Growth of a human intestinal Desulfovibrio desulfuricans in continuous cultures containing defined populations of saccharolytic and amino acid fermenting bacteria

Ecological and physiological effects of the sulphate-reducing bacterium (SRB) Desulfovibrio desulfuricans on other intestinal organisms were investigated in anaerobic chemostats (dilution rate approximately 0.2 h-1). Reproducible defined bacterial communities were used in these experiments, comprising 14 different saccharolytic and amino acid fermenting species: Bifidobacterium longum, Bif. adolescentis, Bif. pseudolongum, Bif. infantis, Bacteroides thetaiotaomicron, Bact. vulgatus, Lactobacillus acidophilus, Enterococcus faecalis, Ent. faecium, Escherichia coli, Clostridium perfringens, Cl. butyricum, Cl. innocuum, Cl. bifermentans. Lactobacillus and Cl. bifermentans populations never rose above minimum detection limits (log10 2.0 and 4.0, respectively) under the experimental conditions employed in these studies. Inclusion of Des. desulfuricans in bacterial cultures (c. log10 8.4 viable cells ml-1) resulted in marked reductions (i.e. greater than 1 log) in planktonic cell population densities of several species, particularly Bif. longum, Cl. perfringens and Bif. pseudolongum. The two bacteroides species were unaffected by Des. desulfuricans, while numbers of Cl. butyricum increased. Extensive wall growth developed in the SRB culture, consisting mainly of Des. desulfuricans (log10 9.2 viable cells ml-1), Bact. thetaiotaomicron and Bact. vulgatus, with lesser numbers of facultative anaerobes, Cl. perfringens and Bif. longum. Wall growth was associated with a reduction in planktonic cell mass and increased acid production by the cultures. Chemotaxonomic study of chemostat microbiotas, on the basis of cellular fatty acid methyl ester (FAME) analyses, showed the existence of characteristic bacteroides (C15) and bifidobacterial (C18) markers, but desulfovibrio markers (i-C15:0, C16:0, i-C17:1) could be identified. The metabolic activities of saccharolytic organisms were altered in the SRB chemostat, including synthesis of a number of hydrolytic enzymes involved in carbohydrate breakdown, such as alpha-galactosidase, alpha-glucosidase and beta-galactosidase, together with several mucinolytic enzymes. High concentrations of sulphide (8.2 mmol 1-1) were detected in the SRB chemostat, suggesting that this metabolite may have been inhibitory to some species. Saccharolytic organisms growing in the SRB fermenter utilized more starch, but less galactose-containing polymers, which correlated with the observed glycosidase activities. Profound differences were also recorded with respect to fermentation product formation in the chemostats, where a major switch to acetate production occurred in the SRB culture, with concomitant reductions in propionate, butyrate and lactate, which is an important electron donor for desulfovibrios.     

Single amino acid substitutions can further increase the stability of a thermophilic L-lactate dehydrogenase

Lactate dehydrogenases are of considerable interest as stereospecificcatalysts in the chemical preparation of enantiomerically pure-hydroxyacid synthons. For such applications in synthetic organicchemistry it would be desirable to have enzymes which tolerateelevated temperatures for prolonged reaction times, to increaseproductivity and to extend then applicability to poor substrates.Here, two examples are reported of significant thermostabilizations,induced by sitedirected mutagenesis, of an already thermostableprotein, the L-lactate dehydrogenase (EC 1.1.1.27 [EC] , 35 kDa permonomer subunit) from Bacillus stearothermophilus. Thermal inactivationof this enzyme is accompanied by irreversible unfolding of thenative protein structure. The replacement of Argl71 by Tyr stabilizesthe enzyme against thermal inactivation and unfolding. Thisstabilizing effect appears to be based on improved interactionsbetween the subunits in the core of the active dimeric or tetramericforms of the enzyme. The thermal stability of L-lactate dehydrogenasevariants with an active site Arg residue, either in the 171(wild-type) or in the 102 position, is further increased bysulfate ions. The two stabilizing effects are additive, as foundfor the Argl71Tyr/ Gln1O2Arg double mutant, for which the stabilityof the protein in 100 mM sulfate solution reaches that of L-lactatedehydrogenases from extreme thermophiles. All mutant proteinsretain significant catalytic activity, both in the presenceand absence of stnhilfoing salts, and are viable catalysts inpreparative scale reactions.     

