In vitro effect of benzimidazole drugs on the enzymes of lipid metabolism in Trichuris globulosa

The effects of 2 benzimidazole drugs, fenbendazole and thiabendazole, on the enzymes of lipid metabolism of an intestinal nematode parasite, Trichuris globulosa, have been studied. In vitro addition of these drugs inhibited the activity of these enzymes in both male and female worms. Inhibition is competitive in the case of the lipogenic enzymes, malate dehydrogenase and glucose-6-phosphate dehydrogenase; whereas, it is noncompetitive in the case of the lipolytic enzyme, triacylglycerol lipase. Cholesterol biosynthesis is, however, not affected.     

Characterization of glucose uptake by Trichuris globulosa (Nematoda) in vitro

14C-glucose uptake by adult Trichuris globulosa is found to be a non-linear function of time and limiting substrate concentration. The uptake is a two component process, an initial rapid burst is followed by a lower steady state, implying a mediated process. The uptake is dependent on Na+ ions which cannot be replaced by K+, Li+ or choline. The uptake is also dependent on pH, being maximal at pH 7.4. 14C-glucose absorption is markedly inhibited by glucose, phlorizin, ouabain and to a smaller extent by a number of monosaccharides and other sugar-phosphates, nucleosides and metabolic inhibitors like p-nitrophenyl phosphate and iodoacetate. The inhibition constant for glucose, phlorizin and ouabain has been found to be 8 mM, 5 mM and 7 mM, respectively. A modified Dixon-plot shows that glucose is a completely competitive inhibitor for 14C-glucose uptake while the nature of competitive inhibition of phlorizin and ouabain are found to be partial.     

Effects of mebendazole on the absorption of low molecular weight nutrients by Ascaris suum

Van Den Bossche H. and De Nollin S. 1973. Effects of mebendazole on the absorption of low molecular weight nutrients by Ascaris suum. International Journal for Parasitology3: 401–407. The effect of the anthelmintic drug, mebendazole, on the uptake and/or transport of glucose, fructose, 3-O-methylglucose, glycine, proline, methionine and palmitic acid was studied on in vitro incubated Ascaris suum. The experiments presented indicate that mebendazole inhibits the uptake and/or transport of glucose by A. suum. This inhibition is followed by a marked decrease in the glycogen content of the ascaris muscle. The addition of glucose to the incubation medium significantly enhanced the rate of uptake and/or transport of 3-O-methylglueose, glycine, methionine, proline and palmitic acid indicating that the absorption mechanisms depend on energy.Therefore, the inhibitory effect of mebendazole on the glucose uptake also results in a decreased uptake of 3-O-methylglucose and of the amino acids and fatty acid studied. The fructose uptake was not affected by the addition of glucose.Although mebendazole decreased the uptake of the hexoses and of the amino acids whether or not glucose was added, the uptake of palmitic acid was not affected when glucose was omitted from the medium. Mebendazole failed to exhibit an effect on the uptake, transport and/or utilization of glucose in rat.     

Effect of molecular weight on the antioxidant property of low molecular weight alginate from Laminaria japonica

In this study, three alginate fractions with different molecular weights and ratios of mannuronic acid (M) to guluronic acid (G) were prepared by enzymatic hydrolysis and ultrafiltration to assess the antioxidant property of alginates from Laminaria japonica with molecular weight below 10 kDa. The antioxidant properties of different molecular weight alginates were evaluated by determining the scavenging abilities on superoxide, hydroxyl, and hypochlorous acid and inhibitory effect on Fe²⁺-induced lipid peroxidation in yolk homogenate. The results showed that low molecular weight alginates exhibited high scavenging capacities on superoxide, hydroxyl, and hypochlorous acid radicals and good inhibition of Fe²⁺-induced lipid peroxidation in yolk. By comparison, alginate A1 with molecular weight below 1 kDa and M/G of 1.84 had better scavenging activity on superoxide, hydroxyl, and hypochlorous acid radicals in vitro than A2 (1–6 kDa), A3 (6–10 kDa), ascorbic acid, and carnosine. With similar M/G ratio, A2 exhibited better antioxidant activity on superoxide and hypochlorous acid radicals than A3. However, fraction A3 with molecular weight of 6–10 kDa exhibited higher inhibitory ability on lipid peroxidation in yolk in vitro than A1 and A2. The results indicated that molecular weight played a more important role than M/G ratio on alginate to determine the antioxidant ability. By comparison, low molecular weight alginates composed of guluronic acid and mannuronic acid exhibited better antioxidant ability on oxygen free radicals than sulfated polysaccharides from L. japonica in our previous study and represent a good source of marine polysaccharide with potential application as natural antioxidant.     

