Acta Pathol Microbiol Immunol Scand {B}, 1987 Feb, 95(1), 41 - 7 Phenotypic characterization of Flavobacterium meningosepticum strains identified by DNA-DNA hybridization; Bruun B et al.; Fifty-two strains found to belong to Flavobacterium meningosepticum on the basis of DNA-DNA hybridization analyses were characterized and found to be phenotypically homogeneous . All strains were oxidase, catalase, indole, gelatinase and beta-galactosidase positive, and produced acid from glucose, mannose, fructose, maltose and mannitol; nitrate was not reduced . F . meningosepticum could be differentiated from Flavobacterium group IIb by its ability to produce beta-galactosidase, and by the latter taxon's ability to produce a bright yellow pigment in contrast to the weak or non-existing pigmentation of F . meningosepticum . Phenotypic characteristics that could differentiate between the two main DNA relatedness groups of F . meningosepticum were not discovered, wherefore a subdivision of the present species into two species cannot be recommended . Strains from the DNA relatedness groups comprising 19 of 20 CSF isolates were found to be able to grow at 40 degrees C and to produce a weak yellow pigment; in contrast, strains from the three other DNA relatedness groups were unable to grow at 40 degrees C and produced no pigment. Acta Pathol Microbiol Immunol Scand {B}, 1987 Feb, 95(1), 33 - 9 Genetic heterogeneity of Flavobacterium meningosepticum demonstrated by DNA-DNA hybridization; Ursing J et al.; DNA-DNA reassociation studies on 52 strains of Flavobacterium meningosepticum showed two main hybridization groups about 40-55% interrelated and comprising 4 and 48 strains respectively . The larger group could be further divided into four subgroups if differences in thermal stability of the reassociated duplexes were taken into consideration . Of a total of 20 strains isolated from cerebrospinal fluid, 18 were found in one subgroup of 28 strains, which also contained seven blood isolates . The results indicate that genetically defined subgroups within the species might differ with regard to pathogenic significance. Med Microbiol Immunol (Berl), 1987, 176(2), 103 - 11 Flavobacterium group IIb bacteremia: report of a case and review of Flavobacterium infections; Siegman-Igra Y et al.; A case of nonfatal Flavobacterium CDC group IIb bacteremia in a hospitalized woman is presented . The portal of entry of the organism was either an intravenous line or an open, eroded wound associated with bilateral breast carcinoma . Flavobacterium infections in the post-neonatal age group are reviewed. J Appl Bacteriol, 1987 Jan, 62(1), 29 - 41 Identification and distribution of Flavobacterium meningosepticum in clinical material; Holmes B; During the 19 year period ending December 1984, 82 (1.7%) of 4840 strains of Gram-negative non-fermentative bacteria submitted to the National Collection of Type Cultures (NCTC) for computer-assisted identification were Flavobacterium meningosepticum . These figures suggest either that F . meningosepticum occurs only rarely in clinical material in the UK, or that when it does occur it is not always recognized . The sources from which the NCTC strains were isolated are reported and also the characteristics by which the species may be identified . The clinical significance of F . meningosepticum is reviewed, particularly its role in neonatal meningitis, of which there have been but two reported cases in the UK, both imported . The susceptibility of the species to antimicrobial agents is discussed and also antimicrobial therapy of neonatal meningitis due to this species. Scand J Infect Dis Suppl, 1987, 52, 26 - 31 Intramuscular imipenem/cilastatin for treatment of mild and moderately severe bacterial infections; Keller CA et al.; The efficacy and safety of intramuscularly administered imipenem/cilastatin was studied in 70 patients with mild or moderately severe bacterial infections (skin and soft tissue infections, respiratory tract infections, urinary tract infections and pelvic infections) . Doses of imipenem/cilastatin ranged from 0.5 to 0.75 g twice daily . Fifty-five patients were evaluable for bacteriological efficacy; in the remaining 15 patients no pathogens were isolated or susceptibility data were lacking . MIC50 and MIC90 of imipenem were 0.12 mg/l and 0.5 mg/l, respectively, for Gram-negative pathogens isolated and 0.25 mg/l and 0.5 mg/l, respectively, for Gram-positive pathogens . Only one strain (a Flavobacterium odoratum) was resistant to imipenem . Clinical cure and bacteriological elimination was achieved in 94% of evaluable patients while 3% showed marked clinical improvement . Two patients were considered therapeutic failures . No clinical adverse effects were noted . Abnormal liver transaminases were recorded in 23% of the patients and 11% developed eosinophilia . In no patient was imipenem/cilastatin discontinued due to adverse effects . It is concluded that intramuscular imipenem/cilastatin in these patients was well tolerated and efficacious. Biochem Biophys Res Commun, 1986 Dec 15, 141(2), 825 - 30 Catabolism of pentachlorophenol by a Flavobacterium sp; Steiert JG et al.; The pathway employed for pentachlorophenol (PCP) degradation by an aerobic, chlorophenol-utilizing Flavobacterium sp . was initiated by conversion of PCP to tetrachloro-p-hydroquinone (TCH) . 18O labelling experiments demonstrated that the first dechlorination, where a hydroxyl replaced the chlorine at PCP ring position number 4, involved a hydrolytic reaction . Then two reductive dechlorinations of TCH followed to yield firstly trichlorohydroquinone (TrCH) and then 2,6-dichlorohydroquinone (DCH) . Thus, the initial steps in catabolism of PCP by the Flavobacterium were: PCP----TCH----TrCH. FEBS Lett, 1986 Dec 15, 209(2), 235 - 7 Specificity of prolyl endopeptidase; Nomura K; A series of tetrapeptides, Cbz(Bz)-Gly-X-Leu-Gly, were synthesized and the kinetic parameters, kcat and kcat/Km, determined for their hydrolyses by prolyl endopeptidase from Flavobacterium . The peptides with X = N-Me-Ala, Sar and Ala as well as the standard substrate (X = Pro) were found to be good substrates, while those with X = alpha-aminobutyryl, Hyp, Ser and Gly were poor substrates, and those with X = pipecolyl, alpha-aminoisobutyryl, N-Me-Val, N-Me-Leu, Hyp(O-Bzl) and Ser(O-Bzl) were not cleaved at all . These results suggest that the specificity-determining site or S1 subsite of the enzyme is designed to fit exactly the proline residue of the substrate with allowance for the residues carrying substituents at the N and/or C alpha which must not exceed the size of the pyrrolidine ring of proline. Thromb Res, 1986 Dec 1, 44(5), 599 - 610 Removal of the anticoagulant activities of the low molecular weight heparin fractions and fragments with flavobacterial heparinase; Yang VC et al.; Recently, the development of low molecular weight heparin fractions and fragments (LMHF) as potential antithrombotic agents has gained increased attention . However, the lack of antagonists to neutralize the anticoagulant effects of these drugs may seriously exclude them from possible uses in extracorporeal therapy . This is mainly because of the concern that the high dosage of the drugs employed in extracorporeal therapy could lead to serious bleeding risks . Our earlier work has demonstrated that immobilized heparinase can remove polydisperse heparin both in vitro and in vivo . To examine whether such a system may be used as a novel approach to neutralize the anticoagulant effects of LMHF, different LMHF were tested using heparinase . In vitro data showed that both the APTT and anti-FXa activities of the LMHF including Kabi 2165, PK 10169, Cy 216 and CY 222 were nearly completely eliminated by heparinase in less than 20 min . This study suggests that an immobilized heparinase system may be an useful element for the acceptance of the LMHF for their use in extracorporeal therapy. J Appl Bacteriol, 1986 Nov, 61(5), 421 - 6 A note on starch hydrolysis and beta-glucuronidase activity among flavobacteria; Petzel JP et al.; Most flavobacteria tested with the fluorogenic substrate 4-methylumbelliferyl-beta-D-glucuronide possessed beta-glucuronidase (GUD), but when some of the same strains were tested with the API ZYM gallery, all were negative for GUD . Conflicting reports also appear in the literature about starch hydrolysis among flavobacteria . We observed that the results obtained can depend on the medium used and the length of incubation . Our results indicate that GUD activity and starch hydrolysis are more widely distributed in the genus Flavobacterium than previously reported. Atherosclerosis, 1986 Nov, 62(2), 151 - 8 Effect of very low molecular weight heparin-derived oligosaccharides on lipoprotein lipase release in rabbits; Merchant ZM et al.; Oligosaccharide fragments of heparin were prepared using flavobacterial heparinase . Following sizing, these oligosaccharide fractions were administered (i.v.) to rabbits and were examined for their ability to release lipoprotein lipase . The decasaccharides (dp = 10, Mr avg = 2,800) were the smallest oligosaccharides which resulted in substantial lipase release . The plasma lipase levels obtained with decasaccharides were comparable to low molecular weight heparin and one-third those obtained when heparin was administered at an equivalent dose . The peak plasma lipase concentration was observed 10 min following heparinization and fell off rapidly over the 60-min time course . The lipase release activity paralleled the in vivo pharmacokinetics of the heparin and decasaccharide sample as determined by monitoring their anti-Factor Xa activity . No activation of purified bovine milk lipoprotein lipase or plasma lipase was detectable at the concentrations studied, indicating that the increase in circulating lipolytic activity was due entirely to release . Lipoprotein lipase accounted for a major portion of the released activity with hepatic triglyceride lipase representing the remainder of the lipolytic activity . The sized decasaccharide sample was characterized with regards to its structure and anticoagulant activity . The decasaccharides exhibited reduced anticoagulant activity possibly making it a better drug candidate in the treatment of atherosclerosis. Arch Microbiol, 1986 Nov, 146(2), 125 - 9 Biodegradation of polyethylene glycol by symbiotic mixed culture (obligate mutualism); Kawai F et al.; Neither Flavobacterium sp . nor Pseudomonas sp . grew on a polyethylene glycol (PEG) 6000 medium containing the culture filtrate of their mixed culture on PEG 6000 . The two bacteria did not grow with a dialysis culture on a PEG 6000 medium . Flavobacterium sp . grew well on a dialysis culture containing a tetraethylene glycol medium supplemented with a small amount of PEG 6000 as an inducer, while poor growth of Pseudomonas sp . was observed . Three enzymes involved in the metabolism of PEG, PEG dehydrogenase, PEG-aldehyde dehydrogenase and PEG-carboxylate dehydrogenase (ether-cleaving) were present in the cells of Flavobacterium sp . The first two enzymes were not found in the cells of Pseudomonas sp . PEG 6000 was degraded neither by intact cells of Flavobacterium sp . nor by those of Pseudomonas sp., but it was degraded by their mixture . Glyoxylate, a metabolite liberated by the ether-cleaving enzyme, inhibited the growth of the mixed culture . The ether-cleaving enzyme was remarkably inhibited by glyoxylate . Glyoxylate was metabolized faster by Pseudomonas sp . than by Flavobacterium sp., and seemed to be a key material for the symbiosis. J Biochem (Tokyo), 1986 Oct, 100(4), 1049 - 55 Purification and properties of glucoside 3-dehydrogenase from Flavobacterium saccharophilum; Takeuchi M et al.; A membrane-bound glucoside 3-dehydrogenase {EC 1.1.99.13}, which oxidizes validoxylamine A to the 3-keto derivative, was solubilized from the membrane fraction of Flavobacterium saccharophilum by Triton X-100 and purified about 280-fold with an overall yield of 30% from the membrane fraction by column chromatography on DEAE- and CM-Sepharose CL-6B and gel filtration on Sephacryl S-300 . The purified enzyme exhibited a single protein band on disc gel electrophoresis, and FAD was shown to be the prosthetic group . The enzyme had a molecular weight of 270,000 as determined by gel filtration on Sephacryl S-300 and consisted of 4 identical subunits each with a molecular weight of 66,000 . The enzyme reacted with various artificial electron acceptors such as 2,6-dichlorophenolindophenol (DCIP), phenazine methosulfate, and ferricyanide . The optimum pH for DCIP reductase activity was 6.0 . The enzyme was inhibited by Hg2+ and p-chloromercuribenzoate . D-Glucose and methyl-alpha- and beta-D-glucoside showed the highest susceptibility to the enzyme, and were converted to the corresponding 3-keto sugars. J Gen Microbiol, 1986 Oct, 132 ( Pt 10), 2723 - 32 Characterization of Flavobacterium species by analysis of volatile fatty acid production; Rasoamananjara D et al.; Seventy-four Flavobacterium strains were characterized by gas-liquid chromatographic analysis of volatile fatty acids produced in the culture medium . Principal components analysis permitted the graphic representation of the relative positions of the different strains, and aggregation according to the variance enabled a hierarchical classification to be established . The study revealed three subgroups each for F . meningosepticum and F . odoratum . Our F . breve, Flavobacterium sp . group IIb and F . multivorum strains appeared to be homogeneous . These results tallied with those of previous studies on DNA base composition and reassociation, electrophoretic protein profiles and cellular fatty acid composition. J Biol Chem, 1986 Sep 15, 261(26), 12000 - 5 Transfer of glycerol by Endo-beta-N-acetylglucosaminidase F to oligosaccharides