RESUMO
Isotopic composition of (87) Sr:(86) Sr and natural elemental tracers (Sr, Ba, Mg, Mn and Ca) were quantified from otoliths in juvenile and adult Chinook salmon Oncorhynchus tshawytscha to assess the ability of otolith microchemistry and microstructure to reconstruct juvenile O. tshawytscha rearing habitat and growth. Daily increments were measured to assess relative growth between natal rearing habitats. Otolith microchemistry was able to resolve juvenile habitat use between reservoir and natal tributary rearing habitats (within headwater basins), but not among catchments. Results suggest that 90% (n = 18) of sampled non-hatchery adults returning to the Middle Fork Willamette River were reared in a reservoir and 10% (n = 2) in natal tributary habitat upstream from the reservoir. Juveniles collected in reservoirs had higher growth rates than juveniles reared in natal streams. The results demonstrate the utility of otolith microchemistry and microstructure to distinguish among rearing habitats, including habitats in highly altered systems.
Assuntos
Ecossistema , Membrana dos Otólitos/química , Salmão/crescimento & desenvolvimento , Animais , Água Doce/química , Oregon , Membrana dos Otólitos/crescimento & desenvolvimento , RiosRESUMO
Body mass and sympathetic activity increase with aging and might underlie blood pressure (BP) elevation. Increased body mass index (BMI) may elevate BP by increasing sympathetic activity. Glutathione (GSH) can decrease BP, and declines with aging. We measured systolic (SBP) and diastolic BP, BMI, plasma (NE(pl)) and urine norepinephrine (NEu), and plasma GSH in n=204 twins across the age spectrum. BP correlated directly with BMI, NEpl, and NEu, but inversely with GSH. Age correlated with BP, BMI, NEpl, and NEu. BP, BMI, NEpl, and NEu were higher in older subjects than younger subjects, whereas GSH was lower with aging. In older subjects with high (above median) NEpl, SBP was 8 mmHg higher than in those of comparable age with low NE. In younger subjects with high GSH, BP was significantly lower than in younger subjects having low GSH. NEu was significantly reduced in young high-BMI subjects vs young low-BMI subjects. The heritability (h2) of NEpl, NEu, and GSH ranged from approximately 50 to approximately 70%, and these biochemical quantities were considerably more heritable than BP. We conclude that increases in sympathetic activity contribute to aging-induced SBP elevations, especially in older females. GSH reductions apparently participate in aging-induced BP elevations, most strongly in males. BMI increases contribute to BP elevations, particularly in younger subjects. BMI elevations apparently raise BP mainly by peripheral mechanisms, with generally little sympathetic activation. Substantial h(2) for plasma GSH, NE, and urine NE suggests that such traits may be useful 'intermediate phenotypes' in the search for genetic determinants of BP.
Assuntos
Envelhecimento/fisiologia , Pressão Sanguínea , Índice de Massa Corporal , Glutationa/fisiologia , Hipertensão/genética , Sistema Nervoso Simpático/fisiologia , Adulto , Distribuição por Idade , Fatores Etários , Feminino , Humanos , Hipertensão/sangue , Hipertensão/epidemiologia , Masculino , Fatores SexuaisRESUMO
Hyperhomocysteinemia is common in Alzheimer's disease and is negatively correlated with cognitive function. Hyperhomocysteinemia can increase S-adenosylhomocysteine (SAH), a potent methyltransferase inhibitor. This study investigates the role of brain SAH in the cognitive and neurological disruption in Alzheimer's disease. SAH was significantly (26%) higher in prefrontal cortex of Alzheimer patients than normals. Brain homogenates from Alzheimer patients inhibited an exogenous methyltransferase 15% more than normal homogenates (P <.001). Brain SAH levels correlated (r=.508) with methyltransferase inhibition by brain homogenates. Methyltransferase inhibition by Alzheimer brain homogenates correlated inversely with cognitive function as determined by MMSE (r=-0.36). Phenylethanolamine N-methyltransferase (PNMT) and catechol O-methyltransferase (COMT) activities were more than 30% lower (P<0.001) in Alzheimer than normal brains. Brain PNMT activity correlated significantly with cognitive function (r=0.243), age of Alzheimer's onset (r=0.272), and choline acetyltransferase activity (r=0.333), but negatively with neurofibrillary tangles (r=-0.332). COMT activity also correlated significantly with cognitive function (r=0.324), age of disease onset (r=0.209), choline acetyltransferase activity (r=0.326), levels of synaptophysin (r=0.506), and negatively with tangles (r=-0.216 P=0.039). Elevated SAH in Alzheimer brain inhibits methyltransferases and is related to markers of disease progression and cognitive impairment.