Effect of benzimidazole drugs on tubulin in benzimidazole resistant and susceptible strains of Caenorhabditis elegans

An in vitro assay was used to determine efficacy and if side resistance was present to benzimidazole anthelmintics tested against Caenorhabditis elegans after selection with albendazole. Side resistance was present to all the benzimidazoles tested, except for oxibendazole and parbendazole. At a concentration of 1 mM, all of the drugs, except thiabendazole, were effective in killing 100% of the albendazole susceptible worms. Tubulin from albendazole resistant and susceptible C. elegans was isolated and run on polyacrylamide gels. Western blots with anti-tubulin antibody showed that the albendazole resistant strain had an altered tubulin. Electron microscopy of albendazole-treated drug resistant worms showed microtubules throughout the intestinal cells. Microtubules were not observed in albendazole-treated drug susceptible worms.     

Effect of the low molecular weight thymus factor on T-cells in human blood]

Lowmolecular factor of polypeptide nature (thymarin) was extracted from calf thymus. This factor differed by physical and chemical characteristics from thymosin. The percentage content of "active" T-lymphocytes increased under the effect of thymarin in the cultures of lymphoid cells obtained from healthy individuals. The total T-cell count remained unchanged. Thymarin favoured increase of the total T-lymphocyte and "active" T-lymphocyte count in the cultures of cells from patients with chronic inflammatory processes. Cellular immunity reactions were restored and the course of the main disease improved under the effect of parenteral administration of thymarin. It is supposed that T-immunity system incompetence was associated with the inadequate production of the thymus factor capable of restoring the activity of the thymus-dependent lymphocyte population.     

Zinc uptake by isolated rat enterocytes: effect of low molecular weight ligands

The luminal phase of zinc intestinal absorption has not been well characterized. This study was intended to elucidate the possible role of low molecular weight (LMW) ligands in zinc intestinal transport in an isolated rat enterocyte system. Under these in vitro conditions, zinc uptake by the isolated enterocytes was rapid, leveling off within 1 min. Kinetic analysis revealed that both a mediated and diffusion component were involved in zinc uptake in the absence of LMW ligands by the cells. For the mediated component of zinc transport, the Kt and Vmax were 64.1 microM and 13.9 nmol/20 sec/mg protein, respectively. Zinc uptake was not affected by the addition of metabolic inhibitors. In the presence of histidine or cysteine (2:1 ligand:zinc molar ratio), zinc uptake was greatly reduced and occurred solely via mediated transport. Zinc uptake was also significantly decreased upon the addition of EDTA to the assay media. Other amino acids tested had no effect on zinc uptake by the cells. Albumin markedly reduced zinc uptake by the cells. Histidine and other potential LMW ligands were unable to facilitate albumin-inhibited zinc uptake. The results of this study suggest that the intestinal absorption of zinc may not be effected in the form of chelates with LMW ligands. Amino acids such as histidine and cysteine significantly reduce the uptake of the metal by isolated rat enterocytes, making questionable their putative role as necessary vehicles in the luminal phase of zinc absorption.     

Human alpha-L-iduronidase. I. Purification and properties of the high uptake (higher molecular weight) and the low uptake (processed) forms