during chitobiose core cleavage; Trimble RB et al.; N-Linked oligosaccharides, when hydrolyzed by glycerol-containing preparations of endo-beta-N-acetylglucosaminidase (Endo) F from Flavobacterium meningosepticum were found to have glycerol attached to their reducing ends . The absence of a reducing end was confirmed by high-field 1H NMR spectroscopy, and the incorporated glycerol was verified through mass spectrometry and collisionally activated decomposition fast atom bombardment/mass spectrometry/mass spectrometry techniques . Periodate oxidation of {1(3)-14C}glycerol-labeled oligosaccharides indicated glycerol was glycosidically linked via its 1(3) carbon to the C1 of the reducing end N-acetylglucosamine . In a second, less favored reaction, the glycerol glycoside was hydrolyzed by Endo F using water as the terminal nucleophile, thus regenerating the N-acetylglucosamine reducing end . Glycerol could be removed from Endo F preparations without affecting enzyme stability, and chitobiosyl core hydrolysis in its absence provided intact oligosaccharides with normal N-acetylglucosamine reducing ends . The incorporation of labeled glycerol may provide a useful method for monitoring of Endo F release of oligosaccharides. Quad Sclavo Diagn, 1986 Sep, 22(3), 318 - 29 {Isolation of Flavobacterium odoratum from human matter}; Andreoni S; Described the most important taxonomical, bacteriological, clinical and epidemiological aspects of microorganisms of Flavobacterium genus, the author refer about 8 strains of Flavobacterium odoratum isolated from urine (6), sputum (1), surgical exudate (1) . In some cases, etiopathogenetic possible correlation between isolates and clinical disease discussed, the author emphasizes the large antibiotic resistance kind of this species of microorganism non fermenter. Mikrobiologiia, 1986 Sep-Oct, 55(5), 872 - 4 {Coagulation of bacteria by the action of montmorrillonite}; Nikovskaia GN et al.; Conditions were studied for the coagulation of aggregation-stable Bacillus subtilis, Flavobacterium rigens and Escherichia coli cell dispersions . Their aggregative stability decreased when montmorillonite, a clay mineral, was added to a weakly acid medium or in the presence of Al3+ ions . Bacterial heterocoagulation under the action of montmorillonite can be used as a universal strategy for biomass isolation from aqueous dispersion media. J Biochem (Tokyo), 1986 Sep, 100(3), 773 - 80 Substrate specificity of endo-beta-galactosidases from Flavobacterium keratolyticus and Escherichia freundii is different from that of Pseudomonas sp; Ito M et al.; The substrate specificity of endo-beta-galactosidase of Pseudomonas sp . was found to differ from that of Flavobacterium keratolyticus or Escherichia freundii, based on the following experimental results . The endo-beta-galactosidases from these three bacteria released 6-O-sulfo-GlcNAc beta 1-3Gal as one of the major products from keratan sulfates from different sources . In addition to the sulfated disaccharide, Flavobacterium and Escherichia enzymes produced GlcNAc beta 1-3Gal, which is also an integral repeating unit of keratan sulfate, whereas the Pseudomonas enzyme did not release any non-sulfated disaccharide . Tetrasaccharides were prepared from the teleost skin keratan sulfate by digestion with Pseudomonas enzyme followed by gel filtration on Sephadex G-50 chromatography . A part of the tetrasaccharide fraction was hydrolyzed by Flavobacterium enzyme to produce 6-O-sulfo-GlcNAc beta 1-3Gal and GlcNAc beta 1-3Gal, whereas the fraction was completely resistant to retreatment with the Pseudomonas enzyme . Endo-beta-galactosidases from F . keratolyticus and E . freundii hydrolyzed the internal beta-1,4-galactosyl linkage of various neolacto-type glycosphingolipids to produce glucosylceramides . However, these glycosphingolipids were completely resistant to the Pseudomonas enzyme . These findings clearly show that the sulfation on the N-acetylglucosamine adjacent to galactose in the lactosaminoglycans is essential for expression of the Pseudomonas enzyme, but not for that of the Flavobacterium or Escherichia enzyme. Biochimie, 1986 Sep, 68(9), 1087 - 96 Isolation and some properties of an arylamidase from Flavobacterium IIb; Milliere JB et al.; A constitutive L-leucylarylamidase (EC 3.4.11) hydrolase able to cleave L-aminoacyl-beta naphthylamide and L-aminoacyl-4 nitroanilide substrates, was isolated from sonicated cells of Flavobacterium IIb and partially purified with a 0.9% yield and a 159-fold recovery . Its molecular weight was estimated to be about 170,000 +/- 10% . This arylamidase exhibited optimum activity at pH 7.0 and 28 degrees C for the hydrolysis of L-leucine-4NA and is inhibited strongly by metal chelating agents, and to a weaker extent, by some sulfhydryl and reducing agents . Heavy metal ions: Cd2+, Zn2+, Cu2+, Hg2+ and Co2+, markedly inhibit it, and Zn2+ is a competitive inhibitor . This metalloenzyme, free of carboxypeptidase, proteinase and L-leucine aminopeptidase (L-leucylglycine substrate) activities, hydrolyzes aminoacyl-beta NA, aminoacyl-4NA and some dipeptides with unsubstituted amino groups of the L-configuration . The lowest Km values are associated with substrates having neutral or basic residues, with large side chains. Appl Environ Microbiol, 1986 Jul, 52(1), 92 - 7 Pentachlorophenol degradation: a pure bacterial culture and an epilithic microbial consortium; Brown EJ et al.; The steady-state growth of a Flavobacterium strain known to utilize pentachlorophenol (PCP) was examined when cellobiose and PCP simultaneously limited its growth rate in continuous culture . A concentration of 600 mg of PCP per liter in influent medium could be continuously degraded without affecting steady-state growth . We measured specific rates of PCP carbon degradation as high as 0.15 +/- 0.01 g (dry weight) of C per h at a growth rate of 0.045 h-1 . Comparable specific rates of PCP degradation were obtained and maintained by PCP-adapted, natural consortia of epilithic microorganisms . The consortium results suggest that a fixed-film bioreactor containing a PCP-adapted natural microbial population could be used to treat PCP-contaminated water. J Biochem (Tokyo), 1986 Jun, 99(6), 1571 - 7 Chemical modification by diethylpyrocarbonate of an essential histidine residue in 3-ketovalidoxylamine A C-N lyase; Takeuchi M et al.; 3-Ketovalidoxylamine A C-N lyase of Flavobacterium saccharophilum is a monomeric protein with a molecular weight of 36,000 . Amino acid analysis revealed that the enzyme contains 5 histidine residues and no cysteine residue . The enzyme was inactivated by diethylpyrocarbonate (DEP) following pseudo-first order kinetics . Upon treatment of the inactivated enzyme with hydroxylamine, the enzyme activity was completely restored . The difference absorption spectrum of the modified versus native enzyme exhibited a prominent peak around 240 nm, but there was no absorbance change above 270 nm . The pH-dependence of inactivation suggested the involvement of an amino acid residue having a pKa of 6.8 . These results indicate that the inactivation is due to the modification of histidine residues . Substrates of the lyase, p-nitrophenyl-3-ketovalidamine, p-nitrophenyl-alpha-D-3-ketoglucoside, and methyl-alpha-D-3-ketoglucoside, protected the enzyme against the inactivation, suggesting that the modification occurred at or near the active site . Although several histidine residues were modified by DEP, a plot of log (reciprocal of the half-time of inactivation) versus log (concentration of DEP) suggested that one histidine residue has an essential role in catalysis. Acta Pathol Microbiol Immunol Scand {B}, 1986 Jun, 94(3), 145 - 52 An investigation of a collection of yellow-pigmented Pseudomonas; Sogaard P et al.; Thirty-six strains of yellow-pigmented Pseudomonas from clinical as well as non-clinical material and 11 reference strains of Pseudomonas were investigated by means of conventional bacteriological methods (a total of 53 different tests) . Eighteen of the 36 yellow-pigmented strains could be classified as P . paucimobilis . Apart from the presence of lipid inclusions on beta-hydroxybutyrate, hydrolysis of DNA, and Tween 80 our results showed a high degree of accordance with other investigations . Eight strains showed characteristics compatible with inclusion in the CDC VE group; one orange-yellow strain showed the characteristics of P . vesicularis, and one was a pyoverdin negative, yellow P . putida . Eight strains remained unidentified . Strains of P . paucimobilis were most often resistant to antibiotics used for P . aeruginosa infections (viz . piperacillin, cefsulodin, ceftazidime) while the strains of the CDC VE group were often susceptible . Most strains were susceptible to the aminoglycosides . The difficulties in distinguishing yellow-pigmented strains of Pseudomonas from Flavobacterium spp . or Xanthomonas spp . are discussed. Am J Physiol, 1986 May, 250(5 Pt 2), H879 - 88 Anticoagulantly active heparin-like molecules from mast cell-deficient mice; Marcum JA et al.; To assess the contribution of mast cells to the maintenance of blood fluidity, the hindlimb vasculature of mast cell-deficient mice (W/Wv) and littermates containing normal levels of mast cells (+/+), were perfused with purified human thrombin and antithrombin . Enzyme-inhibitor complex generation within the vasculature was enhanced to a comparable extent for W/Wv and +/+ mice over the uncatalyzed rate, that level of complex produced within a similar time interval in the absence of heparin . Perfusion of purified Flavobacterium heparinase prior to infusion of the hemostatic components, or perfusion of antithrombin modified at the heparin-binding domain, reduced W/Wv and +/+ hindlimb thrombin-antithrombin complex formation to the uncatalyzed rate . To further define the cellular source of the vascular-associated heparin-like molecules, endothelial cells isolated from epididymal fat pads of W/Wv and +/+ mice were grown in vitro . The acceleration of thrombin-antithrombin interactions in the presence of endothelial cell-derived glycosaminoglycans was similar for W/Wv and +/+ mice, was abolished with purified bacterial heparinase, and was expressed to only a minor extent when utilizing modified antithrombin . The biologically active mucopolysaccharides appear to be present on the cell surface. Appl Environ Microbiol, 1986 May, 51(5), 926 - 30 Identification of a plasmid-borne parathion hydrolase gene from Flavobacterium sp . by southern hybridization with opd from Pseudomonas diminuta; Mulbry WW et al.; Parathion hydrolases have been previously described for an American isolate of Pseudomonas diminuta and a Philippine isolate of Flavobacterium sp . (ATCC 27551) . The gene which encodes the broad-spectrum organophosphate phosphotriesterase in P . diminuta has been shown by other investigators to be located on a 66-kilobase (kb) plasmid . The intact gene (opd, organophosphate-degrading gene) from this degradative plasmid was cloned into M13mp10 and found to express parathion hydrolase under control of the lac promoter in Escherichia coli . In Flavobacterium sp . strain ATCC 27551, a 43-kb plasmid was associated with the production of parathion hydrolase by curing experiments . The M13mp10-cloned fragment of the opd gene from P . diminuta was used to identify a homologous genetic region from Flavobacterium sp . strain ATCC 27551 . Southern hybridization experiments demonstrated that a genetic region from the 43-kb Flavobacterium sp . plasmid possessed significant homology to the opd sequence . Similar hybridization did not occur with three other native Flavobacterium sp . plasmids (approximately 23, 27, and 51 kb) present within this strain or with genomic DNA from cured strains . Restriction mapping of various recombinant DNA molecules containing subcloned fragments of both opd plasmids revealed that the restriction maps of the two opd regions were similar, if not identical, for all restriction endonucleases tested thus far . In contrast, the restriction maps of the cloned plasmid sequences outside the opd regions were not similar . Thus, it appears that the two discrete bacterial plasmids from parathion-hydrolyzing soil bacteria possess a common but limited region of sequence homology within potentially nonhomologous plasmid structures. J Nutr Sci Vitaminol (Tokyo), 1986 Apr, 32(2), 137 - 45 Menaquinone (vitamin K2)-6 production by mutants of Flavobacterium meningosepticum; Tani Y et al.; Flavobacterium meningosepticum IFO 12535, a menaquinone (MK) producer, was mutagenized to improve productivity . A mutant, which was resistant to 1-hydroxy-2-naphthoate (HNA), was found to produce MK more abundantly: 34 mg/liter of culture broth and 5.5 mg/g of dry cells . The mutant was less sensitive to inhibition by HNA on MK biosynthesis than the wild-type strain . MK was isolated from cells of the mutant and identified as MK-6 based on its physicochemical characteristics . Mutants, which were given each KCN resistance, aromatic amino acid auxotrophy and no carotenoid productivity, did not show further increase of productivity. South Med J, 1986 Apr, 79(4), 518 - 9 Sepsis from Flavobacterium meningosepticum, an uncommon pathogen with unusual susceptibility patterns; Burnakis TG et al.; Flavobacterium meningosepticum, because of its unusual susceptibility patterns and predilection for debilitated or immunocompromised hosts, can be a dangerous pathogen . Identification of the organism and proper susceptibility studies, which are essential in designing antibiotic therapy, helped in the selection of a regimen that proved curative in this case. Infect Control, 1986 Apr, 7(4), 216 - 9 Microbial examination of kidney lithotripter tub water and epidural anesthesia catheters; Cooper GL et al.; Kidney lithotripsy patients