Structural difference of the insulin receptors from circulating monocytes and erythrocytes

We compared insulin receptors obtained from cells widely used in human studies, the circulating monocytes and erythrocytes. Biochemically, these receptors possess both binding (alpha-subunit) and tyrosine kinase (beta-subunit) activities similar to insulin receptors from other sources. Subtle differences in molecular weight, however, were detected between the alpha-subunits of these two cell types when analyzed by NaDodSO4-PAGE. Crosslinked [125I]insulin-labeled alpha-subunit of the monocyte insulin receptor was of higher apparent molecular weight than the alpha-subunit derived from red cells. Neuraminidase treatment of the alpha-subunits from each cell type indicated more sialic acid residues were present on the monocyte than the red cell alpha-subunit. The structural properties of the insulin receptors of human circulating cells are similar but not identical to insulin receptors of other characterized systems.     

Affinity labeling of dihydrofolate reductase with an antifolate glyoxal

Dihydrofolate reductases from different species contain several highly conserved arginines, some of which have been shown by x-ray crystallography to have their guanido groups near the p-aminobenzoyl glutamate moiety of enzyme-bound methotrexate. The orientation of one of these (Arg-52) appears to be completely reversed in comparing the crystal structures of Escherichia coli with Lactobacillus casei enzyme (Bolin, J. T., Filman, D. J., Matthews, D. A., Hamlin, R. C., and Kraut, J. (1982). J. Biol. Chem. 257, 13650-13662). We synthesized a novel antifolate containing a glyoxal group designed to react specifically with active-site guanido groups which are able to approach the p-aminobenzoyl carbonyl of methotrexate. The binding of this compound to the enzyme was competitive with dihydrofolate (DHF) in ordinary buffers. In borate buffer at pH 8.0 it inactivated dihydrofolate reductases from both E. coli and L. casei at similar maximum rates, while the chicken liver enzyme was more slowly inactivated. The inactivation was stoichiometric, paralleled the loss of the glyoxal chromophore, and showed saturation kinetics. Inhibitor binding and thus inactivation was enhanced by NADPH, while DHF protected the enzyme. This allowed calculation of the Kd for DHF which was found to be identical with its Km. The stoichiometrically inactivated enzyme displayed the 340-nm chromophore characteristic of 4-aminopteridines bound to dihydrofolate reductase confirming active-site labeling with normal orientation of the ligand. The ligand remained covalently bound to inactivated enzyme upon denaturation at low pH but dissociated at neutral pH. Computer graphic modeling of the crystal structures predicted reaction of Arg-31 but not Arg-52 in L. casei dihydrofolate reductase and of only Arg-52 in the E. coli enzyme. Purification of the CNBr fragments from the inactivated enzymes gave a single labeled peptide for each species. The particular peptide tagged in each case was unaffected by the presence of NADPH and was in excellent agreement with the crystallographic predictions.     

An association between a lymphocyte antigen in sheep and the response to vaccination against the parasite Trichostrongylus colubriformis

Lymphocyte antigens were tested in sheep which had been selected for responsiveness to vaccination against the intestinal nematode Trichostrongylus colubriformis. These sheep had been bred in an assortative mating programme which produced offspring designated as either “high responders” or “low responders”, with highly heritable resistance or susceptibility.Ovine lymphocyte antigen (OLA) typing antisera were obtained from parous ewes in the course of matings which produced the high and low responder flocks. A particular antigen (SY1) was found to be present in high frequency on the lymphocytes of high responder (72·2%) and in lower frequency (21·9%) on the lymphocytes of low responder rams. In ewes, the frequency for high responders was 65·7% and for low responders it was 33·5%. A similar association between the SY1 antigen and low faecal egg count was found in random-bred sheep which had been vaccinated with irradiated larvae and challenged with normal larvae. The conclusion was drawn that this lymphocyte antigen was likely to be part of the sheep major histocompatibility complex which influenced the immune response of sheep to vaccination against the parasite.     

Advanced carcinoma of the prostate: treatment with a gonadotrophin releasing hormone agonist.

Ten patients with advanced progressive adenocarcinoma of the prostate were treated with a long acting analogue of gonadotrophin releasing hormone. Eight of these patients responded to treatment in terms of pain relief and clinical regression of tumour. Serum gonadotrophin and testosterone concentrations were significantly suppressed by the end of the second week of treatment, testosterone concentrations being comparable with those achieved by castration. The two patients who failed to respond had both relapsed previously when receiving conventional treatment, and neither showed any endocrine response to the analogue. Superagonists of gonadotrophin releasing hormone may be the treatment of choice in adenocarcinoma of the prostate, but further trials are required to establish long term safety and efficacy.     

Entamoeba histolytica: effect of growth conditions and bacterial associates on isoenzyme patterns and virulence

In xenic culture, isolates of Entamoeba histolytica from asymptomatic carriers are characterized, with rare exception, by possession of a nonpathogenic zymodeme. During the process of axenizing such an isolate, strain CDC:0784:4, a change in the pattern of the isoenzymes from nonpathogenic zymodeme I to pathogenic zymodeme II was observed 40 days after the amebae had been transferred from a medium for xenic cultivation to one used for axenic cultivation, but before axenization of the amebae had actually occurred. Axenization was accomplished by feeding the amebae lethally irradiated bacteria while suppressing and finally eradicating with antibiotics the bacterial flora accompanying the amebae in the original xenic culture. The change in zymodeme was accompanied by a change in virulence as evidenced by the ability of the amebae to produce hepatic abscesses in hamsters and to destroy monolayers of tissue culture cells. Two explanations are offered for the observed changes in zymodeme and virulence: a zymodeme is not a stable inherent property of the ameba. Alternatively, the original isolate consisted of two zymodeme populations and the conditions of growth selected for one or the other of the populations. In either case, our results suggest that the finding of a particular zymodeme in a culture of E. histolytica isolated from an asymptomatic carrier of the parasite cannot be used to predict a clinical condition or serve as a basis for the recommendation of therapy.     