Two major forms of human alpha-L-iduronidase have been individually purified over 175,000-fold to apparent homogeneity by sequential anion exchange, lectin affinity, and gel filtration chromatography. The two forms, initially designated as soluble and membrane-associated, were extracted from human lung in approximately equal amounts. Optimal solubilization of the membrane-associated form was facilitated by use of a non-ionic detergent or mannose 6-phosphate and saponin. Following detergent homogenization, the two forms were separated by anion exchange chromatography and then individually purified. The more electronegative form was membrane-associated, had a pI of approximately 5.9, and was selectively taken up (high uptake) by cultured Hurler syndrome fibroblasts; the more electropositive soluble form had a pI of about 6.6 and was incorporated into Hurler fibroblasts at a markedly lower rate (low uptake). After treatment with alkaline phosphatase, the pI values of both enzymes were about 7.8. Using 4-methylumbelliferyl-alpha-L-iduronide as substrate, the low and high uptake forms were each purified in milligram quantities to specific activities of 284,000 and 202,000 units/mg, respectively, with a combined yield greater than 35%. Each purified enzyme form migrated as a single protein band which also stained for enzymatic activity when electrophoresed in 7% native polyacrylamide disc gels at pH 4.3. By gel filtration, the high uptake form had an Mr = 85,000 whereas the Mr for the low uptake form was 68,000. Molecular weight estimates by analytical polyacrylamide gel electrophoresis were 82,000 and 70,000 for the high and low uptake forms, respectively. Rabbit anti-human low uptake alpha-L-iduronidase antibodies cross-reacted with the high uptake form as demonstrated by both immunotitration and Ouchterlony double immunodiffusion. Amino acid analysis revealed that the high uptake (higher molecular weight) form contained more arginine, glycine, alanine, glutamate or glutamine, leucine, isoleucine, histidine, and proline residues per molecule than the low uptake (lower molecular weight) form. Automated Edman degradation determined that the NH2-terminal residues of both forms were blocked. Both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and high performance liquid chromatography demonstrated that each purified form was composed of several components; each post-high performance liquid chromatographic component retained catalytic activity and was immunologically cross-reactive with antibodies against the low uptake form.(ABSTRACT TRUNCATED AT 400 WORDS)     

Microbial uptake and utilization of low molecular weight organic substrates in soil depend on carbon oxidation state

The fate of low molecular weight organic substances (LMWOSs) in soil is regulated by microbial uptake. However, C oxidation state, the number of C atoms and –COOH groups in the LMWOS can affect their microbial utilization. Thus, the aim of this study was to reveal the effects of substance chemical properties on initial uptake and utilization of sugars, carboxylic and amino acids by microorganisms. Soil solution, spiked with ¹⁴C-labelled glucose, fructose, malate, succinate, formate, alanine or glycine, was added to the soil and ¹⁴C was traced in the soil solution, CO₂, cytosol, and soil organic carbon (SOC) over 24 h. The half-life time of all LMWOS in the soil solution varied between 0.6 min (formic acid) and 5.0 min (sugars), indicating its dependence on C oxidation state of the substances. The half-life time of ¹⁴C in the fast mineralized pool in microorganisms, ranged between 30 (malic acid) and 80 (glycine) min and was independent on either C oxidation state, the number of C atoms, or number of –COOH groups. This suggests that intercellular metabolic pathways are more important for LMWOS transformation in soil than their basic chemical properties. The portion of mineralized LMWOS increased with their C oxidation state (20% for sugars vs. 90% for formic acid) corresponding to the decrease of C incorporated into the cytosol and SOC pools. Concluding, the physicochemical properties of the common LMWOS allow predicting their microbial uptake from soil solution and subsequent partitioning of C within microbial biomass.     

Effect of amino acids and low molecular weight peptides on the dynamics of water

The effect of amino acids and low-molecular-weight peptides on the dynamics of water was studied. Water medium together with molecules of dipeptides and amino acids dissolved in it is considered as a complex of interacting anharmonic oscillators. It was shown that the temperature behaviour of this system is determined by nonlinear resonances, which give rise to both the phenomenon of self-synchronization in the whole system or its part and to the phenomena of phase instability and coherence decay, depending on the store of oscillatory energy. Dissolved molecules are also involved in these oscillations, and if the frequencies and amplitudes of their oscillations are within the range in which nonlinear resonance occurs, they can affect the movement of the whole system.     

Presence of polyphosphate of low molecular weight in zygomycetes.

Polyphosphate of average chain length corresponding to 10 phosphate units was detected in the mycelial extract of zygomycetes. Gel electrophoresis techniques commonly used for the separation and characterization of acidic mucopolysaccharides were successfully used for the detection, purification, and characterization of the polyphosphate.     

