Am J Gastroenterol, 1994 Aug, 89(8), 1226 - 9 Clostridium difficile-associated diarrhea in patients with HIV positivity and AIDS: a prospective controlled study; Lu SS et al.; OBJECTIVE: To compare the clinical manifestations and therapeutic responses of Clostridium difficile infection in HIV-infected and noninfected individuals . METHODS: Patients were identified for this study if they had C . difficile toxin in the stool . The patients were then followed prospectively by the investigators . All patients were treated with a standard regimen, and clinical and laboratory findings were recorded . Persistence and resolution or recurrence of symptoms and complications were recorded . RESULTS: A total of 87 patients were studied, of which 12 were HIV positive, 20 had AIDS, and 55 had no known HIV infection . The AIDS group was younger and had a lower total leukocyte count than the controls . There were no statistically significant differences in temperature, leukocytosis, clinical symptoms, therapeutic response, or recurrence or persistent of symptoms . CONCLUSIONS: Despite the immunosuppression of HIV infection, C . difficile infection behaves no differently in HIV/AIDS patients than it does in controls. J Bacteriol, 1994 Aug, 176(16), 4865 - 74 Characterization and regulation of the NADP-linked 7 alpha-hydroxysteroid dehydrogenase gene from Clostridium sordellii; Coleman JP et al.; A bile acid-inducible NADP-linked 7 alpha-hydroxysteroid dehydrogenase (7 alpha-HSDH) from Clostridium sordellii ATCC 9714 was purified 310-fold by ion-exchange, gel filtration, and dye-ligand affinity chromatography . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the purified enzyme showed one predominant peptide band (30,000 Da) . The N-terminal sequence was determined, and the corresponding oligonucleotides were synthesized and used to screen EcoRI and HindIII genomic digests of C . sordellii . Two separate fragments (4,500 bp, EcoRI; 3,200 bp, HindIII) were subsequently cloned by ligation to pUC19 and transformation into Escherichia coli DH5 alpha-MCR . The EcoRI fragment was shown to contain a truncated 7 alpha-HSDH gene, while the HindIII fragment contained the entire coding region . E . coli clones containing the HindIII insert expressed high levels of an NADP-linked 7 alpha-HSDH . Nucleotide sequence analyses suggest that the 7 alpha-HSDH is encoded by a monocistronic transcriptional unit, with DNA sequence elements resembling rho-independent terminators located in both the upstream and downstream flanking regions . The transcriptional start site was located by primer extension analysis . Northern (RNA) blot analysis indicated that induction is mediated at the transcriptional level in response to the presence of bile acid in the growth medium . In addition, growth-phase-dependent expression is observed in uninduced cultures . Analysis of the predicted protein sequence indicates that the enzyme can be classified in the short-chain dehydrogenase group. Aust N Z J Surg, 1994 Aug, 64(8), 574 - 5 Spontaneous Clostridium septicum myonecrosis in congenital neutropaenia; Keogh G et al.; Spontaneous Clostridium septicum myonecrosis is an uncommon disorder that has been described in association with malignancy, immunosuppression and neutropaenia . Typical clostridial myonecrosis develops without a visible portal of entry and mortality is high . The pathogenesis is not completely understood but the clostridia may gain access to the circulation via areas of ileo-caecal ulceration secondary to enterocolitis, antibiotics or neoplasms . A 5 year old boy with congenital neutropaenia presented with spontaneous Clostridium septicum myonecrosis in the thigh . Limb salvage was achieved using antibiotics, hyberbaric oxygenation and selective debridement . The portal of entry may have been the gastrointestinal tract as colonic ulceration may occur in neutropaenia, and pre-morbid clindamycin administration may have encouraged overgrowth of colonic clostridia. J Cell Biol, 1994 Aug, 126(3), 801 - 10 Inhibition of lysophosphatidate- and thrombin-induced neurite retraction and neuronal cell rounding by ADP ribosylation of the small GTP-binding protein Rho; Jalink K et al.; Addition of the bioactive phospholipid lysophosphatidic acid (LPA) or a thrombin receptor-activating peptide (TRP) to serum-starved N1E-115 or NG108-15 neuronal cells causes rapid growth cone collapse, neurite retraction, and transient rounding of the cell body . These shape changes appear to be driven by receptor-mediated contraction of the cortical actomyosin system independent of classic second messengers . Treatment of the cells with Clostridium botulinum C3 exoenzyme, which ADP-ribosylates and thereby inactivates the Rho small GTP-binding protein, inhibits LPA- and TRP-induced force generation and subsequent shape changes . C3 also inhibits LPA-induced neurite retraction in PC12 cells . Biochemical analysis reveals that the ADP-ribosylated substrate is RhoA . Prolonged C3 treatment of cells maintained in 10% serum induces the phenotype of serum-starved cells, with initial cell flattening being followed by neurite outgrowth; such C3-differentiated cells fail to retract their neurites in response to agonists . We conclude that RhoA is essential for receptor-mediated force generation and ensuing neurite retraction in N1E-115 and PC12 cells, and that inactivation of RhoA by ADP-ribosylation abolishes actomyosin contractility and promotes neurite outgrowth. J Bacteriol, 1994 Aug, 176(15), 4779 - 83 Molecular cloning of two new heat shock genes related to the hsp70 genes in Staphylococcus aureus; Ohta T et al.; We have identified two new heat shock protein genes, orf37 and orf35, in Staphylococcus aureus, located upstream and downstream of grpE(hsp20), dnaK(hsp70), and dnaJ(hsp40) homologous genes in the order orf37-hsp20-hsp70-hsp40-orf35 . The transcripts of both orf37 and orf35 were increased by thermal upshift of the culture from 37 to 46 degrees C . The heat shock promoters were located upstream of orf37 and upstream of hsp40 . The deduced peptide of orf37 showed similarity with those of orfA in Clostridium acetobutylicum and orf39 in Bacillus subtilis . orf35 was unique in S . aureus and has not yet been described in other bacteria. J Clin Microbiol, 1994 Aug, 32(8), 1986 - 91 Application of PCR to a clinical and environmental investigation of a case of equine botulism; Szabo EA et al.; PCR for the detection of botulinum neurotoxin gene types A to E was used in the investigation of a case of equine botulism . Samples from a foal diagnosed with toxicoinfectious botulism in 1985 were reanalyzed by PCR and the mouse bioassay in conjunction with an environmental survey . Neurotoxin B was detected by mouse bioassay in culture enrichments of serum, spleen, feces, and intestinal contents . PCR results compared well with mouse bioassay results, detecting type B neurotoxin genes in these samples and also in a liver sample . Other neurotoxin types were not detected by either test . Clostridium botulinum type B was shown to be prevalent in soils collected from the area in which the foal was raised . Four methods were used to test for the presence of botulinum neurotoxin-producing organisms in 66 soil samples taken within a 5-km radius: PCR and agarose gel electrophoresis (types A to E), PCR and an enzyme-linked assay (type B), hybridization of crude alkaline cell lysates with a type B-specific probe, and the mouse bioassay (all types) . Fewer soil samples were positive for C . botulinum type B by the mouse bioassay (15%) than by any of the DNA-based detection systems . Hybridization of a type B-specific probe to DNA dot blots (26% of the samples were positive) and PCR-enzyme-linked assay (77% of the samples were positive) were used for the rapid analysis of large numbers of samples, with sensitivity limits of 3 x 10(6) and 3,000 cells, respectively . Conventional detection of PCR products by gel electrophoresis was the most sensitive method (300-cell limit), and in the present environmental survey, neurotoxin B genes only were detected in 94% of the samples. J Clin Microbiol, 1994 Aug, 32(8), 1963 - 9 Comparison of restriction endonuclease analysis, ribotyping, and pulsed-field gel electrophoresis for molecular differentiation of Clostridium difficile strains; Kristjansson M et al.; A combined clinical and molecular epidemiologic analysis of 46 strains of Clostridium difficile, including 16 nosocomial isolates from one ward (outbreak ward) plus 17 other nosocomial isolates and 13 community-acquired isolates, was performed . HindIII digests of total cellular DNA were analyzed by restriction enzyme analysis (REA) and ribotyping; SmaI digests were analyzed by pulsed-field gel electrophoresis (PFGE) . Isolates were assigned to typing groups on the basis of the profiles detected; isolates with closely related profiles were assigned to subgroups . The 16 isolates from the outbreak ward were resolved by both REA and PFGE into five distinct groups; 13 isolates represented two REA groups and three PFGE groups and two isolates were resolved as distinct groups by both techniques . DNA obtained from one isolate was persistently partially degraded, precluding analysis by PFGE . Seventeen sporadic nosocomial isolates were resolved by REA and PFGE into comparable numbers of groups (i.e., nine groups) and subgroups (i.e., 15 and 14 subgroups, respectively), with two isolates not evaluable by PFGE . The 13 epidemiologically unrelated community-acquired isolates were assigned to 11 groups by REA and to 12 groups by PFGE . Overall, ribotyping identified only nine groups among the 46 isolates . We conclude that REA and PFGE have comparable discriminatory powers for epidemiologic typing of C . difficile isolates and that ribotyping is appreciably less discriminatory . For a few isolates, partial DNA degradation prevented analysis by PFGE but not by REA or ribotyping; the cause of the degradation is unknown. J Clin Microbiol, 1994 Aug, 32(8), 1911 - 7 Detection of type A, B, and E botulism neurotoxin genes in Clostridium botulinum and other Clostridium species by PCR: evidence of unexpressed type B toxin genes in type A toxigenic organisms; Franciosa G et al.; We studied the effectiveness of the PCR in detecting the type A, B, and E botulism neurotoxin genes in 209 strains of Clostridium botulinum and 29 strains of other Clostridium spp . All 79 strains that produced type A toxin, 77 strains that produced type B toxin, and 51 organisms that produced type E toxin (46 C . botulinum and 5 C . butyricum) were PCR positive in reactions with primers targeting sequences specific for their respective toxin genes . The PCR for type A toxin was positive for one type B toxin-producing strain that produced a small amount of type A toxin in addition to a large amount of type B toxin . Surprisingly, the type B toxin gene was detected in addition to the type A toxin gene in 43 type A toxin-producing strains, only 1 of which could be shown by bioassay to produce biologically active type B toxin in culture . The type B gene was also detected in two strains of C . subterminale, which were determined to be nontoxigenic by bioassay . While the PCR was sensitive and specific in detecting the neurotoxin genes, the discovery of unexpressed toxin genes indicates that PCR results may not be adequate for establishing type B neurotoxigenicity. Am J Infect Control, 1994 Aug, 22(4), 212 - 7 Hospital carpeting and epidemiology of Clostridium difficile; Skoutelis AT et al.; BACKGROUND: Clostridium difficile is the usual and most important cause of antibiotic-associated pseudomembranous enterocolitis . The source of nosocomial acquisition of the organism in nonepidemic settings has not been determined . METHODS: Epidemiologic and microbiologic studies were conducted in a community-teaching hospital complex to assess the impact of carpeting in patient rooms on environmental contamination with C . difficile, along with the prevalence of pseudomembranous enterocolitis . All C . difficile isolates were typed by means of a bacteriophage-bacteriocin typing scheme . RESULTS: No clear evidence of environmental acquisition of C . difficile in a nonepidemic setting of pseudomembranous enterocolitis was found . Carpeted floors were significantly more heavily contaminated for prolonged periods with clinical strains of C . difficile than were noncarpeted floors . CONCLUSION: There was no evidence that contamination of carpeting resulted in increased frequency of pseudomembranous enterocolitis in patients residing in carpeted rooms . Because there is strong evidence of exogenous acquisition of C . difficile during outbreaks, however, room carpeting should be considered a potential reservoir of this organism. Steroids, 1994 Aug, 59(8), 485 - 9 High concentrations of conjugated bile acids inhibit bacterial growth of Clostridium perfringens and induce its extracellular cholylglycine hydrolase; Kishinaka M et al.; To investigate the effects of conjugated bile acid on bacterial growth and cholylglycine hydrolase activity, Clostridium perfringens from human feces was exposed to varying concentrations of taurine- or glycine-conjugated chenodeoxycholic acid . Extracellular enzyme activity was determined by deconjugation of radiolabeled taurocholic acid and viable cells were counted after anaerobic culture at 37 degrees C for 24 h . Viable cells were decreased with more than 1.0 mg of conjugated chenodeoxycholic acid per mL and there were no viable cells with 10.0 mg of bile acid per mL . Although total enzyme activity was decreased according to the bile acid concentration, enzyme activity per bacterium was increased between 1.0 and 4.0 mg/mL . There were no statistically significant differences between the types of conjugation . It was concluded that conjugated bile acids may exert inhibitory