Nippon Juigaku Zasshi, 1989 Jun, 51(3), 582 - 6 Implication of coprophagy in pathogenesis of chicken botulism; Hyun SH et al.; Oral administration of 1 x 10(7) viable spores of Clostridium botulinum type C killed the chickens kept on a board floor to allow them coprophagy, whereas the same dose of the spores failed to develop symptoms in those kept on a wire-net floor not to allow them coprophagy . Type C toxin was detected in the cecal droppings of the chickens of both the groups after feeding the spores and also in serum of symptomatic as well as asymptomatic chickens kept on a board floor . Thus, coprophagy, by which chickens ingest type C toxin (C1 L toxin) and the bacterial cells, seems to be a prerequisite for development of chicken botulism. J Antimicrob Chemother, 1989 Jun, 23(6), 929 - 31 Extended spectrum cephalosporins and Clostridium difficile; Golledge CL et al.; There is little information about how commonly the newer cephalosporins cause diarrhoea due to Clostridium difficile . In this study of 111 patients with C . difficile-associated diarrhoea, 106 had received antimicrobial agents in the four weeks before detection of C . difficile . The relative risk for each antimicrobial agent was greatest with clindamycin, followed by cefotaxime, cephamandole and ceftriaxone . There was no statistically significant difference in risk between the cephalosporins evaluated . Narrower spectrum penicillins, anti-pseudomonal penicillins and aminoglycosides were not potent inciting agents. J Bacteriol, 1989 Jun, 171(6), 2925 - 32 Purification and characterization of a novel form of 20 alpha-hydroxysteroid dehydrogenase from Clostridium scindens; Krafft AE et al.; We have purified a steroid-inducible 20 alpha-hydroxysteroid dehydrogenase from Clostridium scindens to apparent homogeneity . The final enzyme preparation was purified 252-fold, with a recovery of 14% . Denaturing and nondenaturing polyacrylamide gradient gel electrophoresis showed that the native enzyme (Mr, 162,000) was a tetramer composed of subunits with a molecular weight of 40,000 . The isoelectric point was approximately pH 6.1 . The purified enzyme was highly specific for adrenocorticosteroid substrates possessing 17 alpha, 21-dihydroxy groups . The purified enzyme had high specific activity for the reduction of cortisone (Vmax, 280 nmol/min per mg of protein; Km, 22 microM) but was less reactive with cortisol (Vmax, 120 nmol/min per mg of protein; Km, 32 microM) at pH 6.3 . The apparent Km for NADH was 8.1 microM with cortisone (50 microM) as the cosubstrate . Substrate inhibition was observed with concentrations of NADH greater than 0.1 mM . The purified enzyme also catalyzed the oxidation of 20 alpha-dihydrocortisol (Vmax, 200 nmol/min per mg of protein; Km, 41 microM) at pH 7.9 . The apparent Km for NAD+ was 526 microM . The initial reaction velocities with NADPH were less than 50% of those with NADH . The amino-terminal sequence was determined to be Ala-Val-Lys-Val-Ala-Ile-Asn-Gly-Phe-Gly-Arg . These results indicate that this enzyme is a novel form of 20 alpha-hydroxysteroid dehydrogenase. J Gen Microbiol, 1989 Jun, 135 ( Pt 6), 1755 - 62 Cloning and expression of the Clostridium thermohydrosulfuricum alpha-amylase-pullulanase gene in Escherichia coli; Melasniemi H et al.; An alpha-amylase-pullulanase gene from Clostridium thermohydrosulfuricum DSM 3783 was cloned in Escherichia coli on a 7.0 kb EcoRI fragment using a lambda vector . The gene produced, from an indigenous promoter, active thermostable alpha-amylase-pullulanase, seemingly mostly a soluble intracellular enzyme in E . coli . Gel filtration separated the active enzyme produced into three peaks, each having both alpha-amylase and pullulanase activities . Immunoblotting after SDS-PAGE revealed more than ten alpha-amylase-pullulanase specific polypeptides; the biggest of these had an Mr of about 165,000, whereas the smallest enzymically active polypeptide had an Mr of about 100,000 . Despite the marked degeneration of its constituent polypeptides, the apparent temperature optimum of the enzyme (80-85 degrees C) was only some 5 degrees C lower and the heat stability the same as that of the extracellular alpha-amylase-pullulanase produced by the native host . Oligonucleotide probes prepared according to the NH2-terminal amino acid sequences of the enzyme and its satellite polypeptide (a polypeptide associated with the extracellular enzyme of the native host) hybridized to different regions of the 7.0 kb DNA insert. Arch Dis Child, 1989 Jun, 64(6), 871 - 2 Infantile botulism; Smith GE et al.; A 4 month old boy presented with respiratory difficulty and hypotonia . Clostridium botulinum and its toxin were isolated from his faeces and he had electromyographic changes typical of infantile botulism . This is only the second case in the United Kingdom: unfamiliarity with the presentation could result in misdiagnosis. Anal Biochem, 1989 Jun, 179(2), 341 - 6 Determination of ursodeoxycholic acid in serum by a new fluorometric enzymatic method using 7 beta-hydroxysteroid dehydrogenase from Clostridium absonum; Lianidou ES et al.; A fluorometric enzymatic method for the determination of ursodeoxycholic acid (UDCA) and its glycine and taurine conjugates in human serum has been developed . A simple and fast purification and preconcentration procedure using Sep Pak C18 cartridges was employed for the UDCA extraction from human serum . UDCA and its conjugates were determined in the extracted sample by an equilibrium method based on the enzymatic conversion of the 7 alpha-hydroxy group into 7-oxo group by beta-nicotinamide adenine dinucleotide phosphate in the presence of 7 beta-hydroxysteroid dehydrogenase (7 beta-HSD) and the produced NADPH was monitored fluorometrically . The 7 beta-HSD, which is not yet commercially available, was isolated from Clostridium absonum cultures (ATCC No . 27555) and purified by affinity chromatography . The method has a limit of detection of 0.8 microM in serum and the precision varied from 6.1 to 2.0% for low and high concentrations, respectively . The recovery of UDCA from serum samples was about 99% (range 85-105%) . The method was successfully applied to UDCA determination in serum samples from patients treated with UDCA for primary biliary cirrhosis. Appl Environ Microbiol, 1989 Jun, 55(6), 1544 - 8 Regulation of neurotoxin and protease formation in Clostridium botulinum Okra B and Hall A by arginine; Patterson-Curtis SI et al.; Supplementation of a minimal medium with high levels of arginine (20 g/liter) markedly decreased neurotoxin titers and protease activities in cultures of Clostridium botulinum Okra B and Hall A . Nitrogenous nutrients that are known to be derived from arginine, including proline, glutamate, and ammonia, also decreased protease and toxin but less so than did arginine . Proteases synthesized during growth were rapidly inactivated after growth stopped in media containing high levels of arginine . Separation of extracellular proteins by electrophoresis and immunoblots with antibodies to toxin showed that the decrease in toxin titers in media containing high levels of arginine was caused by both reduced synthesis of protoxin and impaired proteolytic activation . In contrast, certain other nutritional conditions stimulated protease and toxin formation in C . botulinum and counteracted the repression by arginine . Supplementation of the minimal medium with casein or casein hydrolysates increased protease activities and toxin titers . Casein supplementation of a medium containing high levels of arginine prevented protease inactivation . High levels of glucose (50 g/liter) also delayed the inactivation of proteases in both the minimal medium and a medium containing high levels of arginine . These observations suggest that the availability of nitrogen and energy sources, particularly arginine, affects the production and proteolytic processing of toxins and proteases in C . botulinum. Helv Paediatr Acta, 1989 Jun, 43(5-6), 521 - 30 {Botulism in infancy}; Gautier E et al.; The authors describe a case of botulism in a 3-month-old infant infected with Clostridium botulinum type A . Symptomatology developed within four days, persisted for two weeks, then regressed . Symptoms were paresis of face muscles, hyporeactive pupils, loss of succion and deglutition, axial hypotonia, weakness of peripheral muscles, lability of the autonomic nervous system with acute episodes of bradycardia and constipation . Anomalies of the electroen-cephalogram and of the auditory evoked responses suggest that the toxin penetrated the central nervous system . Treatment was symptomatic, without need for assisted ventilation . It was not possible to detect the source of infection. Epidemiol Infect, 1989 Jun, 102(3), 467 - 71 The production of Clostridium botulinum toxin in mammalian, avian and piscine carrion; Smith GR et al.; Mice, birds (chicks, quail) and fish (rudd, goldfish) killed shortly after receiving 1300-2000 spores of Clostridium botulinum per os were incubated, usually at 23 degrees C for 7 days . A 10% (w/v) homogenate of each rotting carcass was then prepared, sterilized by membrane filtration, and assayed for toxin . In mouse carcasses a type C strain of C . botulinum usually produced greater than 2 X 10(5) mouse intraperitoneal LD/g; in fish carcasses it usually produced less--often much less--than 2 X 10(4) LD/g . Avian carcasses appeared to be intermediate between those of mice and fish in their ability to support toxigenesis . A type E strain of C . botulinum, unlike type C, produced toxin equally well in fish and mouse carrion, usually at a concentration of between 2 X 10(4) and 2 X 10(5) LD/g. Showa Shigakkai Zasshi, 1989 Jun, 9(2), 97 - 102 Novel method for the preparation of sterile collagen-agarose-plates for isolating collagenolytic bacteria; Takahashi M et al.; An assay method for bacterial collagenase using sterile collagen-agarose plates was developed to isolate collagenolytic bacteria . In order to avoid heat-denaturation of collagen, the plates were made at 37 degrees C mixing microwave-sterilized collagen with low melting point-agarose . Sensitivities to various proteases were tested on the plates; it was shown that whereas the collagen in these plates was digested only by the collagenases, the collagen could also be hydrolyzed by other proteolytic enzymes if it had been either sterilized by ethylene oxide or solidified with conventional agar at a higher temperature . Colonies of Clostridium histolyticum cells grown on the plates had clear zones around them and their collagenase activities could be readily detected. Food Chem Toxicol, 1989 Jun, 27(6), 399 - 402 Phospholipase C-mediated intestinal mucosal damage is ameliorated by quinacrine; Otamiri T; Phospholipase C from Clostridium perfringens, when injected into a closed loop of the rat small intestine in vivo, caused an increase in the activity of intraluminal N-acetyl-beta-glucosaminidase and mucosal permeability to sodium fluorescein, indicating damage to the mucosa . Phospholipase C also caused an influx of granulocytes (neutrophils) into the mucosa, as shown by the myeloperoxidase activity--a granulocyte neutrophil marker, and increased localized lipid peroxidation . Pretreatment of animals with quinacrine, a known inhibitor of phospholipase A2, prevented the increases in the luminal N-acetyl-beta-glucosaminidase activity, mucosal permeability, malondialdehyde and myeloperoxidase activity after deposition of phospholipase C in the gut lumen . It is concluded that phospholipase C might impair the function of the mucosal barrier and increase the permeability of the gut to undesirable molecules and pathogens . Part of its action may be mediated via phospholipase A2 activation since pretreatment with quinacrine afforded protection. Eur J Clin Microbiol Infect Dis, 1989 Jun, 8(6), 550 - 1 Comparative antibacterial activity of the new cephalosporin cefcanel against anaerobic bacteria; Nord CE et al.; The activity of cefcanel against anaerobic cocci, Clostridium perfringens, Bacteroides fragilis, Bacteroides spp . and fusobacteria was determined by the agar dilution method and compared with the activity of cefaclor, cephalexin, cefadroxil, phenoxymethylpenicillin and ampicillin . Cefcanel showed good activity against Clostridium perfringens, Bacteroides spp . and fusobacteria (MIC90 = 1-4 mg/l) . Against anaerobic cocci its MIC90 value was 16 mg/l, and against Bacteroides fragilis, 32 mg/l . Cefcanel has an antibacterial activity that warrants investigation in clinical trials. J Bacteriol, 1989 Jun, 171(6), 3433 - 9 A novel and remarkably thermostable ferredoxin from the hyperthermophilic archaebacterium Pyrococcus furiosus; Aono S et al.; The archaebacterium Pyrococcus furiosus is a strict anaerobe that grows optimally at 100 degrees C by a fermentative-type metabolism in which H2 and CO2 are the only detectable products . A ferredoxin, which functions as the electron donor to the hydrogenase of this organism was purified under anaerobic reducing conditions . It had a molecular weight of approximately 12,000 and contained 8 iron atoms and 8 cysteine residues/mol but lacked histidine or arginine residues . Reduction and oxidation of the ferredoxin each required 2 electrons/mol, which is consistent with the presence of two {4Fe-4S} clusters . The reduced protein gave rise to a broad rhombic electronic paramagnetic resonance spectrum, with gz = 2.10, gy = 1.86, gx = 1.80, and a midpoint potential of -345 mV (at pH 8) . However, this spectrum represented a minor species, since it quantitated to only approximately 0.3 spins/mol . P . furiosus ferredoxin is therefore distinct from other ferredoxins in that the bulk of its iron is not present as iron-sulfur clusters with an S = 1/2 ground state . The apoferredoxin was reconstituted with iron and