host range of specific tet genes . If we are to reverse the trend towards increasingly antibiotic-resistant pathogenic bacteria, we will need to change how antibiotics are used in both human and animal health as well as food production. Nephron, 1997, 75(4), 438 - 43 Detection of peptidoglycan and endotoxin in dialysate, using silkworm larvae plasma and limulus amebocyte lysate methods; Tsuchida K et al.; Silkworm larvae plasma (SLP) reagent is activated by peptidoglycan (PG), a fragment of both the gram-positive and gram-negative bacterial cell wall, as well as beta-glucan (BG), a component of fungi . It is possible to measure contamination of gram-positive bacteria quantitatively by combining the conventional limulus amebocyte lysate (LAL) and PG measurement methods . Therefore, a more highly accurate analysis of dialysate can be made using both SLP and LAL methods to detect endotoxin (ET) and/or PG contamination . We studied the effects of contaminated dialysate on human peripheral blood mononuclear cells (PBMC) by producing various cytokines in vitro . Muramyl dipeptide (MDP) was used as the biologically active minimum constituent of PG . A total of 54 dialysate samples were obtained under sterile conditions from 4 sites: (1) reverse osmosis water unit; (2) proportioning unit; (3) multiple dialysate preparation console, and (4) personal dialysate preparation console, at 9 dialysis facilities . To detect bacterial contamination, the samples were measured with LAL(C), LAL(G) and SLP methods . PBMC were collected from 10 healthy controls and from 10 hemodialysis patients and cultured for 24 h with ET, MDP, ET + MDP and contaminated dialysate . IL-1 receptor antagonist (IL-1Ra), IL-1 beta and TNF-alpha in the culture medium supernatants were measured using the ELISA method . PG was not detected in dialysate from sites 1 or 2 . However, dialysate from the inlet of the dialyzer at the bedside monitor of the central supply and personal console showed 4.1 +/- 6.1 ng/ml for site 3 (in 7 of 18 samples) and 3.3 +/- 4.6 ng/ml for site 4 (in 3 of 18 samples) . Contamination by PG alone and complex contamination by PG and ET were also detected . Furthermore, IL-1Ra, IL-1 beta and TNF-alpha production by PBMC increased in accordance with the concentrations of MDP . Cytokine production was enhanced 5-10 times more where MDP and ET coexisted than where either MDP or ET existed alone, showing the synergic effects of MDP and ET . Based on these results, there is a high possibility that PG may also be a pyrogen in the dialysate prior to this study . ET had been considered the only pyrogen in dialysate . Therefore, it is essential to recognize the existence of both ET and PG in investigating dialysate contamination. Antibiot Khimioter, 1997, 42(2), 33 - 7 {Use of piperacillin/tazobactam as empirical monotherapy in the treatment of bacterial infections in a resuscitation department}; Iakovlev SV et al.; Fifteen patients with severe bacterial infection (12 with pneumonia) that developed in the resuscitation unit were subjected to the empirical monotherapy with piperacillin/tazobactam (P/T) or tazocin under an open randomized controlled experiment . P/T was administered intravenously in a dose of 4.5 g every 8 hours for 5 to 12 days (9.3 days on the average) . When the monotherapy was not sufficiently efficient the patients were additionally treated with amikacin administered intravenously in a dose of 0.5 g every 8-12 hours . The favourable effect was observed in 14 patients (93 per cent) . 7 of them were treated with P/T alone and 7 were treated with P/T in combination with amikacin . The primary pathogens were eradicated in 8 (73 per cent) out of the 11 patients treated with P/T alone . Before the treatment 34 microbial strains were isolated from the patients . 77 per cent of them were susceptible to P/T . The treatment with P/T resulted in eradication of 27 bacterial strains (79 per cent) including 67 per cent of gram-positive organisms and 86 per cent of gram-negative organisms . The adverse effects were recorded in 1 patient on the 6th day of the treatment: skin eruption and pruritis that required the treatment discontinuation . The results showed that the use of P/T in the initial empirical monotherapy of infections in patients under resuscitation conditions could be efficient. Antibiot Khimioter, 1997, 42(2), 21 - 5 {Clinico-laboratory study of piperacillin/tazobactam in the treatment of patients with would infection}; Iakovlev VP et al.; Piperacillin/tazobactam (P/T) or tazocin was used in the treatment of 40 patients with soft tissue purulent necrotic wounds of various genesis and localization . In the majority of the patients the affection was of recurring nature . P/T was administered in a dose of 4.5 g 3 times a day for 1 to 14 days (mainly for 8 to 10 days) as 30-minute intravenous infusions . The clinical effect of the treatment was stated in 36 patients (90 per cent) . In 3 out of 8 patients with diabetes mellitus the treatment failed . In 1 patient the treatment was discontinued after the first dose because of asphyxia and a short-term decrease of the arterial pressure . Among 105 microbial strains isolated from 75 patients 64 were highly susceptible to P/T, 33 were moderately susceptible and 8 were resistant . 76.3 per cent of the isolates produced beta-lactamase, 66.3 per cent of them were susceptible to P/T . 54 gram-positive and gram-negative strains were isolated from 40 patients . 16 out of the 54 strains were isolated as monocultures and the others as associations . After the treatments the pathogen eradication and the pathogen eradication followed by superinfection were stated in 26 patients (66.7 per cent) . In 11 patients (28.2 per cent) the pathogen persistence was observed at the background of the clinical improvement in the majority of the patients . The relapses were recorded in 2 patients (5.1 per cent). J Appl Microbiol, 1997 Jan, 82(1), 7 - 18 Resistance to chlorine of freshwater bacterial strains; Mir J et al.; The disinfectant properties of chlorine have been known for centuries but in the last few years water chlorination has attracted some criticism due to its secondary effects and the increased resistance of bacterial strains to chlorine inactivation . In this paper the kinetics of inactivation by chlorine of different Gram-positive and Gram-negative bacterial strains isolated from chlorinated water is studied . The Gram-positive strains were more resistant to chlorine and the behaviour of some of them in the presence of chloramphenicol suggests either the synthesis of unique proteins or aggregation of the bacteria as mechanisms of resistance to inactivation . The concept of Ki, the inactivation rate constant, by comparison with Ks in Michaelis-Menten enzyme kinetics (considering enzymic saturation), or with Ks in Monod growth kinetics (considering limiting rates of transport and metabolism of substrates), may be an interesting parameter to define microbial resistance to disinfectants and toxics. J Periodontal Res, 1997 Jan, 32(1 Pt 2), 140 - 2 Multiple colonization defects in a cysteine protease mutant of Porphyromonas gingivalis; Kuramitsu H et al.; A cysteine protease mutant, G-102, of Porphyromonas gingivalis 381 defective in the rgp-1 gene has been recently constructed in this laboratory . In order to evaluate the role of the protease in the virulence properties of P . gingivalis, a number of putative periodontopathic properties of the mutant were evaluated . Relative to the parental strain, mutant G-102 was demonstrated to be defective in interacting with Gram-positive bacteria as well as cultured epithelial cells . In addition, the mutant was altered in attaching to the protein components of extracellular matrix as well as to type I collagen . Some of these alterations could result from the decreased autoaggregation displayed by mutant G-102 relative to strain 381 . However, since the epithelial cell attachment assays were carried out at very low bacterial densities, it is unlikely that reduced autoaggregation of the mutant is responsible for its decreased ability to attach to these eucaryotic cells . Electron microscopic examination of the cells also revealed that mutant G-102 was altered in normal fimbrae expression . In addition, reduced expression of the 43 kDa fimbrial subunit in the mutant was detected with both Western and Northern blotting . These results indicated that the rgp-1 gene product can play either a direct or indirect role in the colonization properties of P . gingivalis. Annu Rev Physiol, 1997, 59, 633 - 57 Mechanosensitive channels of Escherichia coli: the MscL gene, protein, and activities; Sukharev SI et al.; Although mechanosensory responses are ubiquitous and diverse, the molecular bases of mechanosensation in most cases remain mysterious MscL, a mechanosensitive channel of large conductance of Escherichia coli and its bacterial homologues are the first and currently only channel molecules shown to directly sense mechanical stretch of the membrane . In response to the tension conveyed via the lipid bilayer, MscL increases its open probability by several orders of magnitude . In the present review we describe the identification, cloning, and first sets of biophysical and structural data on this simplest mechanosensory molecule . We discovered a 2.5-ns mechanosensitive conductance in giant E . coli spheroplasts . Using chromatographies to enrich the target and patch clamp to assay the channel activity in liposome-reconstituted fractions, we identified the MscL protein and cloned the mscL gene . MscL comprises 136 amino acid residues (15 kDa), with two highly hydrophobic regions, and resides in the inner membrane of the bacterium . PhoA-fusion experiments indicate that the protein spans the membrane twice with both termini in the cytoplasm . Spectroscopic techniques show that it is highly helical . Expression of MscL tandems and covalent cross-linking suggest that the active channel complex is a homo-hexamer . We have identified several residues, which when deleted or substituted, affect channel kinetics or mechanosensitivity . Although unique when discovered, highly conserved MscL homologues in both gram-negative and gram-positive bacteria have been found, suggesting their ubiquitous importance among bacteria. J Antibiot (Tokyo), 1997 Jan, 50(1), 18 - 21 Brasiliquinones A, B and C, new benz{alpha}anthraquinone antibiotics from Nocardia brasiliensis . I . Producing strain, isolation and biological activities of the antibiotics; Nemoto A et al.; New benz{alpha}anthraquinone antibiotics (brasiliquinones A, B and C) with an ethyl group at C-3 were isolated . The producer was identified as Nocardia brasiliensis . The antibiotics were active against Gram-positive bacteria including Mycobacterium sp., but not active against Gram-negative bacteria or fungi . They were also active against multiple drug-resistant P388/ADR tumor cells. J Antibiot (Tokyo), 1997 Jan, 50(1), 8 - 12 New anticancer antibiotics pelagiomicins, produced by a new marine bacterium Pelagiobacter variabilis; Imamura N et al.; In the course of our screening for new anticancer compounds produced by marine bacteria, we found that a new genus marine bacterium Pelagiobacter variabilis produced new phenazine antibiotics, pelagiomicins A, B and C . Those compounds were labile in water and alcohols . The absolute structure of the main component, pelagiomicin A, and the structures of the minor ones were determined from the spectroscopic data and by synthesis . Pelagiomicin A exhibits activity against Gram-positive and -negative bacteria and antitumor activity in vitro and in vivo. Jpn J Antibiot, 1997 Jan, 50(1), 12 - 21 {Clinical evaluation of cefpirome sulfate for severe infections in patients with hematological disorders . Hanshin Study Group of Hematopoietic Disorders and Infections}; Hamazaki H et al.; We investigated the therapeutic efficacy and safety of cefpirome sulfate (CPR) in treatment of hematopoietic disorder-associated infections . A total of 219 patients were admitted to 12 hospitals of Hanshin Study Group of hematopoietic disorders and infections between April 1994 and March 1996 and were enrolled in this study . Most patients received intravenously infused CPR at a dose of 1 or 2 g twice a day for 3 days or more . Twenty nine patients dropped out or were excluded and remaining 190 patients were adopted for the evaluation . A overall response rate was 58.4% (111/190) . Among neutropenic patients, the response rate was 50% (8/16) in