effect on bacterial growth and extracellular cholylglycine hydrolase activity in Clostridium perfringens . However, under the physiologic condition in the human intestine, conjugated bile acid might induce production of extracellular cholyglycine hydrolase per bacterium. Infect Control Hosp Epidemiol, 1994 Aug, 15(8), 534 - 5 A common source outbreak of gastroenteritis in a teaching hospital; Khatib R et al.; An outbreak of gastroenteritis at a large teaching hospital affected at least 52 workers . Investigation implicated a tuna salad, and the circumstances suggested Clostridium perfringens as the etiologic agent . The risk of such outbreaks may be reduced by cooling of ingredients prior to mixing and refrigeration in small steel containers. J Pediatr Surg, 1994 Aug, 29(8), 987 - 90; discussion 990-1 Necrotizing enterocolitis in the extremely low birth weight infant; Rowe MI et al.; Improved neonatal management has resulted in an enlarging population of extremely low birth weight (ELBW) infants . These infants have a high incidence of necrotizing enterocolitis (NEC) and a high mortality rate . The authors compared two groups of NEC patients: ELBW infants (< 1,000 g and/or < or = 28 weeks' gestation) and "standard" premature infants (29 to 36 weeks' gestation) . NEC was classified according to the extent of bowel involvement: (1) focal, (2) diffuse, or (3) pan involvement (pan necrosis) . Clinical laboratory, radiological, pathological, and bacteriologic findings, management, and mortality were analyzed . There were no significant differences between the groups with respect to gender, race, delivery mode, or incidence of prenatal or perinatal problems . The most common presenting signs in both groups were abdominal distension, vomiting, and feeding intolerance . The onset of signs and the time of first feedings were significantly later in the ELBW group . Pneumatosis was the most frequent initial radiological finding (60% of the ELBW group, 75% of the premature group) . Portal vein air (PVA) was present in 29% of the ELBW and premature infants . Seventy-one percent of ELBW infants with PVA had pan involvement, versus 40% of premature infants (P < .05) . There were significant differences in the peritoneal cultures between the groups . The premature group had significantly more Escherichia coli (54% v 23%) . The ELBW group had a wider variety of microorganisms (eg, Clostridium sp, Pseudomonas sp, and yeast) . Survival was significantly higher for the premature group (84% v 55%) . The mortality rate was 93% when pan involvement was present in the ELBW group.(ABSTRACT TRUNCATED AT 250 WORDS) J Dermatol, 1994 Aug, 21(8), 539 - 45 Growth-promoting effect of bacterial products from Clostridium perfringens on human keratinocytes; Takada A et al.; Wound healing substance (WHS) from cultured Clostridium perfringens has been reported to be effective in the treatment of wounds . The effects of WHS, now named SNK-863, on proliferation and differentiation of human keratinocytes were examined . The characteristics of WHS are as follows: 1) WHS stimulates human keratinocyte growth and DNA synthesis; 2) WHS and EGF show some additive effects on human keratinocyte growth; 3) WHS does not interfere with the binding of EGF to its receptor; 4) WHS does not counteract the growth inhibitory effects of TGF-beta or vitamin D3 on human keratinocytes; 5) WHS has no significant effect on human keratinocyte differentiation . These results indicate that the growth-promoting effect of WHS on keratinocytes may contribute to the treatment of wound healing. Hepatogastroenterology, 1994 Aug, 41(4), 394 - 6 The role and timing of surgery in the treatment of pseudomembranous colitis . A case complicated by toxic megacolon; Agnifili A et al.; The authors describe a particularly serious case of pseudomembranous colitis due to Clostridium difficile that was complicated by toxic megacolon . It was resolved by surgical intervention, and the reasons why subtotal colectomy is preferable to simple ileostomy are discussed. Am J Hosp Pharm, 1994 Aug 1, 51(15), 1892 - 901; quiz 1958-9 Update on Clostridium difficile-induced colitis, Part 2; Reinke CM et al.; Clostridium difficile is a nosocomial pathogen able to survive unfavorable environments by sporulation; when conditions advantageous for rapid growth appear, the vegetative form is regenerated . Lack of conscientious hand washing and failure of health care providers to use disposable gloves facilitate transmission within institutions . Exposure to certain antimicrobials expedites C . difficile overgrowth within the colon by altering the composition of the normal gut microflora . Antineoplastic agents may also precipitate CDIC . The characteristics of the colonizing strain, the properties of the inciting drug, and individual host factors collectively seem to govern the expression of the disorder . Clinical presentations range from self-limiting diarrhea to severe diarrhea accompanied by abdominal pain, fever, and leukocytosis to potentially life-threatening PMC . A preponderance of data supports the interpretation that oral metronidazole and oral vancomycin are therapeutically equivalent for the treatment of all but the most severe cases of CDIC . Whether the two drugs are equivalent in severe CDIC is controversial and will probably remain so in the absence of a well-designed trial to expand on the findings of the study by Teasley et al . Because of the cost difference and therapeutic equivalence, oral metronidazole should be the preferred routine treatment for CDIC; oral vancomycin should be reserved for severe cases and cases that fail to respond to at least six days of oral metronidazole therapy . Another important argument, albeit a hypothetical one, for limiting institutional use of oral vancomycin is to minimize selective environmental pressure for the emergence and dissemination of vancomycin-resistant enterococci . An epidemic outbreak of CDIC caused by clindamycin-resistant C . difficile in an institution where clindamycin use was extremely high illustrates the possible consequences of such selective pressure . Oral metronidazole 250 mg four times daily will usually provide a satisfactory response, but clinicians may wish to consider increasing the total daily dose for some patients who have symptoms like fever and leukocytosis . For oral vancomycin, 125 mg four times daily is sufficient in virtually all circumstances . Ten days of therapy is usually adequate for either drug . CDIC in a patient unable to take medications orally presents a bit of a therapeutic dilemma . Two approaches that appear effective are rectal administration of vancomycin and intravenous administration of metronidazole, although intravenous metronidazole can fail to work, possibly because the colonic concentrations achieved are inadequate . Clinicians may wish to consider a total daily dose of intravenous metronidazole that is at the upper end of the adult dosage range, if this is feasible.(ABSTRACT TRUNCATED AT 400 WORDS) J Immunoassay, 1994 Aug, 15(3), 293 - 304 Development of an ELISA assay for Clostridium perfringens phospholipase C (alpha toxin); Holdsworth RJ et al.; A new method for the assay of Clostridium perfringens alpha toxin (phospholipase C) is described using a sandwich ELISA . This assay has been shown to be quantitative, to have a high specificity for the toxin and is capable of detecting purified Clostridium perfringens phospholipase C at concentrations of as little as 0.005 units/ml in cooked meat culture medium. Diagn Microbiol Infect Dis, 1994 Aug, 19(4), 227 - 34 Increased in vitro activity of ceftriaxone by addition of tazobactam against clinical isolates of anaerobes; Aldridge KE et al.; A total of 461 clinical strains of anaerobes were tested using a broth microdilution test to determine the activity of the combination of ceftriaxone and tazobactam and other antimicrobials against these isolates . Ceftriaxone was combined with tazobactam in ratios of 1:1, 2:1, 4:1, and 8:1 and twofold dilutions of ceftriaxone in constant concentrations to tazobactam of 2, 4, 8, 16, and 32 micrograms/ml . Against beta-lactamase-producing strains of the Bacteroides fragilis group, B . capillosus, and Prevotella species all combinations of ceftriaxone and tazobactam showed enhanced in vitro activity and were eight- to 2048-fold more active than ceftriaxone alone . By comparison ceftriaxone and tazobactam showed superior or equal activity to ampicillin and sulbactam, piperacillin and tazobactam, amoxicillin and clavulanate, ticarcillin and clavulanate, and metronidazole against these same strains . Against beta-lactamase nonproducing strains of Porphyromonas, Fusobacterium, Clostridium, Eubacterium, Peptostreptococcus, and Veillonella parvula the addition of tazobactam produced no appreciable enhanced ceftriaxone activity . Fixed concentrations of tazobactam at 2 and 4 micrograms/ml appear to be most suitable for susceptibility testing and are within the pharmacologic profile of this inhibitor . Pharmacologic and toxicity studies will be needed to define the role of ceftriaxone and tazobactam in infectious diseases. Acta Ophthalmol (Copenh), 1994 Aug, 72(4), 524 - 8 Gas gangrene panophthalmitis . A case from Greenland; La Cour M et al.; A case of clostridium perfringens gas gangrene panophthalmitis developed after a penetrating eye injury . The affected eye became amaurotic, but the panophthalmitis was controlled by minimal surgical debridement and systemic antibiotic therapy with penicillin, fucidic acid and metronidazole . Elective enucleation was performed 15 days after the trauma for cosmetic reasons . The enucleated eye was examined histopathologically and showed massive retinal necrosis but no signs of bacteriae. Biosci Biotechnol Biochem, 1994 Aug, 58(8), 1496 - 9 Purification and characterization of xylanase A from Clostridium stercorarium F-9 and a recombinant Escherichia coli; Sakka K et al.; Xylanase A encoded by the Clostridium stercorarium F-9 xynA gene was purified to homogeneity from a recombinant clone of Escherichia coli . The N-terminal amino acid sequence and molecular weight (54,000) estimated by SDS-PAGE of the purified enzyme were consistent with those deduced from the nucleotide sequence {Biosci . Biotech . Biochem., 57, 273-277 (1993)} . A xylanase was also purified to homogeneity from a culture supernatant of C . stercorarium F-9 . Its N-terminal amino acid sequence, molecular weight, and enzymatic properties were quite in agreement with those of the recombinant enzyme, indicating that the xynA gene was predominantly expressed as a xylanase gene in C . stercorarium F-9 . The purified enzyme hydrolyzed xylotriose to yield xylobiose and xylose while it was less active toward xylobiose . It was optimally active at 75 degrees C and pH 7.0 Km and Vmax were estimated to be 1.9 mg/ml and 2.8 mumol of xylose equivalent/min/micrograms for oat spelt xylan, respectively. Curr Microbiol, 1994 Aug, 29(2), 69 - 77 Conserved structure of genes encoding components of botulinum neurotoxin complex M and the sequence of the gene coding for the nontoxic component in nonproteolytic Clostridium botulinum type F; East AK et al.; For investigation of the genes of proteins associated in vivo with botulinum neurotoxin (BoNT), polymerase chain reaction (PCR) experiments were carried out with oligonucleotide primers designed to regions of the nontoxic-nonhemagglutinin (NTNH) gene of Clostridium botulinum type C . The primers were used to amplify a DNA fragment from genomic DNA of C . botulinum types A, B, E, F, G and toxigenic strains of Clostridium barati and Clostridium butyricum . The amplified product from all of these strains hybridized with an internal oligonucleotide probe, whereas all nontoxigenic clostridia tested gave no PCR product and showed no reaction with the probe . The NTNH gene was shown to be located upstream of the gene encoding BoNT, thereby revealing a conserved structure for genes encoding the proteins of the M complex of the progenitor botulinum toxin in these organisms . The sequence of the NTNH gene of nonproteolytic C . botulinum type F was determined by PCR amplification and sequencing of overlapping cloned fragments . NTNH/F showed 71% and 61% identity with NTNH of C . botulinum type E and type C respectively. Biochemistry, 1994 Jul 26, 33(29), 8702 - 11 Organization of clusters and internal electron pathways in CO dehydrogenase from Clostridium thermoaceticum: relevance to the mechanism of catalysis and cyanide inhibition; Anderson ME et al.; Cyanide inhibits the CO oxidation activity of carbon monoxide dehydrogenase from Clostridium thermoaceticum by binding tightly to the form of the C-cluster yielding the gav = 1.82 signal (the C1.82 form) . CN- dissociates and the enzyme reactivates upon addition of CO, CO2 plus dithionite, or CS2 plus dithionite . Dithionite slows the inhibition of the enzyme by CN-, but it cannot reactivate the enzyme . This behavior is explained by assuming that binding of CO, CO2, or CS2 at a modulator site accelerates the dissociation of CN- from the C-cluster . With CN- bound at the C-cluster, dithionite, but not CO, can reduce those Fe-S clusters in the enzyme whose redox status can be monitored at 420 nm . The electron pathway used for CO oxidation appears to be as follows: C-cluster-->Fe-S Clusters-->external electron acceptors . The electron used to reduce the NiFe complex originates predominantly from the C-cluster, and this reduction is inhibited when CN- is bound at the C-cluster . The NiFe complex is reduced more slowly (in the absence of CN-) than CO is catalytically oxidized, indicating that this reduction is not part of the catalytic mechanism for CO oxidation . The form of the C-cluster yielding the g(av) = 1.86 signal (C1.86) is proposed to be two electrons more reduced than C1.82 