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1.
J Med Internet Res ; 25: e38713, 2023 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-36729576

RESUMO

BACKGROUND: Alcohol misuse is highly prevalent in the United States and results in a huge financial and public health burden. Current alcohol reduction treatments are underused, and there is a critical need for innovation in the field. Transdermal alcohol biosensors measure alcohol use passively and continuously and may be helpful tools in alcohol interventions. To date, however, alcohol biosensors have not been widely used to directly intervene on alcohol use. There is a new wrist-worn biosensor that could be used to help people reduce their drinking, although it is unclear how best to incorporate such a device into an alcohol intervention. OBJECTIVE: We aimed to identify desired features that would be acceptable and helpful in a wrist-worn biosensor-based alcohol intervention for adults who drink heavily. METHODS: Participants were recruited through an alcohol contingency management study, a contact registry, and participant referral. To qualify, participants had to be aged at least 40 years, report drinking at least twice per week, and indicate interest in reducing their drinking. We conducted a semistructured interview with each participant via Zoom (Zoom Video Communications, Inc). The interview guide addressed general thoughts on the wrist-worn biosensor, how participants thought a wrist-worn biosensor could be used to help people quit or reduce drinking, types of information that participants would want to receive from the biosensor, how they would want to receive this information, and how they thought this information could be used to change their behavior. Interviews were transcribed verbatim and analyzed using thematic analysis. RESULTS: The sample comprised 20 adults (mean age 55.1, SD 6.1 years; 11/20, 55%, women; and 17/20, 85%, Black). Of the 20 participants, 9 (45%) had previous experience with the Secure Continuous Remote Alcohol Monitor continuous alcohol monitoring ankle biosensor from participating in an alcohol contingency management study. The desirable features could be grouped into 5 main themes: features that would influence willingness to use the biosensor (it should look attractive and be both comfortable to wear and accessible), personalized messaging (personalized biosensor-based prompts and feedback could be helpful), preference for time wearing the biosensor (for some, just wearing the biosensor could have an intervention effect), sharing data with others (this was appealing to many but not to all), and mental health support (many felt that mental health support could be incorporated into the biosensor). CONCLUSIONS: Five main themes that would maximize interest in using a wrist-worn biosensor for alcohol intervention were identified. Taken together, the identified themes could inform the development of a just-in-time adaptive intervention that uses a wrist-worn biosensor to help adults who drink heavily reduce their alcohol use.


Assuntos
Técnicas Biossensoriais , Punho , Humanos , Adulto , Feminino , Pessoa de Meia-Idade , Masculino , Etanol , Pesquisa Qualitativa
2.
Respir Res ; 23(1): 177, 2022 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-35780155