patients whose peripheral neutrophil counts (PNC) remained less than 100/microliter throughout the observation period and was 53.7% (22/41) in patients with PNC remained less than 500/microliter . In contrast, in patient whose PNC was below 500 before the treatment but exceeded 501/microliter during of at the end of the treatment, the response rate was as high as 78.4% (29/37) . When G-CSF was combined, the response rate became significantly (P < 0.05) higher, 68.5% (50/73), as compared with that, 52.1% (61/117), in patients without it . In cases in which the causative organisms could be identified, the organisms were eliminated in 81.8% (9/11) of the patients infected with Gram-positive bacteria, whereas in 100% (12/12) in those infected with Gram-negative bacteria . Skin eruption developed in 6 patients during the treatment with CPR, and vascular pain and parosmia in one each other . These symptoms subsided soon after discontinuation or even without discontinuation of CPR . Abnormal laboratory findings, mainly liver dysfunction, i.e . elevation of slight degree of serum transaminase levels, were observed . The values, however, turned to normal immediately after the cessation or completion of the treatment . In conclusion, CPR is considered to be an antibiotic of value with high efficacy and safety in treatment of hematopoietic disorder-associated infections. Infection, 1997 Jan-Feb, 25(1), 22 - 6 Native and prosthetic valve endocarditis caused by Rothia dentocariosa: diagnostic and therapeutic considerations; Binder D et al.; Three cases, one each of native valve, prosthetic valve and composite graft endocarditis caused by Rothia dentocariosa are described . The first patient presented with multiple brain abscesses and severe congestive heart failure due to destructive endocarditis with large vegetations on the mitral valve . He died shortly after emergency valve replacement . Gram-positive coccoid rods were identified in the vegetations of the excised mitral valve . The second patient had a R . dentocariosa endocarditis of a prosthetic aortic valve that was treated empirically with netilmicin and teicoplanin, due to an allergy to penicillin . Both antibiotics were replaced according to susceptibility testing in vitro with rifampin and ciprofloxacin, and the endocarditis was cured within 9 weeks . The third patient presented with a circular root abscess of an aortic composite graft that was successfully treated with rifampin and ceftriaxone without surgery . All patients had extensive periodontal disease which was thought to be responsible for hematogenic spread and seeding of the microorganism . The microbiological identification and antibiotic resistance pattern of the isolates, as well as therapeutic implications are discussed. Gastrointest Endosc, 1997 Jan, 45(1), 52 - 8 Analysis of occluded pancreatic stents and juices in patients with chronic pancreatitis; Smits ME et al.; BACKGROUND: Pancreatic stents may occlude with time, and there is little information available on the nature of the clogging process . METHODS: We analyzed the contents of occluded pancreatic polyethylene stents in nine patients with chronic pancreatitis . In the same patients, the protein patterns in the corresponding pancreatic juices were analyzed . The stents had been in place for a mean of 9 weeks (range 2 to 17) . RESULTS: All stents were occluded at both ends, especially around side holes, with thick creamy-white precipitate . The average dry weight of occluding debris was 3 mg per 3.25 cm 10F stent . Total protein content was 50% (SD 16.3) and total calcium 0.8% of dry weight (SD 0.6) . Light microscopy showed that proteinaceous material completely filled the stent lumen . Yeasts and plant material were seen in two stents . A variable number of bacteria of mixed species, sometimes in clumps, were patchily scattered in the protein matrix . Cultures of stent contents grew several species of Gram-positive and negative bacteria . Scanning and transmission electron microscopy showed an amorphous protein matrix in all stents, arranged as a network in some areas, but in layers in other areas . Sodium dodecylsulfate polyacrimide gel electrophoresis showed that protein patterns of stent contents were remarkably different from the protein patterns of the juice samples of the same patient . A 66 kD band, identified as albumin, appeared in the protein patterns of stent content, whereas it was lacking in most juice samples . CONCLUSIONS: Adherence of protein, especially albumin, plays an important role in the process of pancreatic stent clogging . Other factors, such as bacteria, refluxed duodenal contents, and calcium seem to be of less importance. Clin Immunol Immunopathol, 1997 Jan, 82(1), 18 - 25 Blocking of TNF-alpha and IL-1 inhibits leukocyte infiltration at early, but not at late stage of S . aureus-induced arthritis and the concomitant cartilage destruction in rabbits; Kimura M et al.; We investigated the involvement of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in the pathogenesis of heat-killed S . aureus-induced arthritis . TNF-alpha and IL-1beta peaked at 2 and 24 hr after the injection, respectively . Leukocyte infiltration within 12 hr of the inflammation was significantly inhibited (80%) by coinjection of anti-TNF-alpha mAb and IL-1 receptor antagonist (IL-1Ra) with S . aureus; however, leukocyte infiltration at 24 hr and thereafter was not inhibited by these agents . The loss of proteoglycan in S . aureus-induced arthritis was also unchanged either by anti-TNF-alpha mAb, IL-1Ra, or their combination . These results indicate that direct participation of TNF-alpha and IL-1 in the pathogenesis of S . aureus-induced arthritis may be limited to the early stage of inflammation and blocking of these cytokines did not result in diminishing the severity of inflammation . Thus, therapeutic approaches with the objective to suppress TNF-alpha and IL-1 may not be effective in the clinical treatment of gram-positive bacteria-induced arthritis. J Bacteriol, 1997 Jan, 179(2), 507 - 13 Characterization of dapB, a gene required by Pseudomonas syringae pv . tabaci BR2.024 for lysine and tabtoxinine-beta-lactam biosynthesis; Liu L et al.; The dapB gene, which encodes L-2,3-dihydrodipicolinate reductase, the second enzyme of the lysine branch of the aspartic amino acid family, was cloned and sequenced from a tabtoxin-producing bacterium, Pseudomonas syringae pv . tabaci BR2.024 . The deduced amino acid sequence shared 60 to 90% identity to known dapB gene products from gram-negative bacteria and 19 to 21% identity to the dapB products from gram-positive bacteria . The consensus sequence for the NAD(P)H binding site {(V/I)(A/G)(V/I)XGXXGXXG)} and the proposed substrate binding site (HHRHK) were conserved in the polypeptide . A BR2.024 dapB mutant is a diaminopimelate auxotroph and tabtoxin negative . The addition of a mixture of L-,L-, D,D-, and meso-diaminopimelate to defined media restored growth but not tabtoxin production . Cloned DNA fragments containing the parental dapB gene restored the ability to grow in defined media and tabtoxin production to the dapB mutant . These results indicate that the dapB gene is required for both lysine and tabtoxin biosynthesis, thus providing the first genetic evidence that the biosynthesis of tabtoxin proceeds in part along the lysine biosynthetic pathway . These data also suggest that L-2,3,4,5-tetrahydrodipicolinate is a common intermediate for both lysine and tabtoxin biosynthesis. J Bacteriol, 1997 Jan, 179(2), 430 - 8 A novel member of the subtilisin-like protease family from Streptomyces albogriseolus; Suzuki M et al.; We previously isolated three extracellular endogenous enzymes from a Streptomyces albogriseolus mutant strain which were targets of Streptomyces subtilisin inhibitor (SSI) (S . Taguchi, A . Odaka, Y . Watanabe, and H . Momose, Appl . Environ . Microbiol . 61:180-186, 1995) . In the present study, of the three enzymes the largest one, with a molecular mass of 45 kDa (estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis), termed SAM-P45, has been characterized in detail . The entire gene encoding SAM-P45 was cloned as an approximately 10-kb fragment from S . albogriseolus S-3253 genomic DNA into an Escherichia coli host by using a shuttle plasmid vector . The amino acid sequence corresponding to the internal region of SAM-P45, deduced from the nucleotide sequence of the gene, revealed high homology, particularly in three regions around the active-site residues (Asp, His, and Ser), with the amino acid sequences of the mature domain of subtilisin-like serine proteases . In order to investigate the enzymatic properties of this protease, recombinant SAM-P45 was overproduced in Streptomyces coelicolor by using a strong SSI gene promoter . Sequence analysis of the SAM-P45 gene and peptide mapping of the purified SAM-P45 suggested that it is synthesized as a large precursor protein containing a large C-terminal prodomain (494 residues) in addition to an N-terminal preprodomain (23 and 172 residues) . A high proportion of basic amino acids in the C-terminal prodomain was considered to serve an element interactive with the phospholipid bilayer existing in the C-terminal prodomain, as found in other membrane-anchoring proteases of gram-positive bacteria . It is noteworthy that SAM-P45 was found to prefer basic amino acids to aromatic or aliphatic amino acids in contrast to subtilisin BPN', which has a broad substrate specificity . The hydrolysis by SAM-P45 of the synthetic substrate (N-succinyl-L-Gly-L-Pro-L-Lys-p-nitroanilide) most preferred by this enzyme was inhibited by SSI, chymostatin, and EDTA . The proteolytic activity of SAM-P45 was stimulated by the divalent cations Ca2+ and Mg2+ . From these findings, we conclude that SAM-P45 interacts with SSI and can be categorized as a novel member of the subtilisin-like serine protease family. Gene, 1996 Dec 12, 183(1-2), 175 - 82 Plasmid vectors for gram-positive bacteria switching from high to low copy number; Renault P et al.; A set of vectors for Gram-positive bacteria was constructed with a new feature which enables the switching down of their copy number per cell . These vectors carry the replication region of pAM beta 1, containing a gene essential for replication, repE, and its regulator, copF . The latter gene was inactivated by inserting a linker into its unique KpnI site . Since copF downregulates the expression of repE, its inactivation leads to an increase in the plasmid copy number per cell . The original low copy state can be restored by removal of the linker via KpnI cleavage and ligation . The new replicon was used to build (i) vectors for studying gene regulation by transcriptional or translational fusion with the bacterial luciferase gene, (ii) vectors for gene expression, and (iii) cassettes of the replicon with different multiple cloning sites, which would facilitate construction of vectors for novel purposes. Micron, 1996 Dec, 27(6), 467 - 79 Bacteriocins: nature, function and structure; Daw MA et al.; Bacteriocins are extracellular substances produced by different types of bacteria, including both Gram positive and Gram negative species . They can be produced spontaneously or induced by certain chemicals such as mitomycin C . They are biologically one of the important substances, and have been found to be useful in membrane studies and also in typing pathogenic microorganisms causing serious nosocomial infections . Bacteriocins are a heterogeneous group of particles with different morphological and biochemical entities . They range from a simple protein to a high molecular weight complex: the active moiety of each molecule in all cases seems to be protein in nature . The genetic determinants of most of the bacteriocins are located on the plasmids, apart from few which are chromosomally encoded . These bactericidal particles are species specific . They exert their lethal activity through adsorption to specific receptors located on the external surface of sensitive bacteria, followed by metabolic, biological and morphological changes resulting in the killing of such bacteria . This review summarises the classification, biochemical nature, morphology and mode of action of bacteriocins as well as their genetic determinants and the microbiological relevance of these bactericidal agents. Lupus, 1996 Dec, 5(6), 593 - 7 Induction of anticardiolipin antibody and/or lupus anticoagulant in rabbits by immunization with lipoteichoic acid, lipopolysaccharide