Inhibition of follicle-stimulating hormone-induced ovulation by indomethacin in the perfused rat ovary

In isolated, perfused ovaries of rats treated with pregnant mare's serum gonadotropin (PMSG), purified preparations of ovine follicle-stimulating hormone (FSH) (oFSH-211B) and rat FSH (rFSH-I-6), 100 ng/ml, were found to induce ovulations (4.8 +/- 0.9, n = 4, and 6.4 +/- 2.0, n = 5, ovulations per ovary, respectively). Indomethacin (5 micrograms/ml) added to the perfusate inhibited this ovulatory effect and exogenous prostaglandin F2 alpha (PGF2 alpha) (1 microgram/ml), or prostaglandin E2 (PGE2) (0.5 microgram/ml), reversed the blockade. Ovine FSH and rFSH had only a weak stimulatory effect on estradiol release, and only rFSH caused a significant increase in progesterone accumulation. Indomethacin reduced the stimulatory effect of rFSH on progesterone release, and this effect was reversed by PGE2 but not by PGF2 alpha. In a 6-h incubation experiment with preovulatory rat follicles, we tested the biological activity of gonadotropins used to induce oocyte maturation. The concentration of FSH used in the perfusion experiments induced oocyte maturation in more than 88% of the oocytes studied. The data confirm earlier findings that FSH can induce ovulations and show that prostaglandins are involved in this process. The data also indicate that prostaglandins might be involved in the FSH-induced increase of progesterone levels.     

Binding of F-protein (phosphofructokinase) to F-actin

By the use of sedimentation method the interaction of F-protein (phosphofructokinase) with F-actin has been studied in 0.1 M KCl, 10 mM K-phosphate buffer, pH 6.5. The calculated value of the constant of F-protein binding to F-actin is (7.3 +/- 1.0) X 10(5) M-1. The number of the F-protein binding sites calculated per chain weight of 80,000 is 0.43. The binding of F-protein to F-actin was found to be affected by pH-value.     

Interaction specificity of violamycin BI and BII with DNA using the hyperchromic spectra method

Interaction specificity of the anthracycline antibiotics violamycin BI and violamycin BII in respect to A.T and G.C pairs was investigated. For comparison denaturation of complexes with A.T and G.C specific ligands distamycin A and actinomycin D are presented. Making use of the least squares hyperchromic spectra measured in the course of thermal denaturation were partitioned into the components corresponding to the melting of A.T and G.C base pairs and dissociation of ligand. The mutual dependence of AT and GC denaturation allows one to draw conclusions about specificity of interaction. In case of both violamycins only slight preference of interaction with AT-rich regions was detected. The dissociation of violamycin BII in the latest stage of thermal denaturation was found to be cooperative.     

Cellular localization and age-dependent changes in mRNA for cyclic adenosine 3',5'-monophosphate-dependent protein kinases in rat testis

Gonadotropin activation of cyclic adenosine 3',5'-monophosphate (cAMP)-dependent protein kinases plays an important role in the regulation of testicular function. This study was undertaken to establish the expression of various subunits of cAMP-dependent protein kinases in different testicular cell types as well as during sexual maturation. RNA was extracted from cultured Sertoli cells, cultured peritubular cells, germ cells (pachytene spermatocytes, round spermatids), tumor Leydig cells, as well as whole testis from rats of various ages. Messenger RNA levels were studied by Northern analysis using available cDNA probes. The regulatory subunit (R) designated RII51 was found to be predominantly expressed in cAMP-stimulated Sertoli cells and tumor Leydig cells. Much lower levels were found in cultured peritubular cells and germ cells. A 2.9- and 3.2-kb mRNA for the RI subunit were found at about similar levels in all cell types, whereas the smaller 1.7-kb mRNA was expressed in high levels in germ cells. Also, the catalytic subunit (C) of cAMP-dependent protein kinase, designated C alpha, was expressed in all cell types; the highest mRNA levels for this subunit were found in germ cells and in tumor Leydig cells. The 1.7-kb mRNA for androgen-binding protein (ABP) was abundant in cAMP-stimulated Sertoli cells and was not present in other cell types of the testis. Furthermore, the cellular localization of the cAMP-dependent protein kinase subunits was also supported by developmental studies. The mRNA level of the RII51 3.2-kb species was relatively constant until Day 30, after which there was a tendency to decrease. A 1.6-kb message first appeared at greater ages. The mRNA for the smaller 1.7-kb species of RI, as well as the C alpha, showed a significant increase during development, supporting an enrichment of these mRNAs in germ cells. Messenger RNA levels for ABP were not detected in testis from 5- to 10-day-old rats but increased up to Day 30. After this age, mRNA for ABP revealed an age-dependent decrease, which parallels the relative increase of germ cells in the testis. In summary, these results demonstrate a clear pattern of cellular localization of the various mRNA species for subunits of the cAMP-dependent protein kinase in the rat testis.     

