Cyclic nucleotide phosphodiesterases in the cricket, Acheta domesticus.

Exceptionally high levels of guanosine 3'-5'-cyclic monophosphate (cyclic GMP) in the accessory reproductive gland of the male house cricket, Acheta domesticus, led to an investigation of cyclic nucleotide phosphodiesterase (EC 3.1.4.--) as a possible regulatory enzyme. Cricket cyclic nucleotide phosphodiesterase activity with cyclic GMP or cyclic AMP as substrate had a pH optimum around 9.0, required Mg2+ or Mn2+ for maximal activity, and was inhibited by EDTA and methylxanthines. Cyclic GMP phosphodiesterase occurred mainly in the soluble fraction of homogenates of accessory glands or whole crickets, but cyclic AMP phosphodiesterase in the accessory gland was primarily particulate. Kinetic analysis indicated three forms of cyclic GMP phosphodiesterase, with Km values at 2.9 muM, 71 muM and 1.5 mM. Chromatography of whole cricket or accessory gland extracts on DEAE cellulose gave an initial peak having comparable activity with either cyclic GMP or cyclic AMP, and a second peak specific for cyclic AMP. There were no appreciable changes in the specific activity or kinetic properties of accessory gland cyclic GMP phosphodiesterase during a developmental period over which cyclic GMP levels rise more than 500-fold. Thus, the accumulation of cyclic GMP in the accessory gland is probably not associated with concomitant developmental modulation of phosphodiesterase activity.     

Reduction of REM sleep by a tryptophan-free amino acid diet.

The acute administration of a tryptophan-free amino acid diet to rats has previously been reported to produce a marked reduction in brain serotonin concentrations. The present study examined the effects of such a diet on electroencephalographic sleep measures. There was a decrease in REM sleep and a small increase in nonREM sleep, with no change in total sleep time. In view of these and other observations, the hypothesis that the serotonergic system plays an important role in the maintenance of nonREM sleep should be carefully reevaluated.     

Vimentin is transiently co-localized with and phosphorylated by cyclic GMP-dependent protein kinase in formyl-peptide-stimulated neutrophils.

The effects of cGMP-dependent protein kinase (G-kinase), a major cellular receptor of cGMP, were investigated in activated human neutrophils. Immunocytochemistry demonstrated that G-kinase translocated from a diffuse localization in the cytoplasm to the cytoskeleton and nucleus after stimulation with N-formyl-methionyl-leucyl-phenylalanine (fMLP), and transiently co-localized with the intermediate filament protein, vimentin. During this time period, the most remarkable co-localization of G-kinase and vimentin was observed between 1-2.5 min stimulation with fMLP. At that time co-localization of G-kinase and vimentin was predominantly confined to filaments which extended from regions adjacent to the nucleus into the uropod. Distinctive localization for only G-kinase was observed at the microtubule organizing center and euchromatin of the nucleus. The filamentous staining pattern for G-kinase and vimentin was enhanced in the presence of 8-Br-cGMP. Coincident with co-localization of G-kinase and vimentin in adherent neutrophils was a transient increase in cGMP levels and an increase in the phosphorylation of vimentin in fMLP-stimulated cells. The increase in cGMP levels was dependent upon cell adherence, was enhanced by preincubating neutrophils with L-arginine (the precursor for nitric oxide synthesis), and attenuated with the nitric oxide synthase inhibitor, NG-monomethyl-L-arginine. Phosphorylation of vimentin in the fMLP-stimulated neutrophil was observed in the presence or absence of exogenous cGMP, although in the presence of low concentrations of 8-Br-cGMP a more rapid phosphorylation of vimentin was observed that correlated with the enhanced co-localization of G-kinase and vimentin. Phosphorylation of vimentin was not observed in non-activated cells treated with 8-Br-cGMP, suggesting that phosphorylation only occurs when G-kinase is co-localized with vimentin. The presence of the protein kinase C inhibitors, staurosporine or H-7, did not inhibit vimentin phosphorylation during fMLP stimulation, while 8-Br-cGMP enhanced phosphorylation in fMLP-treated cells. This suggests that neither protein kinase C nor cAMP-dependent protein kinase catalyze the phosphorylation of vimentin in neutrophils activated by fMLP. These results indicate that vimentin and G-kinase are co-localized in neutrophils and that vimentin is phosphorylated by G-kinase in response to the co-localization of the two proteins. A model for the targeting of G-kinase and vimentin is presented which hypothesizes that the transient redistribution of G-kinase may regulate neutrophil activation.     