and lipid A; Gotoh M et al.; It still remains unclear how anti-phospholipid antibody develops in a specific patient group, however, it is possible that certain microorganism(s) may cause anti-cardiolipin antibody (aCL) development since aCL is frequently detected in patients with Treponema pallidum (TP) and/or other infectious diseases . Accordingly, we conducted an investigation to clarify whether or not anticardiolipin antibody (aCL) and/or lupus anticoagulant (LA) can be induced in rabbits by immunization with Gram-positive or -negative microorganism derivatives, such as lipoteichoic acid, lipopolysaccharide and lipid A . We detected the induction of SLE type-aCL (beta 2GPI-dependent) and LA in some rabbits immunized with lipid A and lipoteichoic acid, thereby suggesting that some microorganisms may contribute to even the production of pathogenic (SLE-type) antiphospholipid antibody. Cytokine, 1996 Dec, 8(12), 933 - 7 Lymphotoxin-alpha (TNF-beta) during sepsis; Sriskandan S et al.; T cell release of lymphotoxin-alpha (LT-alpha, or TNF-beta) is stimulated by pyrogenic exotoxins of Gram-positive bacteria and mitogens . In contrast to TNF-alpha, it is unknown whether LT-alpha plays any role in the pathogenesis of sepsis and, in particular, the pathogenesis of Gram-positive sepsis . Sera from patients with sepsis were examined for LT-alpha and compared with normal volunteers and pregnant women . LT-alpha was detected in 33% of sepsis sera (mean 608.4 pg/ml SE 306), 16% of normal sera (mean 167 pg/ml SE 87) and 23% of sera from pregnant women (mean 714 pg/ml SE 191) . These differences were not significant and there were no differences within sepsis sera when grouped by the type of causative organism, or disease severity . LT-alpha detected by immunoassay in serum was not bioactive, in contrast to that produced in cell culture . Recombinant soluble TNF receptors (rSTNFR) neutralized the bioactivity of recombinant LT-alpha at rSTNFR concentrations which did not interfere with immunoreactivity and which are known to prevail in vivo . Hence, LT-alpha is unlikely to have a critical role in the pathogenesis of sepsis . Much of the potential bioactivity of this lymphokine may be abrogated by TNFR in serum. Nippon Kyobu Geka Gakkai Zasshi, 1996 Dec, 44(12), 2221 - 4 {Rupture of the aortic arch due to bacterial aortitis--a case report of a patient undergoing successful surgical therapy}; Sudo Y et al.; A 59-year-old man came to our hospital because of hoarseness and chest pain . On initial examination, a chest roentgenogram showed no abnormality . Three weeks later, while the patient was waiting to undergo a computed tomographic (CT) scan, shock suddenly developed . A chest roentgenogram revealed an extremely wide mediastinal shadow . A CT scan revealed rupture of the aortic arch near the base of the left subclavian artery, although there was no aneurysm . An emergency operation was performed . The ruptured aortic arch was replaced with an artificial graft during cardiopulmonary bypass with selective cerebral perfusion . A diagnosis of infectious aortitis was not made until a pathologist discovered colonies of gram-positive cocci in the resected specimen of the aortic arch . Postoperatively, antibiotics were administered for only four days . The patient is well about two years after the operation . Infectious aortitis had a very low prevalence and rarely involves the thoracic aorta . However, we should be aware of that it may cause aortic rupture even in the absence of aneurysmal dilatation. Biosci Biotechnol Biochem, 1996 Dec, 60(12), 2059 - 63 Efficient lipase-catalyzed preparation of long-chain fatty acid esters of bile acids: biological activity and synthetic application of the products; Sugai T et al.; A highly regioselective (3-position) and efficient (quantitative yield) acylation of bile acids catalyzed by immobilized Candida antarctica lipase was established . Methyl cholate derivatives acylated with long-chain fatty acids (C12-C16) showed an inhibitory effect on the growth of some strains of Gram-positive and -negative bacteria (27-400 micrograms/ml) . The anti-bacterial activity was slightly weaker than has been observed for methyl cholate, while the increased lipophilicity and lower melting points of the present derivatives are well suited for a potential germicide which would be safe and be topically applied . This enzyme-catalyzed transesterification is also demonstrated as an expeditious route to ursodeoxycholic acid, in respect of the regioselective introduction of acyl protecting groups on the hydroxyl groups of the intermediates . 7-Ketolithocholic acid, a known direct precursor of ursodeoxycholic acid, was obtained from cholic acid via chenodeoxycholic acid in a 46% yield and 9 steps. Protein Sci, 1996 Dec, 5(12), 2566 - 74 Kinetic and crystallographic studies of Escherichia coli UDP-N-acetylmuramate:L-alanine ligase; Emanuele JJ Jr et al.; Uridine diphosphate-N-acetylmuramate:L-alanine ligase (EC 6.3.2.8, UNAM:L-Ala ligase or MurC gene product) catalyzes the ATP-dependent ligation of the first amino acid to the sugar moiety of the peptidoglycan precursor . This is an essential step in cell wall biosynthesis for both gram-positive and gram-negative bacteria . Optimal assay conditions for initial velocity studies have been established . Steady-state assays were carried out to determine the effect of various parameters on enzyme activity . Factors studies included: cation specificity, ionic strength, buffer composition and pH . At 37 degrees C and pH 8.0, kcat was equal to 980 +/- 40 min-1, while K(m) values for ATP, UNAM, and L-alanine were, 130 +/- 10, 44 +/- 3, and 48 +/- 6 microM, respectively . Of the metals tested only Mn, Mg, and Co were able to support activity . Sodium chloride, potassium chloride, ammonium chloride, and ammonium sulfate had no effect on activity up to 75 mM levels . The enzyme, in appropriate buffer, was stable enough to be assayed over the pH range of 5.6 to 10.1 . pH profiles of Vmax/K(m) for the three substrates and of Vmax were obtained . Crystallization experiments with the enzyme produced two crystal forms . One of these has been characterized by X-ray diffraction as monoclinic, space group C2, with cell dimensions a = 189.6, b = 92.1, c = 75.2 A, beta = 105 degrees, and two 54 kDa molecules per asymmetric unit . It was discovered that the enzyme will hydrolyze ATP in the absence of L-alanine . This L-alanine independent activity is dependent upon the concentrations of both ATP and UNAM; kcat for this activity is less than 4% of the biosynthetic activity measured in the presence of saturating levels of L-alanine . Numerous L-alanine analogs tested were shown to stimulate ATP hydrolysis . A number of these L-alanine analogs produced novel products as accessed by HPLC and mass spectral analysis . All of the L-alanine analogs tested as inhibitors were competitive versus L-alanine. J Bacteriol, 1996 Dec, 178(23), 6778 - 81 Menaquinone (vitamin K2) biosynthesis: overexpression, purification, and properties of o-succinylbenzoyl-coenzyme A synthetase from Escherichia coli; Kwon O et al.; The coenzyme A (CoA)- and ATP-dependent conversion of o-succinylbenzoic acid {OSB; 4-(2'-carboxyphenyl)-4-oxobutyric acid}, to o-succinylbenzoyl-CoA is carried out by the enzyme o-succinylbenzoyl-CoA synthetase . o-Succinylbenzoyl-CoA is a key intermediate in the biosynthesis of menaquinone (vitamin K2) in both gram-negative and gram-positive bacteria . The enzyme has been overexpressed and purified to homogeneity . The purified enzyme was found to have a native molecular mass of 185 kDa as determined by gel filtration column chromatography on Sephacryl S-200 . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis established a subunit molecular mass of 49 kDa . Thus, the enzyme is a homotetramer . The enzyme showed a pH optimum of 7.5 to 8.0 and a temperature optimum of 30 to 40 degrees C . The Km values for OSB, ATP, and CoA were 16, 73.5, and 360 microM, respectively . Of the various metal ions tested, Mg2+ was found to be the most effective in stimulating the enzyme activity . Studies with substrate analogs showed that neither benzoic acid nor benzoylpropionic acid (succinylbenzene) is a substrate for the enzyme . Thus, it appears that both the benzoyl carboxyl group and the succinyl side chain are required for activation of the aliphatic carboxyl group. Curr Opin Biotechnol, 1996 Dec, 7(6), 659 - 66 Gram-positive commensal bacteria for mucosal vaccine delivery; Fischetti VA et al.; To avoid the use of engineered pathogens for vaccine delivery, systems have been developed that allow the expression of heterologous antigens in commensal Gram-positive bacteria . In some cases, both a serum IgG and secretory IgA response are induced to the recombinant protein after vaccination, verifying the validity of the approach . These live recombinant bacteria may be used in the future to introduce a protective immune response to pathogenic microorganisms after mucosal colonization. Gene, 1996 Nov 7, 179(1), 9 - 19 Bacterial resistances to toxic metal ions--a review; Silver S; Bacterial plasmids encode resistance systems for toxic metal ions, including Ag+, AsO2-, AsO4(3-), Cd2+, Co2+, CrO4(2-), Cu2+, Hg2+, Ni2+, Pb2+, Sb3+, TeO3(2-), Tl+ and Zn2+ . The function of most resistance systems is based on the energy-dependent efflux of toxic ions . Some of the efflux systems are ATPases and others are chemiosmotic cation/proton antiporters . The Cd(2+)-resistance ATPase of Gram-positive bacteria (CadA) is membrane cation pump homologous with other bacterial, animal and plant P-type ATPases . CadA has been labeled with 32P from {alpha-32P} ATP and drives ATP-dependent Cd2+ (and Zn2+) uptake by inside-out membrane vesicles (equivalent to efflux from whole cells) . Recently, isolated genes defective in the human hereditary diseases of copper metabolism, namely Menkes syndrome and Wilson's disease, encode P-type ATPases that are more similar to bacterial CadA than to other ATPases from eukaryotes . The arsenic resistance efflux system transports arsenite {As(III)}, alternatively using either a double-polypeptide (ArsA and ArsB) ATPase or a single-polypeptide (ArsB) functioning as a chemiosmotic transporter . The third gene in the arsenic resistance system, arsC, encodes an enzyme that converts intracellular arsenate {As(V)} to arsenite {As(III)}, the substrate of the efflux system . The triple-polypeptide Czc (Cd2+, Zn2+ and Co2+) chemiosmotic efflux pump consists of inner membrane (CzcA), outer membrane (CzcC) and membrane-spanning (CzcB) proteins that together transport cations from the cytoplasm across the periplasmic space to the outside of the cell. Int J Immunopharmacol, 1996 Nov, 18(11), 661 - 8 Occurrence and specificity of human natural and in vitro induced antibodies to Nocardia opaca antigens; Prokesova L et al.; Nocardia opaca, a Gram-positive bacterium, is a potent source of immunostimulatory substances . Screening of sera of adult human donors revealed that all sera tested contained antibodies reactive with isolated Nocardia fractions (Nocardia delipidated cell mitogen, NDCM; Nocardia lysozyme digest, NLD; Nocardia water-soluble mitogen, NWSM; and fraction B) . The respective values of reciprocal titres for IgM and IgG were in the range of 100 to 12,800, and 10 to 320 for IgA antibody isotypes, when NLD or fraction B were used as antigens in enzyme-linked immunosorbent assay (ELISA) tests . The level of antibodies directed to NDCM, a potent polyclonal B cell activator, was found to be the lowest . In vitro spontaneous as well as NDCM-induced production of antibodies to NDCM by human peripheral blood lymphocytes involved mainly the IgM class . Western-blot analysis demonstrated that antibodies in normal human sera react with nocardial antigens of molecular mass approximately 60, 40, 20 and 15-10 kDa . The same antigens were also recognized by rabbit and mouse hyperimmune sera, also confirming the immundominancy of these nocardial antigens in other species . The presence of anti-nocardia antibodies in human sera and their production by both stimulated and non-stimulated lymphocytes points to the natural sensitization of humans either by ubiquitous no-cardial components or by cross-reactive bacterial or food antigens. J Chir (Paris), 1996 Nov, 133(7), 348 - 52 {Abdominal actinomycosis . Apropos of a case}; Kabwa L et al.; Abdominal actinomycosis is a chronic suppurating granulomatosis caused by a Gram positive germ, Actinomyces . Manifestations of this rare disease are usually pseudotumoral syndrome leading to surgical exeresis . The diagnosis is obtained from the pathology report . We report a pseudotumor of the transverse colon which invaded the greater omentum and the anterior abdominal wall . From a review of the recent literature, we observed that most papers deal essentially with a relatively new entity, pelvic actinomycosis related to prolonged use of an intra-uterine device without medical surveillance. J Antibiot (Tokyo), 1996 Nov, 49(11), 1101 - 4 Maltophilin: a new antifungal compound produced by Stenotrophomonas maltophilia R3089; Jakobi M et al.; Strains of Stenotrophomonas maltophilia R3089, isolated from the rhizosphere of rape plants (Brassica napus L.), produced a novel antifungal compound, named maltophilin, which inhibited the growth of various saprophytic, human-pathogenic and phytopathogenic fungi but was inactive against Gram-positive and Gram-negative bacteria . Maltophilin is a novel macrocyclic lactam antibiotic with a molecular mass of 510 mu . The compound was isolated from the culture filtrate by ethyl acetate extraction and gel filtration on Sephadex LH20 and purified by preparative HPLC on reversed phase . The structure of maltophilin was elucidated by electrospray mass spectrometry and 1H and 13C NMR spectroscopy. J Clin Periodontol, 1996 Nov, 23(11), 1025 - 31 Predominant cultivable supragingival plaque in Chinese "rapid" and "slow" plaque formers; Zee KY et al.; The aim of this study was to compare the predominant cultivable bacterial flora in supragingival plaque samples of Chinese "rapid" and "slow" plaque formers, using the experimental gingivitis model . 