and able to bind and reduce CO2 . CO is proposed to be oxidized by C1.82 . Neither CO or CN- appears to bind C1.86. Biochim Biophys Acta, 1994 Jul 21, 1222(3), 331 - 8 Tissue-specific variations in the expression and regulation of the small GTP-binding protein Rho; Fritz G et al.; Rho proteins are involved in the regulation of the assembly of the microfilamental cellular network and are known to be specific substrates for the ADP-ribosyltransferase C3 from Clostridium botulinum . Here, we studied the distribution of Rho and Rho-regulating proteins in extracts from various rabbit tissues . The highest amounts of {32P}ADP-ribosylated proteins were detected in cell extracts from lung and kidney . Compared to these tissues, 50-95% reduced labeling of Rho proteins was observed in extracts from liver, spleen, brain, heart and muscle . The level of the C3-mediated {32P}ADP-ribosylation of Rho did not correlate with the amount of RhoA proteins detected by Western analysis . The relative amounts of {32P}ADP-ribosylated proteins located in cytosolic or membrane fractions, respectively, depended on the type of tissue investigated, indicating a tissue-specific variation in the subcellular distribution of Rho proteins . The same was true for the complexation of Rho with other factors and the expression of diverse Rho species . In respect to Rho-regulating proteins, extracts from lung and brain contained the highest amounts of guanine nucleotide dissociation-inhibitor proteins (Rho-GDI) . The association of Rho with Rho-GDI however showed tissue specificity and did not correlate with Rho-GDI amounts . The highest Rho-GAP (GAP = GTPase-activating protein) activities were observed in extracts from lung, kidney and spleen, the lowest ones in extracts from muscle and heart . In total, our data demonstrate tissue-specific differences in the expression of RhoA, {32P}ADP-ribosylated proteins and Rho-regulating factors, indicating a tissue-specific variation in the activity and regulation of Rho proteins. FEMS Microbiol Lett, 1994 Jul 15, 120(3), 297 - 301 Roles of the carboxy-terminal region of Clostridium perfringens alpha toxin; Nagahama M et al.; Treatment of Clostridium perfringens alpha toxin with aminopeptidase resulted in no effect on various activities of the toxin . Aminopeptidase did not hydrolyze the native toxin or the toxin treated with urea in the presence of EDTA . Treatment with carboxypeptidase for 30 min resulted in a 75% decrease in these activities . Incubation of the native toxin with carboxypeptidase for 30 min released approximately 15 amino acids from the C-terminus of the toxin . The biological activities of a mutant toxin lacking 20 C-terminal residues of the toxin (AT1-350) showed about 59-87% of the activity of native toxin . The mutant toxin showed partial antigenic identity with the native toxin . These data suggest that the C-terminal domain contributes to maintaining the active form of the toxin. Biochem Biophys Res Commun, 1994 Jul 15, 202(1), 591 - 5 Some structural features of cluster-coordinating cysteines of Clostridium pasteurianum ferredoxin are revealed by 2D TOCSY 1H NMR on the oxidized protein; Acquotti D et al.; Different sets of geminal J coupling constants for the eight beta-CH2 protons in the iron-coordinating cysteines in Clostridium pasteurianum ferredoxin were detected by 2D TOCSY 1H NMR experiments on the oxidized protein . Four resonances were characterized by quite similar high values of J, two more resonances had a J value about half of the former ones, while the last two had extremely low J values . These findings suggest that the cysteines required for cubane symmetry around the iron atoms are constrained into different geometries . The simplified model used for fine tuning of tau m in these TOCSY experiments is also presented and discussed. Am J Hosp Pharm, 1994 Jul 15, 51(14), 1771 - 81 Update on Clostridium difficile-induced colitis, Part 1; Reinke CM et al.; Recent findings on the epidemiology, pathogenesis, clinical manifestations, diagnosis, and treatment of Clostridium difficile-induced colitis (CDIC) are discussed . CDIC is a gastrointestinal disorder that results from colonization by and overgrowth of C . difficile . Among patients in the community who are treated with an oral antimicrobial, only 1 to 3 individuals per 100,000 develop CDIC, compared with as many as 1 per 100 hospitalized patients treated with an antimicrobial . The requirements for CDIC are (1) a readily available source of C . difficile; (2) exposure to drugs, most commonly certain antimicrobials, that disrupt the normal colonic microflora; (3) production of the requisite cytotoxins by the C . difficile strain colonizing the colon; and (4) the presence of individual risk factors, including advanced age, severe underlying illness, and a prolonged hospital stay . Among the varied clinical manifestations of CDIC, diarrhea is predominant and is often the sole symptom . In more severe cases, fever and leukocytosis are also present . The formation of pseudomembranous plaques is pathognomonic but relatively infrequent . Presumptive diagnosis is usually based on a positive cytotoxin assay result in the symptomatic patient . Patients who respond to discontinuation of the inciting drug or drugs should not be treated indiscriminately with antimicrobials . Asymptomatic carriers should not be treated, and a period of watchful waiting may be advisable in mild cases . When treatment is necessary, oral metronidazole is the agent of choice in all but the most severe cases . Whether oral metronidazole is therapeutically equivalent to oral vancomycin in severe CDIC is controversial . Regardless of the antimicrobial used, some patients suffer a recurrence of CDIC and a few have several relapses . There have been no comparative studies of treatment for relapsing CDIC . Of the investigational treatments, the tiacumicin macrolides and the yeast Saccharomyces boulardii appear most promising . Diagnostic assays based on the polymerase chain reaction should allow more timely intervention . Health care professionals who improve their understanding of CDIC will be better able to recognize the disorder, select the best treatment, and perhaps reduce the frequency of CDIC in hospitalized patients by working to alter patterns of antimicrobial use. Biochim Biophys Acta, 1994 Jul 13, 1193(1), 155 - 64 Modulation of the activity of Clostridium perfringens neuraminidase by the molecular organization of gangliosides in monolayers; Perillo MA et al.; The activity of Clostridium perfringens neuraminidase against gangliosides GM3, GD1a and GM1 was studied in lipid monolayers at the air-buffer solution interface . The enzyme activity assay against pure ganglioside monolayers is based on the markedly different molecular packing areas of the substrate gangliosides and the resulting product glycosphingolipids . This allows to control and monitor the surface pressure and the ganglioside intermolecular organization (cross-sectional packing areas and dipole potentials) in a continuous manner during the catalytic process . It was found that the rate and the extent of the enzymatic reaction depended markedly on the lateral surface pressure . In general, the activity of neuraminidase against GM3 and GD1a was higher at lower surface pressure . This corresponded to larger intermolecular spacings among the ganglioside molecules . Both the activity and the extent of the reaction against GM3 were higher than toward GD1a . GM1 could not be degraded by the enzyme, irrespective of the surface pressure but the enzyme could interact with this ganglioside . A latency period, longer for GM3 than for GD1a, was observed prior to the onset of rapid degradation; this indicates that pre-catalytic steps are occurring at the interface before effective ganglioside degradation takes place . The latency period, the total amount of ganglioside degraded, and the velocity of the reaction varied with the surface pressure in different manners . Our data indicate that the different steps of the catalytic reaction occurring at the surface (i.e., substrate recognition and interfacial adsorption, catalysis, maximum extent of substrate conversion) are independently regulated by the molecular organization of the substrate gangliosides. Can J Microbiol, 1994 Jul, 40(7), 592 - 6 Features of the cellodextrinase gene from Fibrobacter succinogenes S85; Iyo AH et al.; The nucleotide sequence of a 2.3-kb DNA fragment containing a cellodextrinase gene (cedA) from the ruminal anaerobe Fibrobacter succinogenes S85 was determined . Activity was expressed from this fragment when it was cloned in both orientations in pBluescript KS+ and SK-, indicating a functional F . succinogenes promoter in Escherichia coli . Promoter sequences (TTGAACA and AATAA) were identified upstream of the ATG initiation codon preceded by a putative ribosome binding site . The cedA open reading frame of 1071 base pairs encoded a protein of 357 amino acid residues with a calculated molecular mass of 41.9 kDa, similar to the 40-kDa size of the native protein as determined by gel filtration chromatography . CedA is proposed to belong to family 5 (family A) of the glycosyl hydrolases . The primary structure of the cellodextrinase showed over 40% similarity with endoglucanase 3 from F . succinogenes S85 . Short regions of similarity were also demonstrated with endoglucanase C from Clostridium thermocellum, CelA from Ruminococcus flavefaciens, and two exoglucanases from yeast. J Infect Dis, 1994 Jul, 170(1), 227 - 30 Risk factors for Clostridium difficile stool cytotoxin b among critically ill patients: role of sucralfate; Jensen GL et al.; To identify risk factors other than antimicrobial exposure for Clostridium difficile stool cytotoxin b, subjects admitted to critical care units over 18 months and who had stool cytotoxin assays were evaluated . Twenty-two cases (cytotoxin b-positive) were compared with 125 controls (cytotoxin b-negative) . Cases and controls were similar with respect to age, sex, therapeutic index severity score, duration of hospitalization before cytotoxin b testing, and antimicrobial exposure . Adjusted odds ratios (OR) revealed white blood cell count of > 12,000 on the day of stool sampling (OR, 4.0; 95% confidence interval {CI}, 1.3-12.4) and sucralfate exposure (OR, 0.15; 95% CI, 0.05-0.42) as significant independent positive and negative risk factors, respectively . Sucralfate exposure may decrease risk for C . difficile stool cytotoxin b by interfering with its detection, altering toxin production, or inhibiting colonization by the organism . Additional evaluation is needed to elucidate the mechanisms involved. Antimicrob Agents Chemother, 1994 Jul, 38(7), 1671 - 4 Increased activity of a new chlorofluoroquinolone, BAY y 3118, compared with activities of ciprofloxacin, sparfloxacin, and other antimicrobial agents against anaerobic bacteria; Aldridge KE; A total of 435 clinical isolates of anaerobes were tested with a broth microdilution method to determine the activity of BAY y 3118 compared with those of other agents against anaerobic bacteria . All strains of Bacteroides capillosus, Prevotella spp., Porphyromonas spp., Fusobacterium spp., Clostridium spp., Eubacterium spp., Peptostreptococcus spp., and Veillonella parvula were susceptible (MICs of < or = 2 micrograms/ml) to BAY y 3118 . Against the 315 strains of the Bacteroides fragilis group, five strains required elevated MICs (> or = 4 micrograms/ml) of BAY y 3118 . Only imipenem and metronidazole were active against all anaerobes . Overall, BAY y 3118 was more active than ciprofloxacin, sparfloxacin, piperacillin, cefotaxime, and clindamycin against the test isolates. Ann Vasc Surg, 1994 Jul, 8(4), 387 - 9 Clostridial aortic graft infection; Holland FW et al.; Aortic graft infection represents one of the most formidable challenges encountered by the vascular surgeon . Current principles of treatment are based on experience primarily derived from infection with Staphylococcus and enteric bacteria . Anaerobic prosthetic infection is a case event . Infection with Clostridium has heretofore been reported only twice . An additional case of clostridial infection of an aortic prosthesis is presented with review of the literature . Its clinical significance and management are discussed. Arzneimittelforschung, 1994 Jul, 44(7), 859 - 62 In vitro activity of meropenem compared with imipenem, metronidazole, ampicillin, and ampicillin/sulbactam against anaerobes; Schumacher U et al.; The aim of the present study was to compare the in vitro activity of meropenem (ICI 194660, CAS 96036-03-2) with imipenem, metronidazole, clindamycin, ampicillin and ampicillin/sulbactam against a variety of anaerobic bacteria using an agar dilution method . 