The Plastids and Pigments of Fresh and Dried Chinese Gooseberries (Actinidia chinensis)

The pericarp of Chinese gooseberries is green due to the presenceof low concentrations of chlorophylls. On a f. wt basis thereis about 1.5 times more tetrapyrollic pigments (chlorophyllsand related compounds) in the outer pericarp than in the innerpericarp, whereas the carotenoid pigment values only showed1.25 times more in the outer than in the inner part of the fruit.The drying of Chinese gooseberries at 40 °C for 40 h resultedin the loss of at least half the tetrapyrollic pigments andof carotenoids. Chlorophylls a and b were converted to chlorophyllides,pheophytins and pheophorbides. Chloroplasts in the outer pericarp are clustered closely aroundthe nucleus and have a well-defined grana and inter-granal membranesystem. In the inner pericarp the chloroplasts are again clusteredaround the nucleus and there is a proliferation of inter-granalmembranes. In dried tissue the limiting membrane of chloroplastswas completely dispersed whereas some of the internal membranesremained intact. Actinidia chinensis Planch., Chinese gooseberry, Kiwi fruit, pigments, chloroplast structure     

Influence of prey species and stages on predatory efficiency and development of two phytoseiid mites

The effect of prey species and the different stages of prey on the predatory efficiency and biology of the phytoseiid mites,Amblyseius gossipi Elbadry andTyphlodromus mangiferus sp. n. was studied. It was found that feeding either predator onTetranychus cucurbitacearum (Sayed) promoted faster development and a higher rate of oviposition than rearing on the twospotted spider mite,T. urticae (Koch). Different stages of both prey species also produced different responses in the biological activities of these predaceous mites.

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Résumé On a étudié l'effet de l'espèce de la proie et de ses différents stades sur l'efficacité et la biologie des acariens phytoseiides,Amblyseius gossipi Elbadry etTyphlodromus mangiferus sp. n. Il a été constaté que l'alimentation des 2 prédateurs avecTetranychus cucurbitacearus (Sayed) assure un dévelopment plus rapide et une fécondité plus élevée que leur élevage surT. urticae (Koch). Les différents stades de ces 2 proies produisent des réponses différentes dans les activités biologiques des 2 acariens prédateurs.

     

The distribution and abundance of species of Danthonia DC on the New England Tablelands (Australia)

The distribution and percent frequency of the 13 species and one variety of Danthonia recorded on the New England Tablelands were examined in a survey of 110 predominantly pastoral sites. The relationships between site factors and the presence and percent frequency of the different Danthonia species were subjected to canonical correlation analysis. Individual species were found to have particular sets of environmental factors associated with their distribution and percent frequency; a fact not well recognized in the past. The most common factor associated with the abundance of Danthonia was either a long time since the last cultivation or no cultivation at all. Other factors such as altitude, drainage, soil parent material, soil texture and total soil phosphorus status were also of some importance. The six species most commonly recorded throughout the New England Tablelands were D. racemosa RBr, D. pilosa RBr, D. laevis J. Vickery, D. linkii Kunth., D. richardsonii Cashmore and D. tenuior (Steud.) Conert. Of these the first three are ecologically wide species. The first two can coexist with other naturalized species under grazing while D. laevis tends to disappear with disturbance. D. linkii and D. richardsonii require fairly narrow habitat conditions but these conditions and consequently these species are common. Both species respond positively to disturbance. Danthonia linkii increases with cultivation and D. richardsonii colonizes eroded soil surfaces. Danthonia tenuior has more restricted requirements and is not so frequent The other seven species and one variety did not occur often enough for detailed study and the occurrence of two of these species, D. carphoides and D. caespitosa must be considered as rare.     

Purine metabolic enzymes in lymphocytes. I. Adenosine deaminase and purine nucleoside phosphorylase activities in mouse lymphocyte subpopulations

The distribution of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) activities in lymphoid organs and lymphocyte subpopulations in mice, and the effect of phytohemagglutinin P (PHA-P) and concanavalin A (Con A) on the enzyme activities were studied. ADA activity was distributed equally in cells from all organs used and no mouse strain differences were observed. In contrast, PNP activity varied with the mouse strain, being highest in C57BL/6 mice and lowest in BALB/c mice, and with the organ in ICR mice, being high in peripheral blood lymphocytes and spleen lymphocytes, low in mesenteric lymph node cells and absent or very weak in thymus cells. T and B lymphocytes were prepared from spleen of ICR mice. High ADA activity was found in both T and B lymphocytes, whereas PNP activity in the T lymphocytes was about one-third of that in the B lymphocytes. PNP activity in thymus cells was increased to the normal level of T lymphocytes in the spleens by cultivation without stimulant. The development of PNP activity in thymus cells was partially inhibited by Con A but was not affected by PHA-P. ADA activity in thymus cells was enhanced by in vitro stimulation with PHA-P but not with Con A. In contrast, in spleen lymphocytes the development of ADA activity was enhanced by stimulation with PHA-P and Con A, and that of PNP activity was enhanced by PHA-P but not by Con A.