11 Chinese subjects (5 "rapid" and 6 "slow" plaque formers) were selected from 49 healthy young adults . The selection was based on the plaque index on the buccal surfaces of all the canines, premolars and 1st molars after 3-days without plaque control . The 11 subjects began a 14-day on oral hygiene period after prophylaxis . Plaque samples were collected on day 1, day 3, day 7 and day 14 from the buccal surface of upper right canine, 2nd premolar, 1st premolar and 1st molar, respectively . The samples were then dispensed into a tryptic soy broth, and cultured anaerobically to obtain pure isolates which were then identified by conventional means . Results showed similar trends in plaque development in the two groups; gram-positive bacteria were the predominant cultivable species ("rapid": 71-37%; "slow": 53-63%) and gram-negative species increased in proportion to the plaque age ("rapid": 9-47%, "slow": 13-28%) . "Rapid" plaque formers showed a statistically significant higher percentage of gram-negative rods (38%) than the "slow" group (17%) in the 14-day samples . The difference in the proportions of other groups of bacteria between the "rapid" and "slow" plaque formers were also found to be statistically significant using the MANOVA test (p = 0.0162) . Most of the cultivable gram-negative rods belonged to Fusobacterium and Capnocytophaga species . Besides the quantitative difference observed clinically, there seems to be a qualitative difference in the cultivable flora between subjects with different rates of plaque formation. Am J Gastroenterol, 1996 Nov, 91(11), 2418 - 20 Hepatic actinomycosis infiltrating the diaphragm and right lung; Kasano Y et al.; Hepatic actinomycosis is a rare infectious disease caused by an anaerobic gram-positive bacterium of the genus Actinomyces . Herein, we describe an unusual case of hepatic actinomycosis involving the diaphragm and right lung . A 41-yr-old man was admitted to Wakayama Medical School Hospital presenting with right back pain and cough . Computed tomography and magnetic resonance image revealed a 5 x 10 cm tumor in the anterior superior segment of the liver, which extended to the diaphragm and right lung . Angiography demonstrated a hypervascular tumor and the enlarged right inferior phrenic artery feeding around the tumor . The patient underwent a hepatectomy with partial resections of the diaphragm and the right middle pulmonary lobe . Microscopically, the specimen showed sulfur granules and was positive for Gram stain and Grocott stain and negative for Ziehl-Neelsen stain . These findings were consistent with actinomycosis of the liver . His postoperative course was uneventful and no recurrence was observed 1 yr postoperatively . Although there are at least 36 well-documented cases until 1993, no other report has been found infiltrating the diaphragm and lung. Eur J Immunol, 1996 Nov, 26(11), 2686 - 92 Induction of CD14 expression in Lpsn, Lpsd and tumor necrosis factor receptor-deficient mice; Takakuwa T et al.; The involvement of CD14 in lipopolysaccharide (LPS) recognition and signaling has been demonstrated in several studies . For this reason, we investigated whether the resistance of Lpsd mice to LPS might be related to an impaired CD14 expression . We compared the in vivo and in vitro expression of CD14 in Lpsn (LPS sensitive) and Lpsd mice, and its modulation by LPS, killed gram-negative and gram-positive bacteria and double-stranded (ds)RNA . Untreated Lpsn and Lpsd cultured macrophages (M phi), expressed similar amounts of CD14 mRNA and membrane-bound (m)CD14 . LPS enhanced CD14 expression only in Lpsn M phi, while all bacteria, or dsRNA, enhanced CD14 in Lpsn and Lpsd M phi . Similarly, in vivo administration of LPS induced or enhanced CD14 mRNA in different organs of Lpsn mice only, while bacteria or dsRNA in both types of mouse . Furthermore, exogenous recombinant tumor necrosis factor (TNF) induced in vivo and in vitro enhanced CD14 expression in Lpsn, Lpsd and also in TNF receptor 2-deficient (TNFR2-/-) mice, but failed to do so in TNFR1-/- mice, showing that TNFR1 mediates the effect of TNF on CD14 . However, LPS, bacteria and dsRNA induced CD14 in both TNFR2-/- and TNFR1-/- mice to a similar extent, revealing that this induction does not require TNF signaling. Crit Care Med, 1996 Nov, 24(11), 1818 - 23 Prospective, randomized trial of two antiseptic solutions for prevention of central venous or arterial catheter colonization and infection in intensive care unit patients; Mimoz O et al.; OBJECTIVES: To compare the efficacy of a newly available antiseptic solution (composed of 0.25% chlorhexidine gluconate, 0.025% benzalkonium chloride, and 4% benzyl alcohol), with 10% povidone iodine, on the prevention of central venous or arterial catheter colonization and infection . DESIGN: Prospective, randomized clinical trial . SETTING: Surgical-trauma intensive care unit (ICU) in a university hospital . PATIENTS: All patients admitted to the ICU and requiring the insertion of a central venous and/or an arterial catheter from July 1, 1992 to October 31, 1993 . INTERVENTIONS: Patients were randomly assigned to one of two groups according to the antiseptic solution used for insertion and catheter care . The same solution was used for skin disinfection from the time of catheter insertion to the time of removal of each catheter . MEASUREMENTS AND MAIN RESULTS: Catheter distal tips were quantitatively cultured when catheters were no longer necessary, if there was a suspicion of catheter-related infection, and routinely after 7 days of use for arterial catheters, or after 15 days of use for central venous catheters . The rate of significant catheter colonization (i.e., > or = 10(3) colony-forming units {cfu}/mL by quantitative culture), and catheter-related sepsis (as defined by sepsis abating following catheter removal per 1,000 catheter-days), were significantly lower in the chlorhexidine group (12 vs . 31 {relative risk 0.4, 95% confidence interval 0.1 to 0.9, p < .01} and 6 vs . 16 {relative risk 0.4, 95% confidence interval 0.1 to 1, p = 0.5}, respectively) . The rate of central venous catheter colonization and central venous catheter-related sepsis per 1,000 catheter-days were also significantly lower in the chlorhexidine group (8 vs . 31 {relative risk 0.3, 95% confidence interval 0.1 to 1, p = .03} and 5 vs . 19 {relative risk 0.3, 95% confidence interval 0.1 to 1, p = .02}, respectively) . Finally, the rate of arterial catheter colonization per 1,000 catheter-days was significantly lower in the chlorhexidine group (15 vs . 32 {relative risk 0.5, 95% confidence interval 0.1 to 1, p = .05}), whereas the rate of arterial catheter-related sepsis per 1,000 catheter-days was similar for the two study groups (8 in the chlorhexidine group vs . 10 in the povidone iodine group {relative risk 0.8, 95% confidence interval 0.1 to 2.2, p = .6}) . The 0.25% chlorhexidine solution was superior to the 10% povidone iodine solution in preventing catheter colonizations and catheter-related sepsis due to Gram-positive bacteria (5 vs . 20 {p < .001}, and 2 vs . 10 {p < .001}, respectively), whereas the activity of the 0.25% chlorhexidine solution was nonsignificantly superior in preventing Gram-negative infections (7 vs . 4 {p = .5}, and 4 vs . 2 {p = .8}, respectively) . CONCLUSIONS: The 4% alcohol-based solution of 0.25% chlorhexidine gluconate and 0.025% benzalkonium chloride was more effective than 10% povidone iodine for insertion site care of short-term central venous and arterial catheters . This effect appeared related to a more efficacious prevention of infections with Gram-positive bacteria. Mol Pharmacol, 1996 Nov, 50(5), 1065 - 72 Induction of cytochrome P450 2E1 expression in rat and gerbil astrocytes by inflammatory factors and ischemic injury; Tindberg N et al.; Hepatic cytochromes P450 are known to be down-regulated by cytokines, lipopolysaccharide, Gram-positive bacteria, and viruses . Little is known, however, about the regulation by inflammation of cytochromes P450 in other tissues . We have found that lipopolysaccharide and interleukin-1 beta stimulate the expression of catalytically active CYP2E1 (but not CYP1A1 or CYP2B) up to 7-fold in rat brain primary cortical glial cultures . The induction reached a maximum after 24 hr and was accompanied by an increase in CYP2E1 mRNA . Chlormethiazole, a specific inhibitor of hepatic CYP2E1 transcription, completely inhibited the induction of CYP2E1 at the mRNA and enzyme levels . Immunofluorescence studies showed CYP2E1 to be expressed in a subset of astrocytes in the lipopolysaccharide-stimulated cortical glial cultures . Using a model of global ischemic injury in the gerbil, we found CYP2E1 to be induced in vivo in astrocytes in the inflammatory phase, 1-3 weeks after the lesion . Likewise, CYP2E1 was induced in the rat cortex 1 week after a focal ischemic injury . Our results suggest tissue-specific regulation of CYP2E1 by inflammatory factors and that CYP2E1 may play a role in astrocytes during inflammation in the brain. Am J Respir Crit Care Med, 1996 Nov, 154(5), 1261 - 6 Exposure to bacteria in swine-house dust and acute inflammatory reactions in humans; Zhiping W et al.; Inhalation of swine-house dust may cause an acute airway inflammatory condition (organic dust toxic syndrome) . Thirty-eight healthy subjects were exposed to swine dust while weighing swine for 3 h . We studied the correlation between acute health effects and the inhaled bacterial exposure markers peptidoglycan (the main constituent of the cell walls of gram-positive bacteria, but also present in lesser amounts in gram-negative bacteria) and lipopolysaccharides (LPS; present only in gram-negative bacteria) . LPS activity in airborne dust was measured with the Limulus amebocyte lysate assay (LPS(LAL)), and the total LPS was estimated from 3-hydroxy fatty acids, which were measured with gas chromatography-mass spectrometry (GC-MS) (LPS(GC-MS)) . Peptidoglycan was estimated from muramic acid measured with GC-MS . The median (25th to 75th percentile) concentration of inhalable dust was 21 (16 to 25) mg/m3 . LPS(LAL) was 1.2 (0.9 to 1.4) microg/m3; LPS(GC-MS) was 3.9 (2.5 to 4.9) microg/m3; and the peptidoglycan concentration in airborne dust was 6.5 (2.7 to 13) microg/m3 . All exposure markers correlated significantly with an increase in serum interleukin-6 . LPS(LAL) showed the highest correlation (r2 = 0.29) and total inhaled dust the lowest (r2 = 0.09) . LPS(LAL) also correlated with symptoms and with an increase in bronchial responsiveness and decrease in vital capacity (VC) . Peptidoglycan, but not LPS(LAL), correlated with an increase in the blood granulocyte concentration and in body temperature . The results suggest that several microbial agents in inhaled swine-house dust may contribute to acute