ATP and other purine nucleotides stimulate the inactivation of ornithine transcarbamylase by broken lysosomes

Calpain II, a high Ca2+-requiring form of Ca2+-dependent cysteine proteinase (EC 3.4.22.17), isolated from bovine lens was found to cleave actin and vimentin, two major cytoskeletal elements of the lens. Polyacrylamide gel electrophoresis revealed that actin (Mr 43 000) was broken down through intermediary products of approximate Mr 42 000 and 40 000, while vimentin (Mr 57 000) was rapidly cleaved into several fragments ranging from Mr 44 000 to 20 000. The cleavage was dependent on Ca2+ and could be blocked by calpastatin , a calpain-specific inhibitor. These findings suggest that calpain might play a role in age-related degradation of the lens cytoskeleton.     

Electrophoresis '82 : Advanced methods. Biochemical & clinical applications. Proceedings of the International Conference on Electrophoresis,Athens, April 1982 Edited by D. Stathakos Walter de Gruyter; Berlin, 1983 xvi + 867 pages. DM 260.00

A competitive solid-phase immunoassay for the determination of testosterone in serum samples using time-resolved fluorescence is described. The solid phase is a testosterone-3-(O-carboxymethyl)-oxime-ovalbumin conjugate coated to polystyrene microtiter strips. Europium-labelled polyclonal and monoclonal antibodies against testosterone-3-(O-carboxymethyl)-oxime-bovine serum albumin were compared. Their behavior was quite similar although the polyclonal antibody was more sensitive, giving a detection limit of 15 fmol testosterone per assay. Correlation with RIA was very good (r = 0.982 and y = ?0.150 + 0.969x).     

Effect of temperature on the reproduction of Glossina pallidipes, with reference to G. m. morsitans

The reproductive biology of G. pallidipes Austen was studied at 28°, 25° and 22° C. Experiments showed that incubation of puparia at 28° C resulted in sterility of both males and females. Incubation at 22° C resulted in a reduced fecundity of the females due to egg retention; the fertility of the males was not affected.Comparative studies with G. m. morsitans Westw. showed that G. m. morsitans puparia are less affected by a temperature of 28° C than are G. pallidipes puparia.

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Effet de la température sur la reproduction de Glossina pallidipes, avec référence à G. m. morsitans

Résumé Les productivités de G. pallidipes Austen élevés au laboratoire pendant tout leur cycle à 22, 25 et 28° C, ont été comparées.A 28° C, la vie intrapupale est réduite à environ 23 jours, contre 30 jours environ à 25° C; la survie des adultes est plus brève qu'à 25° C et les mouches ne s'accouplent pas. Les ovaires présentent une rétention d'oeufs et seulement 1/3 des mâles contient des spermatozoïdes mobiles. A 22° C, le cycle est considérablement prolongé, la vie intrapupale durant environ 40 jours. Les femelles s'accouplaient environ 14 jours après l'émergence. Les ovaires présentaient une rétention d'oeufs, bien que moins souvent qu'à 28° C. Les mâles contenaient des spermatozoïdes mobiles.Des expériences avec changements de température à différents moments du cycle ont montré que la stérilité des mâles et des femelles est provoquée par l'incubation de pupes de G. pallidipes à 28° C. La mensuration des ovocytes montre à 28° C un effet nocif sur leur maturation. Des observations sur les testicules dans les pupes révèlent, par comparaison avec 25° C, que l'enroulement des testicules et des spermatozoïdes est retardé à 28° C, tandis que la pigmentation des testicules est retardée à 22° C. Les pupes de G. m. morsitans sont moins affectées à 28° C que celles de G. pallidipes.

     

Detection of the messenger RNA coding for preproenkephalin A in bovine adrenal by in situ hybridization

The messenger RNA (mRNA) coding for the adrenal precursor of enkephalins (preproenkephalin-A) has been detected in bovine adrenal medulla cells using in situ hybridization with 32P-labelled preproenkephalin A (PPA) complementary DNA. In formaldehyde- and Carnoy-fixed tissue sections, an intense elective labelling restricted to the cells located at the periphery of the adrenal medulla can be detected after hybridization procedure, using X-ray film and classical autoradiographic procedure. Adequate controls show that this labelling is obtained only using PPA complementary DNA, inserted or not in its vector. Distribution of PPA mRNA appears identical to that of its immunoreactive end products, namely Met-enkephalin and BAM22 peptide, detected by immunohistochemistry. Norepinephrine, detectable using monoamine histofluorescence, appears restricted to the cells of the center of the gland unlabelled for PPA mRNA and its end-products. Cultured bovine adrenomedullary cells that exhibited enkephalin immunoreactivity also contain PPA mRNA located in their cytoplasm.     

Isolation and characterization of axolemma-enriched fractions from discrete areas of bovine CNS

Myelinated axons were isolated by flotation from bovine pons, middle cerebellar peduncle, cervical spinal cord and three regions of the subcortical white matter. The myelinated axons were osmotically and mechanically shocked, followed by fractionation on a linear 15% sucrose to 45% sucrose density gradient. Axolemma-enriched fractions (AEF) found in the 28% to 32% sucrose region of the gradient from brainstem and cord white matter had high acetylcholinesterase (AChE) while little or nil AChE activity was found in corresponding AEF derived from the subcortical white matter. Morphologically, the subcortical white matter from all regions contained a heterogeneous population of well-myelinated to thinly myelinated axons, while brainstem and cord regions contained a more homogeneous population of well-myelinated axons. Histochemical analysis of AChE localized this enzyme to axonal elements. The AEF derived from any white matter source had similar polypeptide compositions. AEF derived from subcortical white matter contained two-fold more myelin basic protein and a three-fold greater content of 2 3 cyclic nucleotide 3 phosphodiesterase (CNP) compared with AEF derived from well myelinated white matter. We conclude that the purity of the AEF is related to the degree of myelination of the white matter from which the AEF is derived. Homogeneously well myelinated white matter (pons, cerebellar peduncle, cervical spinal cord) yields the highest purity AEF, as judged by the low CNP and myelin basic protein content and highest enrichment in AChE specific activity.     