Development and evaluation of terminally epoxidized triglycerides for coatings applications

Epoxidized 10-undecenoic acid triglyceride, an experimental seed oil derivative that has a terminal epoxy group on each of the three acyl glyceride segments, has been found to have good reactivity with amine curatives and allows room temperature cures to be obtained. Coatings based on epoxidized 10-undercenoic acid triglyceride have also shown excellent UV stability. As an example, coatings samples placed in a QUVA chamber exhibit no loss in gloss after 3000 hr of a cycled exposure to high intensity UV lamps and moisture at temperatures of 50–60°C. In comparison, coatings based on standard liquid epoxy resins (LERs) and commercially available hydrogenated LERs lose gloss due to chalking/decomposition within 200–800 hr. Presented at the 81 st Annual Meeting of the Federation of Societies for Coatings Technology, November 12–14, 2003, in Philadelphia, PA.     

Relative Genotoxicities of PAH of Molecular Weight 252 AMU in Coal Tar-Contaminated Sediment

Bioassay-directed chemical fractionation methodology was used to calculate relative mutagenic potencies of polycyclic aromatic hydrocarbons (PAH) of molecular weight 252 amu in coal tarcontaminated sediment from Sydney Harbour, Nova Scotia. A normal phase HPLC technique was used to separate organic solvent extracts into fractions containing isomeric PAH of a single benzologue class. Bioassays with Salmonella typhimurium strain YG1025 with the addition of oxidative metabolism (S9) showed that approximately 50% of the mutagenic activity observed in the sediment extract was associated with PAH of molecular weight 252 amu. Further separation of the 252 PAH fraction using reversed phase HPLC yielded subfractions containing individual compounds; bioassay dose-response curves for these subfractions showed that benzo[a]pyrene was responsible for approximately 75% of the activity of the 252 PAH fraction.     

Identification,Phylogeny, and Comparative Expression of the Lipoxygenase Gene Family of the Aquatic Duckweed,Spirodela polyrhiza,during Growth and in Response to Methyl Jasmonate and Salt

Lipoxygenases (LOXs) (EC 1.13.11.12) catalyze the oxygenation of fatty acids and produce oxylipins, including the plant hormone jasmonic acid (JA) and its methyl ester, methyl jasmonate (MeJA). Little information is available about the LOX gene family in aquatic plants. We identified a novel LOX gene family comprising nine LOX genes in the aquatic plant Spirodela polyrhiza (greater duckweed). The reduced anatomy of S. polyrhiza did not lead to a reduction in LOX family genes. The 13-LOX subfamily, with seven genes, predominates, while the 9-LOX subfamily is reduced to two genes, an opposite trend from known LOX families of other plant species. As the 13-LOX subfamily is associated with the synthesis of JA/MeJA, its predominance in the Spirodela genome raises the possibility of a higher requirement for the hormone in the aquatic plant. JA-/MeJA-based feedback regulation during culture aging as well as the induction of LOX gene family members within 6 h of salt exposure are demonstrated.     