Increased stability of (+/-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo [a]pyrene through interaction with subcellular fractions of rat liver

The rate of hydrolysis of (+/-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo [a]pyrene (BPDE) to tetrahydroxy derivatives (tetrols) in the presence of various subcellular fractions of rat liver was investigated. Microsomes and nuclei increased the half-life of BPDE in a concentration-dependent manner whereas cytosol had no such effect. The presence of 1 mg microsomal protein/ml increased the half-life of BPDE from 4 to 60 min at 22 degrees C and from 1.5 to 20 min at 37 degrees C. Nuclei equivalent of 500 micrograms DNA/ml increased the half-life from 1.9 to 3.6 min at 37 degrees C. Liposomes prepared from microsomal lipids mimicked the effect of microsomes indicating that BPDE is stabilized primarily by interacting with lipids. The significance of these interactions for the stability of BPDE in an intact cell system was evaluated by using isolated hepatocytes. In these cells the half-life of BPDE was substantially shorter (1 min at 5 X 10(6) cells/ml) than in buffer (3 min). However, hydrolysis of BPDE to tetrols was a minor reaction (less than or equal to 3% of added BPDE at a cell density greater than or equal to 5 X 10(6) cells/ml) and the main route of elimination (greater than or equal to 75%) was through conjugation with glutathione.     

The effects of experimental hypo- and hyperthyroidism on blood viscosity and other blood parameters in the rat

Three groups of male Sprague Dawley rats received methimazole without or with Na-thyroxine in drinking water (3 and 0.33 mg T4/l, respectively) to induce characteristic alterations of their thyroid status (hypothyroid, hyperthyroid, euthyroid). A fourth group served as an untreated control without any additive to the drinking water. With respect to the different thyroid status, the following changes in the blood parameters were found: increasing plasma-T3-levels caused a reduction in plasma viscosity, in total plasma protein and in alpha 1-globulin, but an increase in hematocrit, whole blood viscosity, the number of erythrocytes and leukocytes, alpha 2-globulin and beta-globulin. It was concluded that the increase in the plasma viscosity in the hypothyroid status is mainly due to an alteration of the plasma protein pattern, and that the increase in whole blood viscosity in the hyperthyroid rat is a consequence of increased hematocrit.     

Influence of leaf surfaces on movements by the hymenopterous parasitoid Trichogramma exiguum

Walking speeds of female Trichogramma exiguum Pinto & Platner were fastest on maize and soybean (12 cm/min), intermediate on tomato (8 cm/min), and slowest on woolly mullein, Verbascum thapsus (3 cm/min). Similarly, rates of turning along the paths of walking T. exiguum were smallest on maize (median angle=0°±15°), intermediate on soybean and tomato, and greatest on V. thapsus (median angle=30°±15°). Leaf trichome density and morphology influenced walking behavior. Walking was slowed and flight initiation delayed for T. exiguum walking on Amaranthus hybridus leaves compared to either maize or filter paper. When inundative releases are conducted, the effects of plant surfaces on searching rates and arrestment should be considered in determining release rates of Trichogramma spp.

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Résumé La vitesse de marche a été déterminée en lâchant des individus sur chaque substrat végétal et en traçant leur parcours sur une plaque de verre placée à 9 mm au dessus du sujet. Les déviations angulaires de portions successives des tracés, longues de 1 mm, ont été utilisées pour mesurer les taux de changement de direction. Dans une deuxième expérience, des individus ont été lâchés au milieu d'une cercle de 40 mm de diamètre sur chaque substrat et les temps écoulés avant l'envol ou pour atteindre le bord du cercle à la marche ont servi à évaluer respectivement la propension au vol et la vitesse de déambulation.Les vitesses de marche ont été plus rapides sur maïs et soja (12 cm/min), moyennes sur tomate (8 cm/min) et les plus lentes sur Verbascum thapsus (Scrophulariaceae) (3 cm/min). De le même façon, les angles de changement de direction au cours des marches effectuées par T. exiguum ont été plus petits sur maïs, moyens sur soja et tomate, et plus grands sur V. thapsus. Chez T. exiguum marchant sur des feuilles d'Amaranthus hybridus L. (Amaranthaceae), la marche a été plus lente et l'envol plus tardif que sur maîs ou papier filtre.Lors de lâchers inondatifs, les effets des surfaces végétales sur les vitesses d'exploration et d'arrêt devraient être pris en compte pour déterminer les vitesses de lâcher des Trichogramma spp.