systemic health effects. Appl Environ Microbiol, 1996 Nov, 62(11), 3967 - 77 Cloning, sequencing, and expression of isopropylbenzene degradation genes from Pseudomonas sp . strain JR1: identification of isopropylbenzene dioxygenase that mediates trichloroethene oxidation; Pflugmacher U et al.; Pseudomonas sp . strain JR1, recently isolated with isopropylbenzene (IPB) as the inducer substrate for trichloroethene (TCE) oxidation (B . Dabrock, J . Riedel, J . Bertram, and G . Gottschalk, Arch . Microbiol 158:9-13, 1992), is able to degrade IPB via the meta-cleavage pathway . The genes encoding the first three enzymes in the catabolism of isopropylbenzene were isolated from a genomic library with the broad-host-range cosmid vector pWE15 . A 7.6-kb fragment from a 37.7-kb primary cosmid clone was subcloned and sequenced . It contained seven complete open reading frames, designated ipbA1A2orf3A3A4BC . ipbA codes for the three subunits of a multicomponent IPB dioxygenase, ipbB codes for 2,3-dihydro-2,3-dihydroxy-IPB dehydrogenase, and ipbC codes for 3-isopropylcatechol 2,3-dioxygenase . The deduced amino acid sequences of ipbA1A2A3A4BC exhibited the highest homologies with the corresponding proteins of biphenyl-degradative pathways in gram-negative and gram-positive bacteria . The gene products of the ipb genes were identified by an in vitro transcription-translation system on the basis of their expected molecular masses . IPB dioxygenase and 3-isopropylcatechol 2,3-dioxygenase expressed in E . coli oxidized a wide range of alkyl aromatic compounds . Incubation of E . coli cells carrying ipbA1A2A3A4 with IPB and 10O2 yielded reaction products containing both atoms of molecular oxygen, which is in accordance with a dioxygenation reaction . E . coli recombinants harboring and expressing the IPB dioxygenase exhibited the ability to degrade TCE . The ipbA1A2A3A4-carrying E . coli strain required neither IPB nor isopropyl-beta-D-thiogalactopyranoside for induction; the rate of TCE degradation was comparable to that by fully induced Pseudomonas strain JR1. Appl Environ Microbiol, 1996 Nov, 62(11), 3922 - 8 Denaturing gradient gel electrophoresis used to monitor the enrichment culture of aerobic chemoorganotrophic bacteria from a hot spring cyanobacterial mat; Santegoeds CM et al.; Previous studies investigating microbial diversity in the Octopus Spring cyanobacterial mat community (Yellowstone National Park) have shown a discrepancy between bacterial populations observed by molecular retrieval and cultivation techniques . To investigate how selective enrichment culture techniques affect species composition, we used denaturing gradient gel electrophoresis (DGGE) separation of PCR-amplified 16S rRNA gene fragments to monitor the populations contained within enrichment cultures of aerobic chemoorganotrophic bacteria from the ca . 50 degrees C region of the mat community . By varying the degree of dilution of the inoculum, medium composition, and enrichment conditions and duration and by analyzing the cultures by DGGE, we detected 14 unique 16S rRNA sequence types . These corresponded to alpha-, beta-, gamma-, and delta-proteobacteria, Thermus relatives, and gram-positive bacteria with high G + C ratio and, at the highest inoculum dilutions, Chloroflexus aurantiacus relatives, which were estimated to still be approximately 300 times less abundant than cells of the mat primary producer, Synechococcus spp . Only three of these populations were previously cultivated on solidified medium after similar enrichment . Only two of these population have 16S rRNA sequences which were previously cloned directly from the mat . These results reveal a diversity of bacterial populations in enrichment culture which were not detected by either molecular retrieval or strain purification techniques. J Immunol, 1996 Nov 1, 157(9), 4119 - 25 Monocytes can phagocytose Gram-negative bacteria by a CD14-dependent mechanism; Grunwald U et al.; Phagocytosis of bacteria by monocytes and neutrophil granulocytes provides an important first line of defense against bacterial infections . Opsonization of bacteria with complement and phagocytosis by neutrophils is dependent on divalent cations and does not take place in blood that has been anticoagulated with EDTA . Monocytes, however, do carry out phagocytosis even in the presence of EDTA . We show here that this divalent cation-independent phagocytosis pathway requires the presence of the LPS receptor CD14 on the cell surface . This pathway is dependent on the availability of LPS binding protein, can be blocked by anti-CD14 Abs, by an excess of soluble CD14, by excess free LPS, or by an excess of unlabeled Gram-negative bacteria . In contrast, intact Gram-positive bacteria fail to inhibit this process . These experiments define a CD14-dependent phagocytosis pathway for Gram-negative bacteria that operates in monocytes in human whole blood . This pathway may be able to deal with bacterial pathogens that have developed resistance to complement-dependent opsonization and phagocytosis by neutrophils. Microb Ecol, 1996 Nov, 32(3), 305 - 21 Viability and Metabolic Features of Bacteria Indigenous to a Contaminated Deep Aquifer McCarthy CM, Murray L. The quantitation and characterization of indigenous bacteria of a deep aquifer, located in the southwestern United States and contaminated with halogenated aliphatic compounds, was undertaken . Water samples were obtained aseptically from depths of 45 to 151 m from four sites that ranged from 260 to 1,800 m in distance from the location of contaminant release . Sediment samples were also obtained from the proximal and distal sites for analyses . Results for aerobic and anaerobic colony-forming units were obtained on four agar media that were used to retrieve heterotrophs, oligotrophs, and pseudomonads . Most probable number estimates were obtained from a liquid medium favorable for oligotrophs . Representative isolates were tested against Biolog plates (Biolog, Inc., Hayward, Calif.) for patterns of carbon source utilization . Of 103 Gram-negative (GN) isolates, 48 could not be identified and the others were only tentatively identified via the Biolog database, and none of the 35 Gram-positive (GP) isolates were identifiable . However, the metabolic patterns were subjected to average cluster linkage analyses; the GN and GP bacteria were separable into eight and four groups, respectively . The oligotroph group comprised one-third of the GN and one-half of the GP isolates . The consensus carbon source utilization pattern for each group was determined and will be useful in future characterization of additional aquifer bacterial isolates . Although predominantly aerobic and oligotrophic, the microbial community of this aquifer was highly diverse with discernible viability and metabolic features of the microbiota distinctive to each of the four water and two sediment samples. Oral Microbiol Immunol, 1996 Oct, 11(5), 356 - 61 Development and evaluation of a selective and differential medium for the primary isolation of Peptostreptococcus micros; Turng BF et al.; Peptostreptococcus micros, an anaerobic gram-positive coccus, has been associated with periodontal and endodontic lesions, including those refractory to treatment, as well as many human polymicrobial infections in other body locations . A selective and differential medium for the primary isolation of P . micros was developed and evaluated . Columbia CNA agar, a selective medium for gram-positive cocci, was supplemented with glutathione and lead acetate (P . micros medium: PMM) . P . micros has a characteristic of rapidly utilizing the reduced form of glutathione to form hydrogen sulfide, which reacts with lead acetate producing a black precipitate in the medium . When grown on PMM, P . micros can be easily identified by its typical colonial morphology and the presence of a black precipitate directly under the colony . PMM was compared for the growth of P . micros with phenylethyl alcohol agar (PEA) and Columbia base medium (CBM) with 80 strains of P . micros and 30 strains of other gram-positive cocci . All P . micros isolates tested grew and showed the typical morphology of P . micros on PMM . Using colony counts on CBM as controls, there was an average 81.8% recovery in the number of P . micros colonies on PMM, in contrast to an average 6.1% recovery on PEA . Subgingival plaque and tongue samples from 12 adult periodontitis and 6 early-onset periodontitis patients were cultured onto PMM for the isolation of P . micros . P . micros was isolated on PMM and identified biochemically and enzymatically from both adult periodontitis and early-onset periodontitis patients with higher percentages isolated from the diseased periodontal pockets of adult periodontitis patients; furthermore, this is the first isolation of P . micros from tongue samples taken from periodontally diseased patients . This medium in cultural studies will further our understanding and assist future investigations of P . micros involved in disease processes. Avian Dis, 1996 Oct-Dec, 40(4), 894 - 9 Neonatal multifocal encephalomalacia and other lesions in the brains of Georgia chicks: 1991-95; Goodwin MA et al.; Outbreaks of neonatal multifocal encephalomalacia with sepsis have been reported among flocks of very young chicks in Belgium, Scotland, and the United States . The purposes of the present study were to describe intralesional bacterial cocci in chicks with this type of encephalomalacia and to determine its incidence during 1991-95, and to determine the importance of this lesion with respect to the frequency of all other brain lesions/ diseases during the same time period . All laboratory records of broiler chickens examined at the Georgia Poultry Laboratory from Jan . 1, 1991, through Dec . 31, 1995, where the histopathologic diagnoses included the letter string *encephal* were retrieved for further study . The leading etiology for brain disease was nutritional encephalomalacia (57%), followed by neonatal encephalomalacia (22%), septic meningoencephalitis (16%), and Marek's disease (14%), in turn followed by nonpurulent encephalitis (7%), avian encephalomyelitis (3%), and mycotic meningoencephalitis (3%) . Diagnosis of neonatal multifocal encephalomalacia with sepsis in the brains of Georgia chicks is a perennial one . Microscopically, the condition is characterized by mild to maximal multifocal locally extensive fibrin thrombosis of blood capillaries, and necrosis (encephalomalacia, malacia) of surrounding zones of brain stem and/or cerebral hemisphere neuropile . In 44% of cases of neonatal encephalomalacia fibrin thrombosis of blood capillaries was accompanied by intralesional gram-positive coccoid bacteria that were most abundant in medium- and small-sized arterioles or venules . Only gram-positive coccoid bacteria are found in cases of neonatal encephalomalacia (P < 0.05), and only gram-negative rod-shaped bacteria are found in cases of septic meningoencephalitis (P < 0.05) . Therefore, bacterial culture and routine light microscopic histopathology are sufficient for diagnosing the condition when the differential diagnosis for neurologic disease in chicks includes neonatal encephalomalacia. Infect Immun, 1996 Oct, 64(10), 4067 - 73 Role of Porphyromonas gingivalis protease activity in colonization of oral surfaces; Tokuda M et al.; Cysteine proteases, including Arg-gingipain of Porphyromonas gingivalis, have been implicated as important virulence factors in periodontal diseases . These enzymes are also involved in the hemagglutinating activity of the organisms . In order to determine the role of proteases in the colonization of the gingival margin, we have compared the attachment properties of P . gingivalis 381 with those of its Arg-gingipain-defective mutant, G-102 . Interactions with gram-positive bacteria, human oral epithelial cells, extracellular matrix proteins, and type I collagen were evaluated . In all cases, mutant G-102 was deficient in attachment relative to the parental strain . The mutant's defects could be explained, in part, by the weak autoaggregation displayed by the mutant, which appeared to result from altered fimbrial expression . Both Western blot (immunoblot) and Northern (RNA) blot analyses indicated reduced expression of the major 43-kDa fimbrillin subunit in the mutant . These results suggest that Arg-gingipain may play both direct and indirect roles in the colonization of the gingival margin . In addition, fimbriae may play a direct role in interacting