Frequency of resistance to tetracycline and chloramphenicol amongSalmonella strains isolated in the Netherlands in 1962

Out of 12,472 strains ofSalmonella isolated in the Netherlands in 1962, 1365, or 10.94%, were found to be resistant to tetracycline or chloramphenicol or to both. Compared with the findings of the preceding years (1958/59:2.08%, 1960:1.29%, 1961:3.96%) this is a considerable increase. Of these 1365 strains, 1285, or 94.1%, were resistant to tetracycline and 46, or 3.4%, were resistant to chloramphenicol. The remaining 34 strains, or 2.5%, were resistant to both drugs.Among allSalmonella strains isolated in 1962, 5517 belonged to the speciesS. typhi murium. Of these, 1203 or 21.8%, were resistant to tetracycline. The resistance rates of strains originating from human patients, calves, pigs, other animals and other materials were 24.4%, 37.1%, 15.0%, 8.0% and 5.7% respectively.Factors which may possibly have contributed to the greatly increased frequency of drug resistance inS. typhi murium are: (1) the rapid spread of the use of tetracycline for therapeutic, prophylactic and nutritive purposes, and (2) the possibility of an episome-mediated transfer of drug resistance from relatively harmless intestinal bacteria, such asE. coli, toS. typhi murium.     

A Rapid Screening Test for Aflatoxin-synthesizing Aspergilli of the flavus-oryzae Group

A rapid test for the recognition of aflatoxin-synthesizing strains of the Aspergillus flavus–oryzae group is described. For this purpose the strains are cultivated on Czapek–Dox agar enriched with an aqueous extract of groundnuts, and in which sodium nitrate is replaced by ammonium chloride. Toxin production is observed by the production of a bright blue fluorescence in the medium when placed under an ultraviolet lamp.     

Modulation of Histamine Release in the Rat Brain by K-Opioid Receptors

The opioid modulation of histamine release was studied in rat brain slices labeled with L-[3H]histidine. The K(+)-induced [3H]histamine release from cortical slices was progressively inhibited by the preferential kappa-agonists ketocyclazocine, dynorphin A (1-13), Cambridge 20, spiradoline, U50,488H, and U69,593 in increasing concentrations. In contrast, the mu-agonists morphine, morphiceptin, and Tyr-D-Ala-Gly-(NMe)Phe-Gly-ol (DAGO) were ineffective as were the preferential delta-agonists [D-Ala2,D-Leu5]enkephalin (DA-DLE) and [D-Pen2,D-Pen5]enkephalin (DPDPE). Nor-binaltorphimine (nor-BNI) and MR 2266, two preferential kappa-antagonists, reversed the inhibitory effect of the various kappa-agonists more potently than did naloxone, with mean Ki values of 4 nM and 25 nM, respectively. The effects of ketocyclazocine and naloxone also were seen in slices of rat striatum, another brain region known to contain histaminergic nerve endings. We conclude that kappa-opioid receptors, presumably located on histaminergic axons, control histamine release in the brain. However, nor-BNI and naloxone failed, when added alone, to enhance significantly [3H]histamine release from cerebral cortex or striatum, and bestatin, an aminopeptidase inhibitor, failed to decrease K(+)-evoked [3H]histamine release. These two findings suggest that under basal conditions these kappa-opioid receptors are not tonically activated by endogenous dynorphin peptides. The inhibition of cerebral histamine release by kappa-agonists may mediate the sedative actions of these agents in vivo.     

Changes in protein synthesis and phosphorylation patterns during bovine oocyte maturation in vitro

Sequential protein synthesis and protein phosphorylation patterns were generated by radiolabelling bovine cumulus-oocyte complexes after various periods of culture with [35S]methionine and [32P]orthophosphate respectively. The radiolabelled oocytes were assessed for their nuclear status and used individually for gel electrophoresis. Marked changes in the protein synthesis patterns were observed exclusively after germinal vesicle breakdown (GVBD), whereas oocytes which remained in the germinal vesicle stage showed a consistent protein synthesis pattern. The changes were observed after 8 and 16 h or culture, shortly after GVBD and before first polar body extrusion. From 3 h of culture, dominant phosphoprotein bands with apparent molecular weights of 24,000 and two between 50,000 and 60,000 were observed. The latter bands displayed slight molecular weight changes, which were not closely time related. After GVBD, the phosphoprotein band with Mr 19,000 was no longer observed. This study demonstrates that specific changes in protein synthesis and protein phosphorylation are programmed during bovine oocyte maturation.     