frequently receive epidural anesthesia via indwelling epidural catheters . In our hospital, patients are immersed in a tub of warm, continuously-flowing tap water . The epidural catheter-entry site is covered by a transparent occlusive dressing . To determine the risk of microbial colonization of the epidural catheter during lithotripsy, we performed quantitative cultures of tub water and semiquantitative cultures of catheters in 63 lithotripsy procedures . Most of the tub water organisms were typical tap water and skin flora isolates . Total colony counts were generally low with no significant progression during the course of serial procedures . Forty-two epidural catheters were cultured; 34 (81%) were sterile, 8 (19%) were colonized with small numbers of flavobacteria or coagulase-negative staphylococci . Only four catheters had organisms present on catheter segments covered by the transparent occlusive dressing (in each case there was a single colony forming unit per semiquantitative plate) and these organisms were probable contaminants . We conclude that with our current lithotripsy procedures, the risk for the development of epidural catheter-associated infection seems to be low. Appl Environ Microbiol, 1986 Mar, 51(3), 640 - 6 Sulfur regulation of heparinase and sulfatases in Flavobacterium heparinum; Cerbelaud EC et al.; Sulfur regulation of heparinase synthesis and sulfatase synthesis was studied in Flavobacterium heparinum . Heparinase synthesis was strongly repressed by sulfate and L-cysteine, while the activity of this enzyme showed little or no inhibition by these compounds . Heparinase was synthesized in the absence of heparin when L-methionine was used as the sole sulfur source . The sulfatases produced by F . heparinum, which include the sulfatases involved in heparin catabolism, were also studied . At least some of the sulfatase activity was regulated by sulfur compounds in a manner similar to heparinase regulation . L-Cysteic acid and taurine were not suitable sulfur sources to support the growth of F . heparinum. Vet Microbiol, 1986 Mar, 11(3), 261 - 70 Phenotypic and genetic relationships of so-called Moraxella (Pasteurella) anatipestifer to the Flavobacterium/Cytophaga group; Piechulla K et al.; So-called Moraxella (or Pasteurella) anatipestifer and members of the Flavobacterium/Cytophaga group exhibit remarkable common features: lack of flagellation, low guanine + cytosine content of the chromosomal DNA, production of menaquinones and branched-chain fatty acids, absence of carbohydrate fermentation, and similar patterns of hydrolytic enzymes . Using the renaturation method of DNA:DNA hybridization two urease-negative European isolates and the urease-positive type strain (which was isolated in the United States) of M . P . anatipestifer were shown to have about 85% of their genome DNA base sequences in common; they may represent two subspecies . The type strain of this species was neither measurably related to the type species of the genus Moraxella nor to selected members of the family Pasteurellaceae (Pohl 1981) . On the other hand, low but significant degrees of DNA binding between selected strains of so-called M . anatipestifer, Cytophaga marinoflava, Flavobacterium meningosepticum, F . odoratum and F . pectinovorum were observed . On the basis of these findings the transfer of the so-called M . anatipestifer to the Flavobacterium/Cytophaga group (family Cytophagaceae) is proposed . More detailed investigations are required to establish its relationship at the genus level. Carbohydr Res, 1986 Mar 1, 147(1), 87 - 100 Structural differences of dermatan sulfates from different origins; Poblacion CA et al.; The dermatan sulfates from hog, rat, rabbit, and beef liver, hog, rat, beef, and dog spleen, and hog skin were isolated and submitted to structural analysis . All of them migrated as single bands, close to the standard position for dermatan sulfate in agarose-gel electrophoresis . In polyacrylamide gel, however, each dermatan sulfate showed a characteristic electrophoretic migration-pattern: one, two, or three polydisperse bands, corresponding to different molecular weights, were obtained for the dermatan sulfates according to their origins . Chemical analysis showed that all of the dermatan sulfates here described are hybrid polymers composed of D-glucuronic and L-iduronic acid-containing disaccharide units . The relative position of these units in the polymer chains and the presence of 6-sulfated disaccharides were determined with the aid of chondroitinases B and AC from Flavobacterium heparinum . These studies show that each dermatan sulfate has a unique structure as regards the molecular weight, the presence of 6-sulfated disaccharide units, and also the relative amount and position of glucuronic and iduronic acid residues in the chains . These findings suggests a tissue- and species-specificity for the dermatan sulfates. Proc Soc Exp Biol Med, 1986 Mar, 181(3), 432 - 7 Deglycosylation of gonadotropins with an endoglycosidase; Swedlow JR et al.; A commercially available endoglycosidase (N-glycanase, Genzyme, Boston, Mass.) purified from Flavobacterium meningosepticum with a specificity for cleaving asparagine-linked carbohydrate moieties in glycoproteins was tested on several pituitary and chorionic gonadotropins as substrates . All intact hormones tested were resistant to the action of the enzyme as were all beta subunits from the respective gonadotropins . All alpha subunits, however, were susceptible to the enzyme as evidenced by a decrease in molecular size when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) . Preparative experiments with ovine luteinizing hormone subunit (oLH alpha) indicated that only 35-40% of the carbohydrate was removed after N-glycanase treatment, suggesting that perhaps only one of the two carbohydrate moieties was cleavable under the conditions employed . The enzyme-modified subunit (DG-oLH alpha) was able to recombine with untreated oLH beta . An in vitro steroidogenic bioassay (rat Leydig cell) showed that the recombinant (DG-oLH alpha-oLH beta) was about 22% as potent as the native oLH, but in a testicular membrane binding assay for LH, it was equal in potency to the native hormone in competing with the radioligand. J Clin Microbiol, 1986 Feb, 23(2), 267 - 73 Chemical characterization of Flavobacterium odoratum, Flavobacterium breve, and Flavobacterium-like groups IIe, IIh, and IIf; Dees SB et al.; The cellular fatty acid, sphingolipid, and isoprenoid quinone compositions of Flavobacterium odoratum, Flavobacterium breve, and Flavobacterium-like groups IIe, IIh, and IIf were determined, using thin-layer, gas-liquid, and reverse-phase high-performance liquid chromatography . The fatty acid data showed that groups IIe, IIh, and IIf were similar to recognized Flavobacterium species by the presence of relatively large amounts of iso-branched hydroxy and nonhydroxy acids . Groups IIe and IIh were essentially identical in fatty acid composition but were distinguished from group IIf, F . breve, and F . odoratum on the basis of minor qualitative and quantitative differences . All strains tested contained menaquinone 6 as the major isoprenoid quinone, and all lacked sphingolipids . Overall, the chemical data suggest that groups IIe, IIh, and IIf are additional Flavobacterium species and are different from sphingobacteria, which contain sphingolipid and menaquinone 7 as the major quinone. Biochemistry, 1986 Jan 28, 25(2), 405 - 10 Effects of heparan sulfate removal on attachment and reattachment of fibroblasts and endothelial cells; Gill PJ et al.; Human skin fibroblasts and calf aorta endothelial cells were grown as tissue culture monolayers in the presence of {35S}sulfate in order to label the glycosaminoglycan portions of proteoglycans for investigation of their role in cell attachment . The {35S}glycosaminoglycans were then selectively removed from the cell monolayers by the addition of various glycosaminoglycan-degrading enzymes . As previously described, in contrast to trypsin treatment none of these enzymes removed any cells from the culture plates . Incubation with a preparation from Flavobacterium heparinum left only small stubs of {35S}glycosaminoglycans on the cell monolayers, indicating that all the cell-surface proteoheparan {35S}sulfate and proteochondroitin {35S}sulfate was accessible to this enzyme preparation . The treatment did not change the amount or time of incubation with trypsin necessary for release of the cells from the monolayers . Thus, cell attachment was not weakened by removal of heparan sulfate or chondroitin sulfate . In contrast, neither fibroblasts nor endothelial cells in suspension would reattach in the presence of the F . heparinum preparation while reattachment occurred readily in the presence of chondroitin ABC lyase . This provides evidence that heparan sulfate, but not chondroitin sulfate, is involved in the process of cell attachment even though neither is necessary for maintaining attachment. J Biol Chem, 1986 Jan 5, 261(1), 172 - 7 Requirements of cleavage of high mannose oligosaccharides in glycoproteins by peptide N-glycosidase F; Chu FK; Peptide N-glycosidase from Flavobacterium meningosepticum cleaves complex as well as neutral glycoproteins (Plummer, T.H., Jr., Elder, J.H., Alexander, S., Phelan, A.W., and Tarentino, A.L . (1984) J . Biol . Chem . 259, 10700-10704) . Examples of neutral glycoprotein substrates include ribonuclease B (one high mannose oligosaccharide chain) and yeast external invertase (nine chains/invertase subunit) . The rate of deglycosylation by the glycosidase was greatly enhanced if the glycoprotein substrate was denatured prior to enzyme treatment, from a low of 11-fold for external invertase to a high of 844-fold for ribonuclease B . Peptide N-glycosidase F was unable to cleave the asparaginyl-N-acetylglucosamine bond in endo-beta-N-acetylglucosaminidase H-modified external invertase or ribonuclease B, although that in similarly modified glycopeptide substrate was cleaved . Ribonuclease B was digested sequentially with various exoglycosidases to produce an oligosaccharide chain of varied length . Using the resulting forms of ribonuclease B as substrates for peptide N-glycosidase F, the minimum oligosaccharide chain for cleavage was the di-N-acetyl-chitobiosyl core unit. Ann Biol Clin (Paris), 1986, 44(1), 35 - 8 {Phenotype characteristics of 63 strains of Pseudomonas paucimobilis, mostly of hospital origin}; Richard C; Sixty-three strains of Pseudomonas paucimobilis, a Gram-negative, lemon yellow pigmented, glucose nonfermenting bacillus are phenotypically compared to the type strain of this new species (ATCC 29837) . The morphological, cultural, biochemical (ONPG-test, oxidase, aesculin positive) and nutritional (assimilation of many glucides) characteristics and susceptibility or resistance to antimicrobial agents (many antibiotics are active) are reported . The similarities and differences between P . paucimobilis and other yellow pigmented bacteria (Flavobacterium, especially group IIb, Xanthomonas, Pseudomonas maltophilia, stutzeri and cepacia) are presented . The occurrence of P . paucimobilis in diverse aquatic and aqueous sources and hospital environments and its significance in medical bacteriology (many isolates from blood cultures) are discussed. Diagn Microbiol Infect Dis, 1986 Jan, 4(1), 65 - 9 Flavobacterium meningosepticum bacteremia in an adult with acute leukemia . Use of rifampin to clear persistent infection; Hirsh BE et al.; A case of Flavobacterium meningosepticum bacteremia complicating the course of a patient with leukemia is described . The patient was treated successfully when rifampin was added to the antibiotic therapy . Unusual organisms should be considered as agents of infection in immunocompromised hosts and susceptibility testing with drugs not commonly employed for gram-negative rods should be performed in complicated cases. J Biochem (Tokyo), 1985 Dec, 98(6), 1631 - 8 Purification and properties of 3-ketovalidoxylamine A C-N lyase from Flavobacterium saccharophilum; Takeuchi M et al.; 3-Ketovalidoxylamine A C-N lyase was purified about 900-fold from the cell-free extract of Flavobacterium saccharophilum by ammonium sulfate fractionation, column chromatography on CM cellulose and gel filtration on Sephacryl S-200 . The purified enzyme was homogeneous as judged by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis . The molecular weight of the enzyme was estimated to be 36,000 by gel filtration on Sephacryl S-200 and by SDS polyacrylamide gel electrophoresis, indicating that the enzyme is a monomer . The optimum pH was found at 9.0 . The enzyme activity was inhibited by EDTA or ethyleneglycol bis(beta-aminoethylether)-N,N'-tetraacetic acid and the inhibition was reversed by Ca2+ ion . The enzyme was able to eliminate p-nitroaniline or p-nitrophenol from p-nitrophenyl-3-ketovalidamine (IV) or p-nitrophenyl-alpha-D-3-ketoglucoside (VI), but not from p-nitrophenyl-1-epi-3-ketovalidamine or p-nitrophenyl-beta-D-3-ketoglucoside . Apparent Km values for IV and VI were 0.24 mM and 0.5 mM, respectively. Appl Environ Microbiol, 1985 Dec, 50(6), 1512 - 8 Isolation and characterization of Flavobacterium strains that degrade pentachlorophenol; Saber DL et al.; Bacteria able to mineralize 100 to 200 ppm of pentachlorophenol (PCP) were isolated by selective enrichment from PCP-contaminated soils from three geographic areas of Minnesota . Although differing somewhat in their responses to various biochemical and biophysical tests, all strains were assigned to the genus Flavobacterium . Five representative strains were examined in detail . All strains metabolized PCP as a sole source of carbon and energy; 73 to 83% of all carbon in the form of {U-14C}PCP was returned as 14CO2, with full liberation of chlorine as chloride . A comparison between strains in their ability to metabolize PCP showed some strains to be more efficient than others . Guanine-plus-cytosine contents of DNA ranged from 58.8 to 63.8%, and DNA/DNA hybridization studies with total DNA digests suggested substantial genetic homology between strains . All strains were shown to possess an 80- to 100-kilobase plasmid, and evidence suggested the presence of a larger plasmid (greater than 200 kilobases). J Appl Bacteriol, 1985 Dec, 59(6), 561 - 6 A note on a selective agar medium for the enumeration of Flavobacterium species in water; Flint KP; A selective nutrient agar medium containing kanamycin at 50 micrograms/ml was developed for the isolation and enumeration of yellow-pigmented colonies from the River Sowe, Coventry . Such organisms were shown to be members of the heterogeneous genus Flavobacterium . Typically, yellow pigmented colonies constituted less than 10% of the colonies on nutrient agar alone but up to 70% on nutrient agar plus kanamycin . This medium is a useful addition to the range of media available for the isolation and further ecological study of particular species of this important group of micro-organisms. J Biochem (Tokyo), 1985 Dec, 98(6), 1653 - 9 Occurrence of O-glycosidically peptide-linked oligosaccharides of poly-N-acetyllactosamine type (erythroglycan II) in the I-antigenically active Sendai virus receptor sialoglycoprotein GP-2; Suzuki Y et al.; Unique high molecular weight (M.W . 