Selective inactivation of MAO-B by benzyl-dimethyl-silyl-methanamines in vitro

The alpha-silyl amines benzyl-dimethyl-silyl-methanamine and the p-fluoro and p-chloro derivatives are potent time-dependent inhibitors of rat brain MAO-B. The inhibition exhibits saturation kinetics, takes place in the enzyme active-site and is irreversible. The most potent inhibitor in the series is 4-fluorobenzyl-dimethyl-silyl-methanamine (KI = 11 microM, tau 1/2 = 2.3 min). Its selectivity for the B-form relative to the A-form of rat brain MAO is higher than 10(4). Benzyl-dimethyl-silyl-methanamines may represent a new family of anti-Parkinsonian agents.     

Immobilization of microorganisms by adhesion: interplay of electrostatic and nonelectrostatic interactions

The adhesion of three microorganisms (Saccharomyces cerevisiae, Acetobacter aceti, and Moniliella pollinis) to different materials has been studied using various supports (glass, metals, plastics), some of which were treated by an Fe(III) solution. The surface properties of the cells were characterized by the zeta potential and an index of hydrophobicity; characterization of the supports involved surface chemical analysis (XPS) and contact angle measurements. Cell suspensions in pure water at a given pH were left to settle on plates; the latter were then rinsed and examined microscopically, Saccharomyces cerevisiae and A. aceti adhere to metals under certain pH conditions but do not adhere to any of the other materials tested unless it is previously treated by ferric ions; adhesion of these hydrophilic cells is essentially controlled by electrostatic interactions. Moniliella pollinis adhere spontaneously to glass and to polymeric materials, but its attachment is also influenced by cell-cell or cell-support electrostatic repulsions; near the cell isoelectric point, cell flocculation is competing with adhesion to a support.     

Conditions promoting stability of solventogenesis or culture degeneration in continuous fermentations of Clostridium acetobutylicum

Summary The stability of solvent production by Clostridium acetobutylicum has been studied in continuous single-stage and two-stage fermentations. At low dilution rates, metabolic oscillations resulting from product inhibition have been observed especially in the case of fermentations controlled by product accumulation. A second type of instability also observed in product-controlled fermentations, but not in fermentations controlled by nitrogen limitation, was a long-term metabolic drift towards acid production. This acid drift has been shown to be identical to the phenomenon of culture degeneration occurring upon subculturing in batch fermentation. In addition, it was found that acid drift could be reversed by decreases in pH, temperature and dilution rate, by growth limitation in nitrogen-deficient conditions and by the addition of butyric and acetic acids. The existence of two distinct mechanisms, a short-term control (shift) and a long-term control (drift), both triggered by the same physiological conditions, is proposed in the regulation of acid and solvent production.     

Isolation and characterization of butanol-resistant mutants of Clostridium acetobutylicum.

In a wild-type strain of Clostridium acetobutylicum isolated from soil, solvent production appeared limited by butanol toxicity. Butanol-resistant mutants have been obtained which produced significantly higher solvent concentrations (about 30%) than the wild-type strain. Some other physiological differences were observed between a selected resistant mutant and the wild-type strain at the level of solvent resistance and sporulation.     

Negative dominance in gene lamB: random assembly of secreted subunits issued from different polysomes

lamB is the structural gene for the lambda receptor, an oligomeric outer membrane protein from Escherichia coli K12 involved in phage lambda adsorption. We show that, under certain conditions, in a strain diploid for gene lamB, all the missense lamB mutations conferring lambda resistance that we have tested are dominant with respect to wild-type. We propose a model which allows a quantitative interpretation of the data. It is based on negative complementation at the level of oligomerisation. Wild-type and mutant subunits would assemble at random forming homo- and hetero-oligomers. Only wild-type homo-oligomers would be efficient for phage inactivation. For some classes of missense mutations the hetero-oligomers would have the capacity to bind, but not to inactivate the phage. The model confirms that active lambda receptor is a trimer and implies that for this secreted protein there is no preferential assembly of subunits originating from the same polysome.     