sulfide to give a protein that was indistinguishable from the native ferredoxin by its iron content and electron paramagnetic resonance properties, which showed that the novel iron-sulfur clusters were not artifacts of purification . The reduced ferredoxin also functioned as an electron donor for H2 evolution catalyzed by the hydrogenase of the mesophilic eubacterium Clostridium pasteurianum . P . furiosus ferredoxin was resistant to denaturation by sodium dodecyl sulfate (20%, wt/vol) and was remarkably thermostable . Its UV-visible absorption spectrum and electron carrier activity to P . furiosus hydrogenase were unaffected by a 12-h incubation of 95 degrees C. Gene, 1989 May 30, 78(2), 355 - 64 Molecular analysis and nucleotide sequence of the adh1 gene encoding an NADPH-dependent butanol dehydrogenase in the Gram-positive anaerobe Clostridium acetobutylicum; Youngleson JS et al.; The nucleotide sequence of a 2081-bp fragment of Clostridium acetobutylicum DNA containing the adh1 gene was determined . The butanol dehydrogenase gene is referred to as the adh1 gene since it was shown to have activity using butanol and ethanol as substrates . The adh1 gene consisted of 1164 bp and encoded an alcohol dehydrogenase (ADH) enzyme of 388 aa residues with an Mr of 43,274 . The adh1 gene was separated from an upstream open reading frame by an intergenic region of 354 bp . No promoter consensus sequences were identified in the intergenic upstream region and the adh1 gene did not appear to be expressed off its own promoter in Escherichia coli . Three separate types of ADH have been recognized . The ADH1 from C . acetobutylicum exhibited 39% homology with the Fe-containing ADH2 from Zymomonas mobilis and 37% homology with the ADH4 from Saccharomyces cerevisiae, but showed little or no homology with the other characterised types of ADH. Biochemistry, 1989 May 30, 28(11), 4675 - 80 Kinetic characterization of the {3'-32P}coenzyme A/acetyl coenzyme A exchange catalyzed by a three-subunit form of the carbon monoxide dehydrogenase/acetyl-CoA synthase from Clostridium thermoaceticum; Ramer SE et al.; The ability of acetyl coenzyme A synthesizing carbon monoxide dehydrogenase isolated from Clostridium thermoaceticum to catalyze the exchange of {3'-32P}coenzyme A with acetyl coenzyme A is studied . This exchange is found to have a rate exceeding that of the acetyl coenzyme A carbonyl exchange also catalyzed by CO dehydrogenase ({1-14C}acetyl coenzyme A + CO in equilibrium acetyl coenzyme A + 14CO) . These two exchanges are diagnostic of the ability of CO dehydrogenase to synthesize acetyl coenzyme A from a methyl group, coenzyme A, and carbon monoxide . The kinetic parameters for the coenzyme A exchange have been determined: Km(acetyl coenzyme A) = 1500 microM, Km(coenzyme A) = 50 microM, and Vmax = 2.5 mumol min-1 mg-1 . Propionyl coenzyme A is shown to be a substrate (Km approximately 5 mM) for the coenzyme A exchange, with a rate 1/15 that of acetyl coenzyme A, but is not a substrate for the carbonyl exchange . CO dehydrogenase capable of catalyzing both these two exchanges, and the oxidation of CO to CO2, is isolated as a complex of molecular weight 410,000 consisting of three proteins in an alpha 2 beta 2 gamma 2 stoichiometry . The proposed gamma subunit, not previously reported as part of CO dehydrogenase, copurifies with the enzyme and has the same molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as the disulfide reductase previously separated from CO dehydrogenase in a final chromatographic step. Vet Rec, 1989 May 27, 124(21), 558 - 60 Type C botulism in cattle being fed ensiled poultry litter; Neill SD et al.; A botulinum toxin from ensiled poultry litter which caused a major outbreak of bovine botulism was characterised as type C1 . The litter produced transient ataxia when fed to two experimental calves and the clinical signs were accompanied by a transient appearance of serum toxin . Type C1 toxin was demonstrated in muscle tissues which had been taken during the outbreak from an affected animal with high circulating serum toxin, and held frozen for seven months . Clostridium botulinum type C organisms were demonstrated in faeces from another affected animal and also in kidney tissue from a third animal . These observations have implications for the diagnosis and management of future outbreaks of botulism and for the potential health risk from the meat of affected animals. Lancet, 1989 May 27, 1(8648), 1156 - 60 Bacteriotherapy for chronic relapsing Clostridium difficile diarrhoea in six patients; Tvede M et al.; Six patients with chronic relapsing diarrhoea caused by Clostridium difficile were treated with rectal instillation of homologous faeces (one patient) or a mixture of ten different facultatively aerobic and anaerobic bacteria diluted in sterile saline (five patients) . The mixture led to a prompt loss of Cl difficile and its toxin from the stools and to bowel colonisation by Bacteroides sp, which had not been present in pre-treatment stool samples . Strains of Escherichia coli, Cl bifermentans, and Peptostreptococcus productus in the mixture inhibited the in-vitro growth of Cl difficile, which in turn inhibited the growth of Bacteroides ovatus, Bacteroides vulgatus, and Bacteroides thetaiotaomicron . The finding that Bacteroides sp had been absent during the patients' illness but was present after recovery suggests that the absence of Bacteroides sp may result in chronic relapsing Cl difficile diarrhoea, and that its presence may prevent colonisation by Cl difficile. Wien Med Wochenschr, 1989 May 15, 139(9), 202 - 6 {Diarrhea induced by antibiotics}; Mittermayer H; The most frequent cause of antibiotic-associated colitis is Clostridium difficile . This gram-positive, spore-forming anaerobic bacillus releases toxins, which produce diarrhea and damage the colonic mucosa . Endoscopy shows a wide range of alterations, "unspecific colitis" with reddening or edema, ulcerations or at the worst pseudomembranous colitis . Nearly all antibiotics are able to trigger Clostridium difficile colitis . An enhanced risk is exerted by broad spectrum substances, which act also on the anaerobic flora protecting the gastrointestinal tract from unphysiological colonization . Clusters of cases were observed in hospitalized patients . The patients risk factors coincide with the administration of antibiotics . Furthermore Clostridium difficile is likely to be spread as a nosocomial infection in many instances . Less often colitis is observed in connection with oral antibiotics outside the hospital . However, substantial underreporting of cases has to be considered . Clinical symptoms usually start 4 to 10 days after first administration of the antibiotic . Leading symptoms are frequent profuse watery stools . Abdominal cramps and tenderness as well as fever and leukocytosis are common . Intense symptoms can simulate serious conditions like perforation . Upon clinical suspicion the diagnosis is made by endoscopy, stool culture and possibly demonstration of toxin . The predictive value of the stool culture equals that of toxin detection . In adult patients there is a good correlation between positive stool culture and clinical presentation . Infants can carry Clostridium difficile as part of their normal flora, therefore positive stool culture or toxin detection in an infant cannot necessarily be linked to clinical symptoms . In some cases Clostridium difficile has to be regarded as etiologic organism also in infants.(ABSTRACT TRUNCATED AT 250 WORDS) J Chromatogr, 1989 May 5, 490(1), 91 - 100 Clostridium difficile toxin B: characterization and sequence of three peptides; Bisseret F et al.; The cytotoxin, also named toxin B, was isolated from a toxigenic strain of Clostridium difficile, purified to homogeneity and partially characterized . The purification procedure included ultrafiltration followed by anion-exchange chromatography . We noticed that a non-specific nucleic material eluted with the protein during the purification . The presence of these nucleic acids appeared to be important for the toxic activity of the protein . Some characteristics of the cytotoxin were examined, especially the amino acid composition and the sequence of three tryptic fragments. East Afr Med J, 1989 May, 66(5), 319 - 23 Feacal carriage of cytotoxigenic strains of Clostridium difficile by adult Nigerians; Rotimi VO et al.; The isolation rate of Clostridium difficile and the presence of its cytotoxin in stool were studied in adults with diarrhoea and healthy normal individuals over a period of one year . C . difficile was isolated from 23(56%) out of 41 patients with diarrhoea who gave history of antibiotic exposure prior to the development of the disease from 12(31.6%) of the 38 healthy controls with no history of antibiotic exposure . Of the 23(31.6%) C . difficile isolated from the patients, 16(69.6%), and only 8(66.6%) of the 12 isolates from the healthy adults, were cytotoxigenic . Only one stool specimen from a diarrhoeic patient, was positive for cytotoxin assay . The clinical significance of this finding is discussed. J Clin Microbiol, 1989 May, 27(5), 889 - 93 Evaluation of a latex agglutination test for Clostridium difficile in two nursing home outbreaks; Bennett RG et al.; The Culturette Brand Clostridium difficile test (CDT; Marion Laboratories, Inc., Kansas City, Mo.) is a latex agglutination test for C . difficile . The recent controversy involving the identity of antigens detected by CDT has made decisions on its use difficult . We compared the test results with those of selective culture and stool cytotoxin assays in investigations of two nursing home outbreaks of C . difficile-associated disease in order to formulate usage recommendations . Selective culture for C . difficile identified 27 (19%) of 142 subjects as carriers . CDT and the stool cytotoxin assay identified only 52 and 48% of these carriers, respectively . Compared with the stool cytotoxin assay, CDT had a high sensitivity (92%) and specificity (89%) for the detection of C . difficile disease, but the positive predictive value of the test was only 17% when the prevalence of disease was 2% . We conclude that the CDT should not be used to identify carriers but that it is a sufficiently sensitive and specific screening test for diagnosing C . difficile disease . However, since the positive predictive value of the CDT is low when the prevalence of disease is low, positive test results should be confirmed by the stool cytotoxin assay. J Clin Microbiol, 1989 May, 27(5), 806 - 7 Microbiology of postthoractomy sternal wound infection; Brook I; Specimens from 74 patients with postthoractomy sternal wound infection were studied for aerobic and anaerobic bacteria . Bacterial growth was obtained in specimens from 65 patients (88%) . Aerobic or facultative bacteria only were recovered in 50 specimens (77%), anaerobic bacteria only in 6 (9%), and mixed aerobic, facultative, and anaerobic bacteria in 9 (14%) . Eighty-seven isolates were recovered (1.3 per specimen): 68 aerobic or facultative (1.0 per specimen) and 19 anaerobic (0.3 per specimen) . The predominant aerobes were Staphylococcus epidermidis (28 isolates), Staphylococcus aureus (21 isolates), and members of the family Enterobacteriaceae (14 isolates) . The predominant anaerobes were Peptostreptococcus spp . (10 isolates), Bacteroides spp . (4 isolates), and Clostridium spp . (3 isolates) . Polymicrobial infection occurred in 18 instances (28%) . A single organisms was recovered in 47 instances (72%); these included 20 isolates of S . epidermidis, 15 of S . aureus, 5 of Enterobacteriaceae, and 4 of anaerobes . These data highlight the previously unrecognized polymicrobial aerobic and anaerobic bacteriology in a percentage of patients with postthoractomy sternal wound infection. J Clin Microbiol, 1989 May, 27(5), 1137 - 8 Liver abscess caused by Clostridium bifermentans following blunt abdominal trauma; Nachman S et al.; A case of hepatic abscess and subsequent septicemia caused by Clostridium bifermentans is described . The abscess manifested itself on the third day after blunt trauma to the torso . The patient had nausea, vomiting, fever, evidence of hepatic dysfunction, and subphrenic gas . This case illustrates the association of hepatic abscess and blunt trauma to the torso. Br Vet J, 1989 May-Jun, 145(3), 291 - 2 Double intussusception fatally complicated by clostridial infection in a dog (a case report); Okewole PA et al.; A 10-month-old Alsation dog with history of anorexia, diarrhoea, dehydration and vomition developed a double intussusception which affected the distal jejunum and proximal ileum . Necropsy revealed the intussusception to be swollen and congested with fibrinous adhesions between the intussusceptum and intussuscepiens . Two pieces of bone believed to be the inciting cause were found within the intussusceptum . Clostridium welchii and Clostridium bifermentans were isolated from the lesions. J Clin Pathol, 1989 May, 42(5), 511 - 5 Typing of Clostridium difficile causing diarrhoea in an orthopaedic ward; McKay I et al.; In an outbreak of diarrhoeal disease in an orthopaedic ward Clostridium difficile was isolated from all six patients with diarrhoea . Attempts were made to type these isolates by means of antibiogram, detection of pre-formed enzymes, analysis of surface proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting, and plasmid profile analysis . This showed that a single strain (type E) indistinguishable by the four distinct methods of typing, was isolated from all six patients at some time during their episodes of diarrhoea . Relapse was caused by the acquisition of a new strain in two patients, and by re-emergence or reacquisition of the original strain in two patients . The immunochemical method was the most sensitive and discriminatory of the typing strategies adopted. Medicine (Baltimore), 1989 May, 68(3), 151 - 62 Bacteremia caused by non-sporulating anaerobes in cancer patients . A 12-year experience; Fainstein V et al.; The clinical, epidemiological and laboratory characteristics of bacteremia caused by anaerobic organisms other than Clostridium spp . in cancer patients are described and compared to other previously reported series . Of the 315 episodes, 246 (78%) were caused by a single organism and 69 (22%) were polymicrobial . The most common underlying malignancies were genitourinary and gynecological tumors, acute leukemia, and gastrointestinal malignancies . Most patients (94%) were febrile, and septic shock was documented in 24% of monomicrobial episodes and in 58% of those with polymicrobial infection . Soft-tissue infection was present in 44% of the cases, and it presented as tissue necrosis in 11% . The most common sites identified as the portal of entry were intra-abdominal abscesses, soft tissue, and the oropharynx . The most common organisms were Bacteroides fragilis (57%) and other Bacteroides spp . (22%) . Most polymicrobial infections were caused by 2 organisms, the second most commonly another anaerobe or an aerobic gram-negative bacillus . The most active antibiotic in vitro was chloramphenicol . High rates of resistance to penicillin were observed not only among B . fragilis, but also among Bacteroides spp . The frequency of penicillin resistance increased throughout the study years . The overall survival was 70% . The cure rate for monomicrobial bacteremias was 76% vs . 