4,000-9,000) sugar chains termed erythroglycan II have been obtained from alkali/sodium borohydride digests of I-active asialoglycoprotein derived from sialoglycoprotein GP-2, which was isolated recently from bovine erythrocyte membranes as Sendai virus receptor (Suzuki, Y . et al . (1983) J . Biochem . 93, 1621-1633; (1984) ibid, 95, 1193-1200) . It was found that these sugar chains comprise about 40% of total alkali-labile oligosaccharides of asialo GP-2 and contain endo-beta-galactosidase (Flavobacterium keratolyticus)-resistant highly branched and heterogeneous oligosaccharides of poly-N-acetyllactosamine type which are linked O-glycosidically to the peptide backbone through N-acetylgalactosamine . Erythroglycan II also contains endo-beta-galactosidase-susceptible straight terminal polylactosaminyl side chains . A major oligosaccharide released by the enzyme cochromatographed with Gal beta 1-4GlcNAc beta 1-3Gal . Inhibitory activity of Sendai virus-mediated hemagglutination and the receptor activity for the virus were reduced significantly but not completely by the endo-beta-galactosidase . These results indicate that both linear and branched sialosylpolylactosamine sequences in erythroglycan II are important for the reception of the virus into the target cells. Biochim Biophys Acta, 1985 Nov 22, 843(1-2), 1 - 7 Isolation and characterization of a heparin with high anticoagulant activity from Anomalocardia brasiliana; Dietrich CP et al.; The isolation, some structural features, physicochemical properties and pharmacological activities of a heparin from Anomalocardia brasiliana are reported . It is shown that the mollusc heparin is very similar to those present in mammalian tissues with regard to chemical composition, physicochemical properties, pharmacological activities and susceptibility to heparinase and heparitinase II from Flavobacterium heparinum, as well as to the types of products formed by the action of these enzymes . Three significant quantitative differences were observed for the mollusc heparin when compared with the ones from mammalian origin, namely, a higher degree of binding with antithrombin III (45%), higher molecular weight (27-43 kDa) and higher anticoagulant activity (320 I.U./mg) . The possible biological role of heparin is discussed in view of the present findings. J Infect, 1985 Nov, 11(3), 233 - 8 Flavobacterium odoratum ventriculitis treated with intraventricular cefotaxime; Macfarlane DE et al.; A 6-week-old infant admitted to the University Hospital of the West Indies with hydrocephalus later developed ventriculitis . A heavy growth of Flavobacterium odoratum susceptible to gentamicin and cefotaxime was recovered from the ventricular fluid . Since intraventricular therapy was envisaged, a Pudenz reservoir was installed and ventricular fluid aspirated every 24 h to monitor treatment . Initial therapy consisted of intravenous cefotaxime, 50 mg/kg q.i.d . for 4 days . No significant reduction in the number of organisms in the ventricular fluid was achieved with this regimen . Intravenous therapy was therefore discontinued . On day 5 intraventricular therapy began with 5 mg cefotaxime 24 h for 6 days, followed by 1 mg/24 h for 4 days . Daily monitoring of intraventricular fluid indicated a high degree of antibacterial activity with rapid elimination of bacteria . Ventricular fluid remained sterile 10 days after therapy stopped . The Pudenz reservoir was removed, a ventriculoperitoneal shunt installed, and the patient discharged from hospital 4 days later without noticeable sequelae. J Antibiot (Tokyo), 1985 Nov, 38(11), 1476 - 86 Studies on WB-3559 A, B, C and D, new potent fibrinolytic agents . II . Structure elucidation and synthesis; Uchida I et al.; The structures of WB-3559 A, B, C and D, new fibrinolytic agents isolated from Flavobacterium sp . No . 3559, have been elucidated to be as shown in 1, 2, 3 and 4, respectively, on the basis of chemical and spectroscopic evidence . Total synthesis of WB-3559 D (4) was achieved starting from the optically active aldehyde 14. J Antibiot (Tokyo), 1985 Nov, 38(11), 1469 - 75 Studies on WB-3559 A, B, C and D, new potent fibrinolytic agents . I . Discovery, identification, isolation and characterization; Yoshida K et al.; WB-3559 A, B, C and D were produced by a bacterium which was classified as a member of the genus Flavobacterium . These compounds were purified by solvent extraction followed by chromatography on silica gel and then isolated by HPLC . The chemical structures were determined on the basis of chemical and spectroscopic evidence as in the succeeding papers . WB-3559 A, B, C and D stimulated rabbit plasma euglobulin clot lysis time . A chemically synthesized compound (WB-3559 D-syn) stimulated mouse plasma euglobulin clot lysis time when injected intravenously. J Appl Bacteriol, 1985 Oct, 59(4), 311 - 23 A numerical taxonomic study of Flavobacterium-Cytophaga strains from dairy sources; Jooste PJ et al.; Phenotypic data on 203 Gram-negative non-fermentative bacteria of the Flavobacterium-Cytophaga group isolated from milk and butter were analyzed by numerical taxonomic techniques . Twenty reference strains including species of Flavobacterium, Cytophaga and strains of Pseudomonas paucimobilis were included in the study . Using the matching coefficient of Sokal & Michener with antibiotic susceptibility data included, 189 isolates were recovered in nine clusters . Six of these clusters were linked at or above the 85% S level while three were linked at or above the 79% S level . The largest cluster, representing 46.3% of the isolates, could be equated with Flavobacterium sp . Group IIb . Other clusters could be equated with Flavobacterium sp . L 16/1 (22.7% of isolates), F . balustinum (10.8% of isolates), F . breve (4.4%), F . multivorum (3.5%) and Cytophaga johnsonae (1.5%) . The cluster resembling Flavobacterium sp . L 16/1 and a smaller unclassified cluster, were exceptional in being susceptible to the antibiotics cephalothin and penicillin G. Eur J Biochem, 1985 Oct 1, 152(1), 75 - 82 Flavobacterium heparinum 6-O-sulphatase for N-substituted glucosamine 6-O-sulphate; Bruce JS et al.; A specific glyco-6-O-sulphatase has been purified to homogeneity from Flavobacterium heparinum . The enzyme hydrolyses the 6-O-sulphates of 2-deoxy-2-sulphamido-6-O-sulpho-D-glucose (GlcNS-6S), 2-acetamido-2-deoxy-6-O-sulpho-D-glucose (GlcNAc-6S) and 2-amino-2-deoxy-6-O-sulphato-D-glucose (GlcN-6S) . The activity was purified 2100-fold by successive chromatography on CM-Sepharose CL-6B, Sepharose CL-4B, hydroxyapatite and blue-Sepharose CL-6B . Sodium dodecyl sulphate/polyacrylamide gel electrophoresis showed a protein of relative molecular mass 64000 . Four novel assays were developed using 35S-labelled and 14C-labelled monosaccharides . The purified enzyme was free of all other known heparin-degrading enzymes . In particular this was the first resolution of the 6-O-sulphatase from the sulphamidase . Optimal activity was at pH 7.5 . Enzyme activity was virtually unaffected by Na+ and K+ ions . Enhancements of activity of 12% and 30% were effected by Mg2+ and Ca2+ ions respectively . Inorganic phosphate and sulphate (both 0.005 mol dm-3) inhibited activity by 48% and 50% respectively . The Km value for the free amino substrate GlcN-6S was 1.35 mmol dm-3 . In contrast the Km values for the GlcNAc-6S and GlcNS-6S were 54 mumol dm-3 and 16 mumol dm-3 respectively. J Biochem (Tokyo), 1985 Oct, 98(4), 975 - 9 Comparison of inhibitory effects of prolinal-containing peptide derivatives on prolyl endopeptidases from bovine brain and Flavobacterium; Yoshimoto T et al.; The inhibitory effects of proline-containing peptides and their derivatives on prolyl endopeptidases from Flavobacterium meningosepticum and bovine brain were compared . Replacement of the carboxyl terminal proline in N-blocked peptides with prolinal resulted in remarkable decreases in Ki values for both prolyl endopeptidases . Further reduction of the prolinal to prolinol led to a decrease in their inhibitory effects . Z-Pro-, Z-Val-, and Suc-Pro-prolinals were similarly inhibitory for both the enzymes with Ki values of nM order . However, the inhibitory effects of Z-Pyr-prolinal and Boc-Pro-prolinal on these enzymes were significantly distinguished: they strongly inhibited the mammalian prolyl endopeptidase with Ki values of nM order, while the Ki values of these compounds for the microbial enzyme were only of microM order . These results suggest that there are some structural differences in the S2 and S3 subsites between the two enzymes, though their substrate specificities are apparently indistinguishable. Biochemistry, 1985 Sep 24, 24(20), 5587 - 92 Deglycosylation of alpha 1-proteinase inhibitor by endo-beta-N-acetylglucosaminidase F; Steube K et al.; The glycosidase endo-beta-N-acetylglucosaminidase F (endo F) from Flavobacterium meningosepticum was used for the deglycosylation of rat alpha 1-proteinase inhibitor (alpha 1 PI) . alpha 1 PI containing three oligosaccharide side chains of the complex type was isolated from rat serum or from the medium of rat hepatocyte primary cultures . High-mannose-type alpha 1 PI or hybrid-type alpha 1 PI was isolated from the media of hepatocytes treated with 1-deoxymannojirimycin or swainsonine, respectively . The susceptibility of complex-type alpha 1 PI to endo F was studied in the presence of various detergents . 3-{(3-Cholamidopropyl)dimethylammonio}-1-propanesulfonate and octyl glucopyranoside turned out to be most effective . In the absence of detergents, digestion of alpha 1 PI with high concentrations of endo F and/or long times of incubation led to the formation of alpha 1 PI with one and two oligosaccharide side chains . In the presence of 0.5% octyl glucopyranoside, the major cleavage products were unglycosylated alpha 1 PI and alpha 1 PI carrying one carbohydrate side chain . In contrast to the complex-type alpha 1 PI, the high-mannose type can be totally deglycosylated by endo F even in the absence of detergents . The susceptibility of the hybrid-type alpha 1 PI to endo F is between that of the complex and the high-mannose types. Biochemistry, 1985 Aug 13, 24(17), 4665 - 71 Deglycosylation of asparagine-linked glycans by peptide:N-glycosidase F; Tarentino AL et al.; Endo-beta-N-acetylglucosaminidase F (Endo F) and peptide:N-glycosidase F (PNGase F) were purified from cultures of Flavobacterium meningosepticum by ammonium sulfate precipitation followed by gel filtration on TSK HW-55(S) . This system separated the two enzymes and provided PNGase F in a high state of purity, but the basis for the resolution appeared to be hydrophobic interaction and not molecular size . Studies using purified Endo F and PNGase F with defined glycopeptides demonstrated that Endo F was somewhat similar to Endo H in that it hydrolyzed many, but not all, high-mannose and hybrid oligosaccharides, as well as complex biantennary oligosaccharides . PNGase F, in contrast, hydrolyzed all classes of asparagine-linked glycans examined, provided both the alpha-amino and carboxyl groups of the asparagine residue were in peptide linkage . Deglycosylation studies with PNGase F revealed that many proteins in their native conformation were susceptible to this enzyme but that prior denaturation in sodium dodecyl sulfate greatly decreased the amount of enzyme required for complete carbohydrate removal. Biochem J, 1985 Jul 15, 229(2), 369 - 77 Structure of heparin-derived tetrasaccharides; Merchant ZM et al.; The structure of heparin was examined by characterizing a disaccharide and five of the more than a dozen tetrasaccharide components obtained by its depolymerization with flavobacterial heparinase . Enzymic depolymerization of porcine mucosal heparin results in a mixture of di-, tetra-, hexa- and higher oligo-saccharides . The di- and tetra-saccharide components represent 75mol/100mol of these heparin fragments . Ion-exchange chromatography indicates the presence of only one disaccharide, deltaIdu2S(1----4)-alpha-D-GlcNS6S (where Idu is iduronic acid, deltaIdu is 4-deoxy-alpha-L-threo-hex-4-enopyranosyluronic acid, GlcN is glucosamine, GlcA is glucuronic acid and S is sulphate), but results in the isolation of five major and at least seven minor tetrasaccharide components . The structures of the disaccharide and five major tetrasaccharides were determined