The role ofPseudomonas spp. and competition for carbon,nitrogen and iron in the enhancement of appressorium formation byColletotrichum coccodes on velvetleaf

Colletotrichum coccodes is currently being investigated as a mycoherbicide against the weed velvetleaf (Abutilon theophrasti). Two isolates ofPseudomonas spp. (Ps2 and Ps5) reduced the percentage of germ tubes and increased appressorial formation ofC. coccodes on detached leaves of velvetleaf. A study was conducted to see whether this effect could be attributed to competition for nutrients or iron betweenC. coccodes andPseudomonas spp. Ps2 and Ps5 had no effect on early spore germination, but reduced the percentage of germ tubes at 24 and 30 h, compared to the nontreated control. This reduction was diminished by the addition of nutrients but not Fe³⁺. Ps2 and Ps5 stimulated the formation of dark-coloured appressoria without germ tubes (AWGT), but this stimulation was diminished by the addition of nutrients or Fe³⁺. Germ tube branching at 30 h was also inhibited by the bacteria, but was not diminished by the addition of nutrients or iron. EDTA stimulated conidial germination at 10 h, which was reduced by the addition of Fe³⁺. However, EDTA did not stimulate the formation of appressoria (AWGT). These results suggest that the reduction in the percentage of germ tubes and the increase in the percentage of appressoria induced by the bacteria may be due to the competition for carbon or nitrogen. Iron competition may also be involved in the stimulation of appressorial formation, but not in the reduction in germ tube percentage and branching. Phylloplane bacteria may compete for carbon, nitrogen and iron, limiting the saprophytic phase of the pathogen on the phylloplane and accelerating the development of the parasitic phase. This may enhance the field efficacy ofC. coccodes as a biocontrol agent against velvetleaf.     

Effects of phenobarbitone on serum biochemical tests in dogs

OBJECTIVES: To investigate effects of phenobarbitone on serum activities of alanine aminotransferase, alkaline phosphatase and gamma-glutamyl transferase and concentrations of bilirubin, albumin, cholesterol and total protein in dogs. ANIMALS: Ten crossbreed experimental dogs and 10 client-owned dogs of mixed breeds treated chronically with phenobarbitone to control seizures. PROCEDURES: Experimental dogs were allocated to treatment (6 mg/kg oral phenobarbitone, n = 6) and control (no treatment, n = 4) groups in which serum biochemical tests were performed at intervals during a 3-month period. Biochemical tests were performed once on the 10 epileptic dogs. RESULTS: Phenobarbitone caused increased serum alkaline phosphatase activity but did not affect gamma-glutamyl transferase activity or bilirubin, cholesterol, albumin and total protein concentrations. Phenobarbitone had minimal effect on alanine aminotransferase activity. CONCLUSIONS: Individual dogs treated with phenobarbitone may have small increases in serum alanine aminotransferase activity and variable increases in alkaline phosphatase activity but are unlikely to have alterations in gamma-glutamyl transferase activity or bilirubin, cholesterol, albumin or total protein concentrations.     

Alanine aminotransferase apoenzyme in dogs

Unusually low serum activity of alanine aminotransferase (ALT) was detected in a Rottweiler dog with gastric dilatation-volvulus. Activity of ALT in the same sample was found to be much higher (estimated increase of 14,225%) when measured by methods adding the cofactor pyridoxal-5'-phosphate (P5P), indicating that nearly all serum ALT was in the apoenzyme form. An investigation was undertaken to determine the frequency of high serum aminotransferase apoenzyme levels in dogs. Eighty canine serum samples submitted to the Clinical Pathology Laboratory at Cornell University were assayed for ALT and aspartate aminotransferase (AST) in the presence or absence of exogenous P5P. In 79 dogs, inclusion of P5P in the LAT assay resulted in a median decrease in AST activity of -6.3% (range -33.3% to 25.0%) in all 80 dogs. One dog had an increase of 336% in ALT activity after inclusion of P5P in the assay, but lacked a similar increase in AST activity. The reason for the high levels of ALT apoenzyme in the 2 dogs was not determined.     

丙氨酸、半胱氨酸及精氨酸对牛精液冷冻效果的影响

在常用的果糖-柠檬酸钠-卵黄-甘油组合稀释液的基础上,分别添加不同浓度的丙氨酸(0.25、0.50、0.75、1.00 mmol·L-1)、半胱氨酸(0.25、0.50、0.75、1.00 mmol·L-1)和精氨酸(2.50、5.00、7.50、10.00mmol·L-1),以观察3种氨基酸在牛精液冷冻中应用的最佳浓度.再以最佳浓度的3种氨基酸进行两两或三重组合,与3种氨基酸单一应用进行效果对比,检测这些氨基酸对牛精液冷冻后质量是否有协同作用.结果表明,无论单一或合并应用丙氨酸、半胱氨酸及精氨酸,均未能显著提高精液的冻后质量.但相同的处理应用于不同的公牛可能会有显著不同的效果.     