Assuntos
Doença de Alzheimer/metabolismo , Encéfalo/metabolismo , Encéfalo/patologia , Cognição , Metiltransferases/metabolismo , S-Adenosil-Homocisteína/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/psicologia , Feminino , Humanos , Hiper-Homocisteinemia/metabolismo , Masculino , Pessoa de Meia-Idade , Emaranhados Neurofibrilares/patologia , Córtex Pré-Frontal/metabolismo , Córtex Pré-Frontal/patologia , Fatores de TempoRESUMO
Both monoamine oxidase (MAO) A and MAO B in the brain have been implicated in the etiology of Alzheimer's disease. MAO B is elevated in plaque-associated glia in Alzheimer brain. Elevations in MAO A in Alzheimer neurons have been linked to increases in neurotoxic metabolites and neuron loss. We investigated the relationship between cognitive function in Alzheimer patients and post-mortem prefrontal cortex MAO A and B activities. Prefrontal cortex tissue from 92 Alzheimer patients and 74 neurologically normal subjects was obtained at autopsy and analyzed for activities of MAO A and B by radioenzymatic methods. Mini Mental Status Exam was performed on Alzheimer patients within 1 year of death. Alzheimer brains were analyzed for Braak stage, tangles, plaques and choline acetyltransferase activity. Prefrontal cortex MAO B activity was significantly increased by 16% in Alzheimer patients versus normals, whereas MAO A activity was significantly decreased by 17% in these same patients. Neither MAO A nor MAO B activities correlated with cognitive function (MMSE score), choline acetyltransferase activity, plaques, neurofibrillary tangles, Braak stage, or age of disease onset in the Alzheimer patients. With increasing Alzheimer duration or increasing Braak stage, MMSE scores and choline acetyltransferase activity declined, but levels of MAO A and B in prefrontal cortex were unchanged. Patients in the upper quintile for MAO A or B activity did not differ significantly from those in the lowest quintile with respect to MMSE scores or age of Alzheimer disease onset. We conclude that the changes in MAO A and B in the prefrontal cortex occur very early in Alzheimer's disease and remain relatively constant as the disease progresses.
Assuntos
Doença de Alzheimer/enzimologia , Monoaminoxidase/metabolismo , Córtex Pré-Frontal/enzimologia , Idade de Início , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/patologia , Colina O-Acetiltransferase/metabolismo , Cognição/fisiologia , Humanos , Emaranhados Neurofibrilares/patologia , Testes Neuropsicológicos , Placa Amiloide/patologia , Córtex Pré-Frontal/patologiaAssuntos
Catecolaminas/metabolismo , Células Cromafins/metabolismo , Neuropeptídeo Y/metabolismo , Animais , Bovinos , Células Cultivadas , Células Cromafins/efeitos dos fármacos , Galanina/farmacologia , Fármacos Neuroprotetores/farmacologia , Fatores de Tempo , Peptídeo Intestinal Vasoativo/farmacologiaRESUMO
OBJECTIVES: To examine whether per capita income and income inequality are independently associated with teen birth rate in populous U.S. counties. METHODS: This study used 1990 U.S. Census data and National Center for Health Statistics birth data. Income inequality was measured with the 90:10 ratio, a ratio of percent of cumulative income held by the richest and poorest population deciles. Linear regression and analysis of variance were used to assess associations between county-level average income, income inequality, and teen birth rates among counties with population greater than 100,000. RESULTS: Among teens aged 15-17, income inequality and per capita income were independently associated with birth rate; the mean birth rate was 54 per 1,000 in counties with low income and high income inequality, and 19 per 1,000 in counties with high income and low inequality. Among older teens (aged 18-19) only per capita income was significantly associated with birth rate. CONCLUSIONS: Although teen childbearing is the result of individual behaviors, these findings suggest that community-level factors such as income and income inequality may contribute significantly to differences in teen birth rates.
