Histochemical Specificity of β-Galactose Binding Lectins from Arachis Hypogaea (Peanut) And Ricinus Communis (Castor Bean)

Previous histochemical studies have demonstrated disparities in the binding of two lectins with a nominal specificity for terminal β-D-galactose. Biochemical studies have shown that the most complementary structure for binding peanut agglutinin (PNA) is the terminal disaccharide Gal-(β1 → 3)-GalNAc, whereas the most complementary structure for binding Ricinus communis agglutinin I (RCA I) is the terminal disaccharide Gal-(β1 → 4)-GlcN Ac. However, it is not known if only these differences in affinity account for the different histochemical staining reactions observed on tissue sections. In the present study we compared the staining patterns of PNA and RCA I by inhibiting in situ the binding of each lectin conjugated to horseradish peroxidase (HRP) with increasing concentrations of unlabeled PNA or RCA I. The PN A-HRP conjugate did not stain most tissue sites suspected of containing an abundance of glycoconjugates with terminal Gal-(β → 4)-GlcNAc. Moreover, unlabeled PNA failed to significantly inhibit strong RCA 1-HRP staining in these sites. In loci thought to contain variable amounts of glycoconjugates with terminal Gal-(β1 → 3)-GalNAc, unlabeled RCA I decreased PNA-HRP reactivity only slightly or not at all, whereas weak to strong RCA I-HRP staining was diminished or abolished by unlabeled PNA. The results suggest that PNA staining is restricted to glycoconjugates with terminal Gal-(β1 → 3)-GalN Ac. RCA I apparently reacts most strongly with glycoconjugates having the terminal disaccharide Gal-(β1 → 4)-GlcNAc, but also stains sites containing a moderate to abundant amount of glycoconjugates with the terminal Gal-(β→ 3)-GaINAc sequence.     

Variables that influence the activity of captive orangutans

It is well known that the environment significantly influences the behavior of captive animals. However, the specific nature of the cues that promote speciestypical behavior patterns is not usually known. This study extends an earlier investigation by Wilson (Zoo Biology 1: 201–209, 1982). The objective was to identify and quantify specific environmental components that influence activity levels in orangutans. Six enclosure variables were quantified, and activity levels measured, for 29 orangutans housed in nine zoological parks. Multiple regression analysis indicated that the combination of the number of animals, amount of usable surface area, number of movable objects, and enclosure volume was the best predictor of activity levels, accounting for 58% of the variance in activity levels. It was concluded that the provision of large enclosures, containing large numbers of movable objects and providing social opportunities, would promote higher levels of activity in captive orangutans. © 1992 Wiley-Liss, Inc.     

Expression and regulation of Q8 b in a transfected cell line

RNase protection experiments showed that Q8 ^b was actively transcribed in a stably transfected cell line. Moreover, Q8 ^b responded to interferon- (IFN-) treatment with increased levels of mRNA expression. Thus Q8 ^b demonstrates a regulatory response to IFN- characteristic of many other class I genes. Cell surface expression of a Q8^b product could also be detected by flow cytometric analysis with the Qa-2-specific monoclonal antibody D3.262. The expression of the Q8^b cell surface product increased only slightly after cells were treated with IFN-. The Q8^b cell surface product was not sensitive to cleavage by phosphatidylinositol-phospholipase C. These results suggest that the Q8^b product, unlike the predominant forms of Qa-2-bearing molecules, is not anchored via phosphatidylinositol to the cell membrane. These results also suggest that Q8 ^b has the potential to contribute to the Qa-2 phenotype in vivo. Address correspondence and offprint requests to: L. Flaherty.     

Localization of 5′-ectonucleotidase/phosphatase activity within the olfactory sensilla of the spiny lobster,Panulirus argus

Summary Previous electrophysiological studies have shown that the olfactory organ (antennule) of the spiny lobster, Panulirus argus, has external chemoreceptors, which are selectively stimulated by adenosine 5-monophosphate (AMP) when present in seawater. Subsequent biochemical investigations revealed that AMP can be rapidly dephosphorylated by 5-ectonucleotidase/phosphatase activity associated with the olfactory sensilla (aesthetascs). In this study the deposition of cerium phosphate was used to examine the ultrastructural distribution of 5-ectonucleotidase/phosphatase activity in aesthetascs. Utilizing AMP as substrate, we found dephosphorylating activity to be associated with the outer membranes of both dendrites and auxiliary cells. Moreover, this activity was specifically localized to a narrow band that approximately corresponds to the transitional zone where dendrites develop cilia and branch extensively to form the outer dendritic segments. A similar distribution of the cerium phosphate reaction product was found when -glycerol phosphate was substituted for AMP. The alkaline-phosphatase inhibitor, levamisole, had no apparent effect on the deposition of reaction product when either AMP or -glycerol phosphate was used as substrate. The ectoenzymatic activity in the transitional zone may be of importance in clearing exogenous chemoexcitatory nucleotides from this region.Abbreviations ADP adenosine 5-diphosphate - AMP adenosine 5-monophosphate - ATP adenosine 5-triphosphate - EM electron microscopy     