423 clinical isolates were tested belonging to 70 species of 15 anaerobic genera . They included Bacteroides fragilis (n = 62), Bacteroides thetaiotaomicron (n = 45), Prevotella bivia (n = 11), Fusobacterium nucleatum (n = 12), Clostridium perfringens (n = 15) and several rarely isolated species and genera, e.g . Selenomonas sputigena and Clostridium symbiosum . Bacteroides species were inhibited by meropenem at < or = 2.0 micrograms/ml, Clostridium species, including C . difficile, at < or = 4.0 micrograms/ml and all the other anaerobes at < or = 0.5 microgram/ml . Meropenem and imipenem were the most active substances, but often equal to, or only slightly better than, metronidazole, clindamycin or ampicillin/sulbactam, dependent on species . Meropenem was especially active against Bacteroides gracilis (MIC90 0.015 microgram/ml), Prevotella disiens (MIC90 0.03 microgram/ml), Fusobacterium nucleatum (MIC90 0.015 microgram/ml), Clostridium perfringens (MIC90 0.015 microgram/ml) and Veillonella parvula (MIC90 0.03 microgram/ml) . The results obtained indicate that meropenem might be a useful adjunct to chemotherapy of anaerobic and mixed aerobic and anaerobic infections. Proteins, 1994 Jul, 19(3), 269 - 71 Crystallization and preliminary X-ray diffraction study of the green flavoenzyme 5-hydroxyvaleryl-CoA dehydratase/dehydrogenase from Clostridium aminovalericum; Eikmanns U et al.; The bifunctional flavoenzyme 5-hydroxyvaleryl-CoA dehydratase/dehydrogenase has been crystallized from solutions containing ammonium sulfate (form I) or polyethylene glycol (form II) as precipitant . In both cases, the crystals grew in the monoclinic space group C2 . The unit cell dimensions for form I crystals were determined as a = 162.8 A, b = 71.8 A, c = 83.5 A, beta = 109.1 degrees; corresponding values for form II crystals were a = 161.2 A, b = 71.6 A, c = 82.2 A, beta = 109.3 degrees . In both cases most probably there are two monomers per asymmetric unit . The crystals diffract to about 2 A resolution and are rather stable in the X-ray beam. Eur J Clin Microbiol Infect Dis, 1994 Jul, 13(7), 576 - 81 Evaluation of an oligonucleotide probe and an immunological test for direct detection of toxigenic Clostridium difficile in stool samples; Green GA et al.; A 33 basepair oligonucleotide probe, designed from the sequence of the Clostridium difficile toxin B gene, was evaluated for its ability to detect toxigenic Clostridium difficile directly in stool samples, without culture or DNA isolation . Two different labelling techniques were investigated: radiolabelling and digoxigenin-labelling . One hundred ninety-six stools were tested, with a good correlation (96%) obtained between the oligonucleotide probe and the gold standard, the cytotoxicity tissue culture assay . The sensitivity and specificity were 83% and 100%, respectively . In parallel, a new commercially available enzyme immunoassay for the detection of Clostridium difficile toxin A in stool specimens was investigated . In 162 samples tested, a sensitivity of 80% and a specificity of 98% were obtained. J Ind Microbiol, 1994 Jul, 13(4), 258 - 68 Analysis of Tn916-induced mutants of Clostridium acetobutylicum altered in solventogenesis and sporulation; Mattsson DM et al.; The conjugative transposon Tn916 was used for mutagenesis of Clostridium acetobutylicum ATCC 824 . Tetracycline-resistant mutants were screened for loss of granulose synthesis and five classes of granulose mutants, that contained single transposon insertions, were identified on the basis of altered solvent production . Class 1 mutants did not make acetone or butanol, lacked activity of enzymes induced during solventogenesis, and did not sporulate, indicating that they are regulatory mutants . The class 2 mutant strains also did not produce acetone but did form small amounts of butanol and ethanol, while the class 3 mutants produced low amounts of all solvents . Class 4 and 5 mutants produced essentially the same or higher amounts of solvents than the parent strain . Transposon insertions in the class 1 mutants were used as markers for in vitro synthesis of flanking chromosomal DNA using Tn916-specific primers . The DNA fragments were labeled to produce specific probes . Transposon insertion sites in the chromosomes of 13 different class 1 regulatory mutants were compared by hybridization of the specific probes to Southern blots of restriction endonuclease-digested parental chromosomal DNA . Insertions in two mutants appeared to be in the same region of the chromosome . These results predict that multiple regulatory elements are required to induce solvent production and sporulation. Int J Syst Bacteriol, 1994 Jul, 44(3), 591 - 3 Phylogenetic placement of Sarcina ventriculi and Sarcina maxima within group I Clostridium, a possible problem for future revision of the genus Clostridium . Request for an opinion; Willems A et al.; The 16S rRNA gene sequences of Sarcina ventriculi DSM 286T (T = type strain) and Sarcina maxima DSM 316T were determined . Phylogenetic analysis revealed that these two species are closely related to each other and belong to group I Clostridium (sensu Johnson and Francis) . The implications of these phylogenetic findings for future revision of the genus Clostridium are discussed. J Mol Biol, 1994 Jul 1, 240(1), 95 - 101 Identification of a grpE heat-shock gene homolog in the archaeon Methanosarcina mazei; Conway de Macario E et al.; A grpE heat-shock gene was found by sequencing in the genome of the methanogenic archaeon Methanosarcina mazei S-6 . It is the first example of grpE from the phylogenetic domain Archaea . Since the other seven sequenced homologs are from the domain Bacteria, it may be concluded that grpE appeared early in evolution, before the two domains separated . The archaeal grpE is located in the dnaK locus, 431 base-pairs upstream of dnaK, which is followed downstream by the dnaJ gene . The organization of these three genes is known for Bacillus subtilis, Clostridium acetobutylicum, Borrelia burgdorferi and Mycobacterium tuberculosis . The archaeal locus organization, grpE-dnaK-dnaJ, is similar to that of the former three bacteria, but different from that of M . tuberculosis . This, and sequence homologies, suggest that the M . tuberculosis GrpE belongs, together with the Streptomyces coelicolor homolog, to a subgroup of the GrpE proteins . The M . mazei grpE gene encodes a protein of 209 amino acid residues . The deduced amino acid sequence shows 28.2 to 34.6% identities, and 50.3 to 58.9 similarities (identities plus conservative substitutions) with the other six complete GrpE sequences available . These percentages fall within the range observed for the other GrpEs . Two regions in the second and fourth quarters of the GrpE molecule show higher homology, particularly in three stretches of nine, six and nine amino acid residues, respectively . The archaeal gene uses all codons but three, whereas the bacterial homologs lack higher numbers of codons . The M . mazei grpE responded to heat-shock by increasing transcription, in a manner similar to that of the nearby heat-shock gene dnaK. JAMA, 1994 Jun 22-29, 271(24), 1913 - 8 A randomized placebo-controlled trial of Saccharomyces boulardii in combination with standard antibiotics for Clostridium difficile disease; McFarland LV et al.; OBJECTIVE--To determine the safety and efficacy of a new combination treatment for patients with Clostridium difficile-associated disease (CDD) . The treatment combines the yeast Saccharomyces boulardii with an antibiotic (vancomycin hydrochloride or metronidazole) . DESIGN--A double-blind, randomized, placebo-controlled, parallel-group intervention study in patients with active CDD . Patients received standard antibiotics and S boulardii or placebo for 4 weeks, and were followed up for an additional 4 weeks after therapy . Effectiveness was determined by comparing the recurrence of CDD in the two groups using multivariate analysis to control for other risk factors for CDD . SETTING--National referral study of ambulatory or hospitalized patients from three main study coordinating centers . PATIENTS--A total of 124 eligible consenting adult patients, including 64 who were enrolled with an initial episode of CDD, and 60 who had a history of at least one prior CDD episode . Patients who were immunosuppressed due to acquired immunodeficiency syndrome or cancer chemotherapy within 3 months were not eligible . INTERVENTION--Treatment with oral S boulardii (1 g/d for 4 weeks) or placebo in combination with a standard antibiotic . MAIN OUTCOME MEASURE--Recurrence of active CDD . RESULTS--A history of CDD episodes dramatically increased the likelihood of further recurrences . Multivariate analysis revealed that patients treated with S boulardii and standard antibiotics had a significantly lower relative risk (RR) of CDD recurrence (RR, 0.43; 95% confidence interval, 0.20 to 0.97) compared with placebo and standard antibiotics . The efficacy of S boulardii was significant (recurrence rate 34.6%, compared with 64.7% on placebo; P = .04) in patients with recurrent CDD, but not in patients with initial CDD (recurrence rate 19.3% compared with 24.2% on placebo; P = .86) . There were no serious adverse reactions associated with S boulardii . CONCLUSIONS--The combination of standard antibiotics and S boulardii was shown to be an effective and safe therapy for these patients with recurrent CDD; no benefit of S boulardii was demonstrated for those with an initial episode of CDD. J Biol Chem, 1994 Jun 17, 269(24), 16706 - 11 Interaction of Clostridium botulinum C2 toxin with lipid bilayer membranes . Formation of cation-selective channels and inhibition of channel function by chloroquine; Schmid A et al.; Lipid bilayer experiments were performed with the C2-II binding component of the ADP-ribosylating C2 toxin from Clostridium botulinum . The trypsin-activated but not the nonactivated form of the protein was able to increase the specific conductance of artificial lipid bilayer membranes by the formation of ion-permeable channels . The channels had on average a single-channel conductance of 55 pS in 0.1 M KCl and were found to be cation-selective and voltage-dependent . The single-channel conductance was only moderately dependent on the bulk aqueous KCl concentration, which indicated point charge effects on the channel properties . Incubation of the activated C2-II binding component with antibodies against C2-II or with C2-I toxin inhibited channel formation to a large extent . Addition of chloroquine, a known inhibitor of endocytosis in cells, led to a dose-dependent decrease of the C2-II-induced membrane conductance . This result suggested that the activated C2-II component contains a binding site for chloroquine inside the channel . It is discussed that the channels formed by C2-II component are involved in the translocation of C2-I toxin across the target cell membrane. Mol Gen Genet, 1994 Jun 15, 243(6), 631 - 40 Organization of the botulinum neurotoxin C1 gene and its associated non-toxic protein genes in Clostridium botulinum C 468; Hauser D et al.; A 12.3 kb DNA fragment encompassing the botulinum neurotoxin C1 (BoNT/C1) gene and an upstream flanking region was sequenced from Clostridium botulinum C 468 phage 1C . The resulting bont/C1 locus includes six genes which are organized into three transcriptional units . Cluster 1 encompasses the bont/C1 gene and an upstream gene encoding a non-toxic protein associated with the toxin (Antp139/C1) . Transcriptional analysis revealed that these two genes form an operon; the bont/C1 gene can be transcribed alone or co-transcribed with antp139/C1 . Cluster 2 encompasses three genes (antp33/C1, antp17/C1 and antp70/C1), which also form an operon . The corresponding proteins are similar to components of the hemagglutinin complex associated with BoNT/A and BoNT/B of C . botulinum A and B . In addition, Antp33/C1 is identical to HA-33, an hemagglutinin encoded by C . botulinum C-Stockholm phage C-St; Antp70/C1 displays some relatedness to C . perfringens enterotoxin . The third transcriptional unit consists of orf-22, which encodes a basic protein showing 29% identity with the gene product of uviA, a plasmid-encoded protein of 22 kDa which has been identified as a positive regulator of the bacteriocin production in C . perfringens . Orf-22 could be an effector controlling the expression of the bont/C1 and its antp genes in C . botulinum C 468. Cytometry, 1994 Jun 15, 18(2), 103 - 8 Development and clinical evaluation of an amplified flow cytometric fluoroimmunoassay for Clostridium difficile toxin A; Renner ED; A rapid (2 h) amplified flow cytometric fluoroimmunoassay (AFCF) for Clostridium difficile toxin A was developed and compared with the cytotoxin assay (CTA) and culture of the organism from stool specimens from patients with suspected C . difficile-associated gastrointestinal disease (CAD) . For this assay polyclonal antitoxin A was attached to 10-microns diameter and monoclonal antitoxin A was attached to fluorescent 0.1 micron-diameter polystyrene microspheres . The microspheres and sample were reacted together as in a conventional double-antibody sandwich assay . However, laser flow cytometric measurement allowed the omission of separation and washing steps by gating on light scattered by the larger microspheres and measuring only the fluorescence associated with these particles . The amount of fluorescence from the attached 0.1 micron microspheres was dependent on the concentration of toxin A in the sample . The AFCF detected purified toxin A at levels of 1 pg/ml and was linear from 1 to 40 pg/ml . The AFCF was compared with the CTA and culture of C . difficile for clinical use by comparing results from 198 stool specimens from patients with suspected CAD . The AFCF was 85.7% sensitive and 95.8% specific relative to the CTA, and 85.2% sensitive and 98.3% specific compared to the culture assay . If the isolation of toxigenic C . difficile or the patients clinical course was considered indicative of CAD, the sensitivities of the AFCF, CTA, and culture assay were 77.4%, 67.7% and 96.8%, respectively . The AFCF demonstrated a specificity of 98.8%, while both CTA and culture had a specificity of 100%.