RESUMO

BACKGROUND: Respiratory infections such as influenza account for significant global mortality each year. Generating lipid profiles is a novel and emerging research approach that may provide new insights regarding the development and progression of priority respiratory infections. We hypothesized that select clusters of lipids in human sputum would be associated with specific viral infections (Influenza (H1N1, H3N2) or Rhinovirus). METHODS: Lipid identification and semi-quantitation was determined with liquid chromatography and high-resolution mass spectrometry in induced sputum from individuals with confirmed respiratory infections (influenza (H1N1, H3N2) or rhinovirus). Clusters of lipid species and associations between lipid profiles and the type of respiratory viral agent was determined using Bayesian profile regression and multinomial logistic regression. RESULTS: More than 600 lipid compounds were identified across the sputum samples with the most abundant lipid classes identified as triglycerides (TG), phosphatidylethanolamines (PE), phosphatidylcholines (PC), Sphingomyelins (SM), ether-PC, and ether-PE. A total of 12 lipid species were significantly different when stratified by infection type and included acylcarnitine (AcCar) (10:1, 16:1, 18:2), diacylglycerols (DG) (16:0_18:0, 18:0_18:0), Lysophosphatidylcholine (LPC) (12:0, 20:5), PE (18:0_18:0), and TG (14:1_16:0_18:2, 15:0_17:0_19:0, 16:0_17:0_18:0, 19:0_19:0_19:0). Cluster analysis yielded three clusters of lipid profiles that were driven by just 10 lipid species (TGs and DGs). Cluster 1 had the highest levels of each lipid species and the highest prevalence of influenza A H3 infection (56%, n = 5) whereas cluster 3 had lower levels of each lipid species and the highest prevalence of rhinovirus (60%; n = 6). Using cluster 3 as the reference group, the crude odds of influenza A H3 infection compared to rhinovirus in cluster 1 was significantly (p = 0.047) higher (OR = 15.00 [95% CI: 1.03, 218.29]). After adjustment for confounders (smoking status and pulmonary comorbidities), the odds ratio (OR) became only marginally significant (p = 0.099), but the magnitude of the effect estimate was similar (OR = 16.00 [0.59, 433.03]). CONCLUSIONS: In this study, human sputum lipid profiles were shown to be associated with distinct types of viral infection. Better understanding the relationship between respiratory infections of global importance and lipids contributes to advancing knowledge of pathogenesis of infections including identifying populations with increased susceptibility and developing effective therapeutics and biomarkers of health status.


Assuntos
Vírus da Influenza A Subtipo H1N1 , Influenza Humana , Pneumonia , Infecções Respiratórias , Viroses , Teorema de Bayes , Humanos , Vírus da Influenza A Subtipo H3N2 , Lisofosfatidilcolinas , Fosfatidilcolinas , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/epidemiologia , Rhinovirus , Escarro , Viroses/diagnóstico , Viroses/epidemiologia
3.
AIDS Behav ; 26(10): 3242-3253, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35380289

RESUMO

Alcohol use is associated with poor outcomes among people living with HIV (PLWH), but it remains unclear which alcohol use measures best predict future HIV viral non-suppression over time. This study aimed to compare the ability of five alcohol use measures to predict risk of suboptimal HIV viral load trajectories over 36 months. We analyzed data from a cohort of PLWH in Florida including survey data linked to the state HIV surveillance system on prospective HIV viral loads over 36 months (n = 783; 66% male; 55% Black; Mage=46, SD = 11). Four trajectory patterns for HIV viral load were identified: consistently low (65.1%), decreasing (15.9%), increasing (10.6%), and consistently high (8.4%). Past year alcohol use frequency (OR = 2.1, CI:1.0-4.4), drinks consumed on a typical drinking day (OR = 2.2, CI:1.2-4.1), frequency of binge drinking (OR = 2.6, CI:1.3-5.2), and alcohol-related problems score (OR = 1.7, CI:1.1-2.7) were the measures predictive of the risk of future viral non-suppression above specific thresholds.


RESUMEN: El consumo de alcohol está asociado con malos resultados entre las personas que viven con el VIH (PLWH), pero aún no está claro qué medidas de consumo de alcohol predicen mejor la falta de supresión viral del VIH en el futuro con el tiempo. Este estudio tuvo como objetivo comparar la capacidad de cinco medidas de consumo de alcohol para predecir el riesgo de trayectorias subóptimas de la carga viral del VIH durante 36 meses. Analizamos datos de una cohorte de PLWH en Florida, incluidos datos de encuestas vinculadas al sistema estatal de vigilancia del VIH sobre posibles cargas virales del VIH durante 36 meses (n = 783; 66% hombres; 55% afroamericanos; Maños=46, SD = 11). Se identificaron cuatro patrones de trayectoria para la carga viral del VIH: consistentemente baja (65,1%), decreciente (15,9%), creciente (10,6%) y consistentemente alta (8,4%). Frecuencia de consumo de alcohol en el último año (OR = 2,1, IC: 1,0­4,4), bebidas consumidas en un día típico de consumo de alcohol (OR = 2,2, IC: 1,2­4,1), frecuencia de consumo excesivo de alcohol (OR = 2,6, IC: 1,3­5,2), y la puntuación de problemas relacionados con el alcohol (OR = 1,7, IC: 1,1­2,7) fueron las medidas predictivas del riesgo de no supresión viral futura por encima de umbrales específicos.