Assuntos
Etnicidade/psicologia , Renda , Idade Materna , Gravidez na Adolescência/psicologia , Adolescente , Análise de Variância , Escolaridade , Feminino , Humanos , Pobreza , Gravidez , Fatores SocioeconômicosRESUMO
PURPOSE: Previous studies have shown a correlation between measures of social capital and morbidity, mortality, and violent crime. This article examines the association across U.S. states between social capital (as measured by mutual trust and civic engagement) and firearm availability. METHODS: The analysis uses OLS to determine degrees of association across U.S. states. Measures of mutual trust come from responses to questions on the U.S. General Social Survey that "you can't be too careful in dealing with people," and most people "would try to take advantage of you." Measures of formal civic engagement come from responses to Lifestyle Survey questions concerning times volunteered, club meetings attended, community projects worked on, and church services attended. Informal civic engagement measures come from responses to number of times bowled, played cards, entertained at home, and gave or attended dinner parties, and number of greeting cards sent. The Lifestyle Survey also asked whether respondent believed whether "most people are honest." The percentage of suicides from firearms, and the average percentage of suicides and homicides from firearms, are used as proxies for state firearm ownership rates. Control variables are the degree of urbanization, the rates of poverty, and the percentage of nonwhites in the state. RESULTS: Across the U.S. states, higher levels of firearm ownership are associated with significantly lower levels of mutual trust and civic engagement. CONCLUSION: While the analysis cannot show causation, states with heavily armed civilians are also states with low levels of social capital.
Assuntos
Armas de Fogo/estatística & dados numéricos , Meio Social , Estudos Transversais , Humanos , Prevalência , Estados Unidos/epidemiologiaRESUMO
Protein tyrosine phosphatases are a class of enzymes that function to modulate tyrosine phosphorylation of cellular proteins and play an essential role in regulating cell function. PTP1B has been implicated in the negative regulation of the insulin signaling pathway by dephosphorylating the activated insulin receptor. Inhibiting this phosphatase and preventing the insulin-receptor downregulation has been suggested as a target for the treatment of Type II diabetes. A high-throughput screen for inhibitors of PTP1B was developed using a scintillation proximity assay (SPA) with GST-- or FLAG--PTP1B((1-320)) and a potent [(3)H]-tripeptide inhibitor. The problem of interference from extraneous oxidizing and alkylating agents which react with the cysteine active-site nucleophile was overcome by the use of the catalytically inactive C215S form of the native enzyme (GST--PTP1B(C215S)). The GST--PTP1B was linked to the protein A scintillation bead via GST antibody. The radiolabeled inhibitor when bound to the enzyme gave a radioactive signal that was competed away by the unknown competitive compounds. Further utility of PTP1B(C215S) was demonstrated by mixing in the same well both the catalytically inactive GST--PTP1B(C215S) and the catalytically active FLAG--CD45 with an inhibitor. Both a binding and kinetic assay was then performed in the same 96-well plate with the inhibition results determined for the PTP1B(C215S) (binding assay) and CD45 (activity assay). In this way inhibitors could be differentiated between the two phosphatases under identical assay conditions in one 96-well assay plate. The use of a mutant to reduce interference in a binding assay and compare with activity assays is also amenable for most cysteine active-site proteases.
