The role of carbon dioxide in the formation of end-products by Hymenolepis diminuta

The hypothesis that ambient CO₂ levels determine the end-products of energy metabolism excreted by Hymenolepis diminuta was tested by incubating the parasite in a range of CO₂ concentrations and measuring internal concentrations of adenine nucleotides and the excretion of organic acids. The strain of H. diminuta used was found to excrete mainly lactic acid and acetic acid. Succinic acid production was generally less than 5–10% of the total. At high CO₂ concentrations, the rate of excretion of lactic acid decreased while that of succinic acid increased, which conforms with the hypothesis. Acetic acid excretion did not vary significantly over the range of CO₂ concentrations used. Other results did not support the hypothesis. High CO₂ levels reduced the total amounts of acids excreted and the rate of succinic acid excretion was so small as to be ineffective in preventing the accumulation of H⁺ ions. When present in the incubation medium, succinic acid was taken up by H. diminuta. Lactic and acetic acid excretion was always sufficient to limit the accumulation of H⁺ ions. The conditions of incubation were shown not to be responsible for the low rates of succinic acid excreted. Incubation conditions and metabolic end-products were found to affect the rates of excretion of organic acids. There is thus a need, in work of this nature, to regulate and specify experimental conditions and to stipulate the strain of parasite used. The hypothesis was rejected and it was suggested that the energy metabolism of parasitic helminths is adapted to fluctuating O₂ and CO₂ tensions.     

Inborn errors of cobalamin metabolism: Effect of cobalamin supplementation in culture on methylmalonyl CoA mutase activity in normal and mutant human fibroblasts

We have examined the effect of addition of hydroxocobalamin to growth medium on the activity of the adenosylcobalamin-requiring enzyme methylmalonyl CoA mutase in normal human fibroblasts and in mutant human fibroblasts derived from patients with inherited methylmalonicacidemia. The mutant cell lines were assigned to four distinct genetic complementation groups (cbl A, cbl B, cbl C, and cbl D), each deficient in some step in the synthesis of adenosylcobalamin from hydroxocobalamin. After control cells were grown in cobalamin-supplemented medium, mutase holoenzyme activity increased markedly in a time- and concentration-dependent fashion. Growth in cobalamin-supplemented medium had no effect on mutase activity in some mutant lines belonging to the cbl B group, while activity increased severalfold in other cbl B mutants and in all cbl A, cbl C, and cbl D mutants examined, although mutase activity was still <10% of control. Comparison of mutase holoenzyme activity and total propionate pathway activity suggests that enhancement of mutase activity in mutant cells after cobalamin supplementation to values 5–10% of control may be sufficient to overcome the inherited metabolic block and to restore total pathway activity to normal.This work was supported in part by a research grant from the National Institutes of Health (AM 12579). H. F. W. is a recipient of a traineeship from the National Institutes of Health (T01-GM02299).     

Body wall of juvenile and adult Gastromermis boophthorae (Nematoda: Mermithidae): Ultrastructure and nutritional role

During the life cycle of the mermithid nematode Gastromermis boophthorae the main components of the body wall, namely the cuticle, hypodermis and somatic musculature, undergo radical changes in structure and relative proportions. In the non-feeding, free-living stages the cuticle is a well-defined, multilayered structure which lies upon a generally thin hypodermis; the somatic musculature has the extensive and highly organised appearance typical of motile nematodes. In parasitic juveniles the body wall has a completely different structure which is adapted for the suggested purpose of nutrient uptake from the host insect's haemocoelomic fluid; the cuticle is characteristically thin and rather diffuse, the underlying hypodermis becomes very extensive and metabolically active, and there is a corresponding reduction in the proportion of somatic musculature. The sub-cuticular surface of the enlarged hypodermis has a microvillous configuration and has associated alkaline phosphatase activity.     

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.     

Simultaneous tissue profiling of eicosanoid and endocannabinoid lipid families in a rat model of osteoarthritis

We describe a novel LC method for the simultaneous and quantitative profiling of 43 oxylipins including eicosanoids, endocannabinoids, and structurally related bioactive lipids with modified acyl groups. The LC-MS/MS method uses switching at a defined time between negative and positive electrospray ionization modes to achieve optimal detection sensitivity for all the lipids. The validated method is linear over a range of 0.01–5 nmol/g (0.1–50 nmol/g for 2-arachidonoyl glycerol) with intra- and interday precision and accuracy between 1.38 and 26.76% and 85.22 and 114.3%, respectively. The method successfully quantified bioactive lipids in different tissue types in the rat, including spinal cord, dorsal root ganglia (DRGs), knee joint, brain, and plasma. Distinct regional differences in the pattern of lipid measured between tissue types were observed using principle component analysis. The method was applied to analyze tissue samples from an established preclinical rat model of osteoarthritis (OA) pain and showed that levels of 12-hydroxyeicosatetraenoic acid were significantly increased in the OA rat knee joint compared with controls, and that 15-hydroxyeicosatetraenoic acid was significantly increased in the DRGs in the model of OA compared with controls. The developed LC-MS/MS method has the potential to provide detailed pathway profiling in tissues and biofluids where the disruption of bioactive oxylipins may be involved in disease states.     

基因组规模代谢网络模型构建及其应用

微生物制造产业的发展迫切需要进一步提高认识、设计和改造微生物细胞代谢的能力,以推动工业生物技术快速发展。随着微生物全基因组序列等高通量数据的不断积聚和生物信息学策略的持续涌现,使全局性、系统化地解析、设计、调控微生物生理代谢功能成为可能。而基于基因组序列注释和详细生化信息整合的基因组规模代谢网络模型(GSMM)构建为全局理解和理性调控微生物生理代谢功能提供了最佳平台。以下在详述GSMM的应用基础上,描述了如何构建一个高精确度的GSMM,并展望了未来的发展方向。     

Root border cells take up and release glucose-C

BACKGROUND AND AIMS: Border cells are released from the root tips of many plant species, and can remain viable in the rhizosphere for 1 week. Whether border cells are capable of controlled glucose exchange with their environment was investigated. METHODS: Border cells were removed from Zea mays L. root tips, and immersed in (14)C-labelled D-glucose. In one experiment, the hexose transport inhibitor, phlorizin, was used to investigate active glucose uptake from a range of glucose concentrations. In another experiment, glucose efflux from border cells was monitored over time. KEY RESULTS: Glucose uptake by the border cells increased with increasing glucose concentration from 0.2 to 20 mm. At 0.2 mm glucose, uptake was mainly active, as evidenced by the approx. 60 % inhibition with phlorizin. At 2 and 20 mm glucose, however, uptake was mainly via diffusion, as phlorizin inhibition was negligible. Glucose efflux increased with time for live border cells in both 2 and 20 mm glucose. There was no clear efflux/time pattern for heat-killed border cells. CONCLUSIONS: Border cells actively take up glucose, and also release it. Under our experimental conditions, glucose uptake and efflux were of similar order of magnitude. In the rhizosphere net glucose exchange will almost certainly depend on local soil conditions.