51% for polymicrobial episodes . Infection was the cause of death in 20 and 16 episodes, respectively . The response rate for patients in septic shock was 47% in contrast to an 85% recovery rate for those without it . Ninety-five patients had documented abscesses accompanying the bacteremic episode . The most effective antibiotics were clindamycin and chloramphenicol . Overall response to penicillin was only 13% . Suboptimal responses were also observed for the antipseudomonal penicillins . High response rates (82%) were also obtained with cefoxitin, metronidazole, and moxalactam. J Bacteriol, 1989 May, 171(5), 2873 - 5 Electron transport and electrochemical proton gradient in membrane vesicles of Clostridium thermoautotrophicum; Hugenholtz J et al.; Membrane vesicles of Clostridium thermoautotrophicum containing carbon monoxide dehydrogenase generated a proton motive force when exposed to CO . This proton motive force, with a value of -140 mV, consisted of only an electrical potential at pH 7.5 and above and of an electrical potential and pH gradient at a lower pH . The proton motive force drove the uptake of L-alanine by the vesicles to a concentration of 300 times that of the medium. FEMS Microbiol Lett, 1989 May, 50(1-2), 135 - 40 Cloning and expression of a Clostridium acetobutylicum alpha-amylase gene in Escherichia coli; Verhasselt P et al.; A gene library of Clostridium acetobutylicum ATCC824 was constructed in the plasmid vector pEcoR251 . The library was tested for the presence of starch hydrolyzing clones . One clone in which the recombinant plasmid, pVP101, conferred alpha-amylase activity to the Escherichia coli host cell, was detected . The gene is carried on a 3.45-kbp BglII restriction fragment . A detailed physical map of pVP101 is presented. Int J Food Microbiol, 1989 May, 8(2), 121 - 32 Spoilage of an acid food product by Clostridium perfringens, C . barati and C . butyricum; de Jong J; Spoilage of canned pasteurized brined mung bean sprouts, acidified with citric acid to pH 4.0-4.5, was found to be caused by acid tolerant Clostridium spp . including the species barati, perfringens and butyricum . The pH limit for growth in the brine used were estimated 3.7, 3.7 and 4.0 respectively . Some of the isolated C . perfringens strains produced enterotoxins in sporulation media . The spores of the isolated anaerobes appeared to originate from mung beans, but C . barati and C . perfringens strains freshly isolated from dry beans, were unable to grow in acidified brine . During germination and sprouting of mung beans, the oxygen concentration decreased, while carbon dioxide concentration increased considerably, due to respiration of the sprouts and actively growing Enterobacteriaceae and lactobacilli . It was assumed that this allowed C . barati and C . perfringens strains to grow and acquire the observed unusual acid tolerance . After increasing aerobicity during sprouting, no growth of Clostridium spp . was observed, substantiating the assumption. Trans R Soc Trop Med Hyg, 1989 May-Jun, 83(3), 344 - 5 In vitro activity of nalidixic acid and its iron (III) complex on Entamoeba histolytica; Anaya-Velazquez F et al.; The in vitro activity of the antibacterial agent nalidixic acid (HNal) and its iron (III) complex (FeNal) against Entamoeba histolytica HM1 strain trophozoites in axenic or monoxenic (associated with Clostridium symbiosum) cultures was investigated . Using a dilution test with TYI-S-33 medium, this protozoan was found to be susceptible to both drugs, but FeNal showed amoebicidal activity only at concentrations higher than those used with HNal. Biokhimiia, 1989 May, 54(5), 804 - 10 {Study on the effect of some group-specific agents on clostridiopeptidase}; Balaevskaia TO et al.; The interaction of clostridiopeptidase of Clostridium histolyticum with EDC, TNM and MA, the specific reagents for COOH-groups, tyrosine and lysine residues was studied . It was shown that at pH 6.0 EDC inactivates the enzyme . The inactivation process follows the pseudo-first order kinetics and is described by a second order rate constant equal to 1 M-1 min-1 . The synthetic substrate does not prevent, in practical terms, the enzyme inactivation by EDC . At pH 8.0 TNM modifies about 19 tyrosine residues in the clostridiopeptidase molecule which is accompanied by marked inhibition of the enzyme activity (down to 70-90%) . In this case, the inactivation process is not described by simple pseudo-first order kinetics but is characterized by two steps (fast and slow) with second order rate constants of approximately 14 and 3.5 M-1 min-1, respectively . The synthetic substrate partly prevents the inactivation of the enzyme by TNM and protects 11 tyrosine residues . The MA-induced incorporation of 13 +/- 3 maleyl groups into the clostridiopeptidase molecule in partially prevented by the synthetic substrate with protects the enzyme against inactivation . The data obtained suggest that lysine residues are seemingly included into the active center of clostridiopeptidase, whereas tyrosine residues provide for the maintenance of active conformation of the enzyme. Rev Infect Dis, 1989 May-Jun, 11(3), 470 - 3 Postpartum uterine infection with Clostridium perfringens; Dylewski J et al.; Clostridium perfringens is commonly present in the female genital tract . Uterine infection with this organism is a potentially fatal disease infrequently seen in obstetric practice . The manifestations of C . perfringens uterine infection are variable, ranging from endometritis to gas gangrene with fulminant septicemia . The usual precipitating event has been septic abortion, but such infections can also occur spontaneously in uterine tumors and after complicated deliveries requiring mechanical intervention . Diagnosis may be aided by radiologic techniques, and treatment involves high-dose penicillin and possibly surgery . We report two cases and review the clinical presentation and the diagnostic and therapeutic aspects of this disease. FEMS Microbiol Lett, 1989 May, 50(1-2), 221 - 4 Stimulation of growth and sporulation of Clostridium perfringens by its homologous enterotoxin; Dillon ME et al.; C . perfringens enterotoxin shortened the lag phase and time of onset of sporulation of the same organism in a dose-dependent manner . The toxin stimulated macromolecular synthesis of pre-exponential phase cells. FEMS Microbiol Lett, 1989 May, 50(1-2), 173 - 6 Monoclonal antibodies against alpha toxin of Clostridium perfringens; Sato H et al.; Ten distinct monoclonal antibodies (MAbs) against alpha toxin of Clostridium perfringens were produced by the fusion of SP2/O with spleen cells of mice immunized with alpha toxoid, and alpha toxin mixed with or without ethylenediamine-tetraacetate (EDTA) . The antibody activity was evaluated by antigen-binding activity in an enzyme linked immunosorbent assay (ELISA), by phospholipase C (PLC)-neutralizing activity using both egg yolk lecithin and p-nitrophenylphosphoryl-choline (PNPPC) hydrolysis reactions and by anti-lethal activity in mice . Since the toxin-neutralizing activities of each MAb were not parallel, it has been suggested that the three biological activities may not be located in the same site in the toxin molecule . This report also describes the development of a simple purification of the toxin by affinity chromatography and a sensitive immunoassay for quantitation of the toxin using the monoclonal antibody. Arch Pathol Lab Med, 1989 May, 113(5), 534 - 5 Fatal Clostridium perfringens septicemia associated with gastrointestinal arteriovenous malformations (vascular ectasias); Craven CM; Clostridial infections usually occur in association with trauma, malignancy, or intra-abdominal disease . A 72-year-old previously healthy man presented with abdominal distress and fever . He developed a hemolytic anemia, coagulopathy, and fulminant clostridial septicemia . The patient died less than 24 hours after presentation . At autopsy, no malignancy was detected . The patient had an acute clostridial hepatic abscess and multiple arteriovenous malformations (vascular ectasias) of the large and small bowel . The case suggests that these mucosal and submucosal vascular lesions, which usually cause hemorrhage, may also predispose to infection. Scand J Gastroenterol, 1989 May, 24(4), 475 - 84 Phospholipase activation and arachidonic acid release in cultured intestinal epithelial cells (INT 407); Gustafson C et al.; The release of free arachidonic acid (AA) in cultured intestinal epithelial cells (INT 407) was investigated . INT-407 cells were first incubated overnight with radiolabeled 14C-AA, and most of the incorporated 14C-AA esterified into phosphatidylethanolamine, phosphatidylcholine, and phosphatidylinositol . Labeled cells were then exposed to different stimulating agents and the release of free 14C-AA determined . The calcium ionophore A23187 caused a dose-dependent AA release that was preceded by a rapid uptake and a subsequent efflux of 45Ca2+ . By contrast, phospholipase C from Clostridium perfringens caused a great AA release that was accompanied by an apparent uptake and a sustained intracellular accumulation of 45Ca2+ . The cells alos released AA when exposed to the protein kinase C activator, 4 beta-phorbol-12-myristate-13-acetate (PMA), and this agent, like the diacylglycerol 1-oleoyl-2-acetyl-rac-glycerol, significantly potentiated the AA release caused by A23187 . Not only A23187-mediated but also phospholipase C- and PMA-mediated AA release was inhibited by 4-bromophenacyl bromide, a known phospholipase A2 inhibitor . These findings, taken together, indicate that AA release in intestinal epithelial cells can be caused by (i) Ca2+-mediated phospholipase activation, (ii) products of phospholipase C activity, and (iii) stimulation of protein kinase C . It is suggested, therefore, that AA release in intestinal epithelial cells is governed by intracellular Ca2+, protein kinase C-mediated protein phosphorylation, and activation of phospholipase A2. Br J Pharmacol, 1989 May, 97(1), 119 - 24 Contraction of the rat isolated aorta caused by Clostridium perfringens alpha toxin (phospholipase C): evidence for the involvement of arachidonic acid metabolism; Fujii Y et al.; 1 . Alpha toxin produced by Clostridium perfringens contracted the rat isolated aorta and stimulated release of arachidonic acid in the tissue . 2 . Quinacrine did not inhibit contraction caused by the toxin . 3 . Indomethacin blocked contraction caused by the toxin in a dose-dependent manner and markedly increased levels of arachidonic acid released by the toxin . 4 . The toxin-induced contraction was blocked by the thromboxane synthetase inhibitor OKY-046 and the thromboxane A2 (TXA2) antagonist ONO-3708 . 5 . The toxin stimulated production of TXB2 and this was blocked by pretreatment with either indomethacin or OKY-046 . 6 . Toxin-induced contraction was diminished by pretreating aorta with collagenase or by rubbing the intimal surface to remove the endothelium . 