Assuntos
Inibidores Enzimáticos/análise , Inibidores Enzimáticos/metabolismo , Proteínas Tirosina Fosfatases/antagonistas & inibidores , Proteínas Tirosina Fosfatases/metabolismo , Ensaio Radioligante/métodos , Substituição de Aminoácidos/genética , Animais , Anticorpos/imunologia , Ligação Competitiva , Inibidores Enzimáticos/farmacologia , Enzimas Imobilizadas/antagonistas & inibidores , Enzimas Imobilizadas/genética , Enzimas Imobilizadas/imunologia , Enzimas Imobilizadas/metabolismo , Cabras , Concentração Inibidora 50 , Antígenos Comuns de Leucócito/metabolismo , Ligantes , Camundongos , Microesferas , Ligação Proteica , Proteína Tirosina Fosfatase não Receptora Tipo 1 , Proteínas Tirosina Fosfatases/genética , Proteínas Tirosina Fosfatases/imunologia , Coelhos , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/metabolismo , Reprodutibilidade dos Testes , Proteína Estafilocócica A/metabolismoRESUMO
We have studied T-cell protein-tyrosine phosphatase (TCPTP) as a model phosphatase in an attempt to unravel amino acid residues that may influence the design of specific inhibitors. Residues 48--50, termed the YRD motif, a region that is found in protein-tyrosine phosphatases, but absent in dual-specificity phosphatases was targeted. YRD derivatives of TCPTP were characterized by steady-state kinetics and by inhibition studies with BzN-EJJ-amide, a potent inhibitor of TCPTP. Substitution of Asp(50) to alanine or Arg(49) to lysine, methionine, or alanine significantly affected substrate hydrolysis and led to a substantial decrease in affinity for BzN-EJJ-amide. The influence of residue 49 on substrate/inhibitor selectivity was further investigated by comparing subsite amino acid preferences of TCPTP and its R49K derivative by affinity selection coupled with mass spectrometry. The greatest effect on selectivity was observed on the residue that precedes the phosphorylated tyrosine. Unlike wild-type TCPTP, the R49K derivative preferred tyrosine to aspartic or glutamic acid. BzN-EJJ-amide which retains the preferred specificity requirements of TCPTP and PTP1B was equipotent on both enzymes but greater than 30-fold selective over other phosphatases. These results suggest that Arg(49) and Asp(50) may be targeted for the design of potent and selective inhibitors of TCPTP and PTP1B.
Assuntos
Proteínas Tirosina Fosfatases/metabolismo , Linfócitos T/enzimologia , Substituição de Aminoácidos , Sítios de Ligação , Humanos , Proteínas Tirosina Fosfatases/química , Proteínas Tirosina Fosfatases/genética , Especificidade por SubstratoRESUMO
Previous biochemical evidence has yielded conflicting models for the role of protein tyrosine phosphatase-1B (PTP-1B) in the regulation of integrin signaling. Thus, to establish the physiological relevance for such a role, we employed a genetic approach by generating embryonic fibroblasts from PTP-1B knockout mice. Both primary fibroblasts and their derived cell lines were used in this study. Immortalization of wild-type primary cells with the SV40 Large T antigen resulted in a dramatic increase in the endogenous expression of PTP-1B, suggesting a role during transformation. Moreover, the absence of PTP-1B in the transformed cell lines led to a more pronounced effect on different pathways of fibronectin-mediated signaling compared with the untransformed state. Specifically, p130(Cas) phosphorylation, Erk activation as well as cell spreading were delayed in PTP-1B-deficient cells, compared with their wild-type counterparts. Interestingly, this attenuation in integrin-mediated events closely resembles that of Src-deficient fibroblasts. Indeed, PTP-1B deficient, transformed fibroblasts held in suspension do exhibit a hyperphosphorylation of the inhibitory site (Tyr-527) of Src, compared with their wild-type counterparts. These results establish PTP-1B as a positive physiological regulator of integrin signaling in transformed cells, acting upstream of Src Tyr-527 dephosphorylation that leads to several adhesion-dependent events.
Assuntos
Proteínas Tirosina Fosfatases/fisiologia , Transdução de Sinais/fisiologia , Animais , Adesão Celular/fisiologia , Linhagem Celular Transformada , Integrinas/fisiologia , Camundongos , Camundongos Knockout , Fosforilação , Proteína Tirosina Fosfatase não Receptora Tipo 1RESUMO
This paper investigates the different sources of variation between US states in self-rated health using multilevel statistical procedures. The different sources that are considered are based on individual- and state-level factors. Data for the analysis comes from the 1993-94 Behavioral Risk Factor Surveillance System and the 1986-90 General Social Surveys. Results show that individual-level factors (such as low income, being black, smoking) are strongly associated with self-rated poor health. Significant variation, however, remain between states after allowing for individual characteristics. Crucially, between-state variation in self-rated health is different for different income groups. State-level contextual effects are found for per-capita median-income and 'social capital'. While not strong, there seems to be a differential impact of state income-inequality on high-income groups, such that the affluent report better health from living in high inequality states. The paper substantiates the need to connect individual health to their macro socioeconomic context. Importantly, it is argued that without adopting an explicitly multilevel approach, the debate on linkages between individual health and income-inequality/social capital cannot be adequately addressed.
