Phosphorylation of the cyclic AMP response element binding protein mediates transforming growth factor beta-induced downregulation of cyclin A in vascular smooth muscle cells

Transforming growth factor beta (TGFbeta), a multifunctional cytokine associated with vascular injury, is a potent inhibitor of cell proliferation. The current results demonstrate that the TGFbeta-induced growth arrest of vascular smooth muscle cells (VSMCs) is associated with cyclin A downregulation. TGFbeta represses the cyclin A gene through a cyclic AMP (cAMP) response element, which complexes with the cAMP response element binding protein (CREB). The CREB-cyclin A promoter interaction is hindered by TGFbeta, preceded by a TGFbeta receptor-dependent CREB phosphorylation. Induction of CREB phosphorylation with forskolin or 6bnz-cAMP mimics TGFbeta's inhibitory effect on cyclin A expression. Conversely, inhibition of CREB phosphorylation with a CREB mutant in which the phosphorylation site at serine 133 was changed to alanine (CREB-S133A) upregulated cyclin A gene expression. Furthermore, the CREB-S133A mutant abolished TGFbeta-induced CREB phosphorylation, cyclin A downregulation, and growth inhibition. Since we have previously shown that the novel PKC isoform protein kinase C delta (PKCdelta) is activated by TGFbeta in VSMCs, we tested the role of this kinase in CREB phosphorylation and cyclin A downregulation. Inhibition of PKCdelta by a dominant-negative mutant or by targeted gene deletion blocked TGFbeta-induced CREB phosphorylation and cyclin A downregulation. Taken together, our data indicate that phosphorylation of CREB stimulated by TGFbeta is a critical step leading to the inhibition of cyclin A expression and, thus, VSMC proliferation.     

Iron-histidine resonance raman band of deoxyheme proteins: effects of anharmonic coupling and glass-liquid phase transition

Weak anharmonic coupling of two soft molecular vibrations is shown to cause pronounced temperature dependence of the corresponding resonance Raman bands. The developed theory is used to interpret the temperature dependence of the iron-histidine band of deoxyheme proteins and model compounds. It is shown that anharmonic coupling of the iron-histidine and heme doming vibrations must cause pronounced broadening of the band, its asymmetry, and shift of its maximum to the red upon heating. It also can lead to a structured shape of this band at room temperature. Proper consideration of the anharmonic coupling allows simulation of the temperature dependence of the iron-histidine band shape of horse heart myoglobin in the temperature interval of 10-300 K, using the minimum number of necessary parameters. Analysis of this temperature dependence clearly shows that the iron-histidine band of deoxyheme proteins is sensitive to the glass-liquid phase transition in the protein hydration shell, which takes place at 160-190 K.     

Synthesis of calelectrins and calpactin I during cytochalasin mediated cell spreading inhibition

Mammary epithelial cell spreading on collagen gels has previously been shown to be correlated with the synthesis of a group of calcium-binding proteins (CBPs) which we have identified as the calcium-binding proteins termed calelectrins and calpactin I monomer/p36. To determine whether cell spreading per se is required for CBP synthesis, we examined the effect of cytochalasin D on these two events. Concentrations of cytochalasin D that did not reduce total protein synthesis, caused inhibition of cell spreading in a dose-dependent manner, but did not cause inhibition of CBP synthesis. Synthesis of collagen also continued during cytochalasin inhibition of cell spreading. Removal of the inhibitor from the cultures initiated cell spreading and CBP synthesis continued. Membrane-cytoskeleton complexes from control and CD treated cells were identical in regard to binding CBPs in a calcium-dependent manner. Colchicine, which inhibited cell spreading, was shown to be toxic to general protein synthesis at 75 nM. The data clearly indicate that mere inhibition of epithelial cell spreading does not automatically suppress CBP synthesis.     

Peripheral skin temperature effects on muscle oxygen levels

In both hot and cold environments, tissue oxygen saturation levels may affect muscle performance. Using near-infrared spectroscopy, muscle oxygen saturation (StO2) and total hemoglobin levels were measured during exercise. Lowering skin temperature caused a greater StO2 and total hemoglobin decrease by 16% and 15.9%, respectively, compared to controls. Increasing skin temperature resulted in a smaller decrease in both StO2 and total hemoglobin by 12.2% and 8.2%, respectively, compared to controls. These data indicate that warming the skin will cause less of decrease in StO2 and total hemoglobin, while cooling the skin has the opposite effect.     

