Lipid content and carbon assimilation in Collembola: implications for the use of compound-specific carbon isotope analysis in animal dietary studies

In an effort to understand the relationships between both the lipid content and ¹³C values of Collembola and their diet, isotopically labelled (C₃ and C₄) bakers yeasts were cultured and fed to two Collembolan species, Folsomia candida and Proisotoma minuta. The fatty acid composition of Collembola generally reflected that of the diet with the addition of the polyunsaturated components 18:2(n-6), 20:4(n-6) and 20:5(n-3), which appeared to be biosynthesised by the Collembola. Whilst ergosterol was the only sterol detected in the yeast diets, only cholesterol was detected in Collembola, and although the ¹³C values of diet and consumer sterols differed by >2, the ¹³C values indicated that cholesterol was derived entirely from dietary sterol. The bulk ¹³C values of Collembola were similar to those of the diets, but fatty acid ¹³C values did not necessarily reflect those of the dietary fatty acids, indicating significant de novo biosynthesis of fatty acids within Collembola. Switching the Collembola from C₃ to C₄ yeast enabled the determination of the rates of incorporation of dietary carbon into Collembolan lipids, and showed that half-lives of the incorporation of dietary carbon varied between 1.5 and 5.8 days at 20°C. Cholesterol exhibited the slowest rate of incorporation in both species, while bulk carbon in F. candida possessed an intermediate rate. These results demonstrate that an understanding of the sources of isotopic fractionation and the role of biochemistry in regulating the ¹³C values of individual compounds is important in the application of compound-specific isotopic analysis to the study of animal trophic activities.     

Hepatic enzymatic synthesis and hydrolysis of CoA esters of solvent-derived oxa acids

Many ethylene glycol-derived solvents are oxidized to xenobiotic alkoxyacetic acids (3-oxa acids) by hepatic enzymes. The toxicity of these ubiquitous solvents has been associated with their oxa acid metabolites. For many xenobiotic carboxylic acids, the toxicity is associated with the CoA ester of the acid. In this study, related alkoxyacetic acids were evaluated as potential substrates for acyl-CoA synthetases found in mitochondrial, peroxisomal, and microsomal fractions isolated from rat liver. Likewise, chemically synthesized oxa acyl-CoAs were used as substrates for acyl-CoA hydrolases associated with the same rat liver fractions. Activities of the xenobiotic oxygen-substituted substrates were compared with analogous physiologic aliphatic substrates by UV-vis spectrophotometric methods. All of the solvent-derived oxa acids were reasonable substrates for the acyl-CoA synthetases, although their activity was usually less than the corresponding physiologic acid. Acyl-CoA hydrolase activities were decreased compared with acyl-CoA synthetase activities for all substrates, especially for the oxa acyl-CoAs. These studies suggest that these xenobiotic carboxylic acids may be converted to reactive acyl-CoA moieties which will persist in areas of the cell proximal to lipid synthesis, beta-oxidation, protein acylation, and amino acid conjugation. The interaction of these xenobiotic acyl-CoAs with those processes may be important to their toxicity and/or detoxification.     

Isolation,sequencing, and expression of a cDNA for the HXM-A form of xenobiotic/medium-chain fatty acid:CoA ligase from human liver mitochondria

The purification of xenobiotic/medium-chain fatty acid:CoA ligases (XM-ligases) from human liver mitochondria resulted in the isolation of two chromatographically separable forms (HXM-A and HXM-B). These two forms were purified to near homogeneity, cleaved with cyanogen bromide, the resulting peptides separated, and the N-terminus of two of the peptides partially sequenced. Identical sequences were obtained for HXM-A and HXM-B for the two peptides. These sequences were used to design probes for screening a human liver cDNA library. This resulted in the isolation of two overlapping cDNAs. Using these sequences we were able to design PCR primers that resulted in the isolation of a full-length cDNA from a human cDNA library. The cDNA contained 1731 bp of open reading frame and coded for a 64230-Da protein. This protein bears 56.2% amino acid homology to the MACS1 (medium-chain acyl-CoA synthetase) enzyme, 58.7% homology to the bovine XL-III XM-ligase, and 81.5% homology to the bovine XL-I XM-ligase. The cDNA could be expressed in COS cells, and the expressed enzyme had greater benzoate activity than phenylacetate activity, which is consistent with the known substrate specificity of HXM-A.     