by chemical, enzymic, electrophoretic and spectroscopic methods, including 13C, 1H n.m.r . and fast atom bombardment-m.s . The structure of these five tetrasaccharides are: delta Idu2S(1----4)-alpha-D-GlcNS6S(1---4)-alpha-L-Idu2S(1-- --4)-alpha -D-GlcNS6S; delta Idu2S(1----4)-alpha-D-GlcNS6S(1----4)-beta-D-GlcA(1--- -4)- alpha-D-GlcNS6S; delta Idu2S(1----4)-alpha-D-GlcNS(1----4)-beta-D-GlcA delta Idu2S(1----4)-alpha-D-GlcNAc(1----4)-beta-D-GlcA(1----4)- alpha-D-GlcNS6S; and delta Idu2S(1----4)-alpha- D-GlcNAc(1----4)-alpha-L-Idu(1----4)-alpha-D-GlcNS6S . Biological activity for the disaccharide and the five major tetrasaccharides was examined, and none of them were found to possess significant anticoagulant activity. J Cell Physiol, 1985 Jul, 124(1), 113 - 9 Two distinct mechanisms for the interaction of cells with fibronectin substrata; Schwarz MA et al.; Fibronectin (Fn) was adsorbed onto neutral, sulfonated, imine-conjugated or gelatin coated polystyrene latex beads . In all cases, the Fn coated beads bound effectively to Chinese hamster ovary (CHO) cells in suspension . However, the binding of Fn coated neutral or positively charged imine conjugated bead was inhibited by low concentrations of heparin or heparan sulfate or by treatment of the cells with Flavobacterium heparanase . By contrast, binding of Fn coated sulfonated or gelatin beads was insensitive to inhibition by heparin and to heparanase treatment of cells . Adhesion of CHO cells to Fn coated tissue culture plastic was not sensitive to heparin, whereas adhesion of CHO cells to Fn-coated imine-conjugated plastic was sensitive to heparin . These observations imply that the functional status of Fn can be modulated by the nature of the surface to which the Fn is adsorbed . They further imply that, under some circumstances, the heparin/heparan sulfate binding domains of Fn can play a role in the attachment of Fn to the cell membrane via membrane proteoglycans . Under other circumstances, the interaction of Fn with the cell may primarily involve other receptors for Fn, presumably cell surface glycoproteins. Ann Inst Pasteur Microbiol, 1985 May-Jun, 136A(3), 359 - 70 {Isolation and description of strains of Flavobacterium multivorum of telluric origin}; Pichinoty F et al.; Twenty encapsulated strains of Flavobacterium multivorum were isolated from soil by elective culture in minimal medium containing inulin as the sole source of carbon and energy . These strains were compared with the type strain and with 5 other strains (including 4 clinical strains) of F . multivorum . Of 168 substrates tested, 18 carbohydrates were used as the sole carbon and energy source by all 26 strains . A few amino acids were used by some strains . The yellow pigment produced was found to be a carotene and its production was photo-inducible . The presence of a cytochrome c oxidase of the aa3 type was suggested by absorption spectra . The major cell lipids were phosphatidylethanolamine and sphingolipids; the major fatty acids were 13-methyltetradecanoic and 2-hydroxy-13-methyltetradecanoic acids . Soil and clinical strains of F . multivorum showed roughly similar patterns of antibiotic multiresistance . The average G + C content of the DNA of 11 strains was 40.8 +/- 1.5 mol%. Eur J Biochem, 1985 Apr 15, 148(2), 359 - 65 Flavobacterium heparinum 3-O-sulphatase for N-substituted glucosamine 3-O-sulphate; Bruce JS et al.; A novel bacterial sulphatase has been discovered in an extract of Flavobacterium heparinum . The enzyme hydrolyses the 3-O-sulphate from 2-deoxy-2-sulphamido-3-O-sulpho-D-glucose and 2-acetamido-2-deoxy-3-O-sulpho-D-glucose . The activity was purified 10 800-fold by chromatography successively on CM-Sepharose CL-6B, hydroxyapatite, taurine-Sepharose CL-4B and CM-Sepharose CL-6B . Sodium dodecylsulphate/polyacrylamide gel electrophoresis showed the enzyme to be homogeneous and of relative molecular mass 56 000 . Two novel assays were developed using 2-{14C}acetamido-2-deoxy-3-O-sulpho-D-glucose and 2-deoxy-2-sulphamido-3-O-sulpho-D-glucose as respective substrates . The purified 3-O-sulphatase was shown to be free of all other known heparin-degrading enzymes . Optimal activity was at pH 7.5 for the disulphated substrate and pH 8.0 for the N-acetylated substrate . Enzyme activity was virtually unaffected by Na+, K+ or Mg2+ ions . A 1.2-fold enhancement of activity was effected by 0.002 mol dm-3 Ca2+ . Inorganic phosphate and sulphate inhibited 3-O-sulphatase activity . The Km value of the N-acetylated substrate was determined to be 42 mumol dm-3 . No activity was detected with 2-amino-2-deoxy-3-O-sulpho-D-glucose. J Biol Chem, 1985 Apr 10, 260(7), 3915 - 22 Purification and properties of creatinine iminohydrolase from Flavobacterium filamentosum; Esders TW et al.; Creatinine iminohydrolase (EC 3.5.4.21), which catalyzes hydrolysis of creatinine to N-methylhydantoin and ammonia, was purified from Flavobacterium filamentosum . The average molecular weight of the purified enzyme was 272,480, and the subunit molecular weight was 44,300 . Extensive specificity studies indicated that the enzyme utilized cytosine (Km, 0.62 mM; Vm, 20.1 units/mg) as well as creatinine (Km, 5.00 mM; Vm, 40.4 units/mg) as a substrate . Each was a competitive inhibitor toward hydrolysis of the other compound . Dialysis of creatinine iminohydrolase in the presence of 0.01 M Tris phosphate buffer, pH 7.5, containing 1,10-phenanthroline decreased activity by 98% . Reactivation was accomplished by incubating the apoenzyme in the presence of certain divalent metal chlorides, listed in decreasing order of effectiveness: iron(II), zinc, cobalt(II), cadmium, and nickel . The extent of reactivation depended on the substrate and on which metal ion was added to the apoenzyme . Creatinine to cytosine activity ratios varied from 1:3.75 (iron(II) chloride), to 1:0.9 (zinc chloride), to 1:0.06 (nickel chloride) . For different preparations of the holoenzyme that ratio ranged from 1:0.45 to 1:1.10 . Variable but significant quantities of zinc and iron were present in all preparations of the purified enzyme. Appl Environ Microbiol, 1985 Apr, 49(4), 765 - 71 Determination of the concentration of maltose- and starch-like compounds in drinking water by growth measurements with a well-defined strain of a Flavobacterium species; van der Kooij D et al.; The growth kinetics of Flavobacterium sp . strain S12 specialized in the utilization of glycerol, and a number of oligo- and polysaccharides were determined in batch-culture experiments at 15 degrees C in pasteurized tap water supplied with very low amounts of substrates . Kss for the growth on maltotriose, maltotetraose, maltopentaose, and maltohexaose were 0.03 microM or less and below those for glucose (1.5 microM) and maltose (0.16 microM) . Kss for starch, amylose, and amylopectin were 8.4, 25.6, and 11.0 micrograms of C per liter, respectively . A yield of 2.3 X 10(7) CFU/micrograms of C on the oligo- and polysaccharides was calculated from the linear relationships observed between maximum colony counts in pasteurized tap water and the concentrations (usually below 25 micrograms of C per liter) of supplied compounds . The maximum colony counts of strain S12 grown in various types of raw water and tap water revealed that raw water contained only a few micrograms of maltose- and starch-like compounds per liter; in tap water the concentrations were all below 1 microgram of C and usually below 0.1 microgram of C per liter . The application of starch-based coagulant aids gave increased concentrations of maltose- and starch-like compounds in the water during treatment, but these concentrations were greatly reduced by coagulation and sedimentation, rapid sand filtration, and slow sand filtration. Antimicrob Agents Chemother, 1985 Apr, 27(4), 612 - 4 Biochemical properties of beta-lactamase produced by Flavobacterium odoratum; Sato K et al.; A constitutively produced beta-lactamase was purified from Flavobacterium odoratum GN14053 . The purified enzyme gave a single protein band on polyacrylamide gel electrophoresis . The isoelectric point was 5.8, and the molecular weight was estimated to be about 26,000 . The enzyme activity was inhibited by EDTA, iodine, p-chloromercuribenzoate, HgCl2, and CuSO4 but not by clavulanic acid, sulbactam, imipenem, and cephamycin derivatives . The enzyme showed a broad substrate profile, hydrolyzing oxyiminocephalosporins, cephamycins, imipenem, and some penicillins. J Clin Invest, 1985 Apr, 75(4), 1308 - 16 Evidence that cell surface heparan sulfate is involved in the high affinity thrombin binding to cultured porcine aortic endothelial cells; Shimada K et al.; It has been postulated that thrombin binds to endothelial cells through, at least in part, cell surface glycosaminoglycans such as heparan sulfate, which could serve as antithrombin cofactor on the endothelium . In the present study, we have directly evaluated the binding of 125I-labeled bovine thrombin to cultured porcine aortic endothelial cells . The thrombin binding to the cell surface was rapid, reversible, and displaced by enzymatically inactive diisopropylphosphoryl-thrombin . The concentration of thrombin at half-maximal binding was approximately 20 nM . Both specific and nonspecific binding of 125I-thrombin to the endothelial cell surface was partially inhibited in the presence of protamine sulfate, after the removal of cell surface heparan sulfate by the treatment of cells with crude Flavobacterium heparinum enzyme or purified heparitinase . The binding as a function of the concentration of thrombin revealed that the maximal amount of specific binding was reduced by approximately 50% with little alteration in binding affinity by these enzymatic treatments . The reversibility and active-site independence as well as the rate of the binding did not change after heparitinase treatment . Whereas removal of chondroitin sulfates by chondroitin ABC lyase treatment of cells did not affect the binding, identical enzymatic treatments of {35S}sulfate-labeled cells showed that either heparan sulfate or chondroitin sulfate was selectively and completely removed from the cell surface by heparitinase or chondroitin ABC lyase treatment, respectively . Furthermore, proteolysis of cell surface proteins by the purified glycosaminoglycan lyases was excluded by the identical enzymatic treatments of {3H}leucine-labeled or cell surface radioiodinated cells . Our results provide the first direct evidence that heparan sulfate on the cell surface is involved in the high-affinity, active site-independent thrombin binding by endothelial cells, and also suggest the presence of thrombin-binding sites that are not directly related to heparan sulfate. J Biol Chem, 1985 Feb 10, 260(3), 1849 - 57 Purification and characterization of heparinase from Flavobacterium heparinum; Yang VC et al.; Heparinase (EC 4.2.2.7) isolated from Flavobacterium heparinum was purified to homogeneity by a combination of hydroxylapatite chromatography, repeated gel filtration chromatography, and chromatofocusing . Homogeneity was established by the presence of a single band on both sodium dodecyl sulfate and acid-urea gel electrophoretic systems . Amino acid analysis shows that the enzyme contains relatively high amounts of lysine residues (9%) consistent with its cationic nature (pI 8.5) but contains only 4 cysteine residues/polypeptide . The molecular weight of heparinase was estimated to be 42,900 +/- 1,000 daltons by gel filtration and 42,700 +/- 1,200 daltons by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The enzyme is very specific, acting only on heparin and heparan monosulfate out of 12 similar polysaccharide substrates tested . It has an activity maximum at pH 6.5 and 0.1 M NaCl and a stability maximum at pH 7.0 and 0.15 M NaCl . The Arrhenius activation energy was found to be 6.3 kcal/mol . However, the enzyme is very sensitive to thermal denaturation and loses activity very rapidly at temperatures over 40 degrees C . Kinetic studies of the heparinase reaction at 37 degrees C gave a Km of 8.04 X 10(-6) M and a Vm of 9.85 X 10(-5) M/min at a protein concentration of 0.5 microgram/ml . By adapting batch procedures of hydroxylapatite and QAE (quaternary aminoethyl)-Sephadex chromatography, gram quantities of heparinase that is nearly free of catalytic enzyme contaminants can be purified in 4-5 h. Appl Environ Microbiol, 1985 Feb, 49(2), 382 - 7 Quantitative studies of heat-stable proteinase from Pseudomonas fluorescens P1 by the enzyme-linked immunosorbent assay; Birkeland SE et al.; Pseudomonas fluorescens P1 is a psychrotrophic bacterium isolated from milk . Proteinase P1, the main extracellular heat-stable proteinase fraction of P . fluorescens P1, has been purified to homogeneity . A procedure with a sandwich enzyme-linked immunosorbent assay, using microplates and alkaline phosphatase conjugate was shown to detect 0.25 ng of proteinase P1 in 1 ml of reconstituted skim milk or defatted cream . The method offers the combination of sensitivity and specificity for the detection of these enzymes in milk and dairy products . In reconstituted skim milk cultures proteinase P1 was detectable when the CFU approached 10(7)/ml . Cultures in milk diluted 1:10, 1:30, or 1:100 with water showed detectable proteinase at population densities close to 10(6) CFU/ml . Aeration stimulated proteinase production; thus, a skim milk culture with shaking at 5 degrees C had a proteinase level of 36,000 ng/ml after 7 days as compared to 80 ng/ml in a stationary culture . The rate of inactivation of proteinase P1 at 150 and 55 degrees C as expressed by residual antigenic activity determined by the enzyme-linked immunosorbent assay was somewhat different from the rate determined on the basis of residual proteolytic activity . The specificity of the enzyme-linked immunosorbent assay