Envelope diversity, characteristics of V3 region and predicted co-receptor usage of human immunodeficiency viruses infecting north indians

Subtypes of human immunodeficiency virus type 1 circulating in 21 north Indian patients were characterized based on the partial sequence of the gp120 envelope protein. A majority of viruses (85.7%, 18/21) were subtype C, while 14.3% (3/21) were subtype A. Sequence analysis revealed that the V3 region was highly conserved compared with V4 and V5. The predicted use of co-receptors indicated exclusive usage of R5, except for two subtype A viruses (AIIMS279 and AIIMS281). Our results demonstrate conservation within the V3 loop of subtype C viruses, and suggest the emergence of non-clade C viruses in the north Indian population.     

Characterization of monoclonal antibodies to Treponema pallidum

Thirteen hybrid cell lines which produce mouse monoclonal antibodies to Treponema pallidum, the causative agent of syphilis, have been established. All of the monoclonal antibodies react with T. pallidum, Nichols strain, in ELISA and in immunofluorescence assays, but do not react with normal rabbit testicular tissue in the ELISA. Two of these antibodies were demonstrated to react with the nonpathogenic treponemes T. phagedenis, biotype Reiter, T. refringens (Noguchi strain), T. vincentii, and T. denticola (strains 11 and W), as well as with Borrelia recurrentis, Leptospira interrogans, serogroup Canicola, and the swine pathogen T. hyodysenteriae. The remaining 11 antibodies react with four recently isolated strains of T. pallidum, but with none of the related nonpathogens nor with Borrelia or Leptospira. Thus, our results to date indicate that these monoclonal antibodies may identify antigenic determinants that are specific either for T. pallidum alone or for those treponemes which are pathogenic for humans. The molecular specificities of six of the 13 antibodies were determined by Western blotting. We anticipate potential usefulness of these antibodies in the investigation of the antigenic structure of T. pallidum, the taxonomic study of the pathogenic and nonpathogenic treponemes, and in the diagnosis of syphilis.     

Modelling reproductive allocation of dusky salamanders using optimal control theory: Pros,cons and caveats

Summary Optimal control theory has been used to examine the evolution of life history characters in a variety of plant and animal species. In this paper, I examine control theoretic models of reproductive allocation in female dusky salamanders and consider some practical aspects of modelling, including the appropriateness of nonlinear formulations, methods for describing semelparous reproduction, and data needed to parameterize models. The model analysed includes state variables for somatic and reproductive tissue, energy intake and requirements for physiological maintenance, and iteroparous reproduction. It predicts that female salamanders should spend the first part of their lives growing. After reaching sexual maturity, females should either spend the remainder of their lives reproducing at the expense of decreasing body size, possibly resulting in death by starvation, or maintain approximately constant body size at the expense of low reproductive output. This lack of correspondence to the observed biology of dusky salamanders suggests that not all the appropriate biology has been described. In particular, inclusion of a storage variable may be necessary in future modelling efforts.     

The frontal sinus in ancient and modern Greenlandic Inuit

The purpose of this study was to compare the frontal sinus size of ancient Greenlandic Inuit with ancient Inuit of Alaska and Canada, and to compare sinus size between ancient and modern Greenlandic Inuit. Also, it was analyzed whether cranial size was a determinant of frontal sinus size. Frontal sinus size was evaluated in terms of absence frequency and planimetrically. Absence was defined as a frontal sinus not exceeding a line drawn between the supraorbital rims. A significant increase in absence frequency was noted from Alaska over Canada to Greenland (males: p<0.03; females p<0.0001). This is in accordance with earlier studies, indicating that although these Inuit populations once have been commonly related to the Old Bering Sea population, the Greenland Inuit represent an endpoint in an eastward migration. There was a significant increase (p<0.0001) in frontal sinus size from ancient to modern Greenlandic Inuit, probably indicative of a high degree of admixture with non-Inuit after modern colonization. The results regarding craniofacial size parameters and frontal sinus side were inconclusive. No single craniofacial variable showed significant effect on frontal sinus size, but the area displayed sexual dimorphism, females having smaller frontal sinuses.