(ABSTRACT TRUNCATED AT 250 WORDS) Carbohydr Res, 1994 Jun 2, 259(1), 103 - 15 Regioselective transglycosylation in the synthesis of oligosaccharides: comparison of beta-galactosidases and sialidases of various origins; Ajisaka K et al.; N-Acetyl-lactosamine(beta-D-Gal p-(1-->4)-D-Glc pNAc) was synthesized regioselectively with the aid of the transglycosylation activity of beta-galactosidase isolated from Diplococcus pneumoniae using p-nitrophenyl beta-D-galactopyranoside as the donor . Also, transglycosylation of the sialyl group in an alpha-(2-->8)-linked sialic acid dimer or p-nitrophenyl glycoside of sialic acid to N-acetyl-lactosamine was performed using sialidases of various origins . When sialidase from Clostridium perfringens, Arthrobacter ureafaciens, or Vibrio cholerae was used, alpha-(2-->6)-linked sialyl N-acetyl-lactosamine was obtained regioselectively . In contrast, when sialidase from newcastle disease virus was used, the alpha-(2-->3)-linked isomer was obtained regioselectively . The regioselectivity of the transglycosylation reaction using beta-galactosidase and sialidase was compared with hydrolysis specificity toward the same linkages. Postgrad Med, 1994 Jun, 95(8), 111 - 4, 117-20 Antibiotic-induced diarrhea and pseudomembranous colitis; Jacobs NF Jr; Pseudomembranous colitis is commonly associated with the use of antibiotics but may follow administration of other drugs and has occurred in patients who have not received any medication . Cases related to antibiotic administration are thought to be due to changes in normal intestinal flora that allow overgrowth of Clostridium difficile and elaboration of toxin . Clusters of cases in hospitals suggest nosocomial transmission of the bacteria . The stool cytotoxin assay is the most specific test for pseudomembranous colitis . Oral vancomycin (Vancocin) is preferred for the treatment of severe cases . It is recommended that hospital personnel caring for patients infected with C difficile wear gloves and wash their hands carefully after contact. Can J Surg, 1994 Jun, 37(3), 245 - 9 Invasive Clostridium septicum infection in association with colorectal carcinoma; Lorimer JW et al.; The association between invasive Clostridium septicum infection and colorectal carcinoma is examined by the presentation of three cases and a review of the literature . In the first two cases the patients presented with nontraumatic metastatic clostridial gas gangrene . In the third case a patient with chemotherapy-induced myelosuppression from concomitant multiple myeloma had a necrotizing transmural infection of the right colon . The apparent portal of entry of Clostridium septicum was an occult carcinoma of the ascending colon . The increasing evidence for a strong link between this organism and some cases of neutropenic enterocolitis is reviewed. J Infect Dis, 1994 Jun, 169(6), 1291 - 6 Oligosaccharide sequences attached to an inert support (SYNSORB) as potential therapy for antibiotic-associated diarrhea and pseudomembranous colitis; Heerze LD et al.; Toxin A produced by Clostridium difficile, the causative agent of pseudomembranous colitis and antibiotic-associated diarrhea, was shown to bind to synthetic oligosaccharide sequences attached to an inert support (SYNSORB) . The oligosaccharide sequences that bind to toxin A were related to sequences previously identified as potential receptors for the toxin . Various SYNSORBs containing a variety of oligosaccharides were examined for their potential to neutralize toxin A activity from toxin-containing solutions as well as clinical stool samples from patients with either pseudomembranous colitis or antibiotic-associated diarrhea . The results from neutralization experiments suggest SYNSORB can effectively neutralize toxin A activity from stool samples and thus could serve as a potential therapy for C . difficile-associated diarrhea. J Infect Dis, 1994 Jun, 169(6), 1206 - 18 Lessons from diarrheal diseases: demography to molecular pharmacology; Guerrant RL; From diarrheal diseases come profound lessons about health and population growth, microbial pathogenesis, and the molecular pharmacology of signal transduction . Epidemics such as cholera, hemorrhagic colitis, salmonellosis, and cryptosporidiosis remind us of how interdependent we are, sharing enteric microbial flora on a global scale . Diarrhea morbidity and mortality teach us that disease and poverty do not control but are associated with population overgrowth . Great advances are being made in understanding new bacterial, viral, and parasitic causes and treatment of diarrhea, especially persistent diarrhea . In addition, microbial toxins provide unique pharmacologic tools to probe cell signaling pathways . The mechanism of action of cholera toxin, once thought so clear, now appears to involve additional pathways such as platelet-activating factor and prostaglandin synthesis . Escherichia coli ST has opened a whole family of activators of guanylate cyclase, including new mammalian products that regulate sodium transport . Clostridium difficile toxin A provides a novel tool to dissect mediators involved in inflammatory diarrhea . These lessons have both basic implications for science and practical applications for medicine and society. Proteins, 1994 Jun, 19(2), 158 - 60 Characterization of two crystal forms of Clostridium thermocellum endoglucanase CelC; Dominguez R et al.; Endoglucanase CelC from Clostridium thermocellum expressed in Escherichia coli has been crystallized in two different crystal forms by the hanging drop method . Crystals of form I were grown with polyethylene glycol as a precipitant . They are orthorhombic, space group P2(1)2(1)2(1), with cell dimensions a = 51.4 A, b = 84.3 A, and c = 87.5 A . Crystals of form II, obtained in ammonium sulfate solutions, belong to the tetragonal space group P4(1)2(1)2 (or P4(3)2(1)2) with cell dimensions of a = b = 130.7 A and c = 69.6 A . Diffraction data to 2.8 A resolution were observed for both crystal forms with a rotating anode generator . Preliminary oscillation images of the orthorhombic form I crystals using a synchrotron radiation source show diffraction to 2.2 A resolution, indicating that these crystals are suitable for high resolution crystallographic analysis. Clin Infect Dis, 1994 Jun, 18(6), 982 - 4 Antibiotic-associated pseudomembranous enteritis due to Clostridium difficile; Tsutaoka B et al.; Although pseudomembranous colitis is relatively common following antibiotic exposure, there have been few reported cases of pseudomembrane formation involving the small intestine . Herein we report a case of pseudomembranous enteritis of the small and large intestine that occurred after antibiotic exposure . The etiologic organism appears to be Clostridium difficile, as evidenced by the characteristic pseudomembranous lesions and a positive ELISA for toxin A in an ileal tissue specimen. Infect Control Hosp Epidemiol, 1994 Jun, 15(6), 382 - 9 A sustained outbreak of Clostridium difficile in a general hospital: persistence of a toxigenic clone in four units; Nath SK et al.; OBJECTIVE: To evaluate the endemicity and epidemiology of toxigenic Clostridium difficile in a sustained outbreak of antibiotic-associated diarrhea . SETTING: University-affiliated, 465-bed tertiary care teaching hospital with adjacent cancer clinic in Hamilton, Ontario . DESIGN: From August 8, 1991, through August 31, 1993, a total of 187 cases were investigated for epidemiologic analysis of toxigenic C difficile from stool cultures, to identify the endemic clone(s) . To assess the nature of contamination, cultures of inanimate surfaces in the patient environment from the four most affected units (medical teaching, nonteaching medical, hematologic oncology, and the intensive care unit) were processed for C difficile . The 229 clinical strains and 24 environmental strains isolated were typed by numerical analysis of SDS-PAGE protein patterns . RESULTS: A majority (81%) of cases in the epidemiologic analysis were associated with a toxigenic electrophoretic (EP) type 1 C difficile that was identical to the strain first isolated from an index case that occurred 18 months before the start of this study . Culture and typing of the C difficile strains from the inanimate surfaces in the four most affected units showed that the patient environment was contaminated with the toxigenic EP type 1 organism . Six other strains that occurred infrequently among cases also were found in the environment . CONCLUSIONS: A single predominant toxigenic clone has been implicated in a sustained outbreak of antibiotic-associated diarrhea that affected elderly patients . The "endemic" clone transmitted for the 25-month study period was linked to an index case shedding a toxigenic EP type 1 strain that occurred 21 months prior to the initial outbreak on the medical teaching unit . The patient environment in the affected units was found to be contaminated with the same clone, possibly due to shedding of organisms by fecally incontinent symptomatic patients . The extrinsic factors contributing to the endemic transmission of this one clone still are not well understood. Mol Microbiol, 1994 Jun, 12(5), 761 - 77 Identification and molecular analysis of a locus that regulates extracellular toxin production in Clostridium perfringens; Lyristis M et al.; The anaerobic bacterium Clostridium perfringens mediates clostridial myonecrosis, or gas gangrene, by producing a number of extracellular toxins and enzymes . Transposon mutagenesis with Tn916 was used to isolate a pleiotropic mutant of C . perfringens that produced reduced levels of phospholipase C, protease and sialidase, and did not produce any detectable perfringolysin O activity . Southern hybridization revealed that a single copy of Tn916 had inserted into a 2.7 kb HindIII fragment in the C . perfringens chromosome . A 4.3kb PstI fragment, which spanned the Tn916 insertion site, was cloned from the wild-type strain . When subcloned into a shuttle vector and introduced into C . perfringens this fragment was able to complement the Tn916-derived mutation . Transformation of the mutant with plasmids containing the 2.7 kb HindIII fragment, or the 4.3 kb PstI fragment, resulted in toxin and enzyme levels greater than or equal to those of the wild-type strain . The PstI fragment was sequenced and found to potentially encode seven open reading frames, two of which appeared to be arranged in an operon and shared sequence similarity with members of two-component signal transduction systems . The putative virR gene encoded a protein with a deduced molecular weight of 30,140, and with sequence similarity to activators in the response regulator family of proteins . The next gene, virS, into which Tn916 had inserted, was predicted to encode a membrane-spanning protein with a deduced molecular weight of 51,274 . The putative VirS protein had sequence similarity to sensor proteins and also contained a histidine residue highly conserved in the histidine protein kinase family of sensor proteins . Virulence studies carried out using a mouse model implicated the virS gene in the pathogenesis of histotoxic C . perfringens infections . It was concluded that a two-component sensor regulator system that activated the expression of a number of extracellular toxins and enzymes involved in virulence had been cloned and sequenced . A model that described the regulation of extracellular toxin production in C . perfringens was constructed. J Dent Res, 1994 Jun, 73(6), 1133 - 41 Definition of a fundamental repeating unit in streptococcal glucosyltransferase glucan-binding regions and related sequences; Giffard PM et al.; The C-termini of the glucosyltransferases (Gtfs) of oral streptococci are responsible for glucan binding . These glucan-binding domains (GBDs) are composed of a series of repeated sequences that have been classified into four different classes (A-D) by virtue of sequence similarity and which, by inference, have been suggested to be of functional importance . In contrast, we propose that repeat sequences evolve in response to selection for an increase in the number of copies of a particular domain through multiple duplication events occurring at different times . According to this hypothesis, repeats should possess various degrees of similarity, especially if only key residues are of functional importance . Analysis of the GBDs of the Gtfs indicated that a common fundamental repeat, designated the "YG" repeat, could be discerned within the "A", "B", "C", and "D" repeats . Similar elements were also conserved in the ligand-binding repeats of the Clostridium difficile toxins and the lysins and the PspA protein of Streptococcus pneumoniae, suggesting that similar selective pressures had also been imposed on these sequences . Analysis of the "YG" repeats present in the GtfJ and GtfK of Streptococcus salivarius indicated that some of the "YG" repeats in the GBDs of these proteins had arisen as a result of duplication events involving a series of three sequential "YG" repeats. Radiol Med (Torino), 1994 Jun, 87(6), 775 - 82 {Imaging diagnosis in pseudomembranous colitis}; Pieri L et al.; Pseudomembranous colitis (PC) is a dangerous inflammatory disease which arises as a complication of systemic antibiotic therapy . The colon is the preferred localization of PC, which is caused by the alteration in the bacterial population of the bowel which favors the growth and activation of several germ types--e.g., the Clostridium difficile, whose toxins can damage the colonic mucosa deeply . Later, the condition may affect extramucosal structures thus causing an actual parietal alteration . Clinically, PC patients present with diarrhea, abdominal pain, onset or worsening of fever, impairement of the main body functions . The colonic mucosa appears macroscopically edematous and is covered with yellowish plaques, called "pseudomembranes", which adhere strictly to the mucosa . Pseudomembranes are made by fibrin, mucus, leucocytes and epithelial remnants . The diagnosis is made on the basis of laboratory tests--i.e . the demonstration of Clostridium difficile or its toxins in the feces . Endoscopy is the examination of choice when PC is suspected because it can demonstrate the typical mucosal alterations directly . In this paper the main etiologic, pathologic and clinical features of PC are presented and the role of diagnostic imaging examinations is discussed, not only in demonstrating the typical lesions but also in the