Isolation and characterization of the bovine hypothalamic corticotropin-releasing factor

A 41 amino acid peptide with high intrinsic corticotropin-releasing activity was isolated from 1000 bovine hypothalami by means of immunoaffinity chromatography, gel filtration, and two steps of reverse phase HPLC. The primary structure of the amino terminal 39 amino acids was characterized by gas phase sequence analysis. The sequence of the amidated carboxyl terminal dipeptide was established by digestion of the intact natural product with Staphylococcus aureus V8 protease, dansylation of the digest and comparative reverse phase liquid chromatography studies with the synthetic dansylated dipeptides Ile-Ala-NH2, Ile-Ala-OH, Ala-Ile-NH2 and Ala-Ile-OH. The complete structure of the bovine corticotropin-releasing factor was established as: Ser-Gln-Glu-Pro-Pro-Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val- Leu- Glu-Met-Thr-Lys-Ala-Asp-Gln-Leu-Ala-Gln-Gln-Ala-His-Asn-Asn-Arg-Lys-Leu- Leu- Asp-Ile-Ala-NH2 using approximately 650 pmol of material.     

Neuromedin B is a major bombesin-like peptide in rat brain: regional distribution of neuromedin B and neuromedin C in rat brain, pituitary and spinal cord

Neuromedin B and neuromedin C are the novel mammalian bombesin-like peptides isolated from porcine spinal cord. We have developed highly specific and sensitive radioimmunoassays for neuromedin B and neuromedin C, and determined their regional distribution in rat central nervous system. Prior to measurements of the tissue contents, immunoreactive neuromedin B and C were characterized by gel-filtration and high performance liquid chromatography. Neuromedin B and C immunoreactivities have similar regional distribution in rat brain, but the content of immunoreactive neuromedin B is 2-6 times higher than that of immunoreactive neuromedin C in every region. These results indicate that neuromedin B is a major endogenous bombesin-like peptide in rat brain and has specific functions of physiological importance.     

Analysis of protein synthesis in morphologically classified bovine follicular oocytes before and after maturation in vitro

Bovine cumulus oocyte complexes (COCs) were isolated from antral ovarian follicles (4-8 mm). Immature COCs were classified into four categories, based on the homogeneity and clearness of the ooplasm and the transparency and compactness of the cumulus investment. In this study, the incorporation of TCA-precipitable 35S-methionine and the protein synthesis patterns of oocytes of these four categories were examined. Before maturation in vitro, similar incorporation rates and identical protein synthesis patterns were observed between oocytes of categories 1-3. Immature oocytes of category 4 showed reduced incorporation rates and exhibited aberrant protein synthesis patterns. After maturation in vitro, the patterns of category 4 oocytes were identical with the patterns of those in categories 1-3. The incorporation of 35S-methionine into in vitro matured oocytes was lower (P less than .001) in all categories. Based on these results, it is concluded that the initial classification of oocytes into four categories can be reduced to two categories.     

Glutamine synthetase gene of Bacillus subtilis

The glutamine synthetase gene (glnA) of Bacillus subtilis was purified from a library of B. subtilis DNA cloned in phage lambda. By mapping the locations of previously identified mutations in the glnA locus it was possible to correlate the genetic and physical maps. Mutations known to affect expression of the glnA gene and other genes were mapped within the coding region for glutamine synthetase, as determined by measuring the sizes of truncated, immunologically cross-reacting polypeptides coded for by various sub-cloned regions of the glnA gene. When the entire B. subtilis glnA gene was present on a plasmid it was capable of directing synthesis in Escherichia coli of B. subtilis glutamine synthetase as judged by enzymatic activity, antigenicity, and ability to allow growth of a glutamine auxotroph. By use of the cloned B. subtilis glnA gene as a hybridization probe, it was shown that the known variability of glutamine synthetase specific activity during growth in various nitrogen sources is fully accounted for by changes in glnA mRNA levels.     

牛胰多肽对大鼠胰外分泌的抑制作用

在大鼠观察了牛胰多肽(BPP)对由蛋白胨和雨蛙素引起的大鼠胰外分泌的影响。向十二指肠灌入25%蛋白胨后90min 时,对照组的蛋白排出量约为其基础值的10倍,而1、3、5及10ug/kg BPP 剂量组的蛋白排出量分別为对照组的47%、33%、29%及29%。在灌注剂量为3μg/kg 雨蛙素后60min 时,对照组的蛋白排出量约为其基础值的14倍,但3、5及10μg/kgBPP 剂量组的蛋白排出量分別为对照组的74%、68%及55%。各组胰淀粉酶浓度的变化和其蛋白排出量变化相平行。胰液量的增加不受 BPP 影响。结果提示,BPP 能抑制胰酶的大量分泌,并有剂量依从关系,但不抑制胰液量。