with proteinase P1 antibodies was identical for proteinase P1 and for enzymes from six other strains of P . fluorescens, one Chromobacterium strain, and one Flavobacterium strain . Some psychrotrophic strains produced immunologically unrelated proteinase(s) . These preliminary observations indicate that proteinase P1-related enzymes are common among psychrotrophs appearing as spoilage bacteria in milk. Gastrointest Endosc, 1985 Feb, 31(1), 10 - 2 Bacteremia during esophageal variceal sclerotherapy: its cause and prevention; Brayko CM et al.; Eleven consecutive patients underwent a total of 34 esophageal variceal sclerotherapy (EVS) sessions for bleeding esophageal varices . Blood cultures were drawn pre-, intra-, and post-EVS . All pre- and post-EVS blood cultures were negative . Five of the initial nine patients studied were found to have positive blood cultures, drawn after a mean of six injections . Pseudomonas aeruginosa was cultured from the blood in four patients and Flavobacterium from one . The source of contamination was found to be contaminated water used during the sclerotherapy sessions . By instituting simple techniques to eliminate this contamination, patients undergoing the remaining 25 EVS sessions were culture negative. Biochem Biophys Res Commun, 1985 Jan 16, 126(1), 365 - 72 Heparinlike molecules with anticoagulant activity are synthesized by cultured endothelial cells; Marcum JA et al.; Cultured microvascular endothelial cells isolated from rat epididymal fat pads produce glycosaminoglycans that accelerate thrombin-antithrombin complex formation . The heparinlike nature of these macromolecules was established by complete destruction of their anticoagulant activity employing purified Flavobacterium heparinase . Only 15% of the biologic activity of these complex carbohydrates was expressed when the heparin binding domain on the protease inhibitor was chemically modified at the Trp 49 residue . The anticoagulantly active species contains disaccharides which constitute the unique antithrombin binding region of the mucopolysaccharide . Removal of the biologically active heparinlike components from endothelial cells with 0.05% trypsin suggests that these molecular species are present on the cell surface. Arzneimittelforschung, 1985, 35(8), 1215 - 9 Fractionation and structural features of two heparin families with high antithrombotic, antilipemic and anticoagulant activities; Bianchini P et al.; 15 heparin preparations from bovine intestine, pancreas and lung and hog intestine were fractionated in two main components by selective barium precipitation . The ones that precipitated at room temperature with barium (slow moving (SM)-heparins) had a high anticoagulant activity measured by the USP and APTT (activated partial thromboplastin time) assay and low antithrombotic activity by the Yin and Wessler method . The fractions precipitated at 5 degrees C with barium (fast moving (FM)-heparins) had a low anticoagulant action and high antithrombotic activity . The maximum anti-Xa activity (chromogenic method) was present in heparins with molecular weights around 12-15 X 10(3) daltons whereas high APTT and LPL releasing activities were present in SM-heparins with molecular weights of 30-40 X 10(3) and 15-25 X 10(3) daltons, respectively . FM-heparins had a higher anti-Xa activity and lower lipoprotein lipase (LPL)-releasing activity when compared with the SM-heparins with the same molecular weights . Significant structural differences were observed between SM- and FM-heparins by 13C-NMR spectra and enzymatic degradation with heparinase and heparitinase from Flavobacterium heparinum . Also, significant differences were observed for anti-Xa and anticoagulant activities for the two types of heparins depending on the pharmacological assay used. J Infect, 1985 Jan, 10(1), 17 - 24 Septicaemia in a teaching hospital in Kuwait--I: Incidence and aetiology; Elhag KM et al.; During a period of 18 months of study, blood cultures were performed on 3845 patients in hospital with clinical signs of infection . Among these, 214 (5.6%) episodes of septicaemia were diagnosed which correspond to 10.9/1000 hospital admissions . About 80% of the episodes were due to Gram-negative organisms, most common of which were Escherichia coli (19.6%), Salmonella spp (16.5%) and Klebsiella spp (15.1%) . Gram-positive organisms implicated in 20% of episodes were mainly Staphylococcus aureus (9.3%) and enterococci (4.9%) . Of all the septicaemias 62.0% were community-acquired with Salmonella spp . being the organism most commonly implicated . Hospital-acquired infections were mainly due to Serratia spp, Pseudomonas spp and Flavobacterium spp . The antibiotic resistance pattern of the organisms showed that hospital-acquired organisms had relatively high resistance to most antibiotics as compared with community-acquired organisms. J Clin Invest, 1985 Jan, 75(1), 272 - 9 Interaction of antithrombin III with bovine aortic segments . Role of heparin in binding and enhanced anticoagulant activity; Stern D et al.; Bovine antithrombin III (AT III) interaction with the luminal surface of bovine aortic segments with a continuous layer of endothelium was examined . Incubation of 125I-AT III with vessel segments, previously washed free of endogenous AT III, demonstrated specific, time-dependent binding to the protease inhibitor to the endothelium . Half-maximal binding was observed at an added AT III concentration of 14 nM . Binding of 125I-AT III to the vessel wall was reversible (50% dissociated in 4 min), and addition of either heparin or Factor Xa accelerated displacement of 125I-AT III from the vessel segment . Dissociation of 125I-AT III from the vessel segment in the presence of factor Xa coincided with the formation of a Factor Xa-125I-AT III complex . Inactivation of Factor IXa and Factor Xa by AT III was facilitated in the presence of vessel segments . Pretreatment of vessel segments with highly purified Flavobacterium heparinase precluded the vessel-dependent augmentation of AT III anticoagulant activity as well as specific binding of 125I-AT III to the vessel endothelium . In contrast, pretreatment of the vessel segments with chrondroitinases (ABC or AC) had no detectable effect on 125I-AT III binding or on AT III anticoagulant activity . AT III binding to vessel segments was competitively inhibited by increasing concentration of platelet factor 4 . Binding of the protease inhibitor to vessel segments was inhibited by chemical modification of AT III lysyl or tryptophan residues . These AT III derivatives retained progressive inhibitory activity . These data suggest that heparin-like molecules are present on the aortic vessel wall and mediate binding of AT III to the vessel surface, as well as enhancing the anticoagulant activity of AT III at these sites. Int J Biochem, 1985, 17(11), 1179 - 83 Immuno-affinity purification of heparinase; Linhardt RJ et al.; Polyclonal IgG rabbit antibodies were prepared against a purified heparinase from Flavobacterium heparinum . Immuno-affinity purification of crude and partially purified heparinase is described . The resulting enzyme was of comparable purity to that prepared using the standard multistep purification scheme . The antibodies prepared were found to increase the activity of bound heparinase. Dev Biol Stand, 1985, 61, 249 - 54 Characteristic cellular fatty acid composition and an ornithine-containing lipid as a new type of hemagglutinin in Bordetella pertussis; Kawai Y; The fatty acid composition of the total extractable cellular lipids of Bordetella pertussis was very characteristic and was mostly hexadecenoic and hexadecanoic acids (90%) in a ratio of about 1:1 . The fatty acid composition of Bordetella parapertussis and Bordetella bronchiseptica differed from that of B . pertussis . The two species were distinguished by the fatty acid composition of cell-bound lipids . The ornithine-containing lipid was characteristic of the genus Bordetella and its main structure was 3-hydroxyhexadecanoic acid amide-linked to ornithine and esterified to the second hexadecanoic acid . The lipid agglutinated human and some animal erythrocytes . The lipid is a new type of hemagglutinin and we proposed that hemagglutination occurred mainly by the hydrophobic interaction between the lipid moiety of the ornithine-containing lipid and phosphatidylcholine in the cell membrane of the erythrocytes . A relatively high content of ornithine-containing lipid was also found in opportunistic pathogens such as Flavobacterium meningosepticum which causes meningitis in babies and children . As the pathogenicity of the opportunistic pathogens is unclear, the ornithine-containing lipid may have an important role in pathogenicity. Eur J Biochem, 1984 Dec 17, 145(3), 607 - 15 Flavobacterium heparinum 2-O-sulphatase for 2-O-sulphato-delta 4,5-glycuronate-terminated oligosaccharides from heparin; McLean MW et al.; The glycosulphatase which hydrolyses the 2-O-sulphate of the disaccharide, 4-deoxy-2-O-sulphato-alpha-L-threohex-4-enopyranosyl uronic acid-(1----4)-2-deoxy-2-sulphamido-6-O-sulphato-D-glucose (delta UA-2S----GlcNS-6S), has been isolated from the soluble fraction of disrupted Flavobacterium heparinum . The activity was purified 3300-fold by chromatography on CM-Sepharose CL-6B, hydroxyapatite, taurine-Sepharose CL-4B and blue-Sepharose CL-6B . From sodium dodecylsulphate/polyacrylamide gel electrophoresis, the enzyme was homogeneous and of 62000 Mr . A novel assay was devised using the de-N-sulphonated {1-3H}alditol, 4-deoxy-2-O-sulphato-alpha-L-threo-hex-4-enopyranosyl uronic acid-(1----4)-2-amino-2-deoxy-6-O-sulphato-D-{1-3H}glucitol (delta UA-2S----{1-3H}GlcNH2-ol-6S) . This alditol was shown by 13C-NMR to be desulphated in the analogous manner to the original reducing trisulphated disaccharide . The purified 2-O-sulphatase was completely free of heparinase I, heparinase II (heparitinase), chondroitinases AC, chondroitinase B, the delta 4,5-glycuronidase for heparin delta 4,5-disaccharides, the 6-O-sulphatase and the 2-sulphamidase . It was optimally active over the range pH 5.5-6.5 and was practically unaffected by Na, K, Ca or Mg ions . Inorganic phosphate inhibited the activity . The Km value for the alditol substrate was 1.22 mmol dm-3 . Using 13C-NMR, the 2-O-sulphatase was found to hydrolyse the analogous esters of higher delta 4,5-oligosaccharides from heparin . This contrasts with the findings of other authors {Dietrich, C . P., Silva, M . E., and Michelacci, Y . M . (1973) J . Biol . Chem . 248, 6408-6415}. J Gen Microbiol, 1984 Dec, 130 ( Pt 12), 3335 - 8 Use of a synthetic oligonucleotide to identify a chromosomal gene for chloramphenicol acetyltransferase in a plasmid-bearing Flavobacterium; Beschle HG et al.; A micro-organism previously designated Flavobacterium sp . CB60 is resistant to chloramphenicol as a consequence of antibiotic acetylation by the enzyme chloramphenicol acetyltransferase and subsequent degradation of the acetylated product by co-metabolism . Although a 15.6 kb plasmid (pCB60) was demonstrated in this Flavobacterium strain, it did not appear to play a role in chloramphenicol acetylation . DNA hybridization was used to identify a fragment of DNA presumptively carrying the cat gene . The sequence of the synthetic probe was based upon known nucleotide sequences corresponding to the highly-conserved active site region of several chloramphenicol acetyltransferase variants . The structural gene for the enzyme in strain CB60 appeared to be chromosomal since the radioactive probe hybridized with a unique restriction fragment from chromosomal DNA but failed to do so with the DNA of plasmid pCB60. J Trop Med Hyg, 1984 Dec, 87(6), 245 - 8 Meningitis in children of Riyadh; Babiker MA et al.; One hundred and forty cases of meningitis admitted to the Children's Hospital in Riyadh from June 1980 to the end of May 1982 were included in a prospective study . Sixty-seven percent were under 1 year of age and 90% were under 5 years old . One hundred and twenty-four (88.6%) had purulent CSF, 12 (8.6%) had viral and four (2.9%) had tuberculous meningitis . E . coli was the commonest causative organism in the neonates . After that age S . pneumoniae was the predominant isolate . The implication of increasing resistance of H . influenzae to ampicillin therapy were discussed and a further rare case of meningitis caused by Flavobacterium meningosepticum was described . Seventy-three (52%) were cured completely, 39 (28%) survived with immediate neurological deficits and 28 (20%) died . Mortality and neurological sequelae were found to be high in four categories of patients: neonates, patients who were stuporose or comatose on admission, those who were partially treated before admission and those who presented late after the onset of their symptoms. Biochemistry, 1984 Nov 20, 23(24), 5801 - 12 Sequence variation in heparin octasaccharides with high affinity for antithrombin III; Atha DH et al.; We have isolated from nitrous acid cleavage products of heparin two major octasaccharide fragments which bind with high affinity to human antithrombin . Octasaccharide S, with the predominant structure iduronic acid----N-acetylglucosamine 6-O-sulfate----glucuronic acid-----N-sulfated glucosamine 3,6-di-O-sulfate----iduronic acid 2-O-sulfate----N-sulfated glucosamine 6-O-sulfate----iduronic acid 2-O-sulfate----anhydromannitol 6-O-sulfate, is sensitive to cleavage by Flavobacterium heparinase as well as platelet heparitinase and binds to antithrombin with a dissociation constant of (5-15) X 10(-8) M . Octasaccharide R, with the predominant structure iduronic acid 2-O-sulfate----N-sulfated glucosamine 6-O-sulfate----iduronic acid----N-acetylglucosamine 6-O-sulfate----glucuronic acid----N-sulfated glucosamine 3,6-di-O-sulfate----iduronic acid 2-O-sulfate----anhydromannitol 