7 . These data suggest that the contractile response to the toxin is associated with stimulation of TXA2 production from arachidonic acid released by the toxin in the endothelial cells of the aorta. J Biol Chem, 1989 Apr 15, 264(11), 6325 - 33 The regulation of platelet-activating factor production in endothelial cells . The role of calcium and protein kinase C; Whatley RE et al.; Endothelial cells (EC) synthesize platelet-activating factor (PAF) when stimulated with agonists that bind to cell-surface receptors . We examined events that link receptor binding to synthesis of PAF by EC . Bovine EC stimulated with agonists that interact with specific cell-surface receptors accumulated PAF only in the presence of extracellular calcium . Hormonal stimulation of EC resulted in Ca2+ entry characteristic of that seen with receptor-operated calcium channels; Indo-1 measurements demonstrated that this inward flux of Ca2+ caused prolonged elevated levels of intracellular Ca2+ . EC were exposed to melittin or theta toxin from Clostridium perfringens (pore-forming peptides that increase the permeability of the plasma membrane for small molecules) resulting in an inward flux of Ca2+ and accumulation of PAF . Ca2+ appears to be regulatory for PAF production at the level of phospholipase A2-mediated production of the PAF precursor 1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine, as Ca2+ was required for the stimulated hydrolysis of 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine . PAF accumulation in EC is also regulated by protein kinase C . Pretreatment of EC with phorbol esters that activate protein kinase C or with dioctanoylglycerol, followed by stimulation, resulted in a 2-fold increase in stimulated PAF production . The regulatory effect of protein kinase C also appears to be at a phospholipase A2-mediated hydrolysis of 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine. Biochem J, 1989 Apr 15, 259(2), 597 - 600 X-ray-absorption-spectroscopic evidence for a novel iron cluster in hydrogenase II from Clostridium pasteurianum; George GN et al.; Hydrogenase II from Clostridium pasteurianum contains three different iron-sulphur clusters . Two are {4Fe-4S}(2+.1+) clusters, whereas the other, which is thought to be the site of interaction with H2 and is known as the 'H cluster', is of unknown structure and possesses unusual spectroscopic properties . Analysis of the iron e.x.a.f.s . spectra shows that the H cluster contains iron co-ordinated mostly to sulphur and possesses 2.8 A (1 A = 0.1 nm) Fe--Fe separations when oxidized and 3.3 A Fe--Fe separations when reduced with H2 . The data suggest that the reduced H cluster represents a new structural type of iron-sulphur cluster. Biochem Biophys Res Commun, 1989 Apr 14, 160(1), 33 - 9 Cloning and sequencing of a phospholipase C gene of Clostridium perfringens; Okabe A et al.; The gene encoding phospholipase C (alpha-toxin) of Clostridium perfringens was cloned into lambda gt10 . The maximal size of the coding region was 1.4 kb and the minimum was 1.1 kb as determined by subcloning into the vector pBR322 and testing for activity . The nucleotide sequence of this region contained a single open reading frame of 1194 bp corresponding to a protein of Mr 45473 with a possible N-terminal signal sequence of 28 amino acids which when removed, would give a mature protein of Mr 42521 . This is in good agreement with the reported size of 43 kDa . The coding region has a dG + dC content of 33.7%, and the codon usage displays a pronounced preference for codons with the lowest dG + dC content. Am J Gastroenterol, 1989 Apr, 84(4), 379 - 82 Evaluation of antibiotic-associated diarrhea with a latex agglutination test and cell culture cytotoxicity assay for Clostridium difficile; Biddle WL et al.; Diagnosis of Clostridium difficile (C . difficile) by its antigen or toxin has improved treatment for patients who have antibiotic-associated diarrhea and opportunistic colonization of the colon with C . difficile . Unfortunately, results from the tissue culture cytotoxicity assay are not available for 48 h . We prospectively compared a latex agglutination test with the tissue culture cytotoxicity assay in 83 patients (15 with antibiotic-associated diarrhea, 23 with non-antibiotic-associated diarrhea, and 45 without diarrhea) . In the antibiotic-associated diarrhea group, 43% of patients with pseudomembranes and 25% without pseudomembranes tested positive with both tests . In the non-antibiotic groups 92% with diarrhea and 90% without diarrhea were negative with both tests . The number of discordant results illustrates the value of using two tests to identify C . difficile, since additional patients can be identified by using two tests . We found false positives were rare, and C . difficile toxin or antigen could not be detected in more than half the patients with pseudomembranes . The latex agglutination test for C . difficile is reliable, specific, and faster than the tissue culture cytotoxicity assay . The data suggest that other organisms may also be responsible for pseudomembranous colitis, and that negative tests do not obviate the need for visual evaluation of colonic mucosa in suspected cases of antibiotic-associated diarrhea. Can J Microbiol, 1989 Apr, 35(4), 481 - 6 Evidence for proton motive force dependent transport of selenite by Clostridium pasteurianum; Bryant RD et al.; The proton motive force mediated the transport of selenite (SeO3(2-)) in Clostridium pasteurianum cells by proton symport . The proton conductor, carbonyl cyanide m-chlorophenylhydrazone, inhibited SeO3(2-) uptake while N,N'-dicyclohexylcarbodiimide prevented SeO3(2-) uptake by presumably inhibiting the unidirectional ATPase . Acid pulse studies and antibiotic experiments with valinomycin suggest that the transmembrane delta pH component of the proton motive force mediated the transport of SeO3(2-) into the cells . The SeO3(2-) porter system in C . pasteurianum was found to be dependent upon energy source, temperature, and medium pH. Zh Mikrobiol Epidemiol Immunobiol, 1989 Apr, (4), 22 - 4 {Preparation of diagnostic antitoxic serum to Clostridium difficile}; Alchinbaeva VM et al.; The results of the studies on the preparation of diagnostic antitoxic sera to C . difficile, intended for use in biological assays with the aim of the laboratory diagnosis of clostridial enteric infections, are presented . The conditions for the detoxification of C . difficile native toxin have been established, the optimum schedules for the immunization of rabbits have been selected and specific antitoxic sera to C . difficile have been obtained . The neutralizing activity of these sera has been evaluated in the lethality test and in the cytotoxic test on human embryo dermo-muscular fibroblast cells M-19. Appl Environ Microbiol, 1989 Apr, 55(4), 845 - 51 Kinetics of biotransformation of 1,1,1-trichloroethane by Clostridium sp . strain TCAIIB; Galli R et al.; Batch experiments were conducted to examine the effects of high concentrations of 1,1,1-trichloroethane (TCA) on the biotransformation of TCA by Clostridium sp . strain TCAIIB . The biotic dehalogenation of TCA to 1,1-dichloroethane by nongrowing cells was measured at 35 degrees C, and the data were used to obtain the kinetic parameters of the Monod relationship half-velocity coefficient Ks (31 microM) and the coefficient of maximum rate of TCA biotransformation (kTCA; 0.28 mumol per mg per day) . The yield of biomass decreased with an increase in the TCA concentration, although TCA concentrations up to 750 microM did not completely inhibit bacterial growth . Also, kTCA was higher in the presence of high concentrations of TCA . A mathematical model based on a modified Monod equation was used to describe the biotransformation of TCA . The abiotic transformation of TCA to 1,1-dichloroethene was measured at 35 degrees C, and the first-order formation rate coefficient for 1,1-dichloroethene (ke) was determined to be 0.86 per year. Appl Environ Microbiol, 1989 Apr, 55(4), 837 - 44 Biotransformation of 1,1,1-trichloroethane, trichloromethane, and tetrachloromethane by a Clostridium sp; Galli R et al.; A gram-positive, strictly anaerobic, motile, endospore-forming rod, tentatively identified as a proteolytic Clostridium sp., was isolated from the effluent of an anaerobic suspended-growth bioreactor . The organism was able to biotransform 1,1,1-trichloroethane, trichloromethane, and tetrachloromethane . 1,1,1-Trichloroethane was completely transformed (greater than or equal to 99.5%) by reductive dehalogenation to 1,1-dichloroethane (30 to 40%) and, presumably by other mechanisms, to acetic acid (7%) and unidentified products . The reductive dehalogenation of tetrachloromethane led to the intermediate trichloromethane, which was further transformed to dichloromethane (8%) and unidentified products . The biotransformation occurred during the exponential growth phase, as well as during the stationary phase . Tetrachlorethene, trichloroethene, 1,1-dichloroethene, chloroethane, 1,1-dichloroethane, and dichloromethane were not biotransformed significantly by the organism. Antimicrob Agents Chemother, 1989 Apr, 33(4), 560 - 1 Antibacterial activity of the new glycopeptide antibiotic SKF104662; Jorgensen JH et al.; The inhibitory activity of the new glycopeptide antibiotic SKF104662 was generally equivalent (+/- 1 concentration increment) to the activities of vancomycin, teicoplanin, and daptomycin against selected gram-positive bacteria . However, SKF104662 demonstrated greater activity against Staphylococcus epidermidis and S . haemolyticus than did teicoplanin and was more active than the other drugs against Clostridium difficile isolates . SKF104662 possessed bactericidal activity quite similar to that of vancomycin against selected isolates of Staphylococcus, Streptococcus, and Enterococcus species. Am J Infect Control, 1989 Apr, 17(2), 77 - 82 Diagnostic studies of nosocomial diarrhea in children: assessing their use and value; Brady MT et al.; During a 17-month period (01/11/85-05/31/86) 225 cases of nosocomial diarrhea were identified in a children's hospital . Diarrhea was considered to be nosocomial if it began at least 72 hours after the patient's hospital admission or within 3 days after discharge . One or more routine diagnostic studies for identification of a pathogen were performed in 195 (87%) cases . The most commonly performed test was the bacterial stool culture . None of these samples yielded a bacterial pathogen . The only pathogens detected by routine laboratory studies were rotavirus (61/137 {45%} samples were positive for rotavirus by ELISA) and Clostridium difficile (9/54 {17%} positive for toxin) . Of the patients whose tests were positive for rotavirus 56 were younger than 2 years of age, and all were identified in the winter and spring . When multiple stool samples were tested by the diagnostic laboratory, rotavirus was identified in an additional 14 patients whose initial stool samples were negative for rotavirus . All patients whose tests were positive for C . difficile toxin had received antibiotics within the previous 3 months . Ova/parasites were not detected in 53 of the tested stools . We also identified enteric adenovirus in six patients . Viruses were identified in 95 (42%) of the 225 cases of nosocomial gastroenteritis . Nosocomial diarrhea is common in a children's hospital . Rotavirus is the most commonly identified pathogen . Rotavirus testing is valuable in children with nosocomial diarrhea who are younger than 2 years of age, especially in the winter and spring . Multiple samples may be necessary to identify rotavirus . C . difficile toxin assay should be considered for patients who are receiving or who have received antibiotics.(ABSTRACT TRUNCATED AT 250 WORDS) Mayo Clin Proc, 1989 Apr, 64(4), 392 - 9 Antimicrobial susceptibilities of anaerobic bacteria isolated at the Mayo Clinic during 1982 through 1987: comparison with results from 1977 through 1981; Musial CE et al.; Results of antimicrobial susceptibility testing of anaerobic bacteria isolated at the Mayo Clinic were reviewed for 1982 through 1987 and compared with a previous survey during 1977 through 1981 at this institution . Between the earlier and the later period, clindamycin resistance increased in the Bacteroides fragilis group (from 4% of isolates to 8%) . We noted continuing penicillin resistance among Bacteroides species other than B . fragilis and rare penicillin resistance among Fusobacterium organisms, with four isolates during the 1982 through 1987 period being beta-lactamase producers . The high levels of resistance to some agents seen in certain Clostridium species in 1977 through 1981 were not as great in the current survey . No major changes were noted in the susceptibilities of C . perfringens, anaerobic non-spore-forming gram-positive bacilli, and anaerobic gram-positive cocci. J Biol Stand, 1989 Apr, 17(2), 117 - 24 The detection of Clostridium perfringens epsilon antitoxin in rabbit serum by monoclonal antibody based competition ELISA; Sojka MG et al.; A competitive enzyme-linked immunosorbent assay (CELISA) has been developed, standardized and compared with the toxin neutralization (TN) test performed in mice for the measurement of antibody responses in rabbits vaccinated with clostridial vaccines . In CELISA, sera were tested at a single dilution for their ability to compete with the reaction between a monoclonal antibody, which neutralizes epsilon toxin, and epsilon toxoid coated on to a solid phase . The results of the two tests correlated well . CELISA was specific, rapid, reproducible and simple to perform and offered an alternative to the TN test that reduced the requirement for experimental animals in the potency testing of clostridial vaccines. Rev Argent Microbiol, 1989 Apr-Jun, 21(2), 47 - 53 {Type D and A Clostridium botulinum in necropsy samples from bovines with "mal de Aguapey"}; Fernandez RA et al.; Bacteriological studies were carried out on several necropsy samples from five animals whose deaths had been attributed to bovine botulism . This disease, regionally called Mal de Aguapey, enzootically affects animals from a wide area of the north-east of Argentina (Province of Corrientes) with a bovine population estimated at near to 2,500,000 . Either C . botulinum type D, its toxin or both were identified in all animal samples, alternatively in contents of rumen, jejunum, ileum, caecum and in samples of spleen, liver and kidney (Table 1) . C . botulinum type A was isolated respectively from the liver and the kidney of two animals . Cultures of 100 soil samples taken in the enzootic area were positive only for C . botulinum type A (3%) . These results enlarge and confirm previous findings and lend support to the botulinic etiology of the Mal de Aguapey. J Antimicrob Chemother, 1989 Apr, 23(4), 623 - 31 Third generation cephalosporins as a risk factor for Clostridium difficile-associated disease: a four-year survey in a general hospital; de Lalla F et al.; The main clinical features of patients who developed pseudomembranous colitis (PMC) or Clostridium