Immunological identification of uncoupling protein in interscapular "brown" adipose tissue of suckling and adult pipistrelle bats (Pipistrellus pipistrellus).

1. Interscapular adipose tissue of suckling and adult pipistrelle bats was examined for the presence of the 32,000 Mr "uncoupling protein" diagnostic of brown adipose tissue. 2. Following separation by SDS-polyacrylamide gel electrophoresis, mitochondrial proteins were blotted onto nitrocellulose and probed for uncoupling protein with an anti-(ground squirrel uncoupling protein) serum. 3. Immunoreactivity consistent with the presence of uncoupling protein was found in all samples of adipose tissue mitochondria from both suckling and adult bats. 4. It is concluded that interscapular adipose tissue in pipistrelle bats exhibits the critical biochemical criterion for being designated functionally "brown".     

Decrease in insulin-containing secretory granules and mitochondrial gene expression in mouse pancreatic islets maintained in culture following streptozotocin exposure

We have previously described a preferential reduction in the secretory response to nutrient secretagogues in pancreatic mouse islets maintained in culture after in vitro exposure to streptozotocin (SZ). This reduction was associated with an impaired substrate metabolism at the mitochondrial level. To further clarify this issue, mouse pancreatic islets were exposed in vitro to 2.2 mM SZ for 30 min. At 4 h after SZ treatment ultrastructural changes were apparent in the endoplasmic reticulum and Golgi areas of the B-cells. However, 2 and 6 days following SZ exposure the B-cells appeared well preserved, except for a marked decrease in the number of insulin-containing secretory granules. A morphometric analysis of the B-cells 6 days after SZ exposure showed a normal B-cell size and a normal volume fraction of B-cell mitochondria. However, there was a decrease in total islet size and a 13% decrease in the volume fraction of B-cells in the islets. These mouse islets exhibited a decreased content of the mitochondrial DNA-encoded cytochrome b mRNA, as evaluated by dot-blot analysis. As a whole, the data obtained indicate that SZ treatment does not induce a decrease in the number of mitochondria or long-lasting ultrastructural damage to this organelle. However, there is a clear decrease in the cytochrome b mRNA, suggesting that SZ can induce damage to the mitochondrial DNA.     

The sulphydryl groups of ox brain and liver glutamate dehydrogenase preparations and the effects of oxidation on their inhibitor sensitivities

Glutamate dehydrogenase preparations from several sources have been shown to have suffered limited proteolysis during purification. This proteolysis has been previously shown to involve removal of the N-terminal tetrapeptide and to result in changes in the regulatory properties of the enzyme. In the present work the previously unidentified N-terminal residue of the unproteolysed enzyme from ox brain and liver is shown to be cysteine. The thiol group of this residue is masked in the native enzyme but it becomes accessible after reduction. Exposure of solutions of the unproteolysed enzyme to air oxidation causes large changes in its sensitivity to inhibition by the antipsychotic drug perphenazine, GTP and by high concentrations of NADH. No such changes occurred in the behaviour of preparations of the enzyme that had suffered proteolysis during purification under these conditions.Special issue dedicated to Dr. Santiago Grisolia.     

Chrysoperla carnea predation on Heliothis virescens larvae parasitized by Microplitis croceipes

Predation by third instar larvae of Chrysoperla (=Chrysopa) carnea (Stephens) (Chrysopidae) did not alter the ratio of unparasitized Heliothis virescens (F.) (Noctuidae) larvae to H. virescens larvae parasitized by Microplitis croceipes (Cresson) (Braconidae) when these second instar larvae were exposed together to the predator on cotton (Gossypium hirsutum L., Malvaceae) in field cages. This indicates that C. carnea larvae did not prefer either parasitized or unparasitized larvae.

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Prédation par Chrysopa carnea des chenilles d'Heliothis virescens parasitées par Microplitis croceipes

Résumé Les prédations de chenilles d'Heliothis virescens, parasitées ou saines, élevées sur coton (Gossypium hirsutum L.), de la variété Stoneville 213, ont été comparées, dans des cages de 10 m² chacune placées dans la nature. Des chenilles du second stade ont été placées sur des pieds de coton dans 10 cages, à raison de 160 chenilles préalablement exposées à M. croceipes pendant leur premier stade et 40 chenilles saines par cage. Des larves du troisième stade de Chrysopa carnea ont été ajoutées dans 6 cages, à raison de 500/cage, et 4 cages ont servi de témoins pour évaluer les autres causes de mortalité. L'expérience a été répétée 2 fois. Les chenilles d'H. virescens ont été retirées au bout d'un jour dans une expérience et au bout de 2 jours dans l'autre. C. carnea n'a fait aucun choix entre chenilles parasitées ou non; la fréquence moyenne de chenilles parasitées n'a pas présenté de différence significative entre les cages avec ou sans C. carnea. Qui qu'il en soit, C. carnea a réduit significativement la survie des chenilles d'H. virescens parasitées ou non.