spatial evaluation of the condition and in its follow-up. J Appl Bacteriol, 1994 Jun, 76(6), 539 - 45 Polymerase chain reaction for detection of Clostridium botulinum types A, B and E in food, soil and infant faeces; Szabo EA et al.; The application of the polymerase chain reaction (PCR) for detection of Clostridium botulinum types A, B and E in foods, environmental and clinical samples was evaluated and compared to the mouse bioassay . Samples inoculated with 10, 100 and 1000 spores of Cl . botulinum types A and B included pasteurized milk, UHT milk, infant formula, infant faeces, meat juice, canned tuna, mushrooms, blood sausage and soil . Clostridium botulinum type E spores were inoculated into fish eggs, canned tuna, picked herring, raw fish and soil at similar levels . Spores were added to 2.5 g of each sample with the exception of soil which was inoculated in 10 g samples . The presence of Cl . botulinum in sample enrichments was determined by both PCR and the bioassay . An overall correlation of 95.6% was observed between PCR results and the mouse bioassay . Of the total of 114 samples tested there was disparity between the mouse bioassay and the PCR in three samples of soil inoculated with 100 type A or E spores and 10 type B spores per 10 g, respectively, and two samples of infant faeces inoculated with 10 type A or B spores per 2.5 g . All of these samples gave negative animal results and positive PCR results. Eur J Biochem, 1994 Jun 1, 222(2), 639 - 44 Reversible and non-denaturing replacement of iron by cadmium in Clostridium pasteurianum ferredoxin; Bonomi F et al.; Incubation of native, reduced Clostridium pasteurianum ferredoxin with different metals gave a range of modifications in the electronic and EPR spectrum of the protein, or made the signals disappear . The reduced protein, isolated after incubation with different metals under identical conditions (50 microM protein, 1 mM metal, 1 h incubation) was found to contain amounts of foreign metals increasing with their thiophylicity, i.e . Cd2+ >> Zn2+ > Co2+ . Little, if any, incorporation was observed for Ni2+, Cu2+, Mn2+ or in the absence of reductant . The activity of substituted ferredoxins in a hydrogenase-coupled assay was proportional to the amount of residual iron, suggesting that the residual iron is present in a population of intact active molecules rather than in partially substituted clusters distributed among individual molecules . The cadmium-substituted ferredoxin did not contain iron, but contained eight cadmium atoms and six labile sulfide atoms/mol . Folding of the isolated, substituted proteins was investigated by CD and 1H-NMR . Both techniques showed retention of the main structural features of the protein upon metal substitution . The rate and extent of the substitution of iron by cadmium were essentially independent of pH, but were found to decrease with increasing ionic strength and to increase with the cadmium concentration . In the cadmium-substituted protein, cadmium was replaced by iron upon incubation with iron and mercaptoethanol in the absence of dithionite . In the presence of dithionite, cadmium was not replaced by iron upon incubation of the cadmium-substituted protein with excess iron and mercaptoethanol . In competition experiments, incubation of iron-containing ferredoxin with stoichiometric amounts of cadmium in the presence of dithionite and excess iron and mercaptoethanol resulted in quantifiable replacement of iron by cadmium . Therefore, substitution of iron by cadmium was only achieved under reducing conditions, and was only reversible in the absence of strong reductants. J Med Microbiol, 1994 Jun, 40(6), 379 - 84 Retrospective study of 108 cases of botulism in Poitiers, France; Roblot P et al.; Botulism, a food-borne toxin-mediated disease caused by Clostridium botulinum is still a common disease, which is most frequent in the rural environment; 108 cases, 66 males and 42 females, average age 32 years, were recorded from 1965 to 1990 in the infectious disease department of the University Hospital of Poitiers (France) . In 83% of patients, the food responsible was home-cured ham . Mean incubation time was 3.4 days; digestive symptoms were observed in 93% of cases, ocular symptoms in 92% and urinary tract dysfunction in 22% . A scale of severity was used to classify the patients into those suffering from severe (6), intermediate (50) and mild (52) forms of the disease . Botulinum toxin type B was found in 36 (52%) of 69 blood samples and in 41 (51%) of 81 samples of the suspected food . From 1965 to 1976, 44 patients were treated with both toxoid and heterologous equine serotherapy . Since 1976, 29 patients have been treated with guanidine hydrochloride (35 mg/kg daily) and 35 patients with guanidine hydrochloride plus heterologous serotherapy . All 108 patients recovered without any sequelae. J Cell Sci, 1994 Jun, 107 ( Pt 6), 1653 - 9 Probing the action of Clostridium difficile toxin B in Xenopus laevis oocytes; Just I et al.; Clostridium difficile toxin B and Clostridium botulinum C3 exoenzyme caused comparable morphological alteration of CHO cells, which was accompanied by disaggregation of the microfilamental cytoskeleton . The cytotoxic effect of toxin B was correlated with a decrease in C3-catalyzed ADP-ribosylation of the low-molecular-mass GTP-binding protein Rho, which is involved in the regulation of the actin cytoskeleton . We used Xenopus laevis oocytes as a model to study the toxin effect on Rho in more detail . Toxin B treatment of oocytes caused a decrease in subsequent ADP-ribosylation of cytoplasmic Rho by C3 . This decrease was observed when toxin B was applied externally or after microinjection . Besides endogenous Rho, microinjected recombinant Rho-glutathione S-transferase fusion protein was affected . Impaired ADP-ribosylation of Rho was neither due to altered guanine nucleotide binding nor to complexation with the guanine nucleotide dissociation inhibitor, which is known to inactivate Rho and to prevent Rho modification by C3 . Proteolytical degradation of Rho was excluded by immunoblot analysis . In intact oocytes toxin B caused neither ADP-ribosylation nor phosphorylation of Rho . The data indicate that C . difficile toxin B acts on Rho proteins in Xenopus oocytes to inhibit ADP-ribosylation by C3 . It is suggested that toxin B mediates its cytotoxic effect via functional inactivation of Rho. Glycobiology, 1994 Jun, 4(3), 367 - 73 Cloning and expression of a soluble sialidase from Chinese hamster ovary cells: sequence alignment similarities to bacterial sialidases; Ferrari J et al.; A cDNA encoding a soluble sialidase from Chinese hamster ovary (CHO) cells has been cloned and expressed . Completely degenerate oligonucleotide primers, which were based on the amino acid sequence of peptides obtained from the purified sialidase (Warner et al., Glycobiology, 3, 455-463, 1993), and the polymerase chain reaction, with single-stranded cDNA template, were employed to generate a unique oligonucleotide probe . The unique probe of 93 bp was used for screening a lambda gt 10 CHO cell cDNA library . A single clone, which contained a 1.4 kb insert, was isolated after screening 450,000 recombinants . The complete coding region of the protein, 1137 nucleotides, was contained in the isolated clone and it predicted a protein of 379 amino acids . The insert had a 186 bp 5' non-coding leader sequence and a 40 bp 3' non-coding region . No signal peptide was identified in the insert, suggesting a cytosolic localization for the protein . No significant primary sequence identities were observed when the deduced amino acid sequence of the CHO cell sialidase was compared with other mammalian proteins or microbial sialidases . However, the protein had significant sequence alignment similarity with several bacterial sialidases . Two 'Asp box' motifs in the CHO cell sialidase had a remarkable alignment positioning in the protein sequence with the similar motifs of the Salmonella LT2 and Clostridium perfringens sialidases . High levels of the enzyme were expressed in Spodoptera frugiperda cells infected with a modified Autographa californica nuclear polyhedrosis virus harbouring the sialidase cDNA. Bioorg Khim, 1994 Jun, 20(6), 627 - 34 {Proteinases with various substrate specificities in structural studies of yeast cell walls}; Kalebina TS et al.; Different proteins are revealed in cell wall of yeast cells Candida utilis by means of specific proteolysis with subtilisins TV and 72, trypsin and purified collagenase of Clostridium histolyticum . Some of them were characterized by resistance to trypsin and sensitivity to subtilisin TV . In young cells this group is represented essentially by a protein of 33 kD, which appears to be one of the structural proteins, binding fibrillae of carbohydrate . Other proteins proved to be sensitive to both trypsin and subtilisin . Among these proteins a protein with mol . mass 80 kD was revealed; its sensitivity to extremely specific hydrolysis by bacterial collagenase suggests it to contain amino acid sequences characteristic for collagens of higher eukaryotes. Protein Eng, 1994 Jun, 7(6), 749 - 60 Structural similarities in glucoamylase by hydrophobic cluster analysis; Coutinho PM et al.; The model of the catalytic domain of Aspergillus awamori var . X100 glucoamylase was related to 14 other glucoamylase protein sequences belonging to five subfamilies . Structural features of the different sequences were revealed by multisequence alignment following hydrophobic cluster analysis . The alignment agreed with the hydrophobic microdomains, normally conserved throughout evolution, evaluated from the 3-D model . Saccharomyces and Clostridium glucoamylases lack the alpha-helix exterior to the catalytic domain . A different catalytic base was found in the Saccharomyces glucoamylase subfamily . The starch binding domain of fungal glucoamylases has identical structural features and substrate interacting residues as the C-terminal domain of models of Bacillus circulans cyclodextrin glucosyltransferases . Three putative N-glycosylation sites were found in the same turns in glucoamylases of different subfamilies . O-Glycosylation is present at different levels in the catalytic domain and in the linker between the catalytic and starch binding domains. J Autoimmun, 1994 Jun, 7(3), 291 - 320 Monoclonal anti-H1 histone autoantibodies from unimmunized Balb/c mice . Specificity and VH and VL domain sequences; Underwood JR et al.; Examination of the nuclear reactivities of monoclonal IgM kappa autoantibodies, secreted by GFM-5 1B12 and NU-6 1F12 hybridomas derived from germ-free and nude mice, respectively, demonstrated homogeneous nuclear immunofluorescence staining patterns consistent with the recognition of histones . Under these conditions, GFM-5 1B12 and NU-6 1F12 mAbs produced species non-specific binding to components within the nuclei of mouse, human and Drosophila melanogaster cells . Immunoblotting confirmed the binding of these two autoantibodies to autologous H1 histones as well as bovine and insect H1 histones . Identification of the epitopes bound by GFM-5 1B12 and NU-6 1F12 mAbs within the D . melanogaster H1 histones was undertaken using 248 overlapping octapeptides encompassing the entire sequence of D . melanogaster H1 histones . GFM-5 1B12 mAbs bound several octapeptides derived from the amino- and carboxyl-terminal regions of D . melanogaster H1 histones with accessible KT, AT or VT amino acids . NU-6 1F12 mAbs, which stained nuclei within sections of D . melanogaster lavae, failed to bind to any of the 248 linear octapeptides, implying recognition of a conformational H1 histone epitope by this autoantibody . ELISA analysis of the polyspecific binding properties of GFM-5 1B12 and NU-6 1F12 mAbs demonstrated that both antibodies exhibited unique polyspecificity profiles . GFM-5 1B12 mAbs recognized bovine carbonic anhydrase and mouse IgG1, while NU-6 1F12 bound bovine cardiolipin, rat cytochrome c, Escherichia coli beta-galactosidase, toxoid from Clostridium tetani, mouse IgG1 and the haptens, DNP and FITC from the 24 antigen test panel . Comparison of the VH and VL domain sequences of GFM-5 1B12 and NU-6 1F12 mAbs demonstrated that the variations in autoreactivity and polyspecificity profiles resulted from amino acid variations in the CDRs of the VH and VL domains of these autoantibodies . Significantly, major differences in the VH domain sequences of the NU-5 1F12 and GFM-5 1B12 mAbs suggest that the VH domains may preferentially contribute to the unique specificities of the two anti-H1 histone autoantibodies. Br J Pharmacol, 1994 Jun, 112(2), 453 - 60 Mediation by bradykinin of rat paw oedema induced by collagenase from Clostridium histolyticum; Legat FJ et al.; 1 . Collagenases are thought to play a major role in the pathology of gas gangrene caused by Clostridium histolyticum, because they can destroy the connective tissue barriers . We investigated possible mediators involved in the oedema formation and plasma protein extravasation which follow the injection of a collagenase (EC 3.4.24.3) from Clostridium histolyticum into one hind paw of anaesthetized rats . 2 . The magnitude of the oedema following a subplantar injection was dependent on the dose of collagenase (30, 100 and 300 micrograms) injected . It reached its maximum within 30 min and remained unchanged for at least 5 h . Plasma protein extravasation into the paw was most pronounced within 20 min of the injection . Heat-inactivated collagenase was ineffective . 