Instrumental assay of microbial lipase at constant pH

A rapid, accurate method with high sensitivity and reproducibility, and having the advantage of a short incubation period under constant pH, has been developed for routine measurement of microbial lipase. Assembled from readily available and economical instrumental components, the apparatus includes a pH meter, a thermoelectric heating and stirring device, a motor-driven burette, and an automatic recorder. The reaction mixture, consisting of 5 ml of a 10% olive oil-gum arabic emulsion, 2 ml of 3 m NaCl, 2 ml of sodium taurocholate (15 mg/ml) of 0.075 m CaCl(2), 5 ml of water, and 1 ml of enzyme solution, was adjusted to pH 8.0 and 37 C. The pH was maintained at a constant value by automatic addition of 0.01 n NaOH during the incubation period, which usually lasted 5 min. A lipase unit, derived from the use of this technique, may be defined as the number of microequivalents of acid liberated per minute under the specified conditions. The method was sensitive to 0.01 units. Various organisms tested produced 0.17 to 1.32 units per ml of the cell filtrate. An Arrhenius plot for staphylococcal lipase yielded 14,500 cal for function A (energy of activation).     

Nephelometric Assay of Bovine Antistaphylocoagulase Serum

A nephelometric assay of staphylococcal coagulase has been utilized to measure coagulase inhibition by bovine anticoagulase serum. Suitably diluted antisera produced maximal inhibition when incubated with purified coagulase at pH 7.3 in phosphate-buffered saline for 15 min at 22 C or 1 hr at 4 C. Neutralization of coagulase activity was measured as the reduction in the clotting rate of a fibrinogen-plasma substrate, and was directly proportional to the concentration of antiserum over a wide range of coagulase activity. A unit of anticoagulase was defined as the amount of inhibitor that neutralized one unit of coagulase. In addition to the heat-stable (56 C, 30 min) antibody contained in the crude gamma-globulin fraction, a heat-labile, nondialyzable coagulase inhibitor was also detected in the sera from 15 of 16 randomly selected dairy cows.     

A comparison of two plating techniques to estimate plasmid stability of a prolonged chemostat culture

Summary A comparison of two plating techniques to estimate the segregational stability ofEscherichia coli RR1 harboring plasmid pBR322 in a chemostat was studied. No significant differences were observed between the spread and replica plating techniques in the beginning of the experiments. However, a noticeable discrepancy between these two methods appeared after approximately 100 hours. This inconsistency can be shown to be statistically significant.     

Fine needle aspiration of peripheral neuroepithelioma of soft tissues.

Peripheral neuroepithelioma of soft tissue is a malignant primitive neuroectodermal tumor that appears both in children and adults and usually has a poor outcome. Fine needle aspiration on two patients with tumors in the lower limbs showed small round cells with unipolar processes and a tendency to form Homer-Wright rosettes. The cells had a round to oval nucleus with fine chromatin, up to four small, conspicuous nucleoli and vacuolated, periodic acid-Schiff-positive cytoplasm. The diagnosis was supported by electron microscopic study of the aspirate, which showed features of neuroblastic differentiation (i.e., neurosecretory granules), and by histologic, immunohistochemical and cytogenetic study of the resected tumors.     

竹上一柄锈新种

本文描述了采自湖南龙山的竹锈一新种:竹生柄锈Puccinia bambusicola S.-x. Wei & J.-y. Zhuang。模式标本保藏在中国科学院微生物研究所真菌标本室.此菌与其它已知的寄生在竹上的柄锈比较,冬孢子很小(25-38 × 14-21 µm),顶壁也不厚(仅3-6 gym),近似于南美产的竹柄锈P. bambusarum Arth.,但后者冬孢子隔膜多为斜生且颜色较淡。附我国竹生柄锈属已知种的检索表。     

柑桔果实中柠檬苦素类化合物的研究现状与展望

柠檬苦素类似物（ｌｉｍｏｎｏｉｄ）是三萜类的植物次生代谢产物,主要存在于芸香科和楝科植物中。柠檬苦素（ｌｉｍｏｎｉｎ）及其类似物存在于柑桔属（Ｃｉｔｒｕｓ）的多种植物中。至今已从柑桔属植物中分离出３６种柠檬苦素类似物及１７种柠檬苦素类似物配糖体。柑桔中苦味原因之一为柠檬苦素类似物。作为其代表具有强烈苦味的柠檬苦素和诺米林如果在柑桔果汁中含量超过６ｍｇ／Ｌ,那么这种柑桔用于饮食业已不适合［１］。近年来,笔者对柑桔果实中的柠檬苦素类似物的生理变化及生物合成作过系统研究［１～９］,并提出将柠檬苦素类似…     

