Evidence for two genetic complementation groups in pyruvate carboxylase-deficient human fibroblast cell lines

We have examined genetic complementation in pyruvate carboxylase deficiency by comparing the enzyme activity in polyethylene glycol-induced heterokaryons with that in unfused mixtures of fibroblasts from three affected children. Complementation, manifested as a three- to sevenfold increase in pyruvate carboxylase activity, was observed in fusions between a biotin-responsive multiple carboxylase (pyruvate carboxylase, propionyl CoA carboxylase, and -methylcrotonyl CoA carboxylase) deficient fibroblast line and two other lines deficient only in pyruvate carboxylase activity. Kinetic analysis of complementing pyruvate carboxylase deficient lines, measured by the rate of restoration of enzyme activity as a function of time, revealed that maximum restoration was achieved within 10–24 hr after fusion. This profile is similar to those observed for fusions between the multiple carboxylase deficient line and two lines deficient in propionyl CoA carboxylase activity that are known to represent different gene mutations. Although the patients with pyruvate carboxylase deficiency had similar clinical findings, our studies indicate that pyruvate carboxylase deficiency is genetically heterogeneous, with at least two distinct, probably intergenic, complementation groups.This work was supported by an NIH research grant (AM 25675) and an A. D. Williams research grant (6-48360). B. Wolf is the recipient of an NIH Research Career Development Award (AM 00677) and is aided by a Basil O'Connor Starter Research Grant from The National Foundation-March of Dimes (5-263). G. Feldman is the recipient of an NIH predoctoral training grant (GM 07492). This article is No. 100 from the Department of Human Genetics at the Medical College of Virginia.     

Comparative sensitivities of electrophoretic assays for human enzymes

The sensitivities of 26 starch gel electrophoretic enzyme assays have been compared using HeLa human cells and A9 mouse cells grown in vitro.This research was supported by National Institutes of Health Grant No. USPHS GM 09966.     

Evolution des activités protéolytiques digestives chez les Jeunes Mulgilidae

Proteolytic activity in the stomach, pyloric caeca and intestine decreases with increasing size in the three fish species Mugil auratus, M. capito and M. saliens. Differences between these species are found mainly in the gastric proteolytic activity which appears to be related to diet. This activity is pronounced in M. saliens whose diet is mainly carnivorous. Comparison of regression lines relating gastric proteolytic activity to size reveals differences which distinguish M. auratus from both of the other species. Total proteolytic activity exhibits high variability depending on the types of diet.     

Free tryptophan pool and tryptophan biosynthetic enzymes in Saccharomyces cerevisiae

The free tryptophan pool and the levels of two enzymes of tryptophan biosynthesis (anthranilate synthase and indoleglycerolphosphate synthase) have been determined in a wild type strain of Saccharomyces cerevisiae and in mutants with altered regulatory properties.The tryptophan pool of wild type cells growing in minimal medium is 0.07 mole per g dry weight. Addition of anthranilate, indole or tryptophan to the medium produces a fifteen- to forty-fold increase in tryptophan pool, but causes no repression of the biosynthetic enzymes. Inclusion of 5-methyltryptophan in the growth medium causes a reduction in growth rate and a derepression of the biosynthetic enzymes, and this is shown here not to be correlated with a decrease in the free tryptophan pool.Mutants with an altered anthranilate synthase showing decreased sensitivity to inhibition by l-tryptophan or by the analogue dl-5-methyltryptophan have a tryptophan pool far higher than the wild type strain, but no repression of indoleglycerolphosphate synthase was observed. Mutants with an anthranilate synthase more sensitive to tryptophan inhibition show a slightly reduced tryptophan pool, but no derepression of indoleglycerolphosphate synthase was found.A mutant with constitutively derepressed levels of the biosynthetic enzymes shows a considerably increased tryptophan pool. Addition of 5-methyltryptophan to the growth medium of non-derepressible mutants causes a decrease in growth rate accompanied by a decrease in the tryptophan pool.Abbreviations CDRP 1-(o-carboxyphenylamino)-1-deoxyribulosephosphate - paba paraaminobenzoic acid - PRA N-(5-phosphoribosyl)-anthranilate - tRNA transfer ribonucleic acid; trp1 to trp5 refer to the structural genes for corresponding tryptophan biosynthetic enzymes     

Mitochondrial abnormalities in human phaeochromocytoma

Summary Mitochondrial abnormalities are reported in four cases of phaeochromocytoma. These abnormalities include swelling and scant cristae, intramitochondrial dense bodies, septate-like junctions, intercristal fusion plus spheroidal bodies, and intramitochondrial rodlets. These structural mitochondrial changes are associated with reduction in activity of the mitochondrial enzymes, monoamine oxidase and succinic dehydrogenase.Supported by a grant-in-aid from the National Health and Medical Research Council     

Enzymes of the tricarboxylic acid cycle in Obeliscoides cuniculi (Nematoda; Trichostrongylidae) during parasitic development

The specific activities of the enzymes of the tricarboxylic acid cycle; citrate synthase, aconitase, isocitrate dehydrogenase, succinate dehydrogenase, fumarase, and malate dehydrogenase, were determined in early fifth-stage, young and mature adult Obeliscoides cuniculi, the rabbit stomach worm. ∝-Ketoglutarate dehydrogenase activity could not be determined in any fraction. Fumarate reductase activity was found only in the mitochondrial fraction while all other enzymes, including an NADP-dependent malic enzyme were localized in the cytoplasm. Glutamate dehydrogenase, acid and alkaline phosphatase activities were also recorded. High levels of those enzymes acting in the “reversed” direction, i.e. MDH and fumarase relative to the enzymes of the “forward” direction, i.e. citrate synthase, aconitase and isocitrate dehydrogenase suggests that under anaerobic conditions a modified tricarboxylic acid cycle can operate. Some variations in specific activities were apparent as the worms matured but no qualitative differences were observed.     