with some host surfaces. Semin Perinatol, 1996 Oct, 20(5), 426 - 38 Antibiotic therapy for preterm premature rupture of the membranes; Mercer BM et al.; Amniotic fluid studies and placental histopathological evaluation have confirmed the association between intrauterine infection and preterm premature rupture of the membranes (pPROM) . This association is increasingly strong with pPROM at early gestational ages . The organisms associated with pPROM include a broad spectrum of aerobic and anaerobic gram-positive and gram-negative bacteria . In many cases, the patient presenting with pPROM will have overt intrauterine infection necessitating delivery . For those amenable to expectant management, the clinical course is usually of brief latency between membrane rupture and delivery . A number of well-designed prospective clinical trials have evaluated the utility of antibiotic treatment during the expectant management of pPROM . Taken together, these studies suggest broad spectrum antibiotic treatment of this population to enhance pregnancy prolongation, and to reduce maternal and neonatal infectious morbidity . There are some data suggesting the potential for a reduction in neonatal gestational age-dependent morbidity . We recommend aggressive adjunctive antibiotic treatment to prolong pregnancy and reduce morbidity in patients with pPROM, at gestations remote from term, when a significant improvement in neonatal outcome can anticipated with expectant management. Clin Infect Dis, 1996 Oct, 23(4), 795 - 805 Prophylaxis with fluoroquinolones for bacterial infections in neutropenic patients: a meta-analysis; Cruciani M et al.; We conducted a two-part meta-analysis to assess the effectiveness of fluoroquinolones for preventing bacterial infections in granulocytopenic patients who are receiving chemotherapy for malignancies . Overall, 19 randomized studies met selection criteria and were included in this meta-analysis of 2,112 patients . Thirteen studies that compared the fluoroquinolones alone with control regimens (co-trimoxazole, oral nonabsorbable antibiotics, or placebo) and six studies that compared the fluoroquinolones plus prophylaxis for bacteremia due to gram-positive bacteria with control regimens (fluoroquinolones or oral nonabsorbable antibiotics) were included in the two meta-analyses . The results of the first meta-analysis indicate that fluoroquinolones alone are effective in preventing gram-negative bacteremia (overall odds ratio {OR}, 0.09; 95% confidence interval {CI}, 0.05-0.16; P < .001), but not gram-positive bacteremia (OR, 1.05; 95% CI, 0.76-1.45; P = .7), fever-related morbidity (OR, 0.76; 95% CI, 0.56-1.04; P = .09), and infection-related mortality (OR, 0.79; 95% CI, 0.47-1.34; P = .4) . The results of the second meta-analysis indicate that a combination of fluoroquinolones plus prophylaxis for gram-positive bacteremia (penicillin, vancomycin, or macrolides) significantly reduces the occurrence of gram-positive bacteremia (OR, 0.46; CI, 0.33-0.63; P < .001) without affecting the incidence of fever-related morbidity (OR, 0.83; 95% CI, 0.62-1.13; P = .2) and infection-related mortality (OR, 0.74; 95% CI, 0.40-1.38; P = .3) Eur J Biochem, 1996 Oct 1, 241(1), 17 - 24 Cloning, purification and characterization of the protein subunit of ribonuclease P from the cyanobacterium Synechocystis sp . PCC 6803; Pascual A et al.; The rnpA gene from the cyanobacterium Synechocystis sp . PCC 6803, which codes for the protein subunit of ribonuclease P (RNase P), has been cloned by functional complementation of an Escherichia coli mutant . This protein had previously been characterized only in proteobacteria and gram-positive bacteria . rnpA and the closely linked rpmH gene, which code for the large subunit ribosomal protein L34, have been sequenced . The Synechocystis 6803 L34 protein is more similar to the homologous protein from some non-green chloroplasts than to the L34 protein from other bacteria . The protein subunit of RNase P from Synechocystis 6803 has been overexpressed in E . coli and purified to homogeneity . Antibodies raised against the Synechocystis 6803 RNase P protein did not recognize the homologous protein from E . coli (C5 protein) . Similarly, antibodies raised against the E . coli C5 protein did not recognize significantly the Synechocystis 6803 protein . In spite of the lack of immunological cross-reactivity and the low level of sequence identity, the E . coli and Synechocystis 6803 proteins are functionally interchangeable . In enzymatic assays using either an E . coli precursor tRNA(Tyr) or a Synechocystis 6803 precursor tRNA(Gln) as substrates, we have detected RNase P activity with holoenzymes reconstituted with the RNA subunit from E . coli and the protein subunit from Synechocystis 6803 or with the RNA subunit from Synechocystis 6803 and the protein subunit from E . coli . The relative efficiency of cleavage of the different substrates is dependent on the origin of the protein subunit used to reconstitute the holoenzyme. Microbiology, 1996 Oct, 142 ( Pt 10), 2863 - 70 Molecular biological evidence for the occurrence of uncultured members of the actinomycete line of descent in different environments and geographical locations; Rheims H et al.; A 16S rDNA based molecular ecological study was performed on a sample taken from a peat bog in Germany . Total DNA was extracted by directly lysing micro-organisms in the peat matrix . The 5' 1400 nucleotides of the bacterial 16S rDNA were amplified using conserved bacterial PCR primers . A clone library was generated by blunt-end cloning and 262 16S rDNA clones were analysed . Of these, 37 were located in the Gram-positive phylum, as determined by hybridization to an oligonucleotide probe specific for Gram-positive bacteria . Analysis of 17 of these clones by sequence analysis and their comparison with published sequences representing all of the main bacterial phyla indicated their membership of the actinomycete line of descent . These peat clones were found to represent three novel lineages, two of which appear to be related to the species Acidimicrobium ferrooxidans, and 'Candidatus Microthrix parvicella' . Clone sequences of the third group are phylogenetically related to Rubrobacter radiotolerans . Comparison with short 16S rDNA clone sequences obtained from DNA isolated from a geothermally heated soil in New Zealand, and from DNA isolated from soil in Australia, Japan and Finland and marine environments from the Atlantic and the Pacific Oceans, suggests that members of these three groups occur in very different environments across the world. J Clin Microbiol, 1996 Oct, 34(10), 2598 - 600 Rapid preparation of bacterial DNA for pulsed-field gel electrophoresis; Matushek MG et al.; A disadvantage of genotyping bacterial strains by pulsed-field gel electrophoresis is that the procedure requires up to 6 days to complete . We modified a standard pulsed-field gel electrophoresis method (B.E . Murray, K.V . Singh, J.D . Health, B.R . Sharma, and G.M . Weinstock, J.Clin . Microbiol . 28:2059-2063, 1990) so that it could be completed in less than 3 days . We successfully applied this method to the analysis of a variety of gram-positive and gram-negative bacteria. Int J Syst Bacteriol, 1996 Oct, 46(4), 967 - 71 Leucobacter komagatae gen . nov., sp . nov., a new aerobic gram-positive, nonsporulating rod with 2,4-diaminobutyric acid in the cell wall; Takeuchi M et al.; A new aerobic, gram-positive, nonsporulating rod-shaped organism is described: Strain IFO 15245T (T = type strain) has the following characteristics: the menaquinone contains a side chain with 11 isoprenyl units; the guanine-plus-cytosine content of the DNA is 66.2 mol%; 2,4-diaminobutyric acid, glutamic acid, alanine, glycine, and gamma-aminobutyric acid are present in the cell wall at a molar ratio of ca . 1:1:2:1:1; and glucose and galactose are also present in the cell wall . A comparison of partial 16S rRNA sequences revealed that IFO 15245T represents a distinct line of descent within the gram-positive bacteria with high guanine-plus-cytosine contents . The taxonomic characteristics of this organism are different from those of previously described aerobic, gram-positive, nonsporulating, rod-shaped bacteria . The name Leucobacter komagatae gen . nov., sp . nov., is proposed for this organism . The type strain is strain IFO 15245. Biochemistry, 1996 Sep 3, 35(35), 11268 - 77 Influence of N-cap mutations on the structure and stability of Escherichia coli HPr; Thapar R et al.; This paper describes the effect of N-capping substitutions on the structure and stability of histidine-containing protein (HPr) . We have used NMR spectroscopy and conformational stability studies to quantify changes in local and global free energy due to mutagenesis at Ser46, the N-cap for helix B in HPr . Previous NMR studies suggested that helix B of Escherichia coli HPr is dynamic as judged by the rate of exchange of amide protons with solvent . Ser46 was chosen because it is the site of regulatory phosphorylation in HPrs from Gram-positive bacteria, and mutation of this residue to an aspartic acid (S46D) in E . coli HPr (Gram-negative) also makes it a poor substrate in the bacterial phosphoenolpyruvate: sugar phosphotransferase system . Therefore, to understand the mechanism of inactivation of E . coli S46D HPr, as well as the effect of mutagenesis on protein stability, we have characterized three mutants of E . coli HPr: Ser46 has been mutated to an Asp, Asn, and Ala in S46D, S46N, and S46A HPrs, respectively . The results indicate that these N-cap replacements have a marked influence on helix B stability . The effect of mutagenesis on local stability is correlated to global unfolding of HPr . The ability of amino acids to stabilize helix B is Asp > Asn > Ser > Ala . In addition, since there are neither large-scale conformational changes nor detectable changes in the active site of S46D HPr, it is proposed that the loss of phosphotransfer activity of S46D HPr is due to unfavorable steric and/or electrostatic interactions of the Asp with enzyme I of the PTS. Clin Oral Implants Res, 1996 Sep, 7(3), 190 - 200 Microbiology of ligature-induced marginal inflammation around osseointegrated implants and ankylosed teeth in cynomolgus monkeys (Macaca fascicularis); Schou S et al.; The microbiota associated with ligature-induced marginal inflammation around osseointegrated dental implants, ankylosed teeth, and normal control teeth was investigated in 8 cynomolgus monkeys (Macaca fascicularis) . Submucosal/subgingival plaque was sampled with paper points on the day of literature placement and after 7 weeks . The samples were evaluated by phase-contrast microscopy and by cultivation on enriched non-selective and various solid media . The submucosal/subgingival flora was changed 7 weeks after ligation . The total number of cultivable bacteria and the proportions of motile rods, anaerobic Gram-negative rods, black-pigmented rods, Porphyromonas gingivalis, and Prevotella intermedia increased significantly around implants, ankylosed teeth, and normal control teeth . Except for a significantly higher proportion of anaerobic Gram-positive cocci around implants compared to ankylosed teeth and normal control teeth at the end of the study, no significant microbiological differences were observed between implants, ankylosed teeth, and normal control teeth neither at baseline nor at the end of the study . Consequently, the microbiota associated with marginal inflammation around implants, ankylosed teeth, and normal control teeth appears to be rather similar in cynomolgus monkeys. Phytochemistry, 1996 Sep, 43(1), 129 - 32 Antibiotic phenol nor-triterpenes from Maytenus canariensis; Gonzalez AG et al.; The new phenols 6-oxo-tingenol, 3-O-methyl-6-oxo-tingenol and 6-oxo-iguesterol were isolated from the root bark of Maytenus canariensis . Their structures were determined by 1H and 13C NMR spectroscopic studies, including HMQC, HMBC, DEPT and ROESY and chemical transformations . The synthesis of 6-oxo-tingenol was achieved from tingenone . These compounds exhibit antibiotic activity against Gram-positive bacteria. Eur J Cancer, 1996 Sep, 32A(10), 1739 - 43 Randomised comparison of ceftazidime and imipenem as initial monotherapy for febrile episodes in neutropenic cancer patients; Aparicio J et al.; With the availability of new, broad-spectrum antibiotics, initial therapy with a single agent has become an alternative to classic combinations in the management of