     

Effects of propoxur exposure on females of the German cockroach, Blattela germanica and their oothecae

Susceptible and insecticide resistant females of Blattella germanica (L.), Dictyoptera, were exposed to propoxur treated surfaces. They carried oothecae that were expected to hatch in 48 to 72 h. Hatch was delayed in females that retained their oothecae throughout the exposure period. Oothecal hatch and nymphal survival were reduced when oothecae hatched on a treated surface but not when oothecae were retained and hatched on an insecticide-free surface. Over half of the susceptible strain females dropped their oothecae during the exposure period while very few resistance females dropped oothecae prematurely. Mortality of susceptible females that dropped their oothecae was higher than among those that retained oothacea. A tendency in this direction was apparent among the few resistant females that dropped their oothecae. The number of nymphs that emerged on the treated surface and also that survived for 24 h was higher in the resistant than in the susceptible strain. Newly hatched nymphs were frequently attached to the oothecae due to an inability to shed the embryonic cuticle.

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Résumé Des femelles de B. germanica, sensibles ou résistantes aux insecticides, ont été mises en présence de surfaces traitées au propoxure. Elles portaient des oothèques dont l'éclosion était attendue entre 48 et 72 h plus tard. Les éclosions ont été retardées, les femelles ayant conservé leurs oothèques pendant toute la période d'exposition. Lex taux d'éclosion des oothèques et de survie larvaire ont été normaux quand il y a eu rétention de l'oothèque et émission sur une surface saine, par contre ils étaient réduits quand l'oothèque a été émise sur une surface traitée. Plus de la moitié des femelles de la souche sensible ont laché leur oothèque pendant la période d'exposition tandis que très peu de femelles de la souche résistante ont abandonné leur oothèque prématurément. La mortalité était plus élevée chez les femelles sensibles qui ont perdu leur oothèque que chez celles qui l'avaient conservée. Une tendance du même type était décelable parmi les quelques femelles de la souche résistante qui perdirent leur oothèque. Le nombre d'écolosions de larves et le nombre de survies larvaires à 24 h étaient plus élevés dans la souche résistante que dans la souche sensible. Les larves néonates étaient souvent fixées à l'oothèque par suite de leur incapacité à se dépouiller de la cuticle embryonnaire.

     

Inhibition of human vitamin-K-dependent protein-S-cofactor activity by a monoclonal antibody specific for a Ca2+-dependent epitope

Protein S is an anticoagulant vitamin-K-dependent plasma protein functioning as a cofactor to activated protein C in the degradation of factors Va and VIIIa. A murine monoclonal antibody, HPS 7, specific for a calcium-stabilized epitope in human protein S, is described. The epitope was available in intact protein S, both in its free form and when protein S was bound to C4b-binding protein. It disappeared upon reduction of disulfide bridges and also after thrombin of chymotrypsin cleavage of protein S. Thrombin cleaves protein S close to the calcium-binding region containing gamma-carboxyglutamic acid (Gla). The cleaved protein still contains the Gla region, linked by a disulfide bridge, but it has a lower affinity for calcium and no protein C cofactor activity. The thrombin-mediated cleavage of protein S could be inhibited by HPS 7. The Ka for the interaction between protein S and the monoclonal was estimated to be approximately 0.7 X 10(8) M-1. Half-maximal binding between HPS 7 and protein S was observed at a calcium concentration of 0.50 mM, indicating that saturation of the Gla region with calcium was required for the interaction. The recently reported Gla-independent high-affinity calcium binding did not induce the epitope. The calcium-dependent binding of protein S to phospholipid vesicles as well as the protein C cofactor activity was inhibited by HPS 7. The data suggests that the epitope for HPS 7 is located in the Gla region of protein S or in the closely positioned thrombin-sensitive region.     

Characteristics of alcohol/polyol dehydrogenases. The zinc-containing long-chain alcohol dehydrogenases

Sixteen characterized alcohol dehydrogenases and one sorbitol dehydrogenase have been aligned. The proteins represent two formally different enzyme activities (EC 1.1.1.1 and EC 1.1.1.14), three different types of molecule (dimeric alcohol dehydrogenase, tetrameric alcohol dehydrogenase, tetrameric sorbitol dehydrogenase), metalloproteins with different zinc contents (1 or 2 atoms per subunit), and polypeptide chains from different kingdoms and orders (mammals, higher plants, fungus, yeasts). Present comparisons utilizing all 17 forms reveal extensive variations in alcohol dehydrogenase, but with evolutionary changes that are of the same order in different branches and at different times. They emphasize the general importance of particular residues, suggesting related overall functional constraints in the molecules. The comparisons also define a few coincidences between intron positions in the genes and gap positions in the gene products. Only 22 residues are strictly conserved; half of these are Gly, and most of the remaining ones are Pro or acidic residues. No basic residue, no straight-chain hydrophobic residues, no aromatic residues, and essentially no branched-chain or polar neutral residues are invariable. Tentative consensus sequences were calculated, defining 13 additional residues likely to be typical of but not invariant among the alcohol dehydrogenases. These show a predominance of Val, charged residues, and Gly. Combined, the comparisons, which are particularly relevant to the data base for protein engineering, illustrate the requirements for functionally important binding interactions, and the extent of space restrictions in proteins with related overall conformations and functions.