     

Copepoda from deep-sea hydrothermal vents and cold seeps

Recent explorations of hydrothermal vents in the eastern Pacific (Juan de Fuca spreading zone, Guaymas Basin in the Gulf of California, East Pacific Rise at 21° N and 13° N, and Galapagos Rift) and on the Mid-Atlantic Ridge have revealed many copepods, mostly siphonostomatoids with few poecilostomatoids. In these habitats in depths from 1 808 to 3 650 m water temperatures may reach nearly 15 ° C. Among more than 22 000 copepods from vents examined two new families, 11 new genera, and 32 new species were represented.In addition, two new copepods were found in 3 260 m at cold seeps at the base of the West Florida Escarpment in the Gulf of Mexico, an environment not thermally active, with water temperatures about 4.39 °C.Some of these copepods were associated with host invertebrates such as a Nuculana-like protobranch bivalve, a polychaete, and two species of shrimps. Others were obtained from washings of bivalves or vestimentiferans or by means of corers or slurp guns.     

Distribution of isoaccepting tRNAs and codons for proline and glycine in collagenous and noncollagenous chicken tissues

The relation between codon usage and tRNA content for proline and glycine, the major constituents of collagen, was studied in two tissues: the magnum of laying hen oviduct and the leg tendons of chick embryo where collagen is produced. Although the relative contents of tRNA(GCCGly) and tRNA(IGGPro) in tendons, as compared to magnum indicate a specialization of the tRNA population for collagen synthesis, the distribution of the preponderant codons in collagen mRNA is correlated but at a lesser extent to that of their cognate tRNAs.     

Relative content of isoaccepting tRNAs for glycine and proline in avian tendon cells with different rates of procollagen synthesis

The relative amounts of iso-tRNAsGly and iso-tRNAsPro existing in chick embryo tendon are indicative of a specialization of the tRNA population for collagen synthesis. These amounts are not modified (i) in primary avian tendon (PAT) cells in culture for which the procollagen production varies from about 10% of total protein synthesis to 60% and (ii) in tendons from immature chicks, which show a 3-fold decrease of procollagen production with increasing age. The characteristic tRNA pattern was not maintained in cells which had lost the ability to make high levels of collagen as observed in the cases of: (i) PAT cells reaching confluency; (ii) virus-transformed PAT cells and (iii) tendon from adult chick. Our data are consistent with the idea that tendon tRNA specialization for collagen synthesis is a differentiation feature independent of the expression level of the collagenic function but related to its maintenance.     

Cementum annulation and age determination in Homo sapiens. II. Estimates and accuracy

The cementum annulation aging technique was evaluated in a sample of 80 clinically extracted premolars (age range 11–70 years). Demineralized thin sections (7μm) stained with hematoxylin were used. The correlation (r) between age and adjusted count (number of annulations added to age of tooth eruption) was 0.78 for the entire sample (N = 73) and 0.86 for a subsample in which teeth with periodontal disease were excluded (N = 55). Standard error of the estimates ranged from 4.7 to 9.7 years depending on sex and health status of the tooth. The technique provided significantly better estimates for females than for males. The overall inaccuracy (mean absolute error) of the technique was 6.0 years, with a bias (mean error) of 0.26 years. Reduced major axis regression of adjusted count on age produced a slope of 0.797 for the entire sample and 0.889 for the nonperiodontal disease subsample. These slopes are consistent with a hypothesis of annual deposition of cementum rings given a decrease in cementogenesis with increasing age.     

Parathyroid hormone-induced changes of the brush border topography and cytoskeleton in cultured renal proximal tubular cells

Summary In order to examine the possibility of parathyroid hormone-mediated ultrastructural rearrangements in target epithelium, isolated canine renal proximal tubular cells were grown on a collagen-coated semipermeable membrane in a defined medium. Scanning and transmission electron microscopy of these monolayers revealed abundant microvilli. Exposure of the proximal tubular cells to parathyroid hormone resulted in a biphasic changes involving: (1) dramatic shortening and rarefaction of microvilli within 1 min; and (2) recovery of microvillar topography after 5 min. A similar shortening of microvilli was observed following exposure to ionomycin, whereas incubation with cyclic AMP resulted in an elongation of microvilli. Parathyroid hormone stimulated cyclic AMP production and increased cytoplasmic free calcium concentration in cultured proximal tubular cells. Pretreatment of cells with a calmodulin inhibitor abolished the effect of parathyroid hormone on brush border topography. Shortening of microvilli was associated with a disappearance of microvillar core filaments. Staining of F-actin with fluoresceinphalloidin showed that parathyroid hormone resulted in fragmentation of stress fibers. It is concluded that parathyroid hormoneinduced cell activation involves cytoplasmic-free calcium, calmodulin, and the cytoskeleton.