Effects of the aglycone of ascaulitoxin on amino acid metabolism in Lemna paucicostata

Ascaulitoxin and its aglycone (2,4,7-triamino-5-hydroxyoctanoic acid, CAS 212268-55-8) are potent phytotoxins produced by Ascochyta caulina, a plant pathogen being developed for biocontrol of weeds. The mode of action of this non-protein amino acid was studied on Lemna paucicostata. Ascaulitoxin is a potent growth inhibitor, with an I₅₀ for growth of less than 1 μM, almost completely inhibiting growth at about 3 μM. Its action is slow, starting with growth inhibition, followed by darker green fronds, and then chlorosis and death. Most amino acids, including non-toxic non-protein amino acids, reversed the effect of the toxin when supplemented in the same medium. Supplemental sucrose slightly increased the activity. d-Amino acids were equally good inhibitors of ascaulitoxin activity, indicating the amino acid effects may not be due to inhibition of amino acid synthesis. Oxaloacetate, the immediate precursor of aspartate, also reversed the activity. LC-MS did not detect interaction of the compound with lysine, an amino acid that strongly reversed the effect of the phytotoxin. Metabolite profiling revealed that the toxin caused distinct changes in amino acids. Reduction in alanine, paralleled by enhanced levels of the branched chain amino acids valine, leucine and isoleucine and nearly unchanged levels of pyruvate, might indicate that the conversion of pyruvate to alanine is affected by ascaulitoxin aglycone. In addition, reduced levels of glutamate/glutamine and aspartate/asparagine might suggest that synthesis and interconversion reactions of these amino group donors are affected. However, neither alanine aminotransferase nor alanine: glyoxylate aminotransferase were inhibited by the toxin in vitro. Our observations might be explained by three hypotheses: (1) the toxin inhibits one or more aminotransferases not examined, (2) ascaulitoxin aglycone affects amino acid transporters, (3) ascaulitoxin aglycone is a protoxin that is converted in vivo to an aminotransferase inhibitor.     

丙氨酸的营养研究进展

丙氨酸是一种非必需氨基酸,近年来,由于其对能量、葡萄糖及其他氨基酸代谢的影响,而越来越受到研究者的重视,文章就丙氨酸对能量、氨基酸、葡萄糖和氮代谢的影响,及其对肠道微生物的影响作一综合阐述。     

慢性乙型肝炎患者血清胆碱酯酶水平与肝组织炎症关系的研究

目的：探讨慢性乙型肝炎患者血清胆碱脂酶（CHE）活性水平与肝组织炎症病理分级（G）的关系。方法：采用RI-1000TM全自动生化分析仪检测106例经病理证实的慢性乙型肝炎患者血清胆碱脂酶（CHE）．丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）活性水平．统计分析血清ALT、AST、CHE水平与肝组织病理炎症分级（G0～G4）的关系。结采：血清CHE水平随着肝组织炎症活动度增高而降低,而ALT、AST水平随着肝组织炎症活动度增高而升高;G4级患者的血清CHE水平明显低于G1、G2、G3级患者,G3级低于G1、G2级．差异有显著性（P〈0．05或0．01）。G4级患者的血清AST水平明显高于G1、G2、G3级患者．但G1、G2、G3级患者AST水平以及各炎症分级之间ALT水平差异无显著性（P〉0．05）。结论：CHE比ALT、AST更能反映肝损害的严重程度、是一项有意义的无创性指标。     

The changes in digestive enzymes and hormones of gilthead seabream larvae (Sparus aurata, L 1758) fed on Artemia nauplii enriched with free methionine

Variations in digestive enzymes and hormones during the larval development of gilthead seabream (Sparus aurata) fed on live prey (Artemia nauplii) enriched with free methionine were investigated for 16 days (from day 24 to day 40). Prior to initiation of the experiment, newly hatched larvae were transferred from incubators to fiber glass tanks (300 l) with black walls and fed with same diets until day 24. Each experiment was performed in triplicate. In the experimental group, the content of the free methionine in the Artemia nauplii was increased by adding a 5.3 mM free methionine solution to the culture water during a 16-h enrichment period. The larvae of both the control and enriched-methionine groups were sampled four times, with 4-day intervals between samplings, during a 16-day period. The larvae in the control group had a significantly lower growth than those of the methionine group at the end of the study (P < 0.05). The highest trypsin activity and leucine aminopeptidase N/leucine–alanine peptidase ratios were observed in the control group. A significant difference between bombesin activities in the treatment groups was not found at 5th minute after the initiation of feeding (P > 0.05), but they were significant at 15th minute post-initiation of feeding (P < 0.05). A significant difference between the cholecystokinin levels of the treatment groups was found (P < 0.05).     

Alanine racemase activity in the microalga <Emphasis Type="Italic">thalassiosira</Emphasis> sp.

ABSTRACT:   In this paper, the authors report the detection of alanine racemase activity in the marine diatom Thalassiosira sp. Since the Thalassiosira sp. was cultured under germ-free conditions, it appeared that D-alanine was not derived from bacteria but was produced through catalysis by algal alanine racemase. The rate of conversion of L-alanine to D-alanine was approximately the same as that for the reverse reaction, and the enzyme catalyzed the equilibration of the D- and L-forms. The crude enzyme preparation obtained from the cells at the stationary phase of the growth cycle had an optimal pH of approximately 9.5. The Lineweaver–Burk analysis showed that the K _m for D- and L-alanine was 16.5 mM and 29.4 mM, respectively. It appears that the enzyme is highly specific for D- or L-alanine because it does not catalyze the racemization of other amino acids. In addition, after gel filtration, the enzyme did not require exogenous pyridoxal 5'-phosphate (PLP) for its activity, however, the effects of several chemicals suggest that the enzyme may be PLP-dependent. The enzyme is more similar to that found in invertebrates when compared with that found in bacteria. This is the first report on the occurrence of alanine racemase activity in the microalga Thalassiosira sp.