difficile-associated diarrhoea (CDAD) during their stay at the S . Anna General Hospital, Como, over the period February 1984 to May 1988, are reported . Forty patients developed either CDAD (ten cases) or PMC (30 cases) . Twenty-seven (65.7%) had undergone surgery and 32 (80.0%) had received prolonged antibiotic treatment . Three patients (7.5%) were given three doses only of ceftriaxone . Five patients (12.5%) had not received any antibiotic treatment; but three were nursed in a bed next to a patient with PMC-CDAD . The number of cases diagnosed were correlated retrospectively with the cumulative consumption of different groups of antibiotics on wards in which PMC or CDAD occurred . A significant difference (P less than 0.01) between third generation cephalosporins (16 cases) and ureidopenicillins (one case), was found . Twenty-five patients were treated with oral vancomycin . Two died of the underlying disease and 23 were cured . The disease recurred clinically in three, and follow-up cultures were positive in another asymptomatic case . Fifteen patients (all PMC cases) were treated with oral teicoplanin . All were clinically cured and remained asymptomatic and all but one were also cleared of C . difficile . No adverse reactions were observed in patients given either drug . Third generation cephalosporins, even when administered as short-term perioperative prophylaxis, but not ureidopenicillins, are significantly associated with C . difficile-related diseases . Teicoplanin proved to be very effective and safe in the treatment of PMC, and should be further evaluated there. Postgrad Med J, 1989 Apr, 65(762), 208 - 10 Effect of treatment with botulinum toxin on spasticity; Das TK et al.; Botulinum toxin, a product of Clostridium botulinum, produces presynaptic neuromuscular block by preventing release of acetylcholine from nerve endings . The toxin was injected directly into the skeletal muscles of six patients with severe spasticity due to stroke-related hemiplegia . It produced both subjective and objective improvement . The toxin injections were well tolerated and no significant side effect was reported. J Hosp Infect, 1989 Apr, 13(3), 309 - 14 Outbreak of Clostridium difficile diarrhoea in an orthopaedic unit: evidence by phage-typing for cross-infection; Degl'Innocenti R et al.; In a three-week period five patients had diarrhoea in an orthopaedic unit . The first case was clinically diagnosed as pseudomembranous colitis but the causative agent was not sought . Of the remaining cases, two were Clostridium difficile positive . The outbreak then apparently ceased, but during the following several days two of seven stool samples taken at random from asymptomatic patients yielded C . difficile . Phage-typing of the isolates showed that all apparently belonged to the same strain. J Bacteriol, 1989 Apr, 171(4), 2148 - 54 Ammonia assimilation pathways in nitrogen-fixing Clostridium kluyverii and Clostridium butyricum; Kanamori K et al.; Pathways of ammonia assimilation into glutamic acid were investigated in ammonia-grown and N2-fixing Clostridium kluyverii and Clostridium butyricum by measuring the specific activities of glutamate dehydrogenase, glutamine synthetase, and glutamate synthase . C . kluyverii had NADPH-glutamate dehydrogenase with a Km of 12.0 mM for NH4+ . The glutamate dehydrogenase pathway played an important role in ammonia assimilation in ammonia-grown cells but was found to play a minor role relative to that of the glutamine synthetase/NADPH-glutamate synthase pathway in nitrogen-fixing cells when the intracellular NH4+ concentration and the low affinity of the enzyme for NH4+ were taken into account . In C . butyricum grown on glucose-salt medium with ammonia or N2 as the nitrogen source, glutamate dehydrogenase activity was undetectable, and the glutamine synthetase/NADH-glutamate synthase pathway was the predominant pathway of ammonia assimilation . Under these growth conditions, C . butyricum also lacked the activity of glucose-6-phosphate dehydrogenase, which catalyzes the regeneration of NADPH from NADP+ . However, high activities of glucose-6-phosphate dehydrogenase as well as of NADPH-glutamate dehydrogenase with a Km of 2.8 mM for NH4+ were present in C . butyricum after growth on complex nitrogen and carbon sources . The ammonia-assimilating pathway of N2-fixing C . butyricum, which differs from that of the previously studied Bacillus polymyxa and Bacillus macerans, is discussed in relation to possible effects of the availability of ATP and of NADPH on ammonia-assimilating pathways. J Vet Diagn Invest, 1989 Apr, 1(2), 170 - 3 A bacteriologic study of scabby-hip lesions from broiler chickens in Texas; Scanlan CM et al.; Broilers from commercial flocks experiencing a 10-60% incidence of scabby-hip lesions at processing were examined, and selected skin lesions were cultured . Over 70% of the lesions were associated with traumatic excoriations, particularly on the caudal dorsal convexity of the birds . Most lesions were observed on birds that were 5 weeks of age or older . From the 27 specimens cultured, Clostridium perfringens was isolated in pure culture from 4 lesions and Staphylococcus species from 10 lesions . Pure cultures of staphylococci were recovered from 4 lesions, and 2-5 different staphylococci were isolated from 6 lesions . Eight staphylococci were identified as S . sciuri, 8 as S . simulans, 2 as S . epidermidis, 2 as S . lentus, 2 as S . warneri, 1 as S . cohnii, and 1 as S . intermedius . Fifty cutaneous specimens from 10 5-week-old normal broilers were cultured . A total of 197 isolates were identified including 65 S . sciuri, 52 S . lentus, 24 S . simulans, 12 S . hyicus, 11 S . warneri, 9 S . cohnii, 9 S . gallinarium, 8 S . xylosus, and 7 S . epidermidis. J Gen Microbiol, 1989 Apr, 135 ( Pt 4), 903 - 9 Cloning in Escherichia coli of the enterotoxin gene from Clostridium perfringens type A; Iwanejko LA et al.; A 26 bp DNA probe has been constructed with minimal degeneracy to the protein sequence for Clostridium perfringens enterotoxin . The probe has been hybridized against a 6-10 kb chromosomal bank from C . perfringens 8239, prepared as a HindIII partial digest in pHG165 . From this survey a clone has been identified containing a 6.8 kb DNA insert with strong hybridization to the probe . Direct plasmid sequencing has identified a translational reading frame within this clone which correlates with the known protein sequence for the type A enterotoxin . DNA sequences 5' to this open reading frame and containing the putative transcriptional control regions show areas of significant homology with regions upstream from the ATG codon of the tetanus toxin gene. Kansenshogaku Zasshi, 1989 Apr, 63(4), 410 - 6 {An outbreak of enterocolitis due to Clostridium perfringens in a hospital for the severely disabled}; Machida Y et al.; We had an outbreak of 14 cases of enterocolitis due to Clostridium perfringens (Cl . perfringens) in a hospital for the severe multiply-disabled, where the 100 disabled were admitted, in summer in 1985 . The signs and symptoms shown by this enterocolitis were primarily diarrhea without fever and loss of appetite . The feces of 10 cases were examined bacteriologically . The test showed 10(3) to 10(6) cells of Cl . perfringens per one gram of their feces and all the strains isolated were untypable by the classification of Hobbs . Nine out of 10 cases were randomly selected and all of the 9 cases were proved to have enterotoxin producing strains . All the strains were highly sensitive to many kinds of antibiotics except kanamycin and gentamicin . Eleven out of the 14 cases were admitted in the same ward and the 7 out of the 11 cases were in the same room of this ward . Considering the spreading route of this infection, it is unlikely that this outbreak occurred due to food supplied from kitchen in this hospital, because all of the disabled, admitted in this hospital, had little chance by which some of the disabled only in a specific ward or room were supplied with bacteriologically contaminated meals from the point of view of cooking and supplying system of this hospital . Adding to this fact, if this outbreak was due to food-born infection, the symptoms of most patients should occur within 1-2 days, because the incubation period of this disease is within a day, however, the patients increased day by day for more than a week.(ABSTRACT TRUNCATED AT 250 WORDS) J Appl Bacteriol, 1989 Apr, 66(4), 263 - 70 Bacterial overgrowth in the jejunum of ICR mice and Wistar rats orally administered with a single lethal dose of fusarenon-X, a trichothecene mycotoxin; Morishita Y et al.; A single oral dose of fusarenon-X (F-X), a trichothecene mycotoxin, resulted in abnormal microflora in the jejunum in ICR mice and Wistar rats with some differences in dose response between the species . In the acute phase, enterobacteria, streptococci, Clostridium perfringens and bacteroides showed remarkably increased counts in the jejunum of mice and rats dosed with F-X while lactobacilli showed a decrease in count . F-X brought an invasion of Pseudomonas aeruginosa into the livers, lungs, kidneys and spleens of ICR mice . Changes in the jejunal microflora appeared after 7 h in ICR mice and after 24 h in Wistar rats after a single oral dose of F-X of 7.5 and 4.0 mg/kg b.w., respectively, and the microflora returned to its normal state at 72 h in mice and 96 h in rats . The changes of intestinal microflora were followed by alterations in the growth curves of both animal species . The pH in the glandular stomach was also greatly enhanced before changes in the jejunal microflora . Acute F-X intoxication may be an involved manifestation of essential cytotoxicity of F-X mycotoxin alone and secondary bacterial overgrowth in the bowel. FEMS Microbiol Lett, 1989 Apr, 49(2-3), 269 - 72 Contraction induced by Clostridium perfringens epsilon toxin in the isolated rat ileum; Sakurai J et al.; Clostridium perfringens epsilon toxin caused contraction of the isolated ileum of the rat in a dose-dependent manner . The contraction caused by the toxin was inhibited by a low Na medium, tetrodotoxin (TTX), atropine, mecamylamine or tetraethylammonium (TEA) . Furthermore, the contractile response induced by the toxin was abolished by incubation in Ca-free medium, and completely restored by and addition of Ca2+ . In addition, verapamil inhibited contraction induced by the toxin in a dose-dependent manner . These data suggest that epsilon toxin induces contraction of the isolated ileum and that the toxin-elicited contraction is the result of an indirect action mediated through the nervous systems. Eur J Cell Biol, 1989 Apr, 48(2), 191 - 202 Mechanism of action of Clostridium difficile toxin B: role of external medium and cytoskeletal organization in intoxicated cells; Ciesielski-Treska J et al.; Toxin B, an exotoxin produced by Clostridium difficile, induces the rounding-up and arborization of cultured mammalian cells, a typical effect which resembles that provoked by cytochalasins . In this study, the effect of toxin B was examined on astroglial cells grown in primary culture . A specific antiserum to toxin B was used to investigate its mechanisms of action . We found that the toxin exerts its effects on cell morphology after its incorporation into cells . The internalization of toxin B requires the presence of calcium ions in the extracellular medium . Replacement of NaCl with sucrose or with potassium glutamate prevents the internalization of the toxin . The direct introduction of calcium ions into cells by the calcium ionophore A23187 stimulates toxin-induced morphological changes . In contrast, toxin-induced morphological transformations were prevented in cells treated with tumor-promoting phorbol . esters or with dibutyryl-cAMP, although such treatment did not abolish the internalization of the toxin . As in the other cell types, the earliest effect of toxin B on astrocyte cytoskeleton is the disruption of actin filaments, without no visible alteration of intermediate filament nor microtubule networks . As astrocytes with toxin-induced stellate morphology survive toxin treatment, the progression of cell morphology and cytoskeleton organization were followed for several weeks . Twenty-six days after exposure to toxin B, stellate astrocytes have processes which were markedly longer and much more branched than those of cells freshly exposed to toxin . At that time, cells are still devoid of F-actin as assessed with rhodamine-conjugated phalloidin and only 70% contain vimentin while all astrocytes present in control cultures express vimentin . Some flat epithelioid astrocytes with prominent bundles of microfilaments reappear during the second week after toxin treatment . Our results show that Clostridium difficile toxin B is internalized into brain astrocytes in culture where it acts by modifying cytoskeletal elements . Its cytopathic effects are reversible . Although actin-related components of the cytoskeleton are the major target of toxin B, other cytoskeletal elements also seem to be affected. Clin Orthop, 1989 Apr, (241), 245 - 7 Clostridium perfringens and Staphylococcus epidermidis polymicrobial septic arthritis . A case report; McAllister CM et al.; Septic arthritis caused by Clostridium perfringens is extremely rare . Previously there has been only one report of Clostridium perfringens in combination with an aerobe causing septic arthritis . This report presents a 23-year-old man with a mixed aerobic/anaerobic septic arthritis treated with intravenous antibiotics and repeated surgical drainage . It is the only report to date of a polymicrobial septic arthritis involving both Staphylococcus epidermidis and Clostridium perfringens . This report and a review of the literature demonstrate the existence of polymicrobial septic arthritis and illustrate its fulminant nature. EMBO J, 1989 Apr, 8(4), 1087 - 92 The mammalian G protein rhoC is ADP-ribosylated by Clostridium botulinum exoenzyme C3 and affects actin microfilaments in Vero cells; Chardin P et al.; Clostridium botulinum C3 is a recently discovered exoenzyme that ADP-ribosylates a eukaryotic GTP-binding protein of the ras superfamily . We show now that the bacterially-expressed product of the human rhoC gene is ADP-ribosylated by C3 and corresponds in size, charge and behavior to the dominant C3 substrate of eukaryotic cells . C3 treatment of Vero cells results in the disappearance of microfilaments and in actinomorphic shape changes without any apparent direct effect upon actin . Thus the ADP-ribosylation of a rho protein seems to be responsible for microfilament disassembly and we infer that the unmodified form of a rho protein may be involved in cytoskeletal control. Am J Physiol, 1989 Apr, 256(4 Pt 1), G767 - 72 Effect of purified Clostridium difficile toxins on intestinal smooth muscle . II . Toxin B; Gilbert RJ et al.; In the companion paper {Am . J . Physiol . 