Effects of stress and beta-funal trexamine pretreatment on morphine analgesia and opioid binding in rats

This study was essentially an in vivo protection experiment designed to test further the hypothesis that stress induces release of endogenous opioids which then act at opioid receptors. Rats that were either subjected to restraint stress for 1 hr or unstressed were injected ICV with either saline or 2.5 micrograms of beta-funaltrexamine (beta-FNA), an irreversible opioid antagonist that alkylates the mu-opioid receptor. Twenty-four hours later, subjects were tested unstressed for morphine analgesia (tail-flick assay) or were sacrificed and opioid binding in brain was determined. [3H]D-Ala2NMePhe4-Gly5(ol)enkephalin (DAGO) served as a specific ligand for mu- opioid receptors, and [3H]-bremazocine as a general ligand for all opioid receptors. Rats injected with saline while stressed were significantly less sensitive to the analgesic action of morphine 24 hr later than were their unstressed counterparts. Beta-FNA pretreatment attenuated morphine analgesia in an insurmountable manner. Animals pretreated with beta-FNA while stressed were significantly more sensitive to the analgesic effect of morphine than were animals that received beta-FNA while unstressed, consistent with the hypothesis that stress induces release of endogenous opioids that would protect opioid receptors from alkylation by beta-FNA. beta-FNA caused small and similar decreases in [3H]-DAGO binding in brain of both stressed and unstressed animals. Stressed rats injected with saline tended to have increased levels of [3H]DAGO and [3H]-bremazocine binding compared to the other groups. This outcome may be relevant to the tolerance to morphine analgesia caused by stress.     

Isolation of intact high-molecular-weight DNA by using guanidine isothiocyanate.

A method for obtaining high-molecular-weight chromosomal DNA from Bacteroides intermedius and Bacteroides gingivalis is described. This technique is a modification of the guanidine isothiocyanate isolation procedure for RNA and should be useful for isolating intact DNA from organisms with high endogenous nuclease activity.     

Assignment of the prealbumin (PALB) gene (familial amyloidotic polyneuropathy) to human chromosome region 18q11.2–q12.1

Summary The assignment of the human prealbumin (PALB) gene to chromosome region 18q11–q12.1 has been achieved using a human genomic probe in the study of human-mouse somatic cell hybrids and by in situ hybridization. Because familial amyloidotic polyneuropathy was reported previously to be due to a mutation in prealbumin, it can be inferred that the gene for this disorder also maps to 18q11.2–q12.1.     

Microbial transformation of 1,6-dichloro-1,6-dideoxy-β,D-fructofuranosyl-4-chloro-4-deoxy-α,D-galactopyranoside (TGS) by soil populations

Summary Soil microorganisms from one site were shown to be consistently capable of the transformation of 1,6-dichloro-1,6-dideoxy-,d-fructofuranosyl-4-chloro-4-deoxy-,d-galactopyranoside (TGS) in laboratory batch cultures. With fresh soils, all of the available chloride ions were released from the molecule. Subcultures of a TGS-dehalogenating bacterial community produced a progressive decline in the dehalogenating capabilities towards the substrate. The soil organisms did not utilise TGS as a carbon source. The transformation was achieved by co-metabolism and was probably supported by an unknown component in the soil. Four bacterial species were isolated from the TGS-dehalogenating soil community: twoBacillus species, anAcinetobacter group isolate and aMicrococcus group isolate. Thin-layer chromatography confirmed the disappearance of the chlorosugar and high-performance liquid chromatography demonstrated that neither of the constituent monosaccharides—1,6-dichlorofructose nor 4-chlorogalactosucrose was accumulated as an intermediate.

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Resumen Microorganismos de suelo de cierto lugar demostraron consistemente ser capaces de realizar la transformación de 1,6-dicloro-1,6 dideoxi--D-fructofuranosil-4-cloro-4-deoxi-,D-alactopiranosa (TGS) in culturas de laboratorio de tipo discontinuo. Con muestras frescas de suelo, todos los iones cloruro fueron liberados de la molecula. Subculturas de una comunidad bacterial capaz de dehalogenizar TGS produjeron una declinación progresiva de la capacidad de dehalogenizar el substrato. Los microorganismos no utilizaron el TGS como fuente de carbono. La transformación se realiza por co-metabolismo y probablemente se base en un componente del suelo, no determinado. Cuatro especies bacteriales fueron aisladas de la comunidad de bacterias de suelo con capacidad de dehalogenar el TGS: dos especies deBacilo, unaAcinelobacteria y unMicrococo. Estudios de cromatografía de capa delgada confirmaron la desaparición del clorosacárido, y estadios de cromatografía liquida demostraron que ninguno de los componentes monoscáridos — 1,6-diclorofructuosa y 4-clorogalactosucrosa — eran acumulados como productors intermedios.