febrile, neutropenic cancer patients . The aims of this study were to compare the efficacy of ceftazidime and imipenem as empirical monotherapy of febrile episodes in neutropenic patients, and to examine the frequency with which second-line antibiotics (amikacin, vancomycin, or both) were required . A prospective clinical trial was carried out in a single centre . Eligible patients with solid tumours or lymphoma were randomised to receive monotherapy with ceftazidime or imipenem . In the event of no response, amikacin and/or vancomycin were added in 48-72 h intervals (sequentially, or according to clinical or microbiological data) . Efficacy was evaluable for 111 assessable episodes . Median neutrophil count at entry was 100 cells/microliters and median duration of neutropenia was 4 days . Febrile episodes were classified as microbiologically (34%) or clinically documented (42%), and fever of unknown origin (24%) . Gram-negative infections (57%) predominated over gram-positive isolates (30%) . The overall success rate with monotherapy (69% versus 70%), or with modification (20% versus 23%) were equivalent for ceftazidime and imipenem (P = 0.75) . The mortality in this series was 5% . Single-agent therapy with either ceftazidime or imipenem is effective for the empirical treatment of febrile episodes in neutropenic patients with solid tumours . Early addition of amikacin and/or vancomycin resolves most failures of the first step. J Photochem Photobiol B, 1996 Sep, 35(3), 149 - 57 Studies on the mechanism of bacteria photosensitization by meso-substituted cationic porphyrins; Merchat M et al.; Cationic porphyrins have been shown to photoinduce the direct inactivation of Gram-positive (G+) and Gram-negative (G-) bacteria, thereby differing from anionic or neutral porphyrins which can photosensitize the G- bacteria only after permeabilization of their outer membrane . The present data show that the differences between these positively and negatively charged porphyrins are not related by a difference in the intrinsic photosensitizing efficiency, as determined by the photo-oxidation of model substrates or the yield of 1O2 generation; moreover, there are only minor differences in the quantum yield of porphyrin photobleaching . Rather, it appears that the positive charge promotes an electrostatic binding of the porphyrin to the outer cell surface inducing an initial limited damage which favours the penetration of the photosensitizer . Actually, the overall photoprocess is inhibited by the preincorporation of the porphyrin into liposomes, while it is enhanced by using amphiphilic dicationic porphyrins which bind to endocellular sites in larger amounts and in a more stable form. J Periodontol, 1996 Sep, 67(9), 846 - 52 Histochemical and immunocytochemical localization of dipeptidyl peptidases II and IV in human gingiva; Kennett CN et al.; Dipeptidyl peptidase (DPP) II and IV activities were demonstrated in unfixed cryostat sections of gingival tissue from chronic periodontitis patients using histochemistry with 2-methoxy-4-naphthylamine (MNA) substrates . In the case of DPP IV, enzyme localization was confirmed by immunocytochemistry with mouse monoclonal antihuman DPP IV (CD26) antibody . Inflammatory cells containing enzyme were identified in adjacent sections with mouse monoclonal antibodies directed against leukocyte differentiation antigens . Lys-Ala-MNA and Ala-Pro-MNA staining in acid buffer for DPP II was only found in a few fibroblasts in superficial tissue . Staining with Gly-Pro-MNA and Ala-Pro-MNA in alkaline buffer for DPP IV was localized in some CD4 and CD8 positive T lymphocytes, CD68 positive macrophages, and fibroblasts and these cells also reacted with the enzyme antibody . DPP IV-containing macrophages and T lymphocytes were seen in the epithelium . In deeper granulomatous tissue Gram positive and negative bacteria stained with the histochemical substrates, but not the DPP IV antibody . Fibroblast DPP II and IV might participate in cellular interactions with collagen, while T lymphocyte DPP IV may be involved in cell signalling. Support Care Cancer, 1996 Sep, 4(5), 364 - 9 Prophylactic antibiotics eliminate bacteremia and allow safe outpatient management following high-dose chemotherapy and autologous stem cell rescue; Meisenberg B et al.; This study examines the effectiveness of prophylactic ciprofloxacin and rifampin following high-dose chemotherapy and autologous stem cell rescue (HDC/ ASCR) . Specific endpoints included the incidence of fever, clinically documented infection, bacteremia, and readmission rates from an outpatient bone marrow transplant setting following infection or fever . A group of 97 patients receiving 134 cycles of HDC/ASCR were studied . Patients were given ciprofloxacin 750 mg p.o . twice daily and rifampin 300 mg p.o . twice daily beginning on the day of stem cell reinfusion (24-48 h after completion of high-dose chemotherapy) . Most patients were either discharged to an outpatient setting following completion of their chemotherapy or received all of their chemotherapy in an outpatient setting . Febrile neutropenia was treated with empirical antibiotics in an outpatient setting unless it was complicated by hypotension, renal failure, severe mucositis or other problems . The median duration of neutropenia (absolute neutrophil count below 500/mm3) was 7 days . Neutropenic fever occurred in 62% of patients but clinically documented bacterial infection occurred in only 2 (1.5%) patients during their neutropenic period . No bacteremia was noted . Readmission to the hospital following fever or infection occurred in 26% of patients maintained in the outpatient setting . There were no deaths from a bacterial infection in this study although 1 patient (0.7%) died from aspergillosis . Prophylactic ciprofloxacin and rifampin is a well-tolerated and highly effective combination that effectively decreases the risk of both gram-positive and gram-negative bacterial infection following HDC/ASCR . It facilitates outpatient management of myelosuppressed patients receiving autologous stem cell rescue. Clin Immunol Immunopathol, 1996 Sep, 80(3 Pt 1), 307 - 10 Increased serum concentration of soluble CD14 is a prognostic marker in gram-positive sepsis; Burgmann H et al.; Increased serum sCD14 concentrations are associated with poor outcome in Gram-negative sepsis and trauma patients . In the present study serum sCD14 concentrations were measured in patients with Gram-positive sepsis and compared with Gram-negative septic and nonseptic intensive care unit patients . Furthermore, serum sCD14 concentration was correlated with patient's outcome . Serum samples of 28 Gram-positive (8 nonsurvivors/20 survivors) and 10 Gram-negative bacteriemic patients (3 nonsurvivors/7 survivors) were obtained at the day they met the sepsis criteria defined by Bone et al . (Day 0) and at Days 4 and 7 and compared with 10 nonseptic ICU patients and 10 healthy volunteers . Serum concentrations of sCD14 were measured by ELISA . Significantly higher sCD14 serum concentrations were found on Days 4 and 7 in Gram-positive nonsurvivors than in Gram-positive survivors (Day 4: 5.85 +/- 0.48 vs 4.07 +/- 0.43 microgram/ml, P < 0.05; Day 7: 6.12 +/- 0.46 vs 3.53 +/- 0.33 microgram/ml, P < 0.01) . In addition, sCD14 concentrations of Gram-positive nonsurvivors were significantly higher than those of nonseptic ICU patients and healthy volunteers at any time of observation . However, no significant difference was calculated between Gram-positive and Gram-negative patients . Summarizing our results, the serum level of sCD14 could be proven to be a good prognostic marker in the course of Gram-positive sepsis . Increased levels are associated with a high mortality. FEMS Microbiol Lett, 1996 Sep 1, 142(2-3), 215 - 21 Taxonomic identification of Streptomyces exfoliatus K10 and characterization of its poly(3-hydroxybutyrate) depolymerase gene; Klingbeil B et al.; The poly(3-hydroxyalkanoate) (PHA) degrading isolate K10 was identified as Streptomyces exfoliatus . This bacterium is distinguished from other PHA-degrading strains by its ability to utilize both poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxyoctanoate) (PHO) . A PHA depolymerase structural gene of S . exfoliatus (phaZ(Sex) was cloned, expressed and partially purified from recombinant Escherichia coli . The depolymerase was specific for PHB and did not hydrolyze PHO . This indicated the presence of at least one additional gene in S . exfoliatus which encodes a PHO depolymerase . 3-Hydroxybutyrate was identified as the only product of PHB hydrolysis . Comparison of the DNA-deduced amino acid sequence revealed high homology to the PHB depolymerase of Comamonas sp . and low to medium homologies to other PHA depolymerases . The PHB depolymerases of S . exfoliatus and Comamonas sp . represent a subgroup within the family of PHA(SCL) depolymerases . To our knowledge, the S . exfoliatus PHB depolymerase is the first briefly characterized PHA depolymerase of a Gram-positive. Anal Chem, 1996 Sep 1, 68(17), 2805 - 10 Characterization of lipid fatty acids in whole-cell microorganisms using in situ supercritical fluid derivatization/extraction and gas chromatography/mass spectrometry; Gharaibeh AA et al.; In situ supercritical fluid derivatization and extraction was used as a sample preparation technique for the classification of bacteria using fatty acid profiling . Addition of a quaternary ammonium salt such as phenyltrimethylammonium hydroxide under static supercritical conditions directly to lyophilized, whole-cell bacteria in an extraction vessel resulted in the saponification of the bacterial lipids and derivatization of their fatty acids . The derivatized fatty acid methyl esters (FAMEs) were then extracted with supercritical CO2 and analyzed without additional treatment using GC/MS . Iso and anteiso C15:0 and C17:0 along with C18:0 were predominant in Gram-positive bacteria, while C16:1, C16:0, C18:1, and cyclopropyl cyC17:0 and cyC19:0 were significant in Gram-negative bacteria . Application of principal components analysis to the FAME GC/MS data resulted in the differentiation between Gram-positive and Gram-negative type bacteria . Differentiation between species among the same genera was also observed. J Bacteriol, 1996 Sep, 178(17), 5222 - 8 An 18-base-pair sequence is sufficient for termination of rolling-circle replication of plasmid pT181; Zhao AC et al.; pT181 and related plasmids of gram-positive bacteria replicate by a rolling-circle mechanism . The replication initiator protein of pT181, RepC, has origin-specific nicking-closing activities . Replication of the plasmid pT181 leading strand initiates by covalent extension of the RepC-generated nick, and the origin of replication contains signals for both initiation and termination of DNA replication . We have investigated the sequence requirements for the initiation and termination steps by using plasmids containing two pT181 origins . In vitro replication experiments showed that 18- and 24-bp synthetic oligonucleotides containing the RepC nick site were active in the termination of replication . However, initiation of replication required a larger region which also includes the RepC binding site . Plasmids containing the 18- and 24-bp region were also found to be nicked by the RepC protein . Our results demonstrate that sequence requirements for initiation and termination of pT181 replication overlap, but while the RepC binding site is required for initiation, it is dispensable for termination. Eur J Biochem, 1996 Aug 15, 240(1), 302 - 6 A member of the arthropod defensin family from edible Mediterranean mussels (Mytilus galloprovincialis) Hubert F, Noel T, Roch P. Plasma from the mussel Mytilus galloprovincialis previously immunized by injecting them with bacteria contains several bactericidal proteins . One protein, MGD-1, was purified by reverse-phase HPLC of supernatant from acidified cell-free hemolymph . Its biological activity is directed against both gram-positive and gram-negative bacteria but it is not cytotoxic towards human erythrocytes nor protozoa . As determined by mass spectrometry, the molecular mass of MGD-1 is 4418 Da . Primary-structure analysis revealed 38 amino acids including 8 cysteines and a modified amino acid residue in position 28 . Computer searches unambiguously recognized the signature of an arthropod defensin, but the presence of two extra cysteines and of one modified amino acid suggest that it is a previously unknown