256 (Gastrointest . Liver Physiol . 19): G759-G766, 1989} we showed that highly purified Clostridium difficile toxin A had a profound effect on intestinal smooth muscle after in vivo but not in vitro exposure . In this study we assessed the effects of in vivo and in vitro exposure to C . difficile toxin B on simultaneous measurements of intracellular membrane potential and contractility in rabbit ileal smooth muscle . Direct exposure of ileal smooth muscle to toxin B (0.1-60 micrograms/ml) in vitro caused membrane depolarization and inhibition of action potential frequency, amplitude, and peak voltage, but no effect on slow wave frequency or amplitude was seen . Toxin exposure also resulted in inhibition of the amplitude of carbachol-induced contractions, with phasic contractions being significantly more sensitive to the effect of toxin B than tonic contractions over the complete dose range . The electromechanical effects of toxin B were not affected by prior administration of tetrodotoxin, atropine, hexamethonium, or phentolamine . In contrast, toxin B administered in vivo into an isolated ileal loop had no effect on spontaneous electromechanical properties of excised smooth muscle strips . Our results indicate that direct exposure in vitro of ileal smooth muscle to C . difficile toxin B causes membrane depolarization in association with inhibition of electromechanical activity . This effect, in combination with the indirect effects of toxin A, may contribute to altered intestinal motility during diarrhea caused by C . difficile. Am J Physiol, 1989 Apr, 256(4 Pt 1), G759 - 66 Effect of purified Clostridium difficile toxins on intestinal smooth muscle . I . Toxin A; Gilbert RJ et al.; In these studies we determined the effects of purified Clostridium difficile toxin A, an enterotoxin, on the electrophysiological and contractile properties of rabbit intestinal circular smooth muscle and correlated these effects with changes of smooth muscle morphology . Simultaneous measurements of intracellular membrane potential and contractility were determined in excised ileal muscle strips after administration of toxin A in vivo (60 micrograms/ml) into an isolated rabbit ileal loop or directly in vitro (0.1-60 micrograms/ml) to a normal muscle strip . Toxin A injection in vivo resulted in membrane depolarization and increased slow wave and action potential frequency . Toxin A injection in vivo also caused increased amplitude of spontaneous and carbachol-induced phasic contractions . The electrophysiological effects of in vivo administration of toxin A were correlated with an inflammatory infiltrate of the lamina propria, but no light or electron microscopic evidence of injury to smooth muscle cells was seen . In contrast to the in vivo studies, direct in vitro exposure of normal ileal muscle strips to toxin A had no effect on either spontaneous or carbachol-induced electromechanical activity . Our results indicate that in vivo administration of C . difficile toxin A into a rabbit ileal loop, but not direct in vitro exposure, causes significant alterations of smooth muscle excitation-contraction coupling that may be mediated by products of local inflammatory cells. J Bacteriol, 1989 Apr, 171(4), 2209 - 15 Purification and partial characterization of the glycine decarboxylase multienzyme complex from Eubacterium acidaminophilum; Freudenberg W et al.; The proteins P1, P2, and P4 of the glycine cleavage system have been purified from the anaerobic, glycine-utilizing bacterium Eubacterium acidaminophilum . By gel filtration, these proteins were determined to have Mrs of 225,000, 15,500, and 49,000, respectively . By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, protein P1 was determined to have two subunits with Mrs of 59,500 and 54,100, indicating an alpha 2 beta 2 tetramer, whereas the proteins P2 and P4 showed only single bands with estimated Mrs of 15,500 and 42,000, respectively . In reconstitution assays, proteins P1, P2, P4 and the previously reported lipoamide dehydrogenase (P3) had to be present to achieve glycine decarboxylase or synthase activity . All four glycine decarboxylase proteins exhibited highest activities when NADP+ was used as the electron acceptor or when NADPH was used as the electron donor in the glycine synthase reaction . The oxidation of glycine depended on the presence of tetrahydrofolate, dithioerythreitol, NAD(P)+, and pyridoxal phosphate . The latter was loosely bound to the purified protein P1, which was able to catalyze the glycine-bicarbonate exchange reaction only in combination with protein P2 . Protein P2 could not be replaced by lipoic acid or lipoamide, although lipoic acid was determined to be a constituent (0.66 mol/mol of protein) of protein P2 . Glycine synthase activity of the four isolated proteins and in crude extracts was low and reached only 12% of glycine decarboxylase activity . Antibodies raised against P1 and P2 showed cross-reactivity with crude extracts of Clostridium cylindrosporum. Gastroenterology, 1989 Apr, 96(4), 981 - 8 Prevention of antibiotic-associated diarrhea by Saccharomyces boulardii: a prospective study; Surawicz CM et al.; Saccharomyces boulardii, a nonpathogenic yeast, has been widely used in Europe to prevent antibiotic-associated diarrhea (AAD) . We performed a prospective double-blind controlled study to investigate AAD in hospitalized patients and to evaluate the effect of S . boulardii, a living yeast, given in capsule form concurrently with antibiotics . Over 23 mo, 180 patients completed the study . Of the patients receiving placebo, 22% experienced diarrhea compared with 9.5% of patients receiving S . boulardii (p = 0.038) . Risk factors found to be associated with AAD were multiple antibiotic combinations (containing clindamycin, cephalosporins, or trimethoprim-sulfamethoxazole) and tube feeding . Clostridium difficile, an anaerobe found in the stools of most patients with pseudomembranous colitis, was variably associated with AAD . We evaluated the role of C . difficile in AAD in the study population and found no significant association between the presence of C . difficile or cytotoxin with AAD . Approximately 33% of the patients without diarrhea harbored at least one C . difficile-positive stool and nearly 50% of these patients had detectable cytotoxin . Similar values were obtained in patients with diarrhea . Of C . difficile-positive patients, 31% (5/16) on placebo developed diarrhea compared with 9.4% (3/32) on S . boulardii; this difference was not statistically significant (p = 0.07) . There were no discernable adverse effects of yeast administration . We conclude that S . boulardii reduces the incidence of antibiotic-associated diarrhea in hospitalized patients. Biochim Biophys Acta, 1989 Mar 16, 995(1), 17 - 20 Coenzyme non-specific glutamate dehydrogenase from Chlorella pyrenoidosa 82T: electron microscopic studies; Shatilov VR et al.; The constitutive coenzyme non-specific glutamate dehydrogenase (GDH) from Chlorella pyrenoidosa 82T was purified to homogeneity by column immunoaffinity chromatography and examined by an electron microscope . The enzyme molecule was found to be a hexameric oligomer composed of monomers arranged in three 2-point group symmetry in two layers slightly twisted round the 3-fold axis . The molecule is 8 +/- 1 nm in diameter and 10 +/- 1 nm in height . The enzyme molecules appear both to dissociate into trimers and to associate along the 3-fold axis forming linear aggregates under certain conditions . A tentative model of the Chlorella GDH molecule is proposed, which is very similar to those described for bovine liver GDH and GDH from Clostridium symbiosum. Biochim Biophys Acta, 1989 Mar 16, 995(1), 59 - 63 Enzymatic cleavage of Clostridium pasteurianum apoferredoxin and reconstitution of the cleaved products; Skjeldal L et al.; Native ferredoxin from Clostridium pasteurianum proved to be resistant to proteolytic cleavage under anaerobic conditions, but was digested in the presence of air . Apoferredoxin was hydrolyzed by the proteinases used, while cobalt-substituted ferredoxin was resistant both under anaerobic and aerobic conditions . These studies indicate that metal binding of the protein stabilizes the folded state, which is extremely resistant to proteolytic attack . Sulfitolyzed apoferredoxin was subjected to specific cleavage by pepsin at pH 3.2, yielding two fragments . The fragments could be reconstituted to an unstable holoprotein with UV-visible absorption features like that of the native form. Biochem Biophys Res Commun, 1989 Mar 15, 159(2), 426 - 31 The ketone cinnamoyl-(1-13C-Phe)-CGly-Pro-Pro is a tetrahedral transition state analog inhibitor of C . histolyticum collagenase; Grobelny D et al.; The ketone cinnamoyl-(1-13C-Phe)-CGly-Pro-Pro {(4-13C-5-cinnamido-4-oxo-6-phenylhexanoyl)-Pro-Pro 2} competitively inhibits a mixture of collagenases from Clostridium histolyticum with a Ki of 40 +/- 6 nM . 13C-nmr spectroscopy of the ketone in the presence of this collagenase shows a bound 13C resonance at 102.6 ppm and the resonance of the free ketone at 212 ppm . Ketone alone shows no trace (less than 0.5%) of a resonance in the region around 100 ppm . The bound resonance is displaceable by another competitive inhibitor . This ketone is thus a transition state analog which is rehybridized from trigonal planar to tetrahedral upon binding to collagenase. J Biol Chem, 1989 Mar 15, 264(8), 4342 - 8 Electron paramagnetic resonance studies of the low temperature photolytic behavior of oxidized hydrogenase I from Clostridium pasteurianum; Kowal AT et al.; The effects of CO and O2 on the EPR spectrum of oxidized Clostridium pasteurianum hydrogenase I have been investigated both before and after prolonged exposure to white light at 8 K and 30 K . Low concentrations of O2 were found to induce analogous changes in the EPR spectrum as CO, i.e . conversion of the rhombic signal with g approximately 2.10, 2.04, 2.00, a characteristic of the novel H2-activating center in oxidized Fe-hydrogenases, to an axial signal with g approximately 2.07, 2.01, 2.01 . The results suggest a common binding site and mode of coordination for CO and O2 and permit rationalization of conflicting reports from different laboratories concerning the EPR properties of oxidized Fe-hydrogenases . The CO- and O2-induced axial EPR signals were found to be light-sensitive at low temperatures . Moreover, they exhibited indistinguishable and unusual photolysis behavior with the dominant photo-product being dependent on the temperature at which illumination was performed . At 8 K, photodissociation of CO or O2 occurs, resulting in an EPR signal identical with that of the oxidized enzyme in the absence of CO or O2 . However, at 30 K, the dominant photoproduct is a rhombic EPR signal with g approximately 2.26, 2.12, 1.89 . While the origin of this new EPR signal is uncertain, the g-value anisotropy and relaxation characteristics resemble those of a low spin Fe(III) center . These two photoproducts cannot be thermally or photolytically interconverted, but both are quantitatively reconverted to the original axial EPR signal on warming in the dark to 200 K . A tentative working hypothesis for the nature of the H2-activating center of Fe-hydrogenases is presented that is consistent with the available physiochemical data and permits rationalization of the novel photolysis behavior. Biochim Biophys Acta, 1989 Mar 14, 1002(1), 37 - 44 NADP-dependent 3 beta-, 7 alpha- and 7 beta-hydroxysteroid dehydrogenase activities from a lecithinase-lipase-negative Clostridium species 25.11.c; Edenharder R et al.; A lecithinase-lipase-negative Clostridium sp . 25.11.c., not fitting in any of the species of Clostridia described so far as judged by morphological, physiological, and biochemical data, was shown to contain NADP-dependent 3 beta-, 7 alpha- and 7 beta-hydroxysteroid dehydrogenases . The three hydroxysteroid dehydrogenases could be demonstrated in the supernatant and in the membrane fraction after solubilization with Triton X-100, suggesting enzymes which were originally membrane bound . The 3 beta-hydroxysteroid dehydrogenase was synthesized constitutively, and the specific enzyme activity was significantly reduced by growth medium supplementation with 3-keto bile acids and trisubstituted bile acids . A pH optimum of 7.5 and a molecular weight of approx . 104,000 were estimated by molecular sieve chromatography . The enzyme reduced the 3-keto group of bile acids; an oxidation of a 3 beta-hydroxyl function could not be demonstrated . The lowest Km values were found for disubstituted bile acids, trisubstituted and conjugated bile acids having higher Km values . 