Dictyocaulus viviparus: The role of pepsin in the exsheathment of infective larvae

Following the report of Silverman and Podger (1964) that pepsin formed an association with larval receptor sites on D. viviparus and that exsheathment had an absolute requirement for pepsin, the role of pepsin was studied in greater detail. A range of enzyme incubation, pepsin labeling, histochemical and electron microscopical techniques were used. Pepsin did cause exsheathment of D. viviparus but, it was not an absolute requirement. Exsheathment occurred in a range of proteolytic enzymes each at its optimum pH. Findings suggest that the area of weakness around the anterior end of the larvae is digested by external protease and that, in vivo, exsheathment is caused by the gut enzymes of the host.     

BIOCHEMICAL TAXONOMY OF MARINE PHYTOPLANKTON BY ELECTROPHORESIS OF ENZYMES. I. THE CENTRIC DIATOMS THALASSIOSIRA PSEUDONANA AND T. FLUVIATILIS1,2

Diatom systematics depends almost entirely upon structure of the silica shell. It is not known to what extent the taxonomic species, as defined by shell structure, corresponds to the genetic species—i.e., to the reproductively isolated population. As an approach to this problem, we report here a comparison of enzymes by electrophoresis. We have examined the genetic constitution of a number of clones of (presumably) the same species for each of 2 closely related, centric diatom species: Thalassiosira pseudonana Hasle and Heimdal and T. fluviatilis Hustedt. The 4 clones of T. fluviatilis form a distinct group, clearly separated from all the T. pseudonana clones. Within T. pseudonana, 4 estuarine clones and one reef clone form a group that is distinctly different from 4 oceanic clones. A single clone of T. pseudonana from the Continental Slope waters is intermediate between these 2 groups and probably shares genes with both groups, indicating that the 2 T. pseudonana groups are not genetically isolated. We conclude that i) within groups, isolates are closely related even though they originated from different continents; and, ii) T. pseudonana is subdivided into ecological races.     

Nitrofuran-Reducing Enzymes of Euglena*

SYNOPSIS. Euglena gracilis strain Z, green, dark-grown, and “bleached”with N-methyl-N-nitro-N-nitrosoguanidine, was found to contain 2 soluble enzymes which reduce nitrofurans. A small amount of activity was demonstrated also in a particulate fraction of sonic extracts, but none in isolated chloroplasts. The reduction of 5 nitrofurans, having widely differing bleaching activities, by each of the 2 enzymes was examined.     

两种麻疯树苗对盐胁迫的生理生态响应

研究两种不同基因型麻疯树苗(南油2、3号)在不同NaCl浓度下生理生态响应特征,并比较不同基因型麻疯树苗的耐盐差异性。结果表明:①用25、50 mmo.lL-1NaCl处理,南油2号全株干重与对照无显著差异,而南油3号全株干重比对照显著降低。用100 mmol.L-1l或以上浓度的NaCl处理,随着盐度增加,两种树苗全株干重皆比对照显著降低,且3号苗降低的幅度大于2号苗。②在用200 mmol.L-1或以下浓度的NaCl处理,南油2、3号叶片相对含水量(RWC)皆与对照无显著差异,而在用300 mmol.L-1NaCl处理,则分别比对照显著降低5%和8%。③用25、50 mmol.L-1NaCl处理,南油2号可溶性糖(SS)含量比对照显著降低,3号与对照无显著差异;用200、300 mmo.lL-1NaCl处理后,两者SS含量均比对照显著降低。同时,2号苗可溶性蛋白(SP)含量比对照显著增加,3号苗SP含量与对照无显著差异。④随着盐度增加,南油2号苗超氧化物歧化酶(SOD)活性先增加后降低。用300 mmol.L-1NaCl处理,比对照显著降低。随着盐度增加,3号苗的SOD活性递减,皆显著低于对照。用25、50 mmo.lL-1NaCl处理,两种树苗的过氧化物酶(POD)活性与对照无显著差异。随着盐度增加,2号苗的POD活性比对照显著增加,而3号苗比对照显著降低。用25、50 mmo.lL-1NaCl处理,两种树苗的过氧化氢酶(CAT)活性皆比对照显著增加,且随着盐度增加,其变化趋势如SOD活性。结果表明,麻疯树幼苗具有较好的耐盐性,且南油2号比南油3号具有更高的耐盐性,因为前者具有更高的保护酶活性、叶片保水能力和叶片SP含量。