6-O-sulfate, is resistant to degradation by both enzymes and binds antithrombin with a dissociation constant of (4-18) X 10(-7) M . The occurrence of a 15-17% replacement of N-sulfated glucosamine 3,6-di-O-sulfate with N-sulfated glucosamine 3-O-sulfate and a 10-12% replacement of iduronic acid with glucuronic acid in both octasaccharides indicates that these substitutions have little or no effect on the binding of the oligosaccharides to the protease inhibitor . When bound to antithrombin, both octasaccharides produce a 40% enhancement in the intrinsic fluorescence of the protease inhibitor and a rate of human factor Xa inhibition of 5 X 10(5) M-1 s-1 as monitored by stopped-flow fluorometry . This suggests that the conformation of antithrombin in the region of the factor Xa binding site is similar when the protease inhibitor is complexed with either octasaccharide. Appl Environ Microbiol, 1984 Nov, 48(5), 936 - 43 Isolation and characterization of a new Cytophaga species implicated in a work-related lung disease; Liebert CA et al.; A yellow-pigmented, gram-negative, gliding bacterium isolated from an industrial water spray air humidification system was implicated as a causative agent in several occurrences of lung disease with hypersensitivity pneumonitis-like symptoms . The bacterium, designated WF-164, lacked microcysts or fruiting bodies and had a DNA base composition of 34.8 mol% of guanine plus cytosine . Gliding, flexing, nonflagellated cells measuring 0.3 by 3.5 to 8.9 micron were observed by using light and electron microscopy . Tests to determine utilization of selected carbohydrates revealed an amylolitic, chitinoclastic, noncellulytic bacterium . A number of additional biochemical and physiological tests were performed . DNA homology studies detected a 77.8% similarity to Cytophaga aquatilis (ATCC 29551) . Comparisons of cellular fatty acid and carbohydrate contents of isolate WF-164 with a Flexibacter sp., several Cytophaga spp., and Flavobacterium reference strains revealed similar patterns to that of C . aquatilis . On the basis of these characteristics, isolate WF-164 was identified as a new Cytophaga sp. J Gen Microbiol, 1984 Nov, 130 ( Pt 11), 2983 - 99 Differentiation between Xanthomonas campestris pv . oryzae, Xanthomonas campestris pv . oryzicola and the bacterial 'brown blotch' pathogen on rice by numerical analysis of phenotypic features and protein gel electrophoregrams; Vera Cruz CM et al.; Thirty-five Xanthomonas campestris pv . oryzae, fourteen X . campestris pv . oryzicola strains and six 'brown blotch' pathogens of rice, all of different geographical origin, were studied by numerical analysis of 133 phenotype features and gel electrophoregrams of soluble proteins, %G + C determinations and DNA:rRNA hybridizations . The following conclusions were drawn . (i) The Xanthomonas campestris pathovars oryzae and oryzicola display clearly distinct protein patterns on polyacrylamide gels and can be differentiated from each other by four phenotype tests . (ii) Both pathovars are indeed members of Xanthomonas which belongs to a separate rRNA branch of the second rRNA superfamily together with the rRNA branches of Pseudomonas fluorescens, Marinomonas, Azotobacter, Azomonas and Frateuria . (iii) 'Brown blotch' strains are considerably different from X . campestris pv . oryzae and oryzicola . They are not members of the genus Xanthomonas, but are more related to the generically misnamed . Flavobacterium capsulatum, Pseudomonas paucimobilis, Flavobacterium devorans and 'Pseudomonas azotocolligans' belonging in the fourth rRNA superfamily . (iv) No correlation was found between the virulence, pathogenic groups or geographical distribution of X . campestris pv . oryzae or oryzicola strains and any phenotypic or protein electrophoretic property or clustering. J Nutr Sci Vitaminol (Tokyo), 1984 Oct, 30(5), 415 - 20 Vitamin B6 biosynthesis and glycolate reductase in Flavobacterium sp . 238-7; Tani Y et al.; An enzyme which reduces glycolate to glycolaldehyde as the first step of the biosynthesis of vitamin B6 was studied using the particulate fraction of Flavobacterium sp . 238-7, a vitamin B6 producer . The enzyme catalyzes the reduction of glycolate in the presence of NADPH, but not the oxidation of glycolaldehyde, and is designated as glycolate reductase . There is a positive correlation between the activity of the enzyme and vitamin B6 biosynthesis . The activity and the formation of the enzyme were not affected by vitamin B6 . This enzyme probably plays a role in the extraordinary accumulation of vitamin B6 by the bacterium. Appl Environ Microbiol, 1984 Oct, 48(4), 690 - 3 Purification and partial characterization of Flavotoxin A; Hu WJ et al.; A heat-resistant, low-molecular-weight toxin was isolated from semisolid potato dextrose agar medium after inoculation with Flavobacterium farinofermentans sp . nov., which was isolated from fermented corn meal that caused some outbreaks of food poisoning in China . The toxin was purified by solvent partition, Sephadex LH-20 gel filtration, and C-18 reversed-phase column chromatography . Thin-layer chromatography and high-pressure liquid chromatographic methods were developed for the identification and analysis of the toxin . The purified toxin exhibited a single spot in thin-layer chromatography and a single peak in high-pressure liquid chromatography and had adsorption maxima at 232 and 267 nm . Mass spectral analysis indicated a molecular weight of 169 with an experimental formula of C9H13O3 . The 50% lethal dose of purified toxin in mice (oral) was less than 6.84 mg/kg, but greater than 0.68 mg/kg . Postmortem examination showed that the mice died of some type of neurological and cardiovascular system toxicity . The name Flavotoxin A is being assigned to the toxin. J Biol Chem, 1984 Sep 10, 259(17), 10700 - 4 Demonstration of peptide:N-glycosidase F activity in endo-beta-N-acetylglucosaminidase F preparations; Plummer TH Jr et al.; Endo-beta-N-acetylglucosaminidase F preparations from Flavobacterium meningosepticum have been found to contain peptide:N-glycosidase activity . Only the second activity, designated as peptide:N-glycosidase F, readily cleaves the beta-aspartylglycosylamine linkage of a fetuin triantennary complex glycopeptide, as shown by the isolation of the corresponding carbohydrate-free peptide containing aspartic acid and of an intact oligosaccharide with a di-N-acetylchitobiosyl moiety at the reducing end . Both activities in the mixture will hydrolyze a high mannose octaglycopeptide from ovalbumin, with the type of product formed being influenced by pH . At pH 4.0, only the endo-beta-N-acetylglucosaminidase F activity is functional, releasing octapeptide-GlcNAc and oligosaccharide-GlcNAc . At pH 9.3, the predominant cleavage is by peptide:N-glycosidase F at the glycosylamine bond, releasing octapeptide and oligosaccharide-GlcNAc-GlcNAc . This latter oligosaccharide is then hydrolyzed by endo-beta-N-acetylglucosaminidase F to oligosaccharide-GlcNAc plus GlcNAc. J Clin Invest, 1984 Aug, 74(2), 341 - 50 Acceleration of thrombin-antithrombin complex formation in rat hindquarters via heparinlike molecules bound to the endothelium; Marcum JA et al.; We have examined the role of heparinlike molecules in the regulation of coagulation by perfusing rat hindquarters with purified human thrombin and with its plasma inhibitor, antithrombin . Our data indicate that contact of the hemostatic components with the endothelium enhances the rate of thrombin-antithrombin complex formation by as much as 19-fold over the uncatalyzed rate of enzyme-inhibitor interaction . Heparinlike molecules are responsible for the antithrombin accelerating activity . The amount of thrombin-antithrombin complex generated within the hindlimb preparation after pretreatment of the vasculature with purified Flavobacterium heparinase or with addition of platelet Factor IV to the hemostatic components, was equal to the uncatalyzed levels . These heparinlike molecules appear to be tightly bound to the luminal surface of the endothelium, since they could not be detected within the physiologic buffer that was perfused through the animal . The above mucopolysaccharides function in a manner similar to commercial heparin, since modification of antithrombin at a site critical for heparin-dependent acceleration of the protease inhibitor resulted in a level of interaction product identical to the uncatalyzed amount . Finally, addition of diisofluorophosphate-thrombin to the enzyme perfusion stream reduced the amount of thrombin-antithrombin complex formed in the animal by 30-40%, which suggested that thrombin bound to the endothelium as well as enzyme free in solution are accessible to antithrombin that has interacted with heparinlike molecules present on the endothelium. J Antibiot (Tokyo), 1984 Aug, 37(8), 859 - 67 Microbial degradation of validamycin A by Flavobacterium saccharophilum . Enzymatic cleavage of C-N linkage in validoxylamine A; Asano N et al.; The enzymatic cleavage of C-N linkage in the degradation of validamycin A by Flavobacterium saccharophilum was examined using N-p-nitrophenyl derivatives of validamine and valienamine as synthetic model substrates for validoxylamine A . Incubation of N-p-nitrophenylvalidamine with the membrane fraction from the organism led to formation of N-p-nitrophenyl-3-ketovalidamine, and succeeding cleavage of C-N linkage . As the products of the cleavage step, one was identified as p-nitroaniline and another keto compound could not be purified enough because of its instability . However, on the basis of its hydrogenation products, the structure of the keto compound could be established as 5D-(5/6)-5-C-(hydroxy-methyl)-2,6-dihydroxy-2-cyclohexen-1-one . The same experiment was carried out with N-p-nitrophenylvalienamine . In this case, N-p-nitrophenyl-3-ketovalienamine could be isolated as an intermediate but the desired keto compound from the cleavage step could not be isolated because of its instability . The participation of two enzymes, that is, a dehydrogenase and a C-N lyase on the cleavage of C-N linkage was assured, and moreover, the analysis of its products, together with those of the previous studies allow us to propose a degradation pathway of validamycin A by Flavobacterium saccharophilum. J Antibiot (Tokyo), 1984 Jul, 37(7), 773 - 80 Bacterial production of 7-formamidocephalosporins . Isolation and structure determination; Singh PD et al.; Two new 7-formamidocephalosporins have been isolated as their acetyl derivatives (SQ 28,516 and SQ 28,517) from fermentations of a Flavobacterium sp . SC 12,154 . Structure 1 was deduced for SQ 28,516 from its spectroscopic properties while structure 2 was proposed for SQ 28,517 . SQ 28,516 exhibits weak antibacterial activity. Arch Dis Child, 1984 Jun, 59(6), 582 - 4 Trimethoprim sulphamethoxazole in neonatal Flavobacterium meningosepticum infection; Linder N et al.; During an outbreak of Flavobacterium meningosepticum septicaemia in a neonatal intensive care unit 9 infants were treated with intravenous trimethoprim sulphamethoxazole . Bacteriological cure was achieved in 8 patients; one infant died of massive intraventricular haemorrhage on the first day of treatment . Apart from prolonged persistence of pre-existing thrombocytopenia there was no evidence of side effects . Trimethoprim sulphamethoxazole should be considered in the treatment of neonatal F meningosepticum sepsis in view of its activity against this organism, good penetration of the blood brain barrier, and the absence of serious side effects. Proc Natl Acad Sci U S A, 1984 Jun, 81(12), 3781 - 5 Segmental homology and internal repetitiousness identified in putative nucleic acid polymerase and human hepatitis B surface antigen of human hepatitis B virus; Ohno S; In a previous paper, it was argued that only those coding sequences descended from oligomeric repeats (the number of bases in the oligomeric unit not being a multiple of 3) can retain sufficiently long alternative open reading frames, and that such alternative open reading frames serve as the reservoir for the sudden generation of new polypeptide chains with novel functions . It was suggested that plasmid-encoded 6-amino hexanoic acid linear oligomer hydrolase that suddenly endowed Flavobacterium sp . K172 with the capacity to live off nylon by-products arose by the above mechanism . A corollary to the above argument is the expectation that those viral base sequences that are known to use two of the three alternative reading frames to encode two different polypeptide chains should invariably contain recognizable remains of the oligomeric tandem repeats, and as a consequence, various oligopeptidic repeats should also be present in the amino acid sequence of each . Furthermore, two polypeptide chains encoded by the same base sequence translated in different reading frames should show segmental homology of the type depicted previously . In the present paper, the base sequence of human hepatitis B virus ayw subtype that encodes an 832 amino acid residue long putative nucleic acid polymerase in one reading frame and a 226 residue long human hepatitis B surface antigen in the other reading frame was examined . All three predictions noted above were satisfied. Infect Control, 1984 May, 5(5), 237 - 9 Flavobacterium meningosepticum; Ratner H; lavobacterium meningosepticum is an opportunistic pathogen of low virulence found in the hospital environment in water-containing equipment . Of primary importance is its role in outbreaks of neonatal meningitis which tends to be severe with a high mortality rate and serious sequelae . Changing all equipment concerned with humidifying or administering gases every 24 hours can help prevent these outbreaks in neonatal nurseries . Treatment is difficult because of the resistance of F . meningosepticum to most antimicrobial agents used to treat meningitis . Sensitivity tests, using a MIC method, are mandatory for infections caused by F . meningosepticum and, pending these results, vancomycin, given intravenously and, if necessary, intrathecally, appears to be the drug of choice for initial therapy. J Bacteriol, 1984 May, 158(2), 419 - 24 DNA-DNA hybridization analysis of nylon oligomer-degradative plasmid pOAD2: identification of the DNA region analogous to the nylon oligomer degradation gene; Negoro S et al.; Fine structure of the gene of 6-aminohexanoic acid cyclic dimer hydrolase, one of the enzymes responsible for the degradation of the nylon oligomer (6-aminohexanoic acid cyclic dimer), on the plasmid pOAD2 harbored in Flavobacterium sp . KI72 was determined by constructing miniplasmids from plasmid pNDH5 (a hybrid plasmid consisting of pBR322 and a 9.1-kilobase-pair HindIII fragment of pOAD2 ) . The 6-aminohexanoic acid cyclic dimer hydrolase produced by cells of Escherichia coli C600 harboring pNDH5 or its miniplasmid was examined immunologically and electrophoretically and was found to be identical to that of Flavobacterium sp . KI72 . A fragment of pOAD2 (17.2- to 19.1-kilobase-pair region on pOAD2 ) was detected as hybridized fragment by Southern blotting experiments, indicating the presence of the DNA region analogous to the 6-aminohexanoic acid cyclic dimer hydrolase gene on the plasmid. Carbohydr Res, 1984 Apr 2, 127(1), 75 - 94 Structural studies on heparin . Tetrasaccharides obtained by heparinase degradation; Linker A et al.; Three tetrasaccharides representing major structural sequences of heparin were isolated in good yield and characterized after degradation of heparin by purified flavobacterial heparinase . N-Desulfation was necessary to achieve good separation of these closely related compounds from each other . One of the tetrasaccharides was shown to be derived from the fully sulfated repeating segments; to contain L-iduronic acid and six sulfate groups, and have the structure delta 4,5- HexpA -(2-SO4)-(1----4)-alpha-D- GlcpN -(N-SO4)-(6-SO4)-(1- ---4)-alpha -L- IdopA -(2-SO4)-(1----4)-D- GlcN -(N-SO4)-(6-SO4) . The second contained a D-glucuronic acid unit that was nonsulfated instead of the L-iduronic acid, and the third, obtained in a fairly low yield, contained five sulfate groups, three of which being located on the disaccharide at the nonreducing end, and having the structure delta 4,5- HexpA -(2-SO4)-(1----4)-alpha-D- GlcpN -(N-SO4)-(6-SO4)-(1- ---4)-alpha -L- IdopA -(2-SO4)-(1----4)-D- GlcN -(N-SO4) . All tetrasaccharides had a sulfated, unsaturated uronic acid unit at the nonreducing end, confirming that the heparinase requires sulfated L-iduronic acid units for activity. J Clin Microbiol, 1984 Apr, 19(4), 568 - 9 Flavobacterium multivorum septicemia in a hemodialyzed patient; Potvliege C et al.; A case of Flavobacterium multivorum septicemia in a hemodialyzed patient is reported . Two blood cultures were positive at 48 h, and the patient became afebrile only after antimicrobial therapy . The origin of the septicemia could not be determined. Proc Natl Acad Sci U S A, 1984 Apr, 81(8), 2421 - 5 Birth of a unique enzyme from an alternative reading frame of the preexisted, internally repetitious coding sequence; Ohno S; The mechanism of gene duplication as the means to acquire new genes with previously nonexistent functions is inherently self limiting in that the function possessed by a new protein, in reality, is but a mere variation of the preexisted theme . As the source of a truly unique protein, I suggest an unused open reading frame of the existing coding sequence . Only those coding sequences that started from oligomeric repeats are likely to retain alternative long open reading frames . Analysis of the published base sequence residing in the pOAD2 plasmid of Flavobacterium Sp . K172 indicated that the 392-amino acid-residue-long bacterial enzyme 6-aminohexanoic acid linear oligomer hydrolase involved in degradation of nylon oligomers is specified by an alternative open reading frame of the preexisted coding sequence that originally specified a 472-residue-long arginine-rich protein. EMBO J, 1984 Mar, 3(3), 581 - 4 Integrity of the pericellular fibronectin matrix of fibroblasts is independent of sulfated glycosaminoglycans; Hedman K et al.; The pericellular matrix fibers of cultured human fibroblasts contain fibronectin, other glycoproteins, and heparan and chondroitin sulfate proteoglycans . In the present study, cell-free pericellular matrices were isolated from metabolically labeled fibroblast cultures . The isolated matrices were digested with heparinase from Flavobacterium heparinum, and then analyzed for sulfated glycosaminoglycans (GAGs) . Nitrous acid degradation was used to distinguish the N-sulfated GAGs (heparan sulfate) from chondroitin sulfate . Fibronectin and the other major matrix polypeptides were studied using gel electrophoresis, enzyme immunoassay and immunofluorescence . Upon heparinase digestion, greater than 95% of sulfated GAGs were degraded in the matrix without detectable release of fibronectin or other matrix polypeptides or alteration of the fibrillar matrix structure . We conclude that in fibroblast cultures the integrity of the fibrillar matrix is independent of sulfated GAGs . Together with earlier observations, this suggests that filamentous polymerization of fibronectin forms the backbone of early connective tissue matrix. Appl Biochem Biotechnol, 1984 Feb, 9(1), 41 - 55 An immobilized microbial heparinase for blood deheparinization; Linhardt RJ et al.; A new medical application of an immobilized microbial enzyme is described . Extracorporeal devices require systemic heparin administration to prevent thrombus formation; however, the use of heparin often leads to serious hemorrhagic complications . Heparinase isolated from Flavobacterium has been immobilized and used in a fluidized bed reactor to eliminate heparin from blood passing through an extracorporeal circuit both in vitro and in vivo . This paper discusses the stepwise development of this heparinase reactor including: (1) improvements in the fermentation resulting in an inexpensive large-scale source of heparinase without the addition of the previously required inducer, heparin; (2) the use of batch processes to adapt previous purification schemes to large-scale heparinase production and the subsequent purification of heparinase to a single SDS-PAGE banding protein; (3) the immobilization of heparinase with a 91% activity recovery and good stability, (4) the design and successful testing of a fluidized bed reactor containing immobilized heparinase in the removal of clinically used quantities of heparin from both human blood in vitro and canine blood in vivo; and (5) the initiation of animal studies focusing on the toxicology of heparinase-derived heparin degradation products and the short and long term effects of exposure to these products and to heparinase. Mol Biochem Parasitol, 1984 Jan, 10(1), 99 - 109 Characterization of polysaccharides of the eggs and adults of Hymenolepis diminuta; Robertson NP et al.; Polysaccharides and other complex carbohydrates were released by proteolysis of the chloroform-methanol insoluble residue of 10 day-old worms and eggs of Hymenolepis diminuta . Gas-liquid chromatographic analysis of alditol acetate derivatives of monosaccharides released from the polysaccharides by hydrolysis revealed that in the 10 day-old worm, glucose was the most abundant sugar, followed by galactose, glucosamine, galactosamine, fucose and possibly rhamnose . Mannose was least abundant and xylose was absent . In the egg, glucose and galactose were equally abundant, followed by the same sugars found in 10 day-old worms, and xylose was present . Uronic acid was detected in both fractions by specific chemical tests . None of the saccharide material from eggs and worms was susceptible to degradation by Streptomyces hyaluronidase, chondroitinase AC, and slightly susceptible to chondroitinase ABC, as shown by electrophoretic analysis on composite 2.2% acrylamide-agarose slab gels and 4.5/12.5% polyacrylamide gels before and after enzymatic treatment . One of the gel-separable bands, however, was degradable by both nitrous acid and Flavobacterium heparinase . Both bands from eggs were degradable by nitrous acid . These results suggest that eggs contain heparin and/or heparan sulfate and perhaps dermatan sulfate and that 10 day-old worms also have these polyglycans but possibly not chondroitin sulfate or hyaluronic acid. Nouv Rev Fr Hematol, 1984, 26(4), 255 - 60 Heparin-like molecules as regulators of atherogenesis; Rosenberg RD et al.; Bovine aortic endothelial cells release a heparin-like substance in the presence of 0.4% fetal calf serum . This substance inhibited the growth of smooth muscle cells in vitro by about 70% . Substitution of platelet-poor plasma for serum resulted in minimal liberation of inhibitory activity from the cells unless at least 10-fold higher concentrations of platelet-poor plasma were utilized . This suggested that a platelet product was involved in the release process . Therefore, we examined the ability of the platelet heparitinase recently isolated in our laboratory to release heparin-like species from cultured endothelial cells . Our results show that when endothelial cells were exposed to serum-free medium containing 1 ng/ml of the purified platelet endoglycosidase, at least as much inhibitory activity was released as was obtained with 0.4% serum . Dose response experiments indicated that only 10 pg/ml of the enzyme were necessary to liberate 50% of the inhibitory activity from endothelial cells . The heparin-like nature of the inhibitory substance was demonstrated by its sensitivity of Flavobacterium heparinase . Utilizing appropriate controls, the release of heparin-like material by the endoglycosidase was shown to be enzyme-specific and was not due to artifacts of experimental manipulations . In addition, this enzyme did not convert prereleased material to an active component, but directly liberated the active heparin-like species from endothelial cells . A simple model describing the possible role of heparin-like components and the endoglycosidase in the normal and injured wall will be presented. Microbiol Immunol, 1984, 28(10), 1083 - 92 Biological activities of partially purified elastase produced by Flavobacterium meningosepticum; Miyazaki S; Injection of viable cells of Flavobacterium meningosepticum at doses of 10(6) colony-forming units/eye caused ophthalmia in rabbits' corneas . Elastase, free of protease activity, was partially purified from broth cultures of F . meningosepticum ATCC12535 and TMS516 . The optimum pH of the elastase activity was about 8.0 . The activity was suppressed by Ni, Zn, Co, EDTA, and o-phenanthroline, and accelerated by Cu and Fe . The minimum dose of the elastase causing ophthalmia in the eyes of rabbits was 5 micrograms/eye (0.065 elastase units) . The minimum lethal dose of the elastase for suckling mice by intracerebral injection was 0.4 micrograms (0.0052 elastase units)/suckling mouse, and its minimum lethal dose for adult mice by intraperitoneal injection was 740 micrograms/mouse (9.52 elastase units). Appl Environ Microbiol, 1983 Dec, 46(6), 1447 - 9 Satellite growth of Legionella pneumophila with an environmental isolate of Flavobacterium breve; Wadowsky RM et al.; Legionella pneumophila serogroup 1 was observed to satellite around colonies of Flavobacterium breve on an L-cysteine-deficient medium which did not support growth of legionellae . Both isolates were recovered from the hot water tanks of hospitals . Ferric PPi stimulated satellite growth between 0.01 and 0.1%. Nature, 1983 Nov 10-16, 306(5939), 203 - 6 Evolutionary adaptation of plasmid-encoded enzymes for degrading nylon oligomers; Okada H et al.; Flavobacterium sp . KI72 metabolizes 6-aminohexanoic acid cyclic dimer, a by-product of nylon manufacture, through two newly evolved enzymes, 6-aminohexanoic acid cyclic dimer hydrolase (EI) and 6-aminohexanoic acid linear oligomer hydrolase (EII) . These enzymes are active towards man-made compounds, the cyclic dimer and linear oligomers of 6-aminohexanoic acid respectively, but not towards any of the natural amide bonds tested . The structural genes of EI (nylA) and EII (nylB) are encoded on pOAD2, one of three plasmids harboured in Flavobacterium sp . KI72 . This plasmid contains two kinds of repeated sequence (RS-I and RS-II); one of the two RS-II sequences, RS-IIA, contains the nylB gene, while the other, RS-IIB, contains a homologous nylB' gene . From comparisons of the nucleotide sequences and gene products of the nylB and nylB' genes, we now conclude that EII enzyme is newly evolved by gene duplication followed by base substitutions on the same plasmid. J Lab Clin Med, 1983 Nov, 102(5), 828 - 37 Heparinase: in vivo activity and immunogenicity in rabbits; Klein MD et al.; Anticoagulation with heparin is required during extracorporeal circulation for hemodialysis and cardiopulmonary bypass as well as during vascular surgery . Reversal of anticoagulation with protamine may be associated with hypotension and rebound anticoagulation and requires stoichiometric doses . Heparinase from Flavobacterium heparinum catalytically degrades heparin and reverses its anticoagulant effect . Heparin was administered to New Zealand White rabbits and plasma levels were assayed with the APTT anticoagulant assay and the azure A chemical assay . Heparinase actively degraded heparin both in vitro in rabbit plasma and in vivo in rabbit blood as determined by both the anticoagulant and chemical assays when compared to control heparin disappearance curves . Antibodies to heparinase were demonstrated by the ELISA technique in rabbits receiving i.v