7 alpha-Hydroxysteroid dehydrogenase, but not 7 beta-hydroxysteroid dehydrogenase, was already present in uninduced cells . The specific activities, however, were greatly enhanced when cells were grown in the presence of chenodeoxycholic acid or 3 alpha-hydroxy-7-keto-5 beta-cholanoic acid . Ursodeoxycholic acid with its 7 beta-hydroxyl group was ineffective as an inducer . Molecular weights of approx . 82,000 and 115,000 were found for the 7 alpha-hydroxysteroid dehydrogenase and the 7 beta-hydroxysteroid dehydrogenase, respectively . In contrast to the in vivo situation, the reaction could only be demonstrated in the reductive direction in vitro . Here, the pH optimum for the overall reaction was 8.5-8.7 . 3 beta-, 7 alpha- and 7 beta-hydroxysteroid dehydrogenase activities were readily demonstrated for at least 48 h when preparations were stored at 4 degrees C, but were found to be heat-sensitive. Orv Hetil, 1989 Mar 12, 130(11), 551 - 6 {Gas gangrene cases in Hungary 1979-1986}; Sere G et al.; The authors evaluate epidemiologic data of 182 patients suffering from gas gangrene during an eight year period from 1979 to 1986 . Of the patients surveyed 127 died; thus lethality reached 69.2% . The average age of the survivors was 49.9 year as opposed to the 66.1 year of the fatal cases . More than half of the illnesses followed amputation of extremities, and a quarter of them was a consequence of an accident . Samples of the bacteriologically examined wound discharges yielded in 93.1% bacteria from the Clostridium genus . Hygiene was poor in operation theatres and in the hospital environment in 1/4-th of the cases . Sixty six patients died within 24 hours after diagnosis . The presented data suggest that in Hungary the number of gas gangrene cases and deaths surpass those of tetanus. J Biol Chem, 1989 Mar 5, 264(7), 4082 - 7 Purification, microheterogeneity, and stability of human lipid transfer protein; Kato H et al.; A method for the purification of lipid transfer protein (LTP) from human plasma was developed with the aid of succinylated low density lipoprotein-Sepharose affinity column chromatography . The purified LTP exhibited a single main band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis . However, upon isoelectric focusing on polyacrylamide gel, the preparations consistently showed nine bands with isoelectric points ranging from 4.6 to 5.4 . The treatment of LTP with Clostridium perfringens neuraminidase shifted these multiple bands toward higher pH regions due to the release of sialic acid . Extensive treatment with neuraminidase resulted in the appearance of a major band with the isoelectric point of 5.6 . The purified LTP was rapidly inactivated upon incubation at 37 degrees C due to the denaturation at the "air"-water interface . Various factors promoting or preventing this interfacial denaturation were elucidated . When purified LTP was stored at 4 degrees C, plasma neuraminidase co-purified with LTP became activated, resulting in the gradual desialylation of LTP . It seemed that the LTP preparations of apparent homogeneity are associated with a trace amount of an inactive form of plasma neuraminidase . The inclusion of 4 mM 2-mercaptoethanol or 0.2% EDTA in the storage media completely prevented the activation of plasma neuraminidase . These agents, however, did not significantly inhibit the already activated neuraminidase . When LTP was stored at -20 degrees C in very low ionic strength media, such as 0.001% EDTA (pH 7.4) and at high protein concentrations, the loss of the activity was minimal even after prolonged storage. J Med Microbiol, 1989 Mar, 28(3), 217 - 21 Recovery of spores of Clostridium difficile altered by heat or alkali; Kamiya S et al.; The effect of heating or alkali-treatment on spore recovery in ordinary growth medium was examined for four strains of Clostridium difficile . Heating spores at 80 degrees C for 10 min produced 95.50-99.95% decreases in the recovery rates . Treatment with 0.1 N NaOH for 15 min produced 99.47 and 99.83% decreases in spore recovery rates for two of the four strains . The influence of either addition of lysozyme after treatment with sodium thioglycollate (thioglycollate-lysozyme method) or addition of sodium taurocholate (taurocholate method) on recovery of heat- or alkali-treated C . difficile spores was also examined . Viable spores of all strains altered by heating at 90 degrees C or 100 degrees C for 10 min could not be recovered at all by the taurocholate method . Nor did this method allow recovery of alkali-altered spores treated with greater than 0.2 N NaOH for 15 min . On the other hand, 10-47% of altered spores heated at 90 degrees C for 10 min were recovered by the thioglycollate-lysozyme method, and alkali-altered spores treated with 0.1-0.3 N NaOH for 15 min were as completely recovered by this method as untreated spores . These results indicate that the thioglycollate-lysozyme method is more effective than the taurocholate method for recovery of the heat- or alkali-altered C . difficile spores. Infect Immun, 1989 Mar, 57(3), 932 - 6 Experimental cecitis in gnotobiotic quails monoassociated with Clostridium butyricum strains isolated from patients with neonatal necrotizing enterocolitis and from healthy newborns; Bousseboua H et al.; Using axenic quails fed a diet containing lactose, we have investigated the potentially pathogenic roles of six Clostridium butyricum strains of human origin . Three strains (CB155-3, CB1002, and CB203-1) isolated from neonatal necrotizing enterocolitis patients and two of three strains (CB19-1 and CB25-2) isolated from healthy newborns led to cecal or crop lesions or both similar to those observed in human neonatal necrotizing enterocolitis: thickening of the cecal wall with gas cysts, hemorrhagic ulcerations, and necrotic areas . The lactose-negative strain (CB46-1) did not develop any lesions . The neuraminidase-producing strain (CB155-3) caused lesions in all monoassociated quails, whereas the other strains caused lesions in 28 to 85% of animals . Removal of dietary lactose suppressed all pathological incidence . These results show that lactose fermentation is a prerequisite in these pathological changes and stress the roles played by both the strain and the host in the expression of C . butyricum enteropathogenicity. Br Poult Sci, 1989 Mar, 30(1), 81 - 9 Influence of peas (Pisum sativum) as a dietary ingredient and flavomycin supplementation on the performance and intestinal microflora of broiler chicks; Brenes A et al.; 1 . Two experiments were carried out to study the effects of diets containing various concentrations of pea meal (Pisum sativum L.), with or without flavomycin supplementation, on the performance and intestinal microflora of broiler chicks . 2 . During the 7 to 28-d period, chicks fed on diets containing 300, 600 and 800 g pea meal/kg consumed more food and gained more weight than chicks receiving a maize-isolated soyabean protein control diet . The addition of flavomycin to the diets had similar effects on the performance of both the control and the pea groups . 3 . Pea diets, with and without supplemental flavomycin, had little influence on the composition of intestinal microflora . The counts of enterococci in the small intestine and Clostridium perfringens and coliforms in the caeca of pea-fed chicks exceeded those of control chicks. Can J Microbiol, 1989 Mar, 35(3), 388 - 98 Morphological changes in putrefactive anaerobe 3679 (Clostridium sporogenes) induced by sorbate, hydrochloric acid, and nitrite; Ronning IE et al.; Putrefactive anaerobe 3679 (Clostridium sporogenes), a gram-positive bacterium, was examined by light and electron microscopy during normal growth and in a medium containing sorbate (50 mM, pH 6.5), hydrochloric acid (pH of medium adjusted from 7 to 5 with HCl), or nitrite (1 mM, pH 7) . During the early exponential growth phase, untreated cells were filamentous and nonseptate, but became septate later and divided when the culture entered the stationary phase . Untreated short and filamentous cells had a double-layered cell wall . Sorbate-treated cells were usually filamentous and nonseptate, but with distorted shapes characterized by numerous bends and bulges . Septation, when present, resulted in minicells . The inner cell wall appeared to be thickened and the outer wall was absent in many areas . Acid-treated cells were similar to sorbate-treated cells but contained septa . Considerable cellular debris was present in the suspension . Nitrite-treated cells were also filamentous, bent, and bulged but the cell wall appeared normal . Considerable cellular debris was also present in suspensions of nitrite-treated cells . Changes in morphology are discussed in relation to possible mechanisms of cell growth regulation and the inhibitory action of sorbate, acid, and nitrite. FEMS Microbiol Lett, 1989 Mar, 49(1), 15 - 20 Construction of shuttle vectors useful for transforming Clostridium acetobutylicum; Truffaut N et al.; Plasmids pIM13, pT127 and pBC16 delta 1, introduced by transformation into Clostridium acetobutylicum N1-4081, were shown to replicate in, and to confer antibiotic resistance upon this new host . Recombinant plasmids were constructed by inserting erythromycin-resistant plasmid pIM13 into the unique ClaI site of pBR322 or by ligating a tetracycline-resistant determinant of plasmid pT127 to HindIII-linearized pIM13 . The hybrid plasmids replicated and expressed erythromycin resistance in C . acetobutylicum strain N1-4081 and in Escherichia coli or Bacillus subtilis, indicating that they might be useful as shuttle vectors for transferring genes between these strains . The efficiency and stability of different replicons in C . acetobutylicum were compared. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1989 Mar, 270(4), 456 - 61 Demonstration of capsules in Clostridium difficile; Strelau E et al.; In four strains of Clostridium difficile the formation of capsules was demonstrated by light and electron microscopy. Biofactors, 1989 Mar, 2(1), 57 - 63 A general method for generation and analysis of defined mutations in enzymes involved in a tetrahydrofolate-interconversion pathway; Barlowe CK et al.; In eukaryotes, 10-formyltetrahydrofolate (THF) synthetase, 5,10-methenyl-THF cyclohydrolase and 5,10-methylene-THF dehydrogenase activities are present on a single polypeptide termed C1-THF synthase . These reactions are generally catalyzed by three separate monofunctional enzymes in prokaryotic cells . In this report a general method for the generation, detection and analysis of specific mutations affecting the catalytic activity of any of the reactions catalyzed by C1-THF synthase or its monofunctional counterparts is described . The method relies on plasmid-borne expression of genes in strains of the yeast Saccharomyces cerevisiae that are missing one or more of the activities of C1-THF synthase . Specific segments of the gene are subjected in vitro to random mutagenesis, the mutant genes expressed in yeast and screened by phenotype for inactivating mutations . Plasmids encoding mutant enzymes are recovered for sequence analysis . One-step purification of C1-THF synthase from the yeast expression system is demonstrated . The feasibility and versatility of the method is shown with the yeast ADE3 gene encoding the cytoplasmic C1-THF synthase and the gene encoding the monofunctional 10-formyl-THF synthetase from Clostridium acidiurici. Biokhimiia, 1989 Mar, 54(3), 387 - 95 {A new type of Clostridium thermocellum endoglucanase produced by the recombinant strain of E . coli . Some properties and identification in donor cells}; Mel'nik MS et al.; The properties of endoglucanase produced by the recombinant strain of E . coli carrying plasmid pCU 104 with a 2.9 kb insert of chromosomal DNA of C . thermocellum encoding the multiple forms of the 35.5 kD polypeptide (pI 4.3-4.7) were studied . The enzyme has a broad pH optimum of activity (6.0-7.5) . The half-inactivation time for different forms of the enzyme at 65 degrees C is similar and is equal to 25-30 minutes . The enzyme is related to endoglucanases weakly adsorbed on cellulose (Kp = 0.065 1/g) . Hydrolysis of microcrystalline cellulose is completed within 7 days (7-9%) and is accompanied by the formation of cellobiose and cellotriose . The enzyme splits dyed lichenan (mixed 1,3-1,4-beta-glucane) at a higher rate than the dyed CM-cellulose . A guinea pig antiserum to enzyme isoforms with a pI of 4.46-4.54 was obtained . Using direct solid phase immunoenzymatic analysis, it was demonstrated that all the enzyme isoforms under study (pI 4.3-4.7) are immunologically related (serum titers for different enzyme isoforms vary from 1:20,000 to 1:50,000) . In the original culture fluid of C . thermocellum, the antigen related to the enzyme isolated from the recombinant strain was unobserved . However, SDS-PAAG electrophoresis of SDS- and mercaptoethanol-treated culture fluids revealed among 11 protein bands at least 4 antigens interacting with antibodies (Mr = 107, 76, 67 and 37 kD), although their antibody titers were far lower and did not exceed 1:300-1:500 . The cumulative data suggest that the endoglucanase under study is not identical to the earlier described enzymes encoded by the cel A- and ceI B-genes of C . thermocellum. Dtsch Tierarztl Wochenschr, 1989 Mar, 96(3), 140 - 3 {Penile inflammation in ganders}; Behr KP et al.; Since 1987 penis-inflammation and -prolapse were observed in north-german breeder-geese . Up to 28% of the ganders showed local symptoms . In the females no clinical signs of cloaca-inflammation were seen . The general condition of the birds was good and there was no increased mortality . Egg production and fertility were not influenced . Bacteriological examinations of the altered penis-tissues revealed in different frequency microorganisms belonging to the genera