Assuntos
Infecções por HIV , Consumo de Bebidas Alcoólicas/epidemiologia , Estudos de Coortes , Feminino , Infecções por HIV/complicações , Infecções por HIV/diagnóstico , Infecções por HIV/epidemiologia , Teste de HIV , Humanos , Masculino , Estudos Prospectivos , Carga Viral
4.
MMWR Morb Mortal Wkly Rep ; 70(6): 208-211, 2021 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-33571175

RESUMO

Approximately 41% of adults aged 18-24 years in the United States are enrolled in a college or university (1). Wearing a face mask can reduce transmission of SARS-CoV-2, the virus that causes coronavirus disease 2019 (COVID-19) (2), and many colleges and universities mandate mask use in public locations and outdoors when within six feet of others. Studies based on self-report have described mask use ranging from 69.1% to 86.1% among adults aged 18-29 years (3); however, more objective measures are needed. Direct observation by trained observers is the accepted standard for monitoring behaviors such as hand hygiene (4). In this investigation, direct observation was used to estimate the proportion of persons wearing masks and the proportion of persons wearing masks correctly (i.e., covering the nose and mouth and secured under the chin*) on campus and at nearby off-campus locations at six rural and suburban universities with mask mandates in the southern and western United States. Trained student observers recorded mask use for up to 8 weeks from fixed sites on campus and nearby. Among 17,200 observed persons, 85.5% wore masks, with 89.7% of those persons wearing the mask correctly (overall correct mask use: 76.7%). Among persons observed indoors, 91.7% wore masks correctly. The proportion correctly wearing masks indoors varied by mask type, from 96.8% for N95-type masks and 92.2% for cloth masks to 78.9% for bandanas, scarves, and similar face coverings. Observed indoor mask use was high at these six universities with mask mandates. Colleges and universities can use direct observation findings to tailor training and messaging toward increasing correct mask use.


Assuntos
Máscaras/estatística & dados numéricos , Máscaras/normas , Saúde Pública/legislação & jurisprudência , Estudantes/psicologia , Universidades/legislação & jurisprudência , Adolescente , COVID-19/epidemiologia , COVID-19/prevenção & controle , Humanos , Estudantes/estatística & dados numéricos , Estados Unidos/epidemiologia , Adulto Jovem
5.
Am J Public Health ; 105 Suppl 1: S83-7, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25706027

RESUMO

The College of Public Health and Health Professions at the University of Florida is composed of five public health departments and four clinical health professions departments, and the college is one of six that make up the university's Health Science Center. These organizational resources, along with the university's explicit emphasis on collaboration across professions, colleges, institutes, and centers and the strong leadership and full support of deans and other academic leaders, provide a strong foundation for educational innovations. Three key areas in which the college has built upon these opportunities are interprofessional education, development of One Health instructional programs, and application of cutting-edge technology to students' educational experiences. These innovations represent the types of creative approaches to preparing the 21st-century workforce that can be developed through collaboration among multiple disciplines in a major university.


Assuntos
Educação Profissional em Saúde Pública/organização & administração , Faculdades de Saúde Pública/organização & administração , Universidades/organização & administração , Instrução por Computador , Currículo , Florida , Humanos , Comunicação Interdisciplinar , Liderança , Desenvolvimento de Programas
6.
Int Arch Occup Environ Health ; 87(5): 557-65, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23897226