3 . The B2 bradykinin (BK) antagonist icatibant (D-Arg-{Hyp3-Thi5-D-Tic7- Oic8} bradykinin, formerly named Hoe-140) reduced oedema formation in a dose-dependent manner with a maximal reduction of around 65% at a dose of 100 nmol kg-1 (s.c.) . A significant effect could already be observed at a dose of 10 nmol kg-1 . The duration of the effect of icatibant (100 nmol kg-1) was found to be at least 3 h . These results demonstrate the high potency and long duration of action of icatibant . Pretreatment of rats with the bradykinin B1 antagonist, des-Arg9-{Leu8}-BK did not affect collagenase-induced paw oedema . Thus, the observed collagenase-induced effects are mainly mediated by BK through activation of B2 receptors.(ABSTRACT TRUNCATED AT 250 WORDS) Southeast Asian J Trop Med Public Health, 1994 Jun, 25(2), 321 - 3 Reactivity of the CD D-1 latex test with Clostridium difficile and other bacteria; Wongwanich S et al.; The reactivity of a commercial latex test with thirty-three species of bacteria was tested . Toxigenic and nontoxigenic strains of Clostridium difficile gave a positive result in the CD D-1 latex test . Cross-reactions were also given by C . putrificum, C . sporogenes and proteolytic C . botulinum. J Clin Microbiol, 1994 Jun, 32(6), 1591 - 3 Use of arbitrary primer PCR to type Clostridium difficile and comparison of results with those by immunoblot typing; Killgore GE et al.; An arbitrarily primed PCR (AP-PCR) assay was used to type Clostridium difficile isolates from a hospital outbreak of antibiotic-associated diarrhea . Forty-one isolates were separated into nine groups, with 66% falling into one group; no other group contained more than 10% . Comparison of AP-PCR grouping with that when the immunoblot technique was used showed agreement for 33 of 34 isolates typed by both techniques, and AP-PCR grouped seven isolates that were not typeable by immunoblotting. Infect Control Hosp Epidemiol, 1994 Jun, 15(6), 371 - 81 Ten years of prospective Clostridium difficile-associated disease surveillance and treatment at the Minneapolis VA Medical Center, 1982-1991; Olson MM et al.; OBJECTIVES: To understand the epidemiology, risks, and management of Clostridium difficile-associated disease (CDAD) and to establish and evaluate reliable methods of surveillance . DESIGN: Case finding was done by daily ward and laboratory rounds . The criteria for CDAD diagnosis were: at least four unformed stools per day for 2 days and a positive culture or cytotoxin for C difficile, or positive endoscopy or autopsy for pseudomembranes . SETTING: The surveillance covered all patients from 1982 through 1991 in the 820-bed Minneapolis Veterans Affairs Medical Center . PARTICIPANTS: The criteria were met by 908 patients . Medical service patients numbered 488; surgical patients, 420 . Frequencies ranged from a high of 149 cases in 1982 to a low of 50 cases in 1989 . RESULTS: Stool specimens were obtained on 898 (99%) of the 908 CDAD patients . Stools were culture-positive in 864 (96%) of 898, cytotoxin-positive in 569 (63%) of 898 . Endoscopy was performed on 196 (22%) of the 908 patients, and 80 (41%) of 196 patients had pseudomembranes . Ten (1%) of the 908 patients were diagnosed by endoscopy without a stool specimen, or at autopsy . No treatment was needed for 135 (15%) of the 908 CDAD patients, and 19 (2%) of the 908 died before treatment was started . Oral metronidazole was the treatment for 632 (70%) of 908 patients (1% intolerance, 2% failure, 7% relapse) and oral vancomycin was given to 122 (13%) of 908 patients (1% intolerance, 1% failure, 10% relapse) . Twelve patients had pseudomembranous colitis at autopsy, and it was the primary cause of death in 5 (0.6%) of 908 . CONCLUSIONS: CDAD usually responds to oral metronidazole or vancomycin but is nonetheless responsible for a high morbidity and occasional mortality in patients even when the diagnosis and treatment are pursued aggressively. Biochem J, 1994 May 15, 300 ( Pt 1), 133 - 9 ADP-ribosylation of Rho proteins by Clostridium botulinum exoenzyme C3 is influenced by phosphorylation of Rho-associated factors; Fritz G et al.; Specific {32P}ADP-ribosylation by Clostridium botulinum exoenzyme C3 was used to study the involvement of phosphorylation in the regulation of the low-molecular-mass GTP-binding protein Rho . Dephosphorylation of CHO cell extracts by alkaline phosphatase treatment resulted in a 80-90% reduction in the C3-catalysed {32P}ADP-ribosylation of Rho proteins in both cytosolic and membrane fractions . Similar results were obtained after dephosphorylation with protein phosphatase type-1 from bovine retina, whereas type-2B and type-2C phosphatases had no effect on the level of subsequent {32P}ADP-ribosylation of Rho by C3 . Incubation of CHO cell lysate under phosphorylation conditions increased the subsequent C3-mediated {32P}ADP-ribosylation of Rho proteins . The protein kinase inhibitors H7 and H9 had no effect on {32P}ADP-ribosylation at concentrations which are specific for inhibition of protein kinase A or C . Recombinant glutathione S-transferase-RhoA fusion protein (GST-RhoA) was phosphorylated by protein kinase A; however, the phosphorylation had no stimulatory effect on the ADP-ribosylation of GST-RhoA by C3 . An approx . 48 kDa phosphoprotein was identified which bound specifically to recombinant GST-RhoA fusion protein . By gel-permeation chromatography, Rho-containing complexes of approx . 50 kDa and 130-170 kDa were detected . The ADP-ribosylation of Rho in the 130-170 kDa complex was reduced by alkaline phosphatase pretreatment . The data suggest that Rho activity is influenced by phosphorylation of Rho-associated regulatory factors . Phosphorylation/dephosphorylation of these Rho-regulating factors appears to alter the ability of Rho to serve as a substrate for C3-induced {32P}ADP-ribosylation. Biochem J, 1994 May 1, 299 ( Pt 3), 775 - 9 ADP-ribosylation of the GTP-binding protein Rho by Clostridium limosum exoenzyme affects basal, but not N-formyl-peptide-stimulated, actin polymerization in human myeloid leukaemic (HL60) cells; Koch G et al.; Treatment of human myeloid leukaemic (HL60) cells with Clostridium limosum exoenzyme, which inactivates the small GTP-binding protein Rho by ADP-ribosylation, decreased the basal F-actin content . Inhibition of F-actin occurred after long-term treatment (24 h) of intact HL60 cells or after introduction of the toxin by electropermeabilization in a toxin-concentration-dependent manner . Concomitantly with the decrease in the basal F-actin content, the GTP-binding protein Rho was ADP-ribosylated in intact cells . However, Clostridium limosum toxin had no inhibitory effect on N-formyl-peptide-induced actin polymerization . Moreover, the relative N-formyl-peptide-stimulated polymerization was substantially enhanced in cells treated with Clostridium limosum transferase . In contrast with Clostridium limosum exoenzyme, component C21 of the Clostridium botulinum C2 toxin, which ADP-ribosylates G-actin, depolymerized basal F-actin and inhibited N-formyl-peptide-induced actin polymerization in electropermeabilized HL60 cells . These findings indicate that Rho proteins are involved in the basal, but not the ligand-evoked, actin polymerization in HL60 cells. Ophthalmology, 1994 May, 101(5), 839 - 42 Clostridium bifermentans panophthalmitis after penetrating eye injury; Rehany U et al.; BACKGROUND: Intraocular and orbital anaerobic infections usually result from penetrating eye injuries with soil-contaminated foreign bodies . The outcome of these infections almost always has been loss of the globe, despite appropriate antibiotic and surgical treatment . The most prevalent etiologic microbe of anaerobic panophthalmitis is Clostridium perfringens . CASE REPORT: To the authors' knowledge, this is the first report of panophthalmitis caused by Clostridium bifermentans after penetrating eye injury . The patient had severe signs and symptoms of intraocular and orbital infection, with early total loss of visual function . Parenteral and intravitreal therapy with penicillin and clindamycin, administered according to antibiotic sensitivity studies of cultures from the anterior chamber and vitreous, did not restore vision . CONCLUSIONS: Due to the early devastating outcome, penetrating eye injuries with soil-contaminated foreign bodies should be regarded as being at high risk for clostridial infection and should be treated promptly with vitrectomy and antibiotic therapy for aerobic and anaerobic infection. J Bacteriol, 1994 May, 176(10), 2828 - 34 Subcellular localization of Clostridium thermocellum ORF3p, a protein carrying a receptor for the docking sequence borne by the catalytic components of the cellulosome; Salamitou S et al.; The ORF3 gene of Clostridium thermocellum encodes a polypeptide (ORF3p) which contains a receptor domain for the docking sequence borne by the catalytic subunits of the cellulosome and a triplicated domain related to some bacterial cell surface proteins . It was thus surmised that ORF3p is a surface protein . In this study, this hypothesis was confirmed . Subcellular fractionation, Western blotting (immunoblotting), and electron microscopy of immunocytochemically labeled cells indicated that ORF3p produced by C . thermocellum was located in the outer surface layer of the bacterium . This layer appeared to consist of a soft matrix shedding off particulate fragments . Nonsedimenting ORF3p derived from sonicated cells was associated with high-molecular-mass fractions (> 20 MDa), probably corresponding to fragments of the outer cell layer . The same high-molecular-mass fractions also contained the cellulosomal marker CipA . Contrary to CipA, however, ORF3p was not associated with 2- to 4-MDa fractions corresponding to individual cellulosomes, and a significant fraction of ORF3p failed to bind to cellulose . It is proposed that ORF3 and ORF3p be renamed olpA and OlpA, respectively (for outer layer protein). J Bacteriol, 1994 May, 176(10), 2822 - 7 Recognition specificity of the duplicated segments present in Clostridium thermocellum endoglucanase CelD and in the cellulosome-integrating protein CipA; Salamitou S et al.; The binding specificity of the duplicated segments borne by Clostridium thermocellum endoglucanase CelD and by the cellulosome-integrating protein CipA was investigated . The fusion protein CelC-DSCelD, in which the duplicated segment of CelD was fused to the COOH terminus of endoglucanase CelC, bound with an affinity of 4.7 x 10(7) M-1 to the fusion protein MalE-RDCipA, in which the seventh receptor domain of CipA was grafted onto the COOH terminus of the Escherichia coli maltose-binding protein MalE . The affinity of CelC-DSCelD for the homologous chimeric protein MalE-RDORF3p, carrying the receptor of the surface protein ORF3p, was 6.9 x 10(6) M-1 . The fusion protein CelC-DSCipA, in which the duplicated segment of CipA was grafted onto the COOH terminus of CelC, did not bind detectably to MalE-RDCipA or MalE-RDORF3p . However, Western blotting (immunoblotting) experiments indicated that the duplicated segment of CipA was able to bind to a set of C . thermocellum proteins which are different from those recognized by the duplicated segment of CelD . These results argue against the hypothesis that ORF3p interacts with the duplicated segment of CipA . More probably, ORF3p binds to individual cellulases and hemicellulases harboring duplicated segments. Arch Surg, 1994 May, 129(5), 557 - 60 Enteritis necroticans with midgut necrosis caused by Clostridium perfringens; Clarke LE et al.; Enteritis necroticans is a necrotizing process manifesting as segmental gangrene of the bowel, triggered by Clostridium perfringens toxins under specific dietary conditions . It is a rare disease in developed countries and is probably underdiagnosed . A case of enteritis necroticans presenting with midgut necrosis with sepsis and hemolysis is reported herein . Bacteriologic culture of blood and peritoneal content revealed C perfringens . Dietary history, including the ingestion of meat together with sweet potatoes, should increase clinical suspicion of enteritis necroticans . Early recognition and timely surgical intervention are required for successful treatment . Clinicians are encouraged to be aware of this clinically fulminant yet rarely recognized surgical entity. Arch Biochem Biophys, 1994 May 1, 310(2), 392 - 6 Comparison of electron transfer kinetics between redox proteins free in solution and electrostatically complexed to a lipid bilayer membrane; Cheddar G et al.; The second-order rate constants obtained in solution for the reduction of horse cytochrome c (cytc; net charge +7) by either Clostridium beijerinckii flavodoxin semiquinone (Fld; net charge -16) or reduced spinach ferredoxin (Fd; net charge -15) decrease monotonically with increasing ionic strength, as expected for reactions between oppositely charged species . Although the rate constant for the Fld reaction is almost two orders of magnitude larger at low ionic strength than that for Fd, the values extrapolated to infinite ionic strength are closely similar, indicating comparable reactivities when electrostatic effects are eliminated . Furthermore, Fld has a much larger value for the electrostatic interaction energy, and thus a larger apparent active site charge, than does Fd, accounting for the rate constant disparity at low ionic strength . Electrostatically binding cytc at low ionic strength to a negatively charged lipid bilayer vesicle (membranes containing mixtures of egg phosphatidylcholine (PC) and cardiolipin (CL)) results in a marked decrease of the observed electron transfer rate constant (k(obs)) for reduction of the cytochrome by both Fld and Fd . The magnitude of this decrease is proportional to the mole percent of CL present in the membrane (10- to 20-fold change over 5-60 mol%) . With Fld, k(obs) decreases monotonically with increasing ionic strength at a fixed CL concentration . With Fd an increase in k(obs) occurs as the ionic strength is increased, which maximizes at intermediate ionic strength at a value larger than that obtained in