Human herpesvirus 7 is a constitutive inhabitant of adult human saliva.

We report the frequent isolation of human herpesvirus 7 from the saliva of healthy adults. Virus isolates recovered from different individuals exhibited minimal restriction enzyme polymorphism, which was mostly confined to heterogeneous (het) sequences in the genome. DNAs of isolates recovered from the same individual over a period of several months showed the same characteristic het fragments, indicating the stability of the het sequences upon virus replication and shedding in vivo. In contrast to the results of previous reports, human herpesvirus 6, the causative agent of roseola infantum, could not be isolated from the saliva specimens, raising questions regarding oral transmission of human herpesvirus 6 and human herpesvirus 7 to young children.     

Both ankyrin and band 4.1 are required to restrict the rotational mobility of band 3 in the human erythrocyte membrane.

A population of band 3 proteins in the human erythrocyte membrane is known to have restricted rotational mobility due to interaction with cytoskeletal proteins. We have further investigated the cause of this restriction by measuring the effects on band 3 rotational mobility of rebinding ankyrin and band 4.1 to ghosts stripped of these proteins as well as spectrin and actin. Rebinding either ankyrin or 4.1 alone has no detectable effect on band 3 mobility. Rebinding both these proteins together does, however, reimpose a restriction on band 3 rotation. The effect on band 3 rotational mobility of rebinding ankyrin and 4.1 are similar irrespective of whether or not band 4.2 is removed from the membrane. We suggest that ankyrin and 4.1 together promote the formation of slowly rotating clusters of band 3.     

EVIDENCE FOR POLLINATION ECOTYPES IN THE YELLOW-FRINGED ORCHID,PLATANTHERA CILIARIS

Platanthera ciliaris is a butterfly-pollinated, terrestrial orchid with a loose terminal raceme of 10–50 orange flowers, characterized by a long nectariferous spur. In the southeastern United States, P. ciliaris occurs in the Appalachian mountains and coastal-plain physiographic provinces, but it is found rarely in the intervening Piedmont. In 1983 and 1984, detailed observations of two populations within these disjunct areas revealed that the butterfly species that serve as the primary pollinators differ sharply. In the mountains, Papilio troilus (spicebush swallowtail) was the most frequent and effective visitor, whereas in the coastal plain, P. palamedes (palamedes swallowtail) was the predominant pollinator. Proboscis lengths of P. troilus (mean = 23.3 mm) were significantly shorter than those of P. palamedes (mean = 28.7 mm). Floral characters, most notably spur length, also differed significantly between mountain (mean = 23.8 mm) and coastal-plain (mean = 25.6 mm) plants. In both 1983 and 1984, levels of pollinator service, as assessed by rates of removal and insertion of pollinia, were higher in the mountains (0.81 and 0.86) than in the coastal plain (0.63 and 0.67). In addition, fruit-set was significantly greater in mountain (83.9% in 1983, 86.5% in 1984) than in coastal-plain (63.8% in 1983, 65.5% in 1984) populations. We hypothesize that selection pressure exerted through pollinator proboscis lengths has resulted in pollination ecotypes of P. ciliaris. The short-spurred mountain plants appear to be in equilibrium with their short-tongued butterfly pollinators, receiving high levels of effective pollination and achieving high levels of fruit-set. Coastal-plain plants produce flowers with longer spurs which, nevertheless, are shorter than optimal for insuring that very long-tongued butterflies make contact with their pollinia. Thus, effective pollinator service and fruit-set are reduced. Correlation analyses in 1984 showed a positive relationship between spur length and fruit-set only in the coastal plain. The presence of long-tongued nectar thieves in the coastal plain may also contribute to lower effective pollinator service and fruit-set. Results of reciprocal-transplant studies attempting to determine the genetic basis of floral characters, including spur length, were inconclusive but suggest that differences between mountain and coastal-plain plants are not due solely to phenotypic plasticity.     