Differential dynamics of receptor down-regulation and tyrosine aminotransferase induction following glucocorticoid treatment

Autoregulation of glucocorticoid receptor (GR) concentration in vivo may be an important determinant of steroid sensitivity. The dynamics of GR regulation were assessed and compared to regulation of tyrosine aminotransferase (TAT) expression in liver tissue taken from rats treated with a single 50 mg/kg i.v. dose of methylprednisolone. Plasma methylprednisolone concentrations were determined by HPLC analysis. Receptor and TAT message levels were determined by quantitative Northern hybridization. Methylprednisolone plasma kinetics showed a half-life of 0.6 h. Receptor occupancy occurred rapidly and cytosolic GR reappeared over 2–12 h. TAT activity rose between 2 and 6 h and then dissipated. Reduction in receptor mRNA levels occurred very rapidly, being detectable by 30 min following steroid administration. A down-regulated steady-state in GR message expression was reached by 2 h post-injection, and was maintained throughout the 18 h examined in this study. Comparison of methylprednisolone kinetics demonstrated that down-regulation was maintained long after drug was eliminated. In contrast, TAT message induction occurred with a sharp peak; maximal induction occurred between 5–6 h and return to baseline at approx. 8–10 h post-induction. This study shows that unlike TAT induction, GR message repression in vivo does not require continual presence of hormone.     

Methanogenesis in Big Soda Lake, Nevada: an Alkaline, Moderately Hypersaline Desert Lake

Incubated sediment slurries from Big Soda Lake, Nevada, produced significant levels of CH₄, and production was inhibited by 2-bromoethanesulfonic acid and by autoclaving. Methane production was stimulated by methanol, trimethylamine, and, to a lesser extent, methionine. Surprisingly, hydrogen, acetate, and formate amendments provided only slight or no stimulation of methanogenesis. Methane production by sediment slurries had a pH optimum of 9.7. A methanol-grown enrichment culture containing a small, epifluorescent coccus as the predominant organism was recovered from sediments. The enrichment grew best when FeS or autoclaved sediment particles were included in the media, had a pH optimum of 9.7, and produced ¹⁴CH₄ from ¹⁴CH₃OH. The methane formed by methanolgrown enrichment cultures was depleted in ¹³C by 72 to 77‰ relative to the methanol.     

Experimental Study of the Pathogenicity of Aspergilli for Mice

The relative virulence was determined for 14 species of aspergilli, by inoculating normal mice intravenously with graded doses of spores. Eleven were found to possess some degree of virulence, whereas three others were avirulent. Members of the Aspergillus flavus group were the only species that consistently killed mice with doses as low as 10(4) viable spores. When the in vivo fate of spores was compared for a virulent and an avirulent strain of Aspergillus, spores of the latter were cleared rapidly from the liver and spleen but grew in the kidneys and brain, producing progressive disease. Mice which inhaled spores did not succumb, but macrophages washed from their lungs contained spores. A relationship of virulence to spore characteristics such as germination time, size, shape, and external markings could not be established. Virulence could not be related to aflatoxin production inasmuch as at least one virulent strain did not produce aflatoxin in vitro.     

Genetic Counseling: An Evaluation of Public Health Genetic Clinics

The geographic distribution of County Health Department clinic facilities in the state of California has made it readily possible to establish a regionalized program for genetic counseling services, using public health nurses as a major source of case-finding. From both consumer and health professional standpoints, regionalized satellite genetic counseling clinics have been successful, and in particular, the effectiveness of public health nurses in identifying clinical genetic problems is readily apparent.Long-term follow-up reinforcement of genetic counseling appears to be an important conclusion from these studies. It is our suggestion that reinforcement of counseling would best be accomplished through the health team member (physician, nurse and so forth) following the patient or family rather than through the consulting geneticist.     

Degradation of Escherichia coli DNA: evidence for limitation in vivo by protein X, the recA gene product.

Summary DNA is more extensively degraded after it is damaged in recA mutants of E. coli than in wild type cells. All data presented here are consistent with the recA gene product, protein X, being an inhibitor of nalidixic acid induced degradation of the bulk DNA (but not of newly replicated DNA). Production of protein X also is correlated with appearance of various S.O.S. repair functions. Evidence was obtained by comparing the rates of protein X synthesis and solubilization of uniformly-labeled DNA in intact cells, incubated in the presence of nalidixic acid. A set of mutants at the lexA locus produced protein X at different rates and degraded their DNA at rates which were inversely correlated to their rates of protein X production. A low concentration of rifampicin quite specifically inhibited protein X production by wild type E. coli, and allowed more rapid DNA degradation. After the DNA was damaged by the incubation of cells in the presence of nalidixic acid, cells preloaded with protein X degraded their DNA more slowly. We propose that protein X could protect DNA against degradation by binding to singlestranded regions, thereby inhibiting nuclease action.