Identifying human cells capable of metabolizing various classes of carcinogens

Human cells that appear capable of metabolizing various classes of carcinogens have been identified using one of two methods: metabolism of tritiated benzo(a)pyrene to aqueous-acetone soluble forms or inhibition of cellular DNA synthesis. Each of the assay systems was optimized and the results on 15 human epithelial cell lines were compared. One or more cell lines were found to activate each of four classes of carcinogens examined: polycyclic hydrocarbons, aromatic amines, heterocyclic hydrocarbons, and nitrosamines. Cells that appeared capable of metabolizing polycyclic hydrocarbons or aromatic amines by these methods were also found to produce metabolites which were cytotoxic to cocultivated human xeroderma pigmentosum fibroblasts after a 48-hr exposure to the carcinogen.     

The use of photosynthesis inhibitor (DCMU) for in situ metabolic and primary production studies on soft bottom benthos

A selective chemical photosynthesis inhibitor, DCMU (Dichorophenyl-dimethylurea), dissolved in DMSO (Dimethyl sulfoxide) was substituted for the dark incubation method commonly used to measure the oxygen consumption in metabolic and primary production studies. We compared oxygen fluxes during light incubations with DCMU and dark incubations procedure, on soft bottom benthos. For this purpose, we studied the effects of different DCMU concentrations. A concentration of 5 · 10^–5 mol l^–1 inside a clear incubation enclosure completely inhibits photosynthesis without affecting the metabolism of soft bottom benthos.     

Temperature effects on bioenergetics of growth, assimilaton efficiency and thyroid activity in juvenile varand lizards

When varanid lizards, Varanus niloticus, were allowed to select their preferred body temperature (c. 34°C) they exhibited swifter growth, larger food intake and superior efficiency of conversion when compared with animals restricted to a maximum body temperature of 24°C. At the higher temperature the animals also exhibited a higher metabolic rate and increased thyroid activity, while those at the lower temperature lostproportionately more energy via respiration. A complete energy budget over a period of 6 weeks is presented.     

Separation of metabolic and non-metabolic steps of Rb+ uptake in spring wheat roots with different K+ status

Uptake of Rb⁺ from a complete nutrient solution with 2.0 mM Rb⁺ was studied in roots of spring wheat seedlings ( Triticum aestivum L. cv. Svenno) with different K⁺ levels. The relationship between Rb⁺ uptake and concentration of K⁺ in the roots indicated a negative feedback mechanism operating through allosteric control. The Rb⁺ uptake process in root cells was divided into two steps: (1) binding of the ion in the free space, and (ii) transmembrane transport into the cytoplasm. Metabolic and non-metabolic components of uptake were separated by addition of the metabolic inhibitor 2,4-dinitrophenol (DNP) to the nutrient solution. It is suggested that metabolic Rb⁺ uptake requires energy in two uptake steps (for binding to the carrier entity in the free space and for transmembrane transport) or in one step only (for transmembrane transport), dependent on the K⁺ status of the roots. The change from metabolic to non-metabolic binding in the free space is accomplished by changing the conformational state of the carrier (slow/fast transitions). There may be a hysteretic effect on metabolic Rb⁺ uptake through a slow transition between carrier states. This is superimposed on the negative cooperativity, strengthening further cooperativity at intermediate K⁺ levels in the roots. Non-metabolic Rb⁺ uptake probably consists of two components, a carrier-mediated (facilitated diffusion) and a parallel diffusive component.     

Cotugnia digonopora: Transport of leucine

The uptake of leucine through the tegument of Cotugnia digonopora, a cestode found in the fowl intestine, occurs by a process of active transport. The K_t of transport is 0.87 mM and the V_max is 0.223 μmol/min/g. Uptake of the amino acid is competitively inhibited by valine (K_t = 1.30 mM). Potassium cyanide and 2,4-dinitrophenol do not completely block the entry of leucine into the parasite.     

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.     

Regulatory properties of a partially purified preparation of pyruvate kinase from Fasciola hepatica

It possesses sigmoid kinetics with PEP; FBP activation changes the relationship to a rectangular hyperbola. The enzyme is inhibited by malate, which competes with PEP; FBP relieves the inhibition slightly. ATP and bicarbonate ions are also inhibitory at high concentrations. ATP inhibition is mixed-competitive with PEP; bicarbonate inhibition is non-competitive. It is suggested that pyruvate kinase may regulate both lactate and acetate production by moderating the size of the cytosolic pyruvate pool.     

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).     

Bioenergetic considerations in the improvement of oil content and quality in oil-seed crops

Summary Production values (PVs), defined as the weight of the end product/weight of the substrate required for carbon skeletons and energy production, were calculated for plant fatty acids. The PVs varied from 0.361 to 0.300 with linolenic acid having the lowest value. In general, the PVs of unsaturated fatty acids were lower than those of saturated fatty acids of similar chain lengths. Using this basic information, PVs of (A) oils from different oilseed crops, based on their standard fatty acid composition and (B) seed biomass with specified oil content and fatty acid composition were calculated. 1/PV gives the glucose required for the biosynthesis of 1 g end product and thus an estimate of the photosynthate requirement for the desired breeding goal can be estimated. Such calculations show that increasing oil percentage in seeds has a maximum energy cost when the increase in oil is associated with a decrease in the amount of carbohydrates where there is no change in protein concentration. Reduction of erucic acid content in the rapeseed oil did not alter its PV. It is inferred that there are no serious bioenergetic constraints in altering the fatty acid composition.