A myosin motor that selects bundled actin for motility

Eukaryotic cells organize their contents through trafficking along cytoskeletal filaments. The leading edge of a typical metazoan cytoskeleton consists of a dense and complex arrangement of cortical actin. A dendritic mesh is found across the broad lamellopodium, with long parallel bundles at microspikes and filopodia. It is currently unclear whether and how myosin motors identify the few actin filaments that lead to the correct destination, when presented with many similar alternatives within the cortex. Here we show that myosin X, an actin-based motor that concentrates at the distal tips of filopodia, selects the fascin-actin bundle at the filopodial core for motility. Myosin X moves individual actin filaments poorly in vitro, often supercoiling actin into plectonemes. However, single myosin X motors move robustly and processively along fascin-actin bundles. This selection requires only parallel, closely spaced filaments, as myosin X is also processive on artificial actin bundles formed by molecular crowding. Myosin X filopodial localization is perturbed in fascin-depleted HeLa cells, demonstrating that fascin bundles also direct motility in vivo. Our results demonstrate that myosin X recognizes the local structural arrangement of filaments in long bundles, providing a mechanism for sorting cargo to distant target sites.     

Socioeconomic status and diagnosed diabetes incidence

AIMS: To investigate the association between socioeconomic status (SES) and incidence of diabetes. METHODS: We investigated three measures of SES and incidence of diagnosed diabetes among women and men in the NHANES I Epidemiologic Followup Study, 1971-1992, who were free of diagnosed diabetes in 1980. RESULTS: Among women, diabetes incidence was inversely associated with income (measured as percent of the poverty level), education, and occupational status, adjusting for age and race/ethnicity. The hazard ratio (HR) for women with > 16 years education was 0.26 (95% confidence interval (CI) 0.13-0.54) relative to those with < 9 years of education. Adjustment for potential mediators, including body size variables, diet, physical activity, and alcohol and tobacco use, substantially attenuated the associations with income and education. Among men a trend toward lower diabetes incidence with higher income and higher education was evident (the HR for men with household income > 5 times the poverty level was 0.44 (95% CI 0.19-0.98) relative to those under the poverty line), but there was no inverse association of diabetes incidence with occupational status. CONCLUSIONS: SES, assessed with any of three common measures, is a risk factor for diagnosed diabetes in women. Among men these associations are less consistent.     

Evidence for capillarity contributions to gecko adhesion from single spatula nanomechanical measurements

The hairy attachment system on a gecko's toes, consisting of one billion spatulae in the case of Gekko gecko [Ruibal, R. & Ernst, V. (1965) J. Morphol. 117, 271-294], allows it to adhere to nearly all surface topographies. The mechanistic basis for gecko adhesion has been intensely investigated, but the lowest hierarchical level, that of the spatula, has become experimentally accessible only recently. This report details measurements of the adhesion force exerted by a single gecko spatula for various atmospheric conditions and surface chemistries. Through judicious choice and modification of substrates, the short- and long-range adhesive forces are separated. In contrast to previous work [Autumn, K., Sitti, M., Liang, Y. C. A., Peattie, A. M., Hansen, W. R., Sponberg, S., Kenny, T. W., Fearing, R., Israelachvili, J. N. & Full, R. J. (2002) Proc. Natl. Acad. Sci. USA 99, 12252-12256], our measurements clearly show that humidity contributes significantly to gecko adhesion on a nanoscopic level. These findings are crucial for the development of artificial biomimetic attachment systems.     

Chimpanzees as an animal model for human norovirus infection and vaccine development

Noroviruses are global agents of acute gastroenteritis, but the development of control strategies has been hampered by the absence of a robust animal model. Studies in chimpanzees have played a key role in the characterization of several fastidious hepatitis viruses, and we investigated the feasibility of such studies for the noroviruses. Seronegative chimpanzees inoculated i.v. with the human norovirus strain Norwalk virus (NV) did not show clinical signs of gastroenteritis, but the onset and duration of virus shedding in stool and serum antibody responses were similar to that observed in humans. NV RNA was detected in intestinal and liver biopsies concurrent with the detection of viral shedding in stool, and NV antigen expression was observed in cells of the small intestinal lamina propria. Two infected chimpanzees rechallenged 4, 10, or 24 mo later with NV were resistant to reinfection, and the presence of NV-specific serum antibodies correlated with protection. We evaluated the immunogenicity and efficacy of virus-like particles (VLPs) derived from NV (genogroup I, GI) and MD145 (genogroup II, GII) noroviruses as vaccines. Chimpanzees vaccinated intramuscularly with GI VLPs were protected from NV infection when challenged 2 and 18 mo after vaccination, whereas chimpanzees that received GII VLPs vaccine or a placebo were not. This study establishes the chimpanzee as a viable animal model for the study of norovirus replication and immunity, and shows that NV VLP vaccines could induce protective homologous immunity even after extended periods of time.     

Age-related changes in superoxide dismutase, glutathione peroxidase, catalase and xanthine oxidoreductase/xanthine oxidase activities in the rabbit cornea

The activities of superoxide dismutase, glutathione peroxidase (GPX) and catalase--the enzymatic scavengers of reactive oxygen species and the activities of xanthine oxidoreductase and xanthine oxidase, an enzyme known to generate reactive oxygen species, were studied in the corneas of normal rabbit eyes of various ages (1 month--young eyes; 4-9.5 months--young adult eyes; 2.0-2.75 years--middle aged eyes; 3.0-5.0 years--aged eyes). The activities of GPX, superoxide dismutase, xanthine oxidoreductase and xanthine oxidase were investigated biochemically in the scraped corneal epithelium. Catalase activity was detected histochemically in the corneal epithelium and endothelium. The results show that young corneas revealed lower activities of all the antioxidant enzymes investigated than did young adult corneas, in which enzymatic activities reached their maximum. In middle-aged corneas, GPX and catalase activities remained approximately at the same levels as seen in young adult corneas, whereas superoxide dismutase activity was decreased. In aged corneas, the activities of all antioxidant enzymes were dramatically decreased or even lost (catalase activity in the corneal endothelium). In contrast, xanthine oxidoreductase activity only slightly decreased with age and the xanthine oxidase proportion of total xanthine oxidoreductase remained unchanged. GPX, superoxide dismutase and catalase are important antioxidant enzymes protecting the cornea against the oxidative damage. Because the activities of these enzymes are lower in young animals and greatly reduced in aged animals, it is suggested that young and particularly aged corneas might be more susceptible to oxidative stress than are young adult corneas. This presumption is supported by the fact that the activities of prooxidant enzymes (xanthine oxidoreductase/xanthine oxidase) are only slightly decreased in aged corneas as compared to young adult corneas so that some imbalance between antioxidant and prooxidant enzymes exists already in the normal aged corneas.     

Electrogastrography in patients with Roux-en-Y reconstruction after previous Billroth gastrectomy

BACKGROUND/AIMS: The aim of this prospective study was to characterize gastric myoelectrical activity in patients with Roux-en-Y reconstruction after previous Billroth gastrectomy. METHODOLOGY: Thirteen patients entered the study (6 men and 7 women, aged 35-57). The mean time from Roux-en-Y reconstruction to electrogastrography (EGG) recording was 5 years. Surface cutaneous EGG was recorded using a Digitrapper EGG in the morning both fasting and after a standard solid test meal. All patients assessed their dyspeptic symptoms at the time of EGG in a semi-quantitative subjective scale. RESULTS: EGG was abnormal in all studied patients (but one postprandial recording). Dyspepsia was not meal-related and was not more severe in Helicobacter pylori positive patients. There was a significant negative correlation between time from Roux-en-Y reconstruction to EGG recording and bradygastria percent activity, both fasted and postprandial (r = -0.576; p = 0.0022). There was an inverse trend between severity of dyspepsia and normal slow-wave rhythm percent activity. Older patients tended to have more severe dyspepsia. CONCLUSIONS: The results of this study suggest that abnormal EGG recording is associated with dyspepsia in patients after Roux-en-Y reconstruction.     

Sites of Replication of Chromosomal DNA in a Eukaryotic Cell

In mouse cells (line P815), newly synthesized DNA labeled for 20-30 sec during exponential growth is found by electron microscope autoradiography at sites throughout the cell nucleus. These sites are relatively more concentrated in the peripheral region of the nucleus (averaged over a random population of S-phase cells), probably reflecting a higher local concentration of DNA in this region. Newly synthesized DNA is not preferentially associated with purified nuclear envelopes, but is found in a fraction of the chromosomal deoxynucleoprotein whose buoyant density in CsCl after formaldehyde treatment is about 1% lower than that of the deoxynucleoprotein peak. Kinetics experiments suggest that this material is a precursor of mature deoxynucleoprotein; it may represent regions of deoxynucleoprotein containing replicating DNA and the additional proteins involved in DNA replication. Other complexes of newly replicated DNA that are found in the interphase after phenol extraction of nuclei are formed during the extraction procedure, probably due to the partially single-stranded nature of replicating DNA, and do not appear to exist in vivo.     

Validity of transhepatic pulp pressure measurements

Cineangiographic studies were undertaken in conjunction with pressure measurements to determine the reliability of transhepatic pulp pressure manometry in assessing portal pressure. Pressure measurements obtained by wedged hepatic vein and/or umbilical-portal vein catheterization were used as a reference. An accurate index to intrasinusoidal pressure was provided when the needle was introduced into the sinusoidal bed and created a vascular “puddle.” Pressures were inaccurate when the needle entered a hepatic venule, the subcapsular space, or a bile duct. Reversible increases in transhepatic pressure were noted with proliferation of sinusoidal-mesenchymal cells attendant to acute liver injury or hyperplasia of the endoplasmic reticulum following administration of drugs that are metabolized in microsomes.     

Genetic evidence for a protein-kinase-A-mediated presynaptic component in NMDA-receptor-dependent forms of long-term synaptic potentiation

The synaptic vesicle protein Rab3A is a small GTP-binding protein that interacts with rabphilin and RIM1alpha, two presynaptic substrates of protein kinase A (PKA). Mice lacking RIM1alpha and Rab3A have a defect in PKA-dependent and NMDA receptor (NMDAR)-independent presynaptic long-term potentiation (LTP) at hippocampal mossy-fiber and cerebellar parallel-fiber synapses. In contrast, the NMDAR-dependent and PKA-independent early phase of LTP at hippocampal CA3-CA1 synapses does not require these presynaptic proteins. Here, we ask whether Rab3A and RIM1alpha participate in forms of LTP that require both PKA and NMDAR activation. We find that Rab3A is necessary for corticoamygdala LTP and late-phase LTP at CA3-CA1 synapses, two forms of LTP that require NMDAR and PKA activation. The latter form of LTP also requires RIM1alpha. These results provide genetic evidence that presynaptic proteins are required in LTP induced through the postsynaptic activation of NMDARs. Thus Rab3A and its effectors are general modules for four distinct types of PKA-dependent LTP in the brain.