RESUMO

PURPOSE: Heat has been known to increase the risk of many health endpoints. However, few studies have examined its effects on stroke. The objective of this case-crossover study is to investigate the effects of high heat and its effect modifiers on the risk of stroke hospitalization in Allegheny County, Pennsylvania. METHODS: We obtained data on first stroke hospitalizations among adults ages 65 and older and daily meteorological information during warm seasons (May-September) from 1994 to 2000 in Allegheny County, Pennsylvania. Using conditional multiple logistic regressions, the effects of heat days (any day with a temperature greater than the 95th percentile) and heat wave days (at least two continuous heat days) on the risk of stroke hospitalization were investigated. The potential interactions between high heat and age, type of stroke, and gender were also examined. RESULTS: Heat day and heat wave at lag-2 day were significantly associated with an increased risk for stroke hospitalization (OR 1.121, 95 % CI 1.013-1.242; OR 1.173, 95 % CI 1.047-1.315, respectively) after adjusting for important covariates. In addition, having two or more heat wave days within the 4 day window prior to the event was also significantly associated with an increased risk (OR 1.119, 95 % CI 1.004, 1.246) compared to having no heat wave days during the period. The effect of high heat on stroke was more significant for ischemic stroke, men, and subjects ages 80 years or older. CONCLUSIONS: Our study suggests that high heat may have adverse effects on stroke and that some subgroups may be particularly susceptible to heat.


Assuntos
Exposição Ambiental/efeitos adversos , Transtornos de Estresse por Calor/epidemiologia , Hospitalização/estatística & dados numéricos , Temperatura Alta/efeitos adversos , Acidente Vascular Cerebral/epidemiologia , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Poluição do Ar/análise , Estudos Cross-Over , Feminino , Humanos , Masculino , Pennsylvania/epidemiologia , Fatores de Risco , Estações do Ano , Fatores Sexuais
7.
Alcohol Clin Exp Res (Hoboken) ; 47(2): 308-318, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36507857

RESUMO

BACKGROUND: Transdermal alcohol biosensors measure alcohol use continuously, passively, and non-invasively. There is little field research on the Skyn biosensor, a new-generation, wrist-worn transdermal alcohol biosensor, and little evaluation of its sensitivity and specificity and the day-level correspondence between transdermal alcohol concentration (TAC) and number of self-reported drinks. METHODS: Participants (N = 36; 61% male, M age  = 34.3) wore the Skyn biosensor and completed ecological momentary assessment (EMA) surveys about their alcohol use over 2 weeks. A total of 497 days of biosensor and EMA data were collected. Skyn-measured drinking episodes were defined by TAC > 5 µg/L. Skyn data were compared to self-reported drinking to calculate sensitivity and specificity (for drinking day vs. nondrinking day). Generalized estimating equations models were used to evaluate the correspondence between TAC features (peak TAC and TAC-area under the curve (AUC)) and number of drinks. Individual-level factors (sex, age, race/ethnicity, body mass index, human immunodeficiency virus status, and hazardous drinking) were examined to explore associations with TAC controlling for number of drinks. RESULTS: Using a minimum TAC threshold of 5 µg/L plus coder review, the biosensor had sensitivity of 54.7% and specificity of 94.6% for distinguishing drinking from nondrinking days. Without coder review, the sensitivity was 78.1% and the specificity was 55.2%. Peak TAC (ß = 0.92, p < 0.0001) and TAC-AUC (ß = 1.60, p < 0.0001) were significantly associated with number of drinks. Females had significantly higher TAC levels than males for the same number of drinks. CONCLUSIONS: Skyn-derived TAC can be used to measure alcohol use under naturalistic drinking conditions, additional research is needed to accurately identify drinking episodes based on Skyn TAC readings.