Sophorose lipid production from lipidic precursors: Predictive evaluation of industrial substrates

Summary Sophorose lipids stand out as biosurfactants with a wide potential for industrial application and which can be produced in good yield from glucose and a lipidic cosubstrate.Candida bombicola CBS 6009 (ATCC 22214) was used in the present study. The influence of the lipidic cosubstrate on various aspects of production performance of these glycolipids (final concentration, yield) and on product composition (in particular, the structure of the hydroxy fatty acid vegetable and animal oils, markedly influenced product composition. In terms of production performance, the best substrates were oils or esters rich in C18:0 and C18:1 fatty acids. Optimal overall performance was obtained with esters (340 g L^–1 sophorose lipids with rapeseed esters). Conclusions drawn from the results allow predictive evaluation of lipidic industrial substrates.     

An Ustilago maydis Mutant Partially Blocked in P45014DM Activity Is Hypersensitive to Azole Fungicides

An Ustilago maydis ergosterol biosynthesis mutant (A14) which is partially blocked in sterol 14alpha-demethylase (P45014DM) activity is described. This mutant accumulated the abnormal 14alpha-methyl sterols, eburicol, 14alpha-methylfecosterol, and obtusifoliol, along with significant amounts of ergosterol. Although the A14 mutant grew nearly as well as the wild type, it was impaired in cell extension growth, which indicated a dysfunction in apical cell wall synthesis. The mutant was also found to be hypersensitive to the azole fungicides penconazole and tebuconazole.     

Effects of caffeine on carotid sinus nerve chemosensory discharge in kittens and cats

Bairam, A., P. De Grandpré, C. Dauphin, and F. Marchal. Effects of caffeine on carotid sinus nervechemosensory discharge in kittens and cats. J. Appl.Physiol. 82(2): 413-418, 1997.Caffeine (C)decreases apneic episodes in premature infants and is thought tostimulate breathing mainly by a central mechanism. While the methylxanthines theophylline and aminophylline are known to alter thecarotid chemoreceptor activity, there are little data on C. The aim ofthe study was to examine the effects of C on the carotid sinus nervedischarge (CSND) in developing animals. Nine kittens 17-21 daysold and six adult cats that were anesthetized and artificially ventilated were studied. They received four consecutive doses of C,each of 10 mg/kg, administered at intervals of 20 min either asintravenous bolus injection (6 kittens, 3 cats) or continuous infusion(3 kittens, 3 cats). Bolus injections of C invariably induced a promptbut transient increase in the CSND from 4.1 ± 0.6 to 8.1 ± 1.0 (SE) impulses/s in kittens (P = 0.01)and from 3.9 ± 0.1 to 7.9 to 1.0 impulses/s in cats (after thefirst injection). This response was associated with a significantdecrease in arterial blood pressure. Continuous infusion of C did notinduce any early change in either CSND or blood pressure in kittens orcats. Fifteen minutes after C injection or infusion was begun, CSNDvalues in air, 8% O₂-balanceN₂, or 100%O₂ were not significantlydifferent from control. Haloperidol administered at theend of the experiment in four cats and four kittens significantlyincreased CSND and did not suppress the early response to C injection.It is concluded that caffeine administered by bolus in the kitteninduces a transient stimulation of the CSND that is associated with adecrease in the arterial blood pressure and is independent of thedopaminergic mechanisms in the carotid body. The lack of sustainedeffect implies the main mechanism to the ventilatory stimulation by Cmust be central.

     

In vitro colonization ability of human colon mucosa by exogenous Lactobacillus strains

Abstract A 5.4 kb Hind III DNA fragment carrying the gene encoding raw starch-digesting α-amylase (RSDA), has been previously cloned from Bacillus circulans F-2 and expressed in Escherichia coli [Kim et al. (1990) Biochim. Biophys. Acta 1048, 2233–2238]. Interestingly, when the cell extract of E. coli harboring a plasmid carrying this fragment was incubated with l M NaCl, it exhibited about 10 times higher enzyme activity than when assayed without NaCl. Differential zymograms showed two different amylase activities: one for RSDA and the other for a salt-dependent a-amylase (SDA). Even though RSDA activity was detected without NaCl, SDA activity was detected only in high concentrations of NaCl. SDA activity was fully detected at above l M NaCl. Results from subcloning of the genes, fractionation analysis of cell extracts, and immunological assays clearly suggested that the two amylases are genetically distinct and that genes for both enzymes are closely linked on the 5.4 kb DNA fragment.