Catalysts for the self-polymerization of adenosine cyclic 2′,3′-phosphate

Summary When adenosine cyclic 2,3-phosphate is evaporated from solution in the presence of simple catalysts such as aliphatic diamines at alkaline pH, and maintained in a dry state at moderate temperatures (25-85°C), self-polymerization to give oligonucleotides of chainlength up to at least 6 is observed. The products contain an excess of [35]-linkages over [25]-linkages. The effects of different catalysts and reaction conditions on the efficiency of the reaction are described. The prebiological relevance of these reactions is discussed.     

Species differences in membrane susceptibility to lipid peroxidation

The susceptibility of liver microsomes to lipid peroxidation was evaluated in seven species: rat, rabbit, trout, mouse, pig, cow, and horse. Lipid peroxidation was measured as thiobarbituric acid reactive substances formed in the presence of either FeCl₃-ADP/ascorbate or FeCl₂/H₂O₂ initiating systems. For rat, rabbit, and trout microsomes, the order of susceptibility to peroxidation was rat > rabbit >> trout. The lack of peroxidation in trout microsomes could be explained by high microsomal vitamin E levels. Membrane fatty acid levels differed between species. Docosahexaenoic acid predominated in the trout, arachidonic acid in the rat, and linoleic acid in the rabbit. The contribution of individual fatty acids to lipid peroxidation reflected the degree of unsaturation with docosahexaenoic > arachidonic >>> linoleic. For all species except trout, the predicted susceptibility to peroxidation, based on the response of individual fatty acids, agreed well with directly measured microsomal peroxidation. With the exception of the trout, vitamin E content ranged from 0.083–0.311 nmol/mg microsomal protein between species, and low levels did not influence susceptibility to peroxidation. Trout microsomes peroxidized only after vitamin E depletion by prolonged incubation. The data indicate that below a vitamin E threshold, species differences in membrane susceptibility to peroxidation can be reasonably predicted based only on content of individual peroxidizable fatty acids.     

The level of free amino acids in erythrocytes of different breeds of hen

Summary The content of free amino acids was determined in erythrocytes of adult Leghorn (Lg, White Rock (WR) and Cornish (Cr) hens, bred under identical conditions. The concentration of total amino acids was twice as high in the erythrocytes as in plasma, amounting to 396 m/100 ml, 424 m/100 ml and 475m/100 ml in White Rock, Cornish and Leghorn hens, respectively.Significant differences were found in the ratio of basic amino acids to acidic amino acids. These values were 0.76, 1.75 and 3.19 in White Rick, Leghorn and Cornish hens, respectively; in the plasma of all 3 breeds the ratio was 1. Statistically significant interbreed differences were expressed more distinctly in erythrocyte than in plasma amino acid concentrations. For absolute concentrations the differences were significant in the case of 9 amino acids.     

DL-苯丙氨酸酶法拆分

ＤＬ－苯丙氨酸的拆分是以Ｎ－乙酰－ＤＬ－苯丙氨酸铵为起始原料,经猪肾酰化酶不对称水解作用,再经离子交换色谱分离,减压浓缩得到Ｌ－苯丙氨酸和Ｎ－乙酰－Ｄ－苯丙氨酸结晶。     

Fat-1基因在n-3多不饱和脂肪酸功能研究中的应用

n-3多不饱和脂肪酸是机体脂肪酸成分之一,维持体内合理的n-3多不饱和脂肪酸比例具有重要的生理意义。但n-3多不饱和脂肪酸在大多数动物体内不能合成,只能从食物中补充。fat-1基因的出现改变了这一现状,它可以将多不饱和脂肪酸从n-6形式转化为n-3形式。文章综述了n-3多不饱和脂肪酸的功能,并介绍了fat-1基因的功能及转fat-1基因动物在n-3多不饱和脂肪酸功能研究上的应用。     

α-氨基酸的立体有择合成

简述了α－氨基酸立体有择合成中几个代表性方法：消旋氨基酸不对称转化、α,β－脱氢氨基酸不对称氢化、甘氨酸等量体不对称烷化和手性自身再生。引用文献１８篇。     

去盲肠鸡和未去盲肠鸡对胱氨酸废液中氨基酸的利用率

用胱氨酸废液提取的复合氨基酸添加剂为试样 ,以去盲肠和未去盲肠的 2 0周龄罗曼公鸡为试鸡 ,测定了两种鸡对胱氨酸废液中氨基酸的利用率。结果表明 :去盲肠鸡和未去盲肠鸡对胱氨酸废液中氨基酸的真利用率(TAAA)都显著高于其表观利用率 (AAAA) ,(P <0 .0 0 1 ) ;而未去盲肠鸡对胱氨酸废液中氨基酸的TAAA和AAAA都显著高于去盲肠鸡 (P均 <0 .0 0 1 )