Assuntos
Técnicas Biossensoriais , Punho , Feminino , Humanos , Masculino , Adulto , Autorrelato , Avaliação Momentânea Ecológica , Etanol , Consumo de Bebidas Alcoólicas/epidemiologia
8.
Disaster Med Public Health Prep ; 17: e176, 2022 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-35492011

RESUMO

The coronavirus disease 2019 (COVID-19) pandemic continues to present unique public health challenges both within the United States and across the globe. Institutions of higher learning are tasked with preventing and responding to COVID-19 on campus while also considering implications for the surrounding communities. The process of re-opening campus, whether at full or partial capacity, has tasked these institutions with overcoming complex challenges associated with balancing the resumption of campus operations while simultaneously protecting university affiliates and surrounding community members from COVID-19 through robust surveillance, contact tracing, and testing efforts. Here, we provide a concise outline related to the development and implementation of the comprehensive and sustainable COVID-19 surveillance program at the University of Florida. We also critically discuss the successes and pitfalls of this program while also providing recommendations for the development of similar programs in the future.


Assuntos
COVID-19 , Estados Unidos/epidemiologia , Humanos , COVID-19/epidemiologia , Universidades , Sudeste dos Estados Unidos/epidemiologia , Saúde Pública , Busca de Comunicante
9.
J Am Coll Health ; : 1-4, 2022 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-35727228

RESUMO

Background: We evaluate the public health surveillance program, Screen, Test, and Protect (STP) designed to control and prevent COVID-19 at a large academic university in the United States. Methods: STP was established at the University of Florida in May 2020. This report details STP's full-time workforce, centralized database, and testing and vaccination programs. We evaluate the program's success in controlling COVID-19 during the 2020-2021 academic school year. Results: COVID-19 cases rose among the campus community in the first few weeks of campus reopening in Fall 2020. Test positivity levels returned to prefall semester levels within one month, however. A few additional, yet smaller, waves occurred during the 2020-2021 school year and were successfully controlled without any campus-wide closures. Conclusions: This program may serve as a framework for other institutions managing the ongoing COVID-19 crisis, in addition to setting the standard for programmatic management of future emerging infectious diseases at universities.

10.
PLoS One ; 17(8): e0271917, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35925972

RESUMO

PURPOSE: This study examined factors associated with TB among persons living with HIV (PLWH) in Florida and the agreement between self-reported and medically documented history of tuberculosis (TB) in assessing the risk factors. METHODS: Self-reported and medically documented data of 655 PLWH in Florida were analyzed. Data on sociodemographic factors such as age, race/ethnicity, place of birth, current marital status, education, employment, homelessness in the past year and 'ever been jailed' and behavioural factors such as excessive alcohol use, marijuana, injection drug use (IDU), substance and current cigarette use were obtained. Health status information such as health insurance status, adherence to HIV antiretroviral therapy (ART), most recent CD4 count, HIV viral load and comorbid conditions were also obtained. The associations between these selected factors with self-reported TB and medically documented TB diagnosis were compared using Chi-square and logistic regression analyses. Additionally, the agreement between self-reports and medical records was assessed. RESULTS: TB prevalence according to self-reports and medical records was 16.6% and 7.5% respectively. Being age ≥55 years, African American and homeless in the past 12 months were statistically significantly associated with self-reported TB, while being African American homeless in the past 12 months and not on antiretroviral therapy (ART) were statistically significantly associated with medically documented TB. African Americans compared to Whites had odds ratios of 3.04 and 4.89 for self-reported and medically documented TB, respectively. There was moderate agreement between self-reported and medically documented TB (Kappa = 0.41). CONCLUSIONS: TB prevalence was higher based on self-reports than medical records. There was moderate agreement between the two data sources, showing the importance of self-reports. Establishing the true prevalence of TB and associated risk factors in PLWH for developing policies may therefore require the use of self-reports and confirmation by screening tests, clinical signs and/or microbiologic data.