Assuntos
Área Programática de Saúde/estatística & dados numéricos , Nível de Saúde , Características de Residência/estatística & dados numéricos , Autoavaliação (Psicologia) , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Fatores Socioeconômicos , Estados Unidos/epidemiologiaRESUMO
OBJECTIVES: Previous studies have linked state-level income inequality to mortality rates. However, it has been questioned whether the relationship is independent of individual-level income. The present study tests whether state-level income inequality is related to individual mortality risk, after adjustment for individual-level characteristics. METHODS: In this prospective, multilevel study design, the vital status of National Health Interview Survey (NHIS) respondents was ascertained by linkage to the National Death Index, with additional linkage of state-level data to individuals in the NHIS. The analysis included data for 546,888 persons, with 19,379 deaths over the 8-year follow-up period. The Gini coefficient was used as the measure of income inequality. RESULTS: Individuals living in high-income-inequality states were at increased risk of mortality (relative risk = 1.12; 95% confidence interval = 1.04, 1.19) compared with individuals living in low-income-inequality states. In stratified analyses, significant effects of state income inequality on mortality risk were found, primarily for near-poor Whites. CONCLUSIONS: State-level income inequality appears to exert a contextual effect on mortality risk, after income is adjusted for, providing further evidence that the distribution of income is important for health.
Assuntos
Renda , Mortalidade , Adolescente , Adulto , Negro ou Afro-Americano/estatística & dados numéricos , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de Risco , Estados Unidos , População Branca/estatística & dados numéricosRESUMO
OBJECTIVES: This study tested the hypothesis that disparities in political participation across socioeconomic status affect health. Specifically, the association of voting inequality at the state level with individual self-rated health was examined. METHODS: A multilevel study of 279,066 respondents to the Current Population Survey (CPS) was conducted. State-level inequality in voting turnout by socioeconomic status (family income and educational attainment) was derived from November CPS data for 1990, 1992, 1994, and 1996. RESULTS: Individuals living in the states with the highest voting inequality had an odds ratio of fair/poor self-rated health of 1.43 (95% confidence interval [CI] = 1.22, 1.68) compared with individuals living in the states with the lowest voting inequality. This odds ratio decreased to 1.34 (95% CI = 1.14, 1.56) when state income inequality was added and to 1.27 (95% CI = 1.10, 1.45) when state median income was included. The deleterious effect of low individual household income on self-rated health was most pronounced among states with the greatest voting and income inequality. CONCLUSIONS: Socioeconomic inequality in political participation (as measured by voter turnout) is associated with poor self-rated health, independently of both income inequality and state median household income.
Assuntos
Educação , Nível de Saúde , Renda , Política , Adulto , Idoso , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Identificação Social , Justiça Social , Fatores Socioeconômicos , Estados Unidos/epidemiologiaRESUMO
OBJECTIVE: To examine the association of state income inequality and individual household income with the mental and physical health of women with young children. DESIGN: Cross sectional study. Individual level data (outcomes, income, and other sociodemographic covariates) from a 1991 follow up survey of a birth cohort established in 1988. State level income inequality calculated from the income distribution of each state from 1990 US census. SETTING: United States, 1991. PARTICIPANTS: Nationally representative stratified random sample of 8060 women who gave birth in 1988 and were successfully contacted (89%) in 1991. MAIN OUTCOME MEASURES: Depressive symptoms (Center for Epidemiologic Studies depression score >15) and self rated health RESULTS: 19% of women reported depressive symptoms, and 7.5% reported fair or poor health. Compared with women in the highest fifth of distribution of household income, women in the lowest fifth were more likely to report depressive symptoms (33% v 9%, P<0.001) and fair or poor health (15% v 2%, P<0. 001). Compared with low income women in states with low income inequality, low income women in states with high income inequality had a higher risk of depressive symptoms (odds ratio 1.6, 95% confidence interval 1.0 to 2.6) and fair or poor health (1.8, 0.9 to 3.5). CONCLUSIONS: High income inequality confers an increased risk of poor mental and physical health, particularly among the poorest women. Both income inequality and household income are important for health in this population.