Functional vasoactive intestinal polypeptide (VIP) receptors in human neuroblastoma subclones that contain VIP precursor mRNA and release VIP-like substances

Three phenotypically distinct subclones (SH-SY-5Y, SH-EP, SH-IN) of the human neuroblastoma cell line SK-N-SH were found to possess vasoactive intestinal polypeptide (VIP) precursor mRNA, release immunoreactive VIP, and express high-affinity VIP receptors coupled to adenylate cyclase. The apparent molecular mass for the receptor polypeptide, as determined by covalent cross-linking of 125I-VIP, was 49 kDa. After 2 days in culture, a concentration of immunoreactive VIP equivalent to the binding affinity of VIP to its receptor was found in the medium in two of these clones (SH-IN and SH-EP). Conditioned medium from SH-IN cells competitively displaced 125I-VIP binding and increased cAMP levels in SH-EP cells, indicating that all of the necessary components for a potential autocrine action of VIP exist in SK-N-SH cells. After numerous cell passages, the SH-EP subclone converted to a distinct phenotype in which VIP precursor mRNA and VIP immunoreactivity in the cell and medium were no longer detectable. In correlation, the VIP receptor number increased, and the EC50 for VIP stimulation of cAMP production shifted to a lower concentration. This points to the possibility that the continuous presence of endogenous VIP in earlier passage SH-EP cells causes a modification in VIP receptor number and cell responsiveness to VIP.     

Purification, properties and immunological detection of a bromoperoxidase-catalase from Streptomyces venezuelae and from a chloramphenicol-nonproducing mutant

A new bromoperoxidase-catalase was purified from the chloramphenicol-producing actinomycete Streptomyces venezuelae ISP 5230. The homogeneous enzyme showed brominating activity, catalase activity and a very low peroxidase activity. The spectral properties and pH dependence of the catalase activity showed similarities to conventional catalases. In contrast to other haem-bromoperoxidases, the bromoperoxidase-catalase was stable when treated with an ethanol/chloroform mixture. Gel filtration gave an estimated Mr of 127,000-136,000. SDS-PAGE showed a single band corresponding in mobility to a species with an Mr of 61,000. The pI was estimated to be 4.5. The bromoperoxidase-catalase was not present in active form in a mutant of S. venezuelae ISP 5230, blocked in the chlorination step of chloramphenicol biosynthesis. However, an inactive species of the enzyme was detected in crude extracts of the mutant by using antibodies. From these results it is concluded that this bromoperoxidase participates in the chlorination step during chloramphenicol biosynthesis.     

Autometallography for histochemical visualization of rat incisor polyanions with cuprolinic blue

Autometallography was applied to semi-thin sections of rat incisors fixed a solution of cuprolinic blue-aldehyde. The resulting reduction of silver ions to metallic silver amplifies the copper sulfide signal of the cationic dye. Silver grains were seen over the cell bodies of ameloblasts and odontoblasts but not over their processes. This was owing to the interaction of cuprolinic blue with the DNA and RNA of these cells. In the extracellular matrix, silver grains were unevenly distributed over the predentin, dentin, and forming enamel. The distal predentin near the mineralization front and a thin band of dentin located near the dentino-enamel junction displayed unexpectedly intense accumulation of silver grains, whereas all other portions of the extracellular matrix exhibited the distribution of glycosaminoglycans expected from previous studies. The present investigation constitutes a new application of autometallography to glycosaminoglycan histochemistry.     

Sexual and site differences in calcium consumption by the Malabar Giant Squirrel Ratufa indica

Summary The calcium ingestion of Malabar Giant Squirrels Ratufa indica (Sciuridae) was examined at two sites, Magod and Bhimashankar, in western India. In females at Magod a positive correlation was found between rates of calcium ingestion from food resources and the contribution of those resources to the daily diet. This relationship was true for all females at Magod irre-spective of their reproductive condition. This relationship was neither significant for males at the same site nor for both sexes at Bhimashankar. A differential requirement for calcium between the sexes and the occurrence of higher rates of calcium ingestion or the probable presence of more easily digestible calcium at Bhimashankar are postulated to explain the observed phenomenon. Mature leaves and bark appear to be reliable sources of calcium at these sites.