Assuntos
Infecções por HIV , Tuberculose , Florida/epidemiologia , Infecções por HIV/complicações , Infecções por HIV/tratamento farmacológico , Infecções por HIV/epidemiologia , Humanos , Prontuários Médicos , Pessoa de Meia-Idade , Fatores de Risco , Autorrelato , Tuberculose/complicações , Tuberculose/epidemiologia
11.
Am J Infect Control ; 48(9): 1028-1031, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-31924388

RESUMO

BACKGROUND: To evaluate risk factors for infection or colonization with carbapenem-resistant Enterobacteriaceae (CRE) to develop an algorithm for targeted CRE screening. METHODS: We conducted a case-control study of 50 CRE-positive cases and 100 CRE-negative controls to identify risk factors that were significant for CRE infection or colonization. The setting was at an acute care academic hospital. Patients who tested positive for CRE or other microbiological laboratory tests during the study period were included. We reviewed medical records of 50 patients who were CRE-positive and 100 matched controls who had a non-CRE culture at a similar anatomic site within the closest time period to the case's culture date. Risk factors were assessed using logistic regression with SAS 9.4, observing the 95% confidence interval (CI) to determine significance. RESULTS: Significant risk factors for CRE infection or colonization included the use of fluoroquinolones (odds ratio [OR], 3.75; 95% CI, 1.35, 10.38) and cephalosporins (OR, 2.37; 95% CI, 1.17, 4.86). In addition, undergoing an invasive procedure with a scope device was also a significant risk factor for our participants (OR, 4.57; 95% CI, 1.31, 16.02). Significance of these risk factors varied within the community-acquired and hospital-acquired cases. CONCLUSIONS: Our results suggest that exposure to certain antimicrobials and invasive procedures with a scope device (endoscopic retrograde cholangiopancreatography, duodenal endoscope) are risk factors for CRE. The findings of significant differences in antimicrobials received highlight the necessity to understand antimicrobial stewardship in the development of CRE colonization and infection. Along with antibiotics, inaccessibility to components within scope devices may be increasing the risk of CRE spread.


Assuntos
Enterobacteriáceas Resistentes a Carbapenêmicos , Infecções por Enterobacteriaceae , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Carbapenêmicos/farmacologia , Estudos de Casos e Controles , Infecções por Enterobacteriaceae/tratamento farmacológico , Infecções por Enterobacteriaceae/epidemiologia , Humanos , Fatores de Risco
12.
Vet Q ; 33(1): 25-9, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23477422

RESUMO

BACKGROUND: Captive elephants infected with tuberculosis are implicated as an occupational source of zoonotic tuberculosis. However, accurate estimates of prevalence and incidence of elephant tuberculosis from well-defined captive populations are lacking in the literature. Studies published in recent years contain a wide range of prevalence estimates calculated from summary data. Incidence estimates of elephant tuberculosis in captive elephants are not available. OBJECTIVE: This study estimated the annual point prevalence, annual incidence, cumulative incidence, and incidence density of tuberculosis in captive elephants within the USA during the past 52 years. ANIMALS AND METHODS: We combined existing elephant census records from captive elephants in the USA with tuberculosis culture results obtained from trunk washes or at necropsy. This data set included 15 years where each elephant was screened annually. RESULTS: Between 1960 and 1996, the annual point prevalence of tuberculosis complex mycobacteria for both species was 0. From 1997 through 2011, the median point prevalence within the Asian elephant population was 5.1%, with a range from 0.3% to 6.7%. The incidence density was 9.7 cases/1000 elephant years (95% CI: 7.0-13.4). In contrast, the annual point prevalence during the same time period within the African elephant population remained 0 and the incidence density was 1.5 cases/1000 elephant years (95% CI: 0.7-4.0). CONCLUSIONS: The apparent increase in new cases noted after 1996 resulted from a combination of both index cases and the initiation of mandatory annual tuberculosis screening in 1997 for all the elephants. This study found lower annual point prevalence estimates than previously reported in the literature. These discrepancies in prevalence estimates are primarily due to differences in terminology and calculation methods. Using the same intensive testing regime, the incidence of tuberculosis differed significantly between Asian and African elephants. CLINICAL IMPORTANCE: Accurate and species specific knowledge of prevalence and incidence will inform our efforts to mitigate occupational risks associated with captive elephants in the