Synthesis of polar vinyl monomer–olefin copolymers by α‐diimine nickel catalyst

Vinyl acetate, methyl methacrylate, acrylonitrile and methyl vinyl ketone were investigated for co‐ and terpolymerization with ethylene and ethylene–propylene. Precursor [bis(N,N ′‐dimesitylimino)acenaphthene]dibromonickel, activated by methylaluminoxane was used as a catalyst system and trialkylaluminium was employed to block the polar groups for these polymerizations. Polymerization activities of the order of magnitude of 10⁶ in the case of vinyl acetate and methyl methacrylate, and 10⁵ in the case of acrylonitrile were achieved. Microanalysis and GPC of acrylonitrile copolymers found about 17 units of acrylonitrile per polymer chain. Copolymers with very different properties from the parent homopolymers were obtained in all cases except that of methyl vinyl ketone. © 2001 Society of Chemical Industry     

Limonene transport and mechanical properties of EVOH and nylon 6,6 films as influenced by RH

The transport properties of d‐limonene through ethylene vinyl alcohol copolymers (EVOH) and nylon 6,6 films as functions of relative humidity (RH) and temperature were studied. Permeation properties of these polymers were strongly influenced by temperature and RH. Compared to the EVOH films, the nylon 6,6 film had much greater limonene permeability. Mechanical property studies indicated that both the tensile modulus and yield strength of the EVOH films decreased with an increase in RH. The polymer changed from being stiff and brittle at low RH to being soft and ductile at high RH. In addition, ethylene content and orientation were found to affect the transport and mechanical properties of limonene through EVOH polymers. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 79: 1949–1957, 2001     

An initial-boundary value problem of structurescopy of a nonferromagnetic metal solid with foreign dielectric inclusions using the residual field of an instantaneously cut-off extraneous current

An initial-boundary value problem of electrodynamics is considered for the structurescopy of a nonferromagnetic metal with foreign dielectric inclusions; it is assumed that structurescopy is carried out using the residual electromagnetic field of an instantaneously cut-off extraneous current. Under the fairly sweeping assumptions on the electric conductivity of the solid, the dielectric permittivity of the foreign inclusions, and the smoothness of the boundaries of the solid and the inclusions, it has been proven that the initial-boundary value problem of electrodynamics under consideration has a unique solution, which depends continuously on the initial conditions.     

Source apportionment of PAH in Hamilton Harbour suspended sediments: comparison of two factor analysis methods

A total of 26 suspended sediment samples collected over a 5-year period in Hamilton Harbour, Ontario, Canada and surrounding creeks were analyzed for a suite of polycyclic aromatic hydrocarbons and sulfur heterocycles. Hamilton Harbour sediments contain relatively high levels of polycyclic aromatic compounds and heavy metals due to emissions from industrial and mobile sources. Two receptor modeling methods using factor analyses were compared to determine the profiles and relative contributions of pollution sources to the harbor; these methods are principal component analyses (PCA) with multiple linear regression analysis (MLR) and positive matrix factorization (PMF). Both methods identified four factors and gave excellent correlation coefficients between predicted and measured levels of 25 aromatic compounds; both methods predicted similar contributions from coal tar/coal combustion sources to the harbor (19 and 26%, respectively). One PCA factor was identified as contributions from vehicular emissions (61%); PMF was able to differentiate vehicular emissions into two factors, one attributed to gasoline emissions sources (28%) and the other to diesel emissions sources (24%). Overall, PMF afforded better source identification than PCA with MLR. This work constitutes one of the few examples of the application of PMF to the source apportionment of sediments; the addition of sulfur heterocycles to the analyte list greatly aided in the source identification process.     

Impact sensitivity of nitroglycerin

Abstract

The impact sensitivity of liquid nitroglycerin, 50% initiation level, was observed to be near 1.8 kg?cm or 3.3 × 10³ j/m². The impact sensitivity of solid nitroglycerin, 50% initiation level, was observed by extrapolation to be near 200 kg?cm or 4.4 × 10⁵ j/m². The impact sensitivity of a mixture of solid and liquid NG was observed to be comparable to solid NG at the two stimulus levels tested. Thus, the presence of solid nitroglycerin reduces the impact sensitivity of liquid nitroglycerin.

The 50% initiation level for Class A cyclo-tetra-methylenetetranitramine was observed to be near 150 kg?cm or 3.3 × 10⁵ j/m² using the same instrument.