The uptake of low molecular weight sulfur-containing compounds by Histoplasma capsulatum and related dimorphic fungi

A number of low molecular weight organic sulfur-containing compounds were tested for their effect on the respiratory activity of yeastlike and mycelialH. capsulatum. Of the compounds tested, L-cyst(e)ine was found to give maximum stimulatory effect on yeastlike phase respiration. The D- and meso isomers of cyst(e)ine as well as substituted derivatives were much less effective in the stimulation of respiratory activity of yeastlikeH. capsulatum. Respiration of homologous mycelial phase cell suspensions was depressed in the presence of L-cystine as substrate, while respiratory activity of yeastlikeB. dermatitidis andS. schenckii was unaffected.Whole cell suspensions of yeastlikeH. capsulatum actively transported S³⁵-labeled L-cystine and methionine but apparently not -mercaptoacetate-S³⁵. Mycelial phaseH. capsulatum and the yeastlike and mycelial phases ofB. dermatitidis andS. schenckii were observed to take up S³⁵-labeled L-cystine to a much lesser degree than yeastlikeH. capsulatum as determined on a dry weight basis. These results suggest significant differences in the transport and subsequent intracellular mechanisms of metabolism of low molecular weight sulfur-containing -amino acids and related compounds by yeastlikeH. capsulatum and its corresponding mycelial phase as well as the dimorphic fungiB. dermatitidis andS. schenckii.

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Zusammenfassung Eine Anzahl organischer, Sulfur-enthaltender Verbindungen mit niedrigem Molekulargewicht sind betreffs ihrer Wirkung an der Atmungsaktivität von hefeähnlichem und myzelialemH. capsulatum untersucht worden. Von den untersuchten Verbindungen gab L-cyst(e)ine die größte Reizwirkung an der Atmung der Hefephase. Die D- und Meso-Isomers von Cyst(e)ine so wie auch die substituierten Derivatives waren in der Reizung der Atmungsaktivität der Hefephase vonH. capsulatum weniger wirksam. Die Atmung der Suspension von Zellen der homologen Myzelphase war in der Gegenwart von L-cystine als Substrat unterdrückt, während die Atmungsaktivität der Hefephase desB. dermatitidis und die desS. schenckii unbeeinflußt blieb. Suspensionen von ganzen Zellen der Hefephase desH. capsulatum transportierten wirksam S³⁵ L-cystine und Methionine, aber anscheinend nicht beta-mercaptoacetate-S³⁵. Myzelphase-H. capsulatum und Hefeund Myzelphasen desB. dermatitidis undS. schenckii nehmen S³⁵-L-Cystine zu einem geringeren Grade auf denn Hefephase-H. capsulatum wie es am Trockengewicht festgestellt worden ist. Diese Ergebnisse legen es nahe, daß wesentliche Unterschiede im Transport und in dem nachfolgenden Intracellularmechanismus des Stoffwechsels von den Sulfurenthaltenden alfa-Aminosäuren mit niedrigem Molekulargewicht und verwandten Verbindingen durch die Hefephase desH. capsulatum und der bezüglichen Myzelphase, so wie auch durch die Doppelphasenpilze:B. dermatitidis undS. schenckii bestehen.

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This study has been supported by Part I VA-8200 Funds.     

Effect of low molecular weight basic proteins on mannose transport in Aspergillus niger microsomes]

Basic proteins of low molecular weight activate the transfer of mannose to endogenous glycoprotein acceptors in microsomal membranes of Aspergillus niger. The enhancement of mannosyltransferase activity is linked to the activation of the transport of mannose across the membrane. The role of these polycationic proteins on the membrane permeability is discussed.     

Effect of low molecular weight regulators on the biosynthesis of rifamycin B by Amycolatopsis mediterranei strains

Formation of differentiation regulators of the A-factor group in representatives of Nocardia and Nocardia-like actinomyces: N. asteroides, N. brasiliensis, Amycolatopsis mediterranei and "Streptomyces listeri" was observed. The effect of the regulators of different nature (barbital, A-factor and B-factor) on biosynthesis of rifamycin B by A. mediterranei strains was studied. It was shown that the A-factor stimulated rifamycin B production in the adifferentiated low active variant isolated from a natural population of the active strain VNIIA 1713 of the rifamycin B-producing culture. B-Factor insignificantly inhibited biosynthesis of rifamycin B in the studied strains of A. mediterranei.