Mycoplasma, Candida, Bacteroides, Clostridium, Streptococcus, Micrococcus, Staphylococcus, Corynebacterium, Escherichia, Proteus and Pseudomonas . Within 70 days only one third of the diseased ganders recovered completely, the others still showed penis-necrosis or -deformation . It is recommended to examine all ganders prior to each sexual season and to eliminate affected birds. Appl Environ Microbiol, 1989 Mar, 55(3), 656 - 60 Combined effect of water activity and pH on inhibition of toxin production by Clostridium botulinum in cooked, vacuum-packed potatoes; Dodds KL; The effects of water activity (aw, 0.955 to 0.970), pH (4.75 to 5.75), and storage time (up to 60 days) on toxin production by Clostridium botulinum in cooked, vacuum-packed potatoes were studied by using factorial design experiments and most-probable-number methodology . Samples were inoculated with 10(3), 10(4), or 10(5) spores of a mixture of five type A and five proteolytic type B strains, incubated at 25 degrees C, and analyzed for toxin production . Toxin was produced at pH levels of greater than or equal to 4.75 when the aw was greater than or equal to 0.970, pH greater than 5.25 when the aw was 0.965, and pH greater than or equal to 5.75 at an aw of 0.960 . No toxin was detected when the aw was 0.955 . The probability of toxigenesis was significantly affected (P less than 0.0001) by storage time, aw, pH, and the interactions aw.pH and aw.storage time . The response to a decrease in pH was linear, while the response to a decrease in aw was curvilinear . Using multiple linear regression, equations were derived which could predict the length of time until toxin production and the probability of toxigenesis by a single spore under defined conditions. Appl Environ Microbiol, 1989 Mar, 55(3), 559 - 67 Occurrence and distribution of Vibrio spp., Listonella spp., and Clostridium botulinum in the Seto Inland Sea of Japan; Venkateswaran K et al.; The distribution of Vibrio species in samples of surface water, bottom water (water 2 m above the sediment), and sediment from the Seto Inland Sea was studied . A simple technique using a membrane filter and short preenrichment in alkaline peptone water was developed to resuscitate the injured cells, followed by plating them onto TCBS agar . In addition, a survey was conducted to determine the incidence of Clostridium botulinum in sediment samples . Large populations of heterotrophs were found in surface water, whereas large numbers of total vibrios were found in bottom water . In samples from various water sampling regions, high counts of all bacterial populations were found in the inner regions having little exchange of seawater when compared with those of the open region of the inland sea . In the identification of 463 isolates, 23 Vibrio spp . and 2 Listonella spp . were observed . V . harveyi was prevalent among the members of the Vibrio genus . Vibrio species were categorized into six groups; an estimated 20% of these species were in the so-called "pathogenic to humans" group . In addition, a significant proportion of this group was hemolytic and found in the Bisan Seto region . V . vulnificus, V . fluvialis, and V . cholerae non-O1 predominated in the constricted area of the inland sea, which is eutrophic as a result of riverine influence . It was concluded that salinity indirectly governs the distribution of total vibrios and analysis of variance revealed that all bacterial populations were distributed homogeneously and the variance values were found to be significant in some water sampling regions.(ABSTRACT TRUNCATED AT 250 WORDS) Vet Clin North Am Food Anim Pract, 1989 Mar, 5(1), 145 - 57 Infectious diseases of New-World camelids (NWC); Thedford TR et al.; Although there are notable infectious conditions that are capable of producing clinical disease in the NWC, overall, these species are quite healthy . Of the bacterial diseases, enterotoxemia caused by Clostridium perfringens types C and D would be deemed the most significant in North America, while type A also would be regarded as important in South America . Other important bacterial infections of potential concern are tuberculosis, Johne's disease, anthrax, malignant edema, actinomycosis, tetanus, and the South American condition referred to as alpaca fever, which, to date, has not been observed in North America . Fungal infections include classical ringworm, principally caused by Trichophyton spp., and the cases of coccidioidomycosis that are associated with the arid desert lands of the southwestern United States . Most notable of naturally occurring viral infections in the NWC would be rabies, ecthyma, and a recently described blindness neuropathy that has been associated with the equine herpesvirus I . NWC can be infected experimentally with agents causing hoof-and-mouth disease and vesicular stomatitis, but naturally occurring cases do not seem to occur . Serological evidence of exposure to many viral agents, including blue tongue, parainfluenza 3, bovine respiratory syncytial virus, bovine herpesvirus I, bovine viral diarrhea, influenza A, and rotavirus, has been demonstrated; however, no clinical disease associated with these agents, as yet, is apparent. Biol Reprod, 1989 Mar, 40(3), 615 - 21 Alteration of the spermatozoal glycocalyx and its effect on duration of fertility in the fowl (Gallus domesticus); Froman DP et al.; The hypothesis that sialic acid has a role in spermatozoal sequestration within the hen's oviduct was tested by treating spermatozoa with Clostridium perfringens neuraminidase . Spermatozoal content of sialic acid ranged from 94 to 135 micrograms per 10(9) spermatozoa (n = 12 roosters) . Spermatozoa contained 80% of total seminal sialic acid (coefficient of variation = 4.6%) . Spermatozoal sialic acid content was reduced by 18% when 10(9) spermatozoa were incubated at pH 6.5 with 10 IU neuraminidase activity (Type V, Sigma Chemical Co.) . Such treatment had no effect on spermatozoal viability as evidenced by ethidium bromide uptake . However, treatment of spermatozoa with neuraminidase prior to intravaginal insemination reduced fertility by 24 percentage units (p less than 0.001) . In contrast, when similarly treated spermatozoa were deposited in the magnum via laparatomy, fertility was not affected (p greater than 0.05) . The preceding work was done with neuraminidase prepared by salt fractionation (Type V, Sigma Chemical Co.) . Type V neuraminidase was absorbed to diethylaminoethyl-Sephacel and then eluted with a stepwise KCl gradient . Treatment of spermatozoa with this preparation of neuraminidase (10 IU/10(9) spermatozoa) prior to intravaginal insemination reduced fertility by 19 percentage units (p less than 0.001) . Decreased fertility could not be attributed to contamination of neuraminidase preparation with proteolytic activity . We conclude that spermatozoal sialic acid has a role in spermatozoal sequestration within the hen's utero-vaginal glands. Mol Microbiol, 1989 Mar, 3(3), 383 - 92 Phospholipase C and haemolytic activities of Clostridium perfringens alpha-toxin cloned in Escherichia coli: sequence and homology with a Bacillus cereus phospholipase C; Leslie D et al.; The Clostridium perfringens alpha-toxin (phospholipase C) gene (cpa) has been cloned and expressed in Escherichia coli . The biological activities of the cloned gene product have been analysed and the complete nucleotide sequence of the cpa gene has been determined . The cloned cpa gene product, which is exported to the periplasm in E . coli, possesses both phospholipase C and haemolytic activities . Haemolysis is not apparent when cell extracts are incubated with isotonic suspensions of sheep erythrocytes, but can be detected and quantified readily when dilutions of the same extracts are placed in wells in sheep-blood agar plates . Like other sequenced clostridial genes, the cpa gene has a high AT content (66.4%), exhibits a strong bias for using codons with A or T in the wobble position, and the 350 base pairs upstream from the gene have a significantly higher AT content (79.5%) than the coding region . The cpa gene encodes a 398 amino acid polypeptide with a deduced molecular weight of 45,481 D . This is very similar to the estimated molecular weight (Mr) of the cpa primary gene product expressed in an in vitro transcription-translation system (Mr 46,000), but larger than the cpa gene product detected in E . coli minicells, E . coli whole cells or in C . perfringens cells (Mr 43,000), suggesting post-translational processing . The 28 N-terminal residues of the deduced alpha-toxin sequence possess the consensus features of a signal peptide and may be removed during secretion . The deduced alpha-toxin sequence shares significant structural homology with the phosphatidylcholine-preferring phospholipase C of Bacillus cereus. J Bacteriol, 1989 Mar, 171(3), 1346 - 54 Isolation of an atypically small lipoamide dehydrogenase involved in the glycine decarboxylase complex from Eubacterium acidaminophilum; Freudenberg W et al.; The lipoamide dehydrogenase of the glycine decarboxylase complex was purified to homogeneity (8 U/mg) from cells of the anaerobe Eubacterium acidaminophilum that were grown on glycine . In cell extracts four radioactive protein fractions labeled with D-{2-14C}riboflavin could be detected after gel filtration, one of which coeluted with lipoamide dehydrogenase activity . The molecular mass of the native enzyme could be determined by several methods to be 68 kilodaltons, and an enzyme with a molecular mass of 34.5 kilodaltons was obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Immunoblot analysis of cell extracts separated by sodium dodecyl sulfate-polyacrylamide or linear polyacrylamide gel electrophoresis resulted in a single fluorescent band . NADPH instead of NADH was the preferred electron donor of this lipoamide dehydrogenase . This was also indicated by Michaelis constants of 0.085 mM for NADPH and 1.1 mM for NADH at constant lipoamide and enzyme concentrations . The enzyme exhibited no thioredoxin reductase, glutathione reductase, or mercuric reductase activity . Immunological cross-reactions were obtained with cell extracts of Clostridium cylindrosporum, Clostridium sporogenes, Clostridium sticklandii, and bacterium W6, but not with extracts of other glycine- or purine-utilizing anaerobic or aerobic bacteria, for which the lipoamide dehydrogenase has already been characterized. Kansenshogaku Zasshi, 1989 Mar, 63(3), 268 - 72 {Infant botulism was confirmed in Ehime Prefecture}; Nabeya T et al.; The third case of infant botulism in Japan is reported . A four-month-old baby boy suddenly had weakness of suckling force, constipation and generalized hypotonicity . He was the product of normal gestation, labor and delivery . Growth and development were normal until he was nineteen weeks old . He received fruit-juice and honey daily . By bacteriological examination, Clostridium botulism type A was isolated from his feces and the honey which he had received . Type A toxin was detected from his feces but not from his serum. Gene, 1989 Feb 20, 75(2), 349 - 54 Nucleotide sequence analysis and expression studies of a chloramphenicol-acetyltransferase-coding gene from Clostridium perfringens; Steffen C et al.; The nucleotide sequence of a CmR determinant, located on the Clostridium perfringens plasmid pIP401, was determined and its gene product was identified as chloramphenicol acetyltransferase (CAT) . The cat structural gene is preceded by transcription-initiation signals characteristic for Escherichia coli sigma 70 or Bacillus subtilis sigma 43 promoters . By promoter probing in the heterologous hosts the direction of transcription of the clostridial cat gene was analysed and the cat mRNA start point was determined in vitro using the RNA polymerases of E . coli and B . subtilis . Comparison of the amino acid sequences of C . perfringens CAT and other CAT proteins of Gram-positive and Gram-negative origin shows a remarkable degree of homology between the various enzymes. Anaesthesist, 1989 Feb, 38(2), 89 - 94 {Monitoring critically ill patients during transport by helicopter using a patient with abdomen apertum as an example}; Mertzlufft FO et al.; Noninvasive continuous monitoring systems are newly emerging as an important means of monitoring during transports in emergency care, e.g . transportation by helicopter . While automatic oscillometric blood pressure monitors have been used in the perioperative area for some time, a similar development can be observed in the field of emergency care and transportation with the availability of light, portable and battery operated systems . For monitoring adequate oxygenation, pulse oximeters have recently been brought into discussion for both the perioperative period and the transport of critically ill patients . In contrast to well-established monitoring techniques during helicopter transports (ECG, inspection, manually measured blood pressure (BP), pulse oximetry reveals an oxygen deficiency due to respiratory and cardiocirculatory problems, enabling precious time to be saved . This concept is illustrated during the helicopter transport of a critically ill patient with abdomen apertum caused by Clostridium perfringens infection . Even with a critical look at the already described mishaps of this method--e.g . overestimation of true O2 saturation (sO2) and additional overestimation caused by Hb-derivatives--pulse oximetry was found to be superior to the established monitoring techniques . Furthermore, oscillometric blood pressure detection was very satisfactory during the 30-min helicopter transport . Based on our results, we believe pulse oximetry and automatic oscillometric BP-measurement to be useful for monitoring during transports in helicopters, thus improving patient safety. Appl Environ Microbiol, 1989 Feb, 55(2), 323 - 9 Coenzyme A transferase from Clostridium a