the absence of lipid vesicles . When Fld is electrostatically bound to a positively charged vesicle composed of 40 mol% dioctadecyldimethylammonium ion (DODAC) and 60 mol% PC, again k(obs) for electron transfer to cytc is decreased over that obtained in solution, and the magnitude is diminished monotonically by increasing ionic strength . In contrast, k(obs) for electron transfer from Fd to cytc is unaffected by the presence of the positively charged membrane . The implications of these results for the role of membrane surface charge in modulating protein-protein interactions is discussed. Infect Immun, 1994 May, 62(5), 1623 - 30 Salmonella typhimurium loci involved in survival within macrophages; Baumler AJ et al.; A set of Tn10 mutants of Salmonella typhimurium which have a diminished capacity to survive in murine macrophages and decreased virulence in mice has been described previously . In this study, we characterized 30 of these mutants and determined map locations of Tn10 insertions for 23 of these strains . In addition, short fragments of transposon-flanking DNA were cloned, and the nucleotide sequence was determined for 23 mutants . Seven mutants carried transposon insertions in known genes, representing six loci: htrA, prc, purD, fliD, nagA, and smpB . The possible roles of these genes in Salmonella virulence are discussed . One insertion was found to be in an unknown gene which shared homology with the open reading frames Bv' and Bv located in the pin inversion system of Shigella boydii . In one mutant, Tn10 was found to be inserted in a gene with significant homology to adhE of Escherichia coli and Clostridium acetobutylicum . The map location and degree of homology indicate that the Salmonella gene encodes a related, but different, dehydrogenase . In 14 of the mutants analyzed, Tn10 was inserted into genes which had no significant homologies to entries in the DNA and protein data bases . In conclusion, 16 insertions define loci, termed ims for impaired macrophage survival, which have not yet been described in S . typhimurium but have been shown previously to be necessary for full virulence in mice . Although most ims loci are distributed randomly throughout the genome, a cluster was found between 75 and 78 min on the Salmonella chromosome. Clin Infect Dis, 1994 May, 18 Suppl 4, S297 - 304 Genetic basis for antibiotic resistance in anaerobes; Sebald M; This review focuses on genetic and molecular data regarding antibiotic resistance in anaerobes, particularly Clostridium perfringens, Clostridium difficile, Bacteroides species, and Prevotella species . The determinants of resistance are frequently transferable through a conjugation-like process; plasmid self-transfer, plasmid mobilization, or (in Bacteroides species) chromosomal conjugative elements can be involved . The determinants can be localized on transposons . At the genetic level, resistance determinants can be highly specific for one or several anaerobes or may exhibit homology with genes from aerobes . The latter observation suggests that anaerobes are able to exchange genetic material from a "gene pool" shared with aerobic organisms. Clin Infect Dis, 1994 May, 18 Suppl 4, S265 - 72 Clostridium difficile: history of its role as an enteric pathogen and the current state of knowledge about the organism; Bartlett JG; Clostridium difficile is the most frequently identified enteric pathogen in patients with antibiotic-associated diarrhea and colitis . It accounts for 10%-25% of all cases of antibiotic-associated diarrhea and virtually all cases of antibiotic-associated pseudomembranous colitis . Clinical features that distinguish infection with C . difficile from that due to many other enteric pathogens are hyperpyrexia, leukemoid reactions, toxic megacolon, pseudomembranous colitis, hypoalbuminemia, and chronic diarrhea . Factors important in the pathogenesis of disease are exposure to antibiotics, the presence of C . difficile in the patient's indigenous flora or acquisition of the organism from an environmental source, production of toxin A, and age-related susceptibility . The criterion standard for testing is the tissue culture assay; alternatives are culture and other methods of antigen detection including EIA, dot blot hybridization assay, and latex agglutination . The optimal drug for treatment is vancomycin; however, metronidazole is often used because it is less expensive . At present the main problems associated with C . difficile infection are treatment of patients with ileus, the management and prevention of nosocomial epidemics, and the management of repeated relapses. Clin Infect Dis, 1994 May, 18 Suppl 4, S250 - 2 Utility of newer techniques for classification and identification of pathogenic anaerobic bacteria; Hofstad T; Results of genetic and biochemical analyses have broadened our understanding of taxonomic relationships among groups of anaerobic bacteria and have led to a better understanding of the pathogenesis of infection . Conventional bacteriologic methods are still of prime importance for the detection and identification of anaerobic pathogens . The use of nucleic acid probes has so far been restricted to research laboratories . A polymerase chain reaction-generated probe would be most useful for the rapid detection of toxigenic Clostridium difficile in feces . Probes are needed for detection of periodontopathogenic bacteria in dental plaque . Use of nucleic acid probes may become a useful adjunct to classic methodology in reference and teaching laboratories. Protein Eng, 1994 May, 7(5), 681 - 7 On the role of conserved proline residues in the structure and function of Clostridium pasteurianum 2{4Fe-4S} ferredoxin; Quinkal I et al.; The widespread occurrence of Pro residues adjacent to Cys ligands in the sequences of {4Fe-4S} electron transfer proteins has not yet found a functional basis . The two such Pro of Clostridium pasteurianum 2{4Fe-4S} ferredoxin have been probed by site-directed mutagenesis . Any one of them, but not both simultaneously, can be substituted without impairing the proper folding of the protein . The reduction potentials of the ferredoxin variants fall in a narrow range of < 20 mV above the potential of the native protein . The biological activities with C . pasteurianum hydrogenase and pyruvate-ferredoxin oxidoreductase do not change significantly, except when Lys replaces Pro . In these cases, the data suggest that the two clusters of 2{4Fe-4S} ferredoxin may not always be equivalent in the interaction with the redox partners . Destabilization of the structure has been observed as the consequence of the Pro19 or Pro48 substitutions . Using 2-D NMR, this effect has been associated with perturbations of both the hydrogen bond network and one amino acid side chain around the {4Fe-4S} clusters . Thus, the conserved Pro found in the binding motif of {4Fe-4S} clusters in proteins strongly stabilizes the active site but does not play an essential role in the mechanism of electron transfer. Ann Pharmacother, 1994 May, 28(5), 581 - 4 Possible red-man syndrome associated with systemic absorption of oral vancomycin in a child with normal renal function; Bergeron L et al.; OBJECTIVE: To report possible red-man syndrome (RMS) associated with oral administration of vancomycin . CASE SUMMARY: A 23-month-old child with acute myeloblastic leukemia developed symptoms compatible with RMS while receiving oral vancomycin for suspected Clostridium difficile colitis . Serum concentrations of vancomycin, measured at the time of the clinical episode, demonstrated significant oral absorption of the drug . Serum concentrations of vancomycin decreased later, implying a possible decrease in absorption, after the patient's neutrophil count returned to normal . The child later experienced another clinical episode compatible with RMS while vancomycin was being administered intravenously for suspected sepsis . DISCUSSION: There is no published report of RMS following oral administration of vancomycin . The reaction described took place while the child was neutropenic . Because of the absence of any significant renal function alteration that could explain the importance of the serum concentrations observed, we assume that C . difficile, neutropenia-, and chemotherapy-associated colitis may have resulted in extensive intestinal lesions, leading to an increased amount of vancomycin being systemically absorbed . This increased absorption during profound neutropenia may have been sufficient to exceed a purported threshold, leading to RMS . CONCLUSIONS: This case demonstrates that significant absorption of vancomycin may occur in neutropenic patients with normal renal function, and that it may be accompanied by RMS, usually associated with rapid infusions or large parenteral doses of the drug. Antimicrob Agents Chemother, 1994 May, 38(5), 1041 - 6 Cloning and sequence analysis of ermQ, the predominant macrolide-lincosamide-streptogramin B resistance gene in Clostridium perfringens; Berryman DI et al.; The erythromycin resistance determinant from Clostridium perfringens JIR100 has been cloned, sequenced, and shown to be expressed in Escherichia coli . An open reading frame with sequence similarity to erm genes from other bacteria was identified and designated the ermQ gene . On the basis of comparative sequence analysis, it was concluded that the ermQ gene represented a new Erm hybridization class, designated ErmQ . Genes belonging to the ErmQ class were found to be widespread in C . perfringens, since 30 of 38 macrolide-lincosamide-streptogramin B-resistant C . perfringens strains, from diverse sources, hybridized to an ermQ-specific gene probe . The ermQ gene therefore represents the most common erythromycin resistance determinant in C . perfringens. Plasmid, 1994 May, 31(3), 320 - 3 Gene cloning in Clostridium difficile using Tn916 as a shuttle conjugative transposon; Mullany P et al.; A pBR322-based vector, pCI195, containing a 4.2-kb region of the conjugative transposon Tn919 was used as a vector for gene cloning in Clostridium difficile . The plasmid was found to integrate into the chromosome of a Bacillus subtilis strain that contained Tn916 delta E . Southern blot analysis of the recombinant demonstrated that pCI195 had inserted into Tn916 delta E by a recombination event . The transposon::plasmid structure could be transferred, by filter mating, from B . subtilis to C . difficile (at a frequency of 10(-8) per donor), where it entered the chromosome at a specific site . Segregation of plasmid and transposon markers was observed on transfer, although the Tn916 delta E::pCI195 was stably maintained in C . difficile . To demonstrate that pCI195 could be used for gene cloning in C . difficile, a 1.1-kb fragment of the C . difficile toxin B gene was cloned into pCI195 to generate pPPM100 . Tn916 delta E::pPPM100 was transferred into a nontoxigenic C . difficile strain by filter mating, where it entered the genome at a specific site . pCI195 should be useful as a general cloning vector for C . difficile, as the transposon::plasmid structure could be transferred to different C . difficile strains . This is the first report of gene cloning in C . difficile. Plasmid, 1994 May, 31(3), 317 - 9 A Clostridium perfringens vector for the selection of promoters; Matsushita C et al.; A promoter selection vector for Clostridium perfringens genes was constructed from a C . perfringens-Escherichia coli shuttle vector, pJIR418 . The plasmid carries a promoterless chloramphenicol acetyltransferase gene (catP), derived from pIP401, downstream of the multiple cloning sites of pUC18 . When a promoter region of the phospholipase C gene was inserted into one of the cloning sites, derivatives of C . perfringens strain 13 carrying the resultant plasmid acquired resistance to chloramphenicol . This plasmid should be useful reporter system for C . perfringens genes. J Clin Microbiol, 1994 May, 32(5), 1142 - 7 Comparison of four commercially available rapid enzyme immunoassays with cytotoxin assay for detection of Clostridium difficile toxin(s) from stool specimens; Merz CS et al.; Rapid (2.5- to 3.5-h) enzyme immunoassays (EIAs) for the detection of Clostridium difficile toxins have been developed . We report the results of simultaneous testing of 700 fresh stool specimens by the tissue culture cytotoxin assay and four EIAs (Bartels Prima System C . difficile Toxin A EIA, Cambridge Biotech Cytoclone A+B EIA, Meridian Diagnostics Premier C . difficile Toxin A EIA, and TechLab C . difficile Tox-A Test EIA) . In cases of disagreement, culturing for toxigenic C . difficile was performed . A total of 61 (8.7%) specimens from 46 patients were positive for C . difficile toxin . The sensitivity of the cytotoxin assay was 87%, and that of culture was 93% . In comparison with the cytotoxin assay results, the sensitivity and specificity of the EIAs were as follows: Bartels, 87 and 96%; Cambridge, 89 and 99%; Meridian, 87 and 98%; and TechLab, 87 and 95%, respectively . In comparison with the cytotoxin assay plus toxigenic culture results, the sensitivity and specificity of the EIAs were as follows: Bartels, 84 and 97%; Cambridge, 85 and 99%; Meridian, 79 and 98%; and TechLab, 80 and 96%, respectively . The EIAs varied in positive predictive values (PPVs) . A high PPV was seen with the Cambridge EIA (96%); lower PPVs were seen with the TechLab (64%), Bartels (72%), and Meridian (80%) EIAs because of high false-positive rates . The negative predictive values (98 to 99%) were excellent with all EIAs . Results were indeterminant with 0.3% of the samples by the Meridian EIA and 3% by all the other EIAs . Although the EIAs were less sensitive than the cytotoxin assay, they provide same-day results and may be useful in laboratories without tissue culture facilities. Am J Perinatol, 1994 May, 11(3), 205 - 7