玉米叶绿体atpB及atpE基因在大肠杆菌中的表达

本实验构建了在大肠杆菌中能表达玉米叶绿体ＣＦ１完整β亚基、缺失Ｎ端２３个氨基酸残基的β亚基以及完整ε亚基的表达质粒。ＳＤＳ－聚丙烯酰胺凝胶电泳及Ｗｅｓｔｅｒｎｂｌｏｔ实验表明,在分别含这些表达质粒的细菌中β、ε亚基蛋白的合成量在诱导３ｈ后即达到饱和,其含量各占细菌体总蛋白的１０％左右。这些表达产物在细菌体中形成不溶性的包含体。可通过对菌体蛋白的分级分高,将表达产物溶于５ｍｏｌ／Ｌ的尿素溶液中。     

Effect of ethanol and fatty acids on malolactic activity ofLeuconostoc oenos

Medium-chain fatty acids (C₆ to C₁₂), produced by yeast metabolism during alcoholic fermentation, are known to be inhibitory to lactic acid bacteria. The purpose of this work was to clarify the effect of both ethanol and decanoic and dodecanoic acids on the growth and malolactic activity of aLeuconostoc oenos strain isolated from Portuguese red wine. Ethanol in concentrations up to 12% had no significant effect on malolactic activity but strongly inhibited cell growth. The fatty acids decanoic acid, in concentrations up to 12.5 mg l^–1, and, dodecanoic acid up to 2.5 mg l^–1 seemed to act as growth factors stimulating also malolactic activity; at higher concentrations they exerted an inhibitory effect. We found clear pH dependence between pH 3.0 and pH 6.0, between decanoic acid concentration and its effect on malolactic activity, indicating that the undissociated molecule is the active form. At pH 3.0 the results can be explained by considering that fatty acids enter the cell as protonated molecules and dissociate in the cytoplasm due to the higher internal pH, leading to increased intracellular hydrogenous concentration. This may be the basis of two different effects that contribute to the observed inhibition: decrease in the intracellular pH and dissipation of the transmembrane proton gradient, thus inhibiting intracellular enzymes and ApH-dependent transport systems.     

Expression of a maize proteinase inhibitor gene is induced in response to wounding and fungal infection: systemic wound-response of a monocot gene

The isolation and characterization of cDNA and genomic clones encoding a proteinase inhibitor protein (MPI) in maize is reported. Accumulation of the MPI mRNA is induced in response to fungal infection in germinating maize embryos. The expression pattern of the MPI gene, in healthy and fungal infected maize tissues, was examined and compared with the expression pattern of a gene that codes for a pathogenesis-related protein (the PRms protein) from maize. These two genes are induced by fungal infection, however different signals trigger their activation. Accumulation of the proteinase inhibitor mRNA is more a consequence of the wound produced by the penetration and colonization of the host tissues by the pathogen, than the result of a direct molecular recognition of the pathogen by the plant, as is the case for the induction of the PRms gene. Wounding, or treatment with abscisic acid or methyl jasmonate, stimulate MPI mRNA accumulation, but not PRms mRNA accumulation. Local and systemic induction of the MPI gene expression in response to wounding occurs in maize plants. To the authors' knowledge, this is the first example of a gene from a monocotyledonous species that clearly shows a systemic wound response. The possible functional implications for the existence of different signal transduction pathways that simultaneously activate a battery of defense mechanisms against potential pathogens are discussed.     

The metabolism of sugar and malic acid by Leuconostoc oenos: effect of malic acid, pH and aeration conditions

The co-metabolism of sugars by Leuconostoc oenos was studied under different environmental conditions. Under aerobic conditions, growth and sugar metabolism were poorer than under CO₂ or N₂ atmosphere and acetic acid accumulated to a larger extent. Glycerol was found in the aerobic cultures while erythritol was detected under N₂ or CO₂. When medium conditions make growth difficult (low pH, aerobic conditions, low nutrients), sugars were only slightly metabolized and growth was very slow while malic acid was rapidly and completely degraded, leading to an increase in the y _ATP. Aeration effects on the malic acid degradation rate depended on the nutrients and carbon source in the medium. Malic acid clearly stimulated bacterial growth, allowing an increase in the molar growth yields and ATP production. The results suggest that under adverse conditions cells are not able to grow and malic degradation supplies additional energy production.