member of that family. Biochim Biophys Acta, 1996 Aug 15, 1296(1), 9 - 12 Cloning of a secA homolog from Streptomyces lividans 1326 and overexpression in both S . lividans and Escherichia coli; Gilbert M et al.; We cloned a gene encoding a SecA homolog from Streptomyces lividans 1326, a Gram-positive bacterium known to produce large amounts of extracellular proteins . A protein sequence alignment with the other bacterial SecA homologs revealed that S . lividans SecA shares from 39.5 to 44% identity with them, while it shares 34.2 to 37.2% identity with SecA homologs from plastids of algae and plants . We overexpressed the secA gene in S . lividans 1326 and Escherichia coli MM52 and in both cases we observed the production of a protein with an apparent molecular mass of 117.4 kDa . Although S . lividans SecA is similar to E . coli SecA, it does not complement a thermosensitive mutation in the E . coli secA gene . However, a hybrid polypeptide consisting of the N-terminal portion (first 242 amino acids) of the S . lividans SecA and the C-terminal portion (657 a.a.) of the wild-type E . coli SecA was able to complement this mutant. FEMS Microbiol Lett, 1996 Aug 15, 142(1), 1 - 10 Site-specific recombination in gram-positive theta-replicating plasmids; Alonso JC et al.; This review summarises current information on the site-specific recombinases encoded by the plasmids of the Gram-positive bacteria that have low guanine and cytosine content in their DNA . It focuses on the peculiar biological features of the recombination systems encoded by the theta-replicating plasmids and compares them with the site-specific recombinases encoded by transposons or plasmids originally isolated from Gram-negative bacteria. Rev Clin Esp, 1996 Aug, 196(8), 515 - 22 {Imipenem combined with teicoplanin or vancomycin in the initial empirical treatment of febrile neutropenia . Analysis of the primary response and of a global sequential strategy in 126 episodes}; Figuera A et al.; The results of empiric antibiotic therapy in 126 episodes of febrile neutropenia in patients with hematologic neoplasms postchemotherapy and bone marrow transplantation are presented . The main objective of this work was the study of the initial control of infection comparing two glycopeptidic antibiotics: vancomycin and teicoplanin combined with imipenem in first line of empiric therapy . The secondary objective was to analyze the overall control of infection during the complete episode of neutropenia using a sequential empiric antibiotic therapy course which included the addition of amikacin followed by intravenous amphotericin B when fever persisted or recurred without microbiological documentation . Both initial courses (no guidelines), imipenem + vancomycin (arm A) and imipenem + teicoplanin (arm B) resulted in a similar percentage of response at 72 hours, both in episodes of fever of unknown origin (FUO) (55% and 68%, respectively; p = NS) and in those microbiologically documented (54% and 34.5%, p = NS); 58% and 79% of these episodes, respectively, were caused by gram-positive organisms . About 60% of patients in both arm ultimately required the empiric addition of amikacin, with or without amphotericin B, because of persistence or recurrence of fever; the percentage of overall responses in both arm did not differ significantly, both in FUO (70% and 86%, p = NS) and in microbiologically documented episodes (71% and 45%, p = NS) . The overall infectious mortality for the whole group was 1.58% . In conclusion, no significant differences were observed in the clinical response or in toxicity between the combination of imipenem with any of the two glycopeptides: vancomycin or teicoplanin, for the initial empiric therapy of febrile neutropenia . The sequential empiric use of amikacin followed by amphotericin B assured an adequate overall control of infection in a group of patients with prolonged severe neutropenia. Nihon Kyobu Shikkan Gakkai Zasshi, 1996 Aug, 34(8), 916 - 20 {Chronic pulmonary nocardiosis with eosinophilia in an immunocompetent host}; Nakagawa Y et al.; A 75-year-old man who worked at a horse racing track, had an abnormal shadow on a chest radiography . He had a history of dilated cardiomyopathy and mitral regurgitation but had not had any respiratory or systemic symptoms for two years . Chest radiographs and CT scans showed slowly increasing consolidation with internal areas of low attenuation, and without cavity formation, at the left S8 . The erythrocyte sedimentation rate, C-reactive protein level, WBC count, percentage of eosinophils, and IgE level were 35 mm/hr, negative, 8400, 17%, and 3628 IU/ml, respectively . Eosinophils were found in samples of sputum . His immunological status was normal . Transbronchial lung biopsy yielded necrotic tissue, along with gram-positive and irregular acid-fast branching filaments that grew in culture . The microorganism was identified as Nocardia asteroides . Occupational inhalation of soil may have caused his disease. Tohoku J Exp Med, 1996 Aug, 179(4), 281 - 9 Transfusion-associated graft-versus-host disease diagnosed by polymerase chain reaction analysis of DNA microsatellites: an autopsy case with immunohistochemical investigation; Kumamoto H et al.; An autopsy case of transfusion-associated graft-versus-host disease (TA-GVHD) is reported with immunohistochemical investigation and polymerase chain reaction (PCR) analysis . In a 58-year-old male, esophagectomy for carcinoma was performed with a transfusion of 4 units of fresh whole blood . Diarrhea, fever, erythematous rash, pain and leukopenia occurred with an onset 11 days after the operation . He died of sudden dyspnea 29 days after the operation . At autopsy, histological examinations revealed lichenoid lesion in the skin, injury of mucosal epithelia in the digestive tract and damage of interlobular bile ducts in the liver . Immunohistochemical investigation suggested the association between these lesions and CD8-positive T lymphocytes . Severe disturbances of bone marrow and lymphoid organs were accompanied with gram-positive cocci infection in the lungs, esophagus and small intestine . PCR analysis of DNA at microsatellite loci, human growth hormone (HGH) and apolipoprotein B (Apo B), showed DNA chimerism and established the definitive diagnosis of TA-GVHD. Pediatr Pol, 1996 Aug, 71(8), 673 - 7 {Experience in the use of Broviac catheters in children with hematologic neoplastic diseases}; Swiatkiewicz V et al.; Between 1990 and 1994, 61 Broviac catheters were implanted in 53 children with haematological neoplastic diseases . The mean duration of catheter function was 206 days (range 14-615 days) . The total observation time was 12544 days, during which 84 catheter-related complications were recorded, i.e . 6.7 per 1000 catheter use days . Infections occurred with a frequency of 3.8 episodes/1000 cath, days, the majority of which were bacteriemias (2.6/1000 cath, days) . Gram-positive and Gram-negative strains were the isolated etiologic agents with equal frequency . Most of infectious episodes (85%) responded well to initial empiric antibiotics treatment without removal of the catheters . Mechanica complications (occlusion, displacement or catheter leakage) occurred significant less frequently and were managed by repairing or replacing the device or clearing the block . No deaths were related to catheter complications . In conclusion indwelling Broviac catheters offer a safe and effective method of long-term venous access . Infections are the most frequent catheter-related complications. Eur J Clin Microbiol Infect Dis, 1996 Aug, 15(8), 688 - 93 In vitro activity of trovafloxacin in combination with ceftazidime, meropenem, and amikacin; Milatovic D et al.; The in vitro activity of trovafloxacin alone and in combination with ceftazidime, meropenem, and amikacin was studied by determining the minimal inhibitory concentrations (MICs) for 111 gram-negative and 71 gram-positive bacteria . In addition, the synergy of these combinations against 46 strains of gram-negative and gram-positive organisms was studied by checkerboard titration and time-kill kinetics . Trovafloxacin exhibited excellent in vitro activity against all strains tested . Synergism was observed in 17% of the gram-negative strains and in 32% of the gram-positive strains . No antagonism was observed with any of the combinations tested. Intensive Care Med, 1996 Aug, 22(8), 735 - 41 Early and late onset bacteremia have different risk factors in trauma patients; Antonelli M et al.; OBJECTIVE: The aim of this study was to identify risk factors and to describe epidemiological patterns for early-(EOB) and late-onset bacteremias (LOB) after trauma . DESIGN: A prospective study conducted on 141 consecutive trauma patients . SETTING: A general intensive care unit (ICU) of a university hospital . PATIENTS: All multiple trauma patients admitted to our general intensive care unit (ICU) from December 1990 to May 1992 were prospectively enrolled in the study . The following information was collected for each patient and recorded in a computer database: demography, severity of trauma according to the Abbreviated Injury Scale (AIS), severity of coma according to the Glasgow Coma Scale (GCS), presence of pneumothorax, pulmonary contusion, rib fractures, hemothorax, and abdominal trauma, use of mechanical ventilation, and placement of central venous catheters . Bacteremias were defined as EOB when onset occurred within 96 h after trauma, and as LOB when appearing after 96 h from trauma . RESULTS: Thirty-seven patients developed bacteremia during their ICU stay (26%): 11 (29.7%) EOB and 26 (70.3%) LOB . Gram-positive cocci were isolated more frequently in EOB than in LOB (chi 2 = 4.1, P = 0.04) . The risk of EOB was significantly increased by the presence of pulmonary contusion {relative risk (RR) 15.0; confidence interval (CI) 1.99-113.25}, pneumonia before the onset of bacteremia (RR 3.56; CI 1.17-10.69), AIS score greater than 32 and an abdominal injury score greater than 9 (RR 3.11; CI 1.02-9.49), while intravascular catheters and mechanical ventilation did not represent risk factors for EOB . LOB had a very different pattern and their risk was significantly increased by exposure to intravascular catheters (RR 4.96; CI 1.23-19.94) and to mechanical ventilation lasting more than 7 days (RR 3.6; CI 1.6-8.1) . CONCLUSIONS: Scoring with the AIS of the abdominal and thoracic trauma at admission to the ICU appears a useful tool for identifying trauma patients at increased risk of EOB . A rigorous policy of catheter placement and maintenance as a means of reducing late bacteremias in trauma patients is essential. Mol Microbiol, 1996 Aug, 21(4), 675 - 82 Two beginnings for a single purpose: the dual-start holins in the regulation of phage lysis; Blasi U et al.; For most large phages of both Gram-positive and Gram-negative bacteria, there appears to be a single pathway for achieving disruption of the host envelope, requiring at least two phage-encoded lysis functions (a holin and an endolysin) . The holin is a small membrane protein which causes a non-specific lesion in the cytoplasmic membrane, which allows the endolysin to gain access to its substrate, the peptidoglycan . The scheduling of host lysis is effected by regulatory mechanisms which govern the synthesis and activity of the holin protein accumulating in the membrane . Accordingly, aspects of expression and function of holin genes are considered here, focusing mainly on the lambdoid S genes . This group of genes, of which lambda S is the prototype, are characterized by a dual-start motif consisting of two Met start codons separated by one or two codons, at least one of which specifies Arg or Lys . Two protein products are elaborated, differing only by two or three N-terminal residues but apparently possessing opposing functions: the shorter polypeptide is the active holin, or lysiseffector, whereas the longer polypeptide apparently acts as an inhibitor of holin function . Models will be considered which may account for the ability of the holin to form a 'hole' in the cytoplasmic membrane at a programmed time, as well as for the inhibitory properties of the longer product . Finally, we discuss recent results suggesting that the dual-start motif can be viewed as a level of regulation superimposed on a timing function intrinsic to the canonical holin structure. Shock, 1996 Aug, 6(2), 112 - 7 ABT-299, a novel PAF antagonist, attenuates multiple effects of endotoxemia in conscious rat