Genetics of lipid deposition in the Japanese quail

Summary The research presented here was designed to investigate the mode of inheritance of fat and lean tissue deposition, and the relationship between them and body weight in Japanese quail. Heterotic effects were found for weight, size, and number of adipocytes in the abdominal fat depots, weight of the sartorial fat depot and percentage carcass fat with means for the hybrids being lower than those for the parental lines. General inferences concerning the importance of nonadditive genetic variation for lean and body weight were precluded due to inconsistencies observed among mating combinations. Thus, although heterosis and recombination effects were general for characteristics associated with fat deposition, the situation for body weight and lean was unique to the populations involved. It may be hypothesized that heterosis in the efficiency of feed utilization is reflected by the heterosis for fat deposition which explains why hybrids utilize feed better than their parental lines.     

Variation and taxonomy of Aesculus pavia L. (Hippocastanaceae) in Texas

The status of Aesculus pavia var. flavescens, a set of yellow-flowered populations endemic to the Edwards Plateau of central Texas, was evaluated on the basis of comparative morphology, geography, flowering phenology, and pollination ecology. Plants of typical A. pavia have red, tubular flowers with exserted stamens and are effectively pollinated by ruby-throated hummingbirds. Plants of var. flavescens have yellow, campanulate flowers with included stamens that can be pollinated effectively by large bees. These ethological reproductive isolating mechanisms are further augmented by a difference in flowering time between the two sets of populations and by geographical restriction of the yellow-flowered plants to localities on the Edwards Plateau. The yellow and red-flowered plants constitute two morphologically distinct groups, despite evidence of limited introgression in counties along the Balcones Escarpment. A yellow-flowered plant discovered in coastal east Texas may indicate the mechanism by which var. flavescens originated, but this plant is much more closely comparable to typical red-flowered var. pavia than to the yellow-flowered plants on the Edwards Plateau. An evolutionary history of section Pavia of Aesculus is presented, and it is concluded that the yellow-flowered plants of central Texas are best regarded as a taxonomic variety.     

Abnormal concentrations of B vitamins and amino acids in plasma, bile, and liver of chicks with aflatoxicosis.

Graded levels of aflatoxin fed to broiler chickens for 3 weeks decreased the levels of most B vitamins in plasma, bile, and liver and decreased all free and hydrolyzed amino acids from peptides in plasma. The levels of thiamine, riboflavin, vitamin B6, pantothenic acid, and choline decreased by more than 60% in bile; vitamin B6, pantothenic acid, and choline decreased by more than 49% in plasma; thiamine, vitamin B6, pantothenic acid, choline, folate, and niacin decreased by more than 19% in liver; and only bile and plasma levels of folate increased (78 and 12%, respectively) with a dietary level of 5 microgram of aflatoxin per g of feed. Plasma levels of cystine and cysteine, methionine, threonine, serine, and aspartic acid decreased by more than 37%, whereas valine, lysine, leucine, histidine, hydroxyproline, and alanine decreased by more than 9% with 2.5 microgram of aflatoxin per g of feed. The data suggest that aflatoxin interferes with the metabolism of B vitamins and amino acids in chicks. However, these changes may be measuring the effects of aflatoxin-induced hepatic necrosis.