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Résumé Les microorganismes du sol d'un certain endroit ont été démontrés être capable, sans exception, de la transformation de 1,6-dichloro-1,6-dideoxy-,D-fructofuranosyl-4-chloro-4-deoxy-,D-galactopyranoside (TGS) en cultures de laboratoire du type discontinu. Avec des prélèvements frais du sol, tous les ions disponibles de chlorure ont été libérés de la molécule. Des souscultures d'une communauté bactérienne capable de déhalogeniser le TGS ont produit un déclin progressif de la capacité de déhalogeniser le substrat. Les microorganismes du sol n'ont pas utilisé le TGS comme source de carbone. La transformation s'est accomplie par cometabolisme et, probablement, s'est basée sur un component indéterminé du sol. Quatre espèces bactériennes ont été isolées de la communauté de bactéries du sol capable de déhalogeniser le TGS: deux espèces deBacillus, unAcinetobacter et unMicrococcus. Des études de chromatographie de couches fines ont confirmées la disparition du chlorosaccharide, tandis que des études de chromatographie liquide de haut rendement ont démontrées que, des monosaccharides constituants, ni 1,6-dichlorofructose ni 4-chlorogalactosucrose, n'ont été accumulés comme produits intermédiaires.

     

Joint report of the Third International Bovine Lymphocyte Antigen (BoLA) Workshop, Helsinki, Finland, 27 July 1986

Two hundred and eighty-two alloantisera were submitted by 20 participating laboratories from 13 countries and tested against lymphocytes of 1298 cattle. The cell panel consisted of samples from 38 Bos taurus breeds, 11 Bos taurus crossbreeds, 4 Bos indicus breeds, 6 Bos taurus x Bos indicus, and a variety of other crossbred populations. Using a standardized lymphocytotoxicity test, all 17 previously identified BoLA specificities were confirmed. The workshop produced agreement on 16 new lymphocyte alloantigenic specificities. Three of the new specificities behaved as splits of previously identified BoLA specificities. Four of the new specificities behaved as alleles at the agreed BoLA-A locus. Seven new specificities are tentatively assigned to the BoLA-A locus but require further definition. Two new specificities may represent products of a second closely-linked BoLA locus.     

Effect of secretagogues on chromogranin A synthesis in bovine cultured chromaffin cells. Possible regulation by protein kinase C.

Chromogranin A is a major component of storage granules in many different secretory cell types. After [35S]methionine labelling of proteins from cultured bovine chromaffin cells, chromogranin A was immunoprecipitated with specific antibodies, and the radioactivity incorporated into chromogranin A was determined and used as an index of its synthesis rate. Depolarization of cells with nicotine or high K+ evoked a Ca2+-dependent increase in chromogranin A synthesis, whereas muscarine, which does not evoke significant Ca2+ influx from bovine chromaffin cells, had no effect on chromogranin A synthesis. Forskolin, an activator of adenylate cyclase, affected neither the basal nor the nicotine-stimulated rate of chromogranin A synthesis. In contrast, 12-O-tetradecanoylphorbol 13-acetate (TPA), an activator of protein kinase C, significantly enhanced the incorporation of radioactivity into chromogranin A. Sphingosine, an inhibitor of protein kinase C, abolished both nicotine-stimulated and TPA-induced chromogranin A synthesis. In addition, long-term treatment of chromaffin cells with TPA decreased protein kinase C activity and inhibited the nicotine-stimulated chromogranin A synthesis. These results suggest that protein kinase C may play an important role in the control of chromogranin A synthesis.     

The taxonomic status and distribution of Bushbabies in Malawi with emphasis on the significance of vocalizations

The debated identity of a small forest bushbaby in Malawi is resolved by a short-term field study of the animals’ behavior. Locomotor styles, calling patterns, and the structure of advertising calls confirm that the species is Galago zanzibaricusrather than G. demidoffor G. thomasi.A detailed comparison of acoustic structure between the Malawi animals and G. zanzibaricusin Kenya demonstrates a degree of between-population variation, although the calls remain conservative in those parameters expected to aid recognition of conspecifics. Distribution records extend the known geographical range of G. zanzibaricusover most of the northern half of Malawi. Further studies are required to link the animals from this region with either of the previously recognized subspecies: G. z. zanzibaricusfrom East Africa or G. z. grantifrom southern Malawi, Mozambique, and Zimbabwe.     

Influence of 2-deoxy-D-glucose upon growth and invertase activity of carrot (Daucus carota L.) cell suspension cultures

Carrot (Daucus carota L.) cell suspension cultures grew well when provided with glucose, fructose, sucrose or raffinose. Galactose and melibiose supported less growth unless supplemented with glucose or fructose. In combination with ten different sugar mixtures, 2-deoxy-D-glucose (dGlc) inhibited culture growth. Inhibitory effects of dGlc were more marked with fructose, melibiose, raffinose or mixtures of these sugars in the culture medium. The presence of glucose or galactose reduced the inhibitory effects of dGlc on culture growth. Experiments with radioactive labelled sugars demonstrated that dGLc uptake was greater in the presence of fructose than glucose, and that growth inhibition of dGlc coincided with its uptake. Reduced protein content was also associated with the inhibitory effects of dGlc. Cultured cells contained lower levels of invertase (EC 3.2.1.26) activity during the active phase of culture growth (up to 25 days after subculture) than when growth had peaked and subsequently declined. Acid and alkaline invertase activities were not greatly reduced by exogenous hexoses. Invertase activity was greatest during periods of low protein content in all cultures and was inhibited by dGlc during the latter phases of the culture period. Free intracellular sugars throughout the culture period consisted mainly of glucose and fructose.