Assuntos
Nível de Saúde , Renda/estatística & dados numéricos , Bem-Estar Materno/estatística & dados numéricos , Saúde Mental/estatística & dados numéricos , Adolescente , Adulto , Estudos Transversais , Depressão/epidemiologia , Escolaridade , Feminino , Inquéritos Epidemiológicos , Humanos , Modelos Logísticos , Casamento , Pessoa de Meia-Idade , Fatores de Risco , Estados Unidos/epidemiologiaRESUMO
A role for protein tyrosine phosphatases in the negative regulation of insulin signaling and a putative involvement in the insulin resistance associated with type 2 diabetes have been postulated since their discovery. The recent demonstration that mice lacking the protein tyrosine phosphatase-1B (PTP-1B) have enhanced insulin sensitivity validates this. Furthermore, when fed a high fat diet, these mice maintained insulin sensitivity and were resistant to obesity, suggesting that inhibition of PTP-1B activity could be a novel way of treating type 2 diabetes and obesity. This commentary reviews our current knowledge of PTP-1B in insulin signaling and its role in diabetes and discusses the development of potent and selective PTP-1B inhibitors.
Assuntos
Diabetes Mellitus/enzimologia , Proteínas Tirosina Fosfatases/metabolismo , Animais , Diabetes Mellitus/metabolismo , Progressão da Doença , Humanos , Imunidade Inata/fisiologia , Camundongos , Obesidade , Fenótipo , Proteína Tirosina Fosfatase não Receptora Tipo 1 , Proteínas Tirosina Fosfatases/antagonistas & inibidores , Proteínas Tirosina Fosfatases/deficiência , Receptor de Insulina/metabolismoRESUMO
PTP-1B is a ubiquitously expressed intracellular protein tyrosine phosphatase (PTP) that has been implicated in the negative regulation of insulin signaling. Mice deficient in PTP-1B were found to have an enhanced insulin sensitivity and a resistance to diet-induced obesity. Interestingly, the human PTP-1B gene maps to chromosome 20q13.1 in a region that has been associated with diabetes and obesity. Although there has been a partial characterization of the 3' end of the human PTP-1B gene, the complete gene organization has not been described. In order to further characterize the PTP-1B gene, we have cloned and determined the genomic organization for both the human and mouse PTP-1B genes including the promoter. The human gene spans >74 kb and features a large first intron of >54 kb; the mouse gene likewise contains a large first intron, although the exact size has not been determined. The organization of the human and mouse PTP-1B genes is identical except for an additional exon at the 3' end of the human that is absent in the mouse. The mouse PTP-1B gene maps to the distal arm of mouse chromosome 2 in the region H2-H3. This region is associated with a mouse obesity quantitative trait locus (QTL) and is syntenic with human chromosome 20. The promoter region of both the human and mouse genes contain no TATA box but multiple GC-rich sequences that contain a number of consensus SP-1 binding sites. The basal activity of the human PTP-1B promoter was characterized in Hep G2 cells using up to 8 kb of 5' flanking sequence. A 432 bp promoter construct immediately upstream of the ATG was able to confer maximal promoter activity. Within this sequence, there are at least three GC-rich sequences and one CCAAT box, and deletion of any of these elements results in decreased promoter activity. In addition, the promoter in a number of mouse strains contains, 3.5 kb upstream of the start codon, an insertion of an intracisternal a particle (IAP) element that possibly could alter the expression of PTP-1B mRNA in these strains.
Assuntos
Genes/genética , Proteínas Tirosina Fosfatases/genética , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Mapeamento Cromossômico , Clonagem Molecular , DNA/química , DNA/genética , Éxons , Expressão Gênica , Regulação da Expressão Gênica , Genes de Partícula A Intracisternal/genética , Humanos , Hibridização in Situ Fluorescente , Íntrons , Masculino , Camundongos , Camundongos Endogâmicos , Dados de Sequência Molecular , Mutagênese Insercional , Regiões Promotoras Genéticas/genética , Proteína Tirosina Fosfatase não Receptora Tipo 1 , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Distribuição Tecidual , Células Tumorais CultivadasRESUMO
We have developed an intact cell assay to be used in the direct quantitation of protein tyrosine phosphatase (PTP) activity. Utilizing the baculovirus expression system, the assay readily allows for a direct activity readout for PTPs such as PTP1B or CD45. Infected Sf9 cells expressing either full-length PTP1B, full-length CD45, CD45 catalytic domain, or hCOX-1 (mock-infected) are harvested 29 hr post-infection, at which time cells are viable and the expressed proteins are processed, as well as localized to their predicted subcellular compartments. Assays are carried out in a 96-well format, with cells expressing the PTP of interest. Cells are preincubated with or without inhibitor and challenged with substrate, and the phosphatase activity is determined spectrophotometrically by monitoring the conversion of p-nitrophenyl phosphate to p-nitrophenol at OD405. Documented PTP inhibitors have been used to validate this assay system. This study demonstrates that a direct readout of PTP activity in intact cells can be achieved, thus providing a useful cell-based screen for determining selective inhibitors of PTPs.
Assuntos
Proteínas Tirosina Fosfatases/análise , Animais , Baculoviridae/genética , Bioensaio/métodos , Western Blotting/métodos , Vetores Genéticos , Antígenos Comuns de Leucócito/análise , Antígenos Comuns de Leucócito/genética , Antígenos Comuns de Leucócito/metabolismo , Proteínas Tirosina Fosfatases/antagonistas & inibidores , Proteínas Tirosina Fosfatases/genética , Controle de Qualidade , Proteínas Recombinantes/análise , Proteínas Recombinantes/antagonistas & inibidores , Reprodutibilidade dos Testes , Spodoptera/citologia , TransfecçãoRESUMO
OBJECTIVES: Social capital consists of features of social organization--such as trust between citizens, norms of reciprocity, and group membership--that facilitate collective action. This article reports a contextual analysis of social capital and individual self-rated health, with adjustment for individual household income, health behaviors, and other covariates. METHODS: Self-rated health ("Is your overall health excellent, very good, good, fair, or poor?") was assessed among 167,259 individuals residing in 39 US states, sampled by the Behavioral Risk Factor Surveillance System. Social capital indicators, aggregated to the state level, were obtained from the General Social Surveys. RESULTS: Individual-level factors (e.g., low income, low education, smoking) were strongly associated with self-rated poor health. However, even after adjustment for these proximal variables, a contextual effect of low social capital on risk of self-rated poor health was found. For example, the odds ratio for fair or poor health associated with living in areas with the lowest levels of social trust was 1.41 (95% confidence interval = 1.33, 1.50) compared with living in high-trust states. CONCLUSIONS: These results extend previous findings on the health advantages stemming from social capital.
Assuntos
Nível de Saúde , Vigilância da População/métodos , Identificação Social , Adulto , Idoso , Análise por Conglomerados , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Análise de Regressão , Fatores Socioeconômicos , Estados UnidosRESUMO
OBJECTIVE: This study examines the factors associated with planning to avoid alcohol-impaired driving and successful avoidance in high-risk young men. METHOD: A targeted telephone survey was conducted with male drivers aged 21-35 years who consume alcohol and live in areas of the country where alcohol-related traffic fatalities occur frequently (N = 750). Heavy episodic drinking drivers (i.e., report driving after consuming five or more drinks) were oversampled (N = 230). Respondents were surveyed to assess their attitudes, behavior and social support regarding drinking-driving. RESULTS: Multiple logistic regression revealed that men who believe they can consume six drinks or more before it is too dangerous for them to drive were 45% less likely to report planning to avoid drinking-driving. Men who believe they can drive safely after heavy episodic drinking were 61% less likely to be successful in avoiding drinking-driving. Having friends who disapprove of driving after heavy episodic drinking and believing a close friend would be successful in preventing drinking-driving were significantly associated with making plans to avoid drinking-driving, although this association was less strong for successful avoidance. Men who had their wife/girlfriend along when they were out drinking were two and one-half times more likely to make plans to avoid drinking-driving. The presence of a wife or girlfriend was an even stronger predictor (multivariate odds ratio = 3.74) of successful avoidance. CONCLUSIONS: Attitude and social factors are associated with drinking-driving avoidance in a group of young men at risk for alcohol-related driving fatalities. Friends and wives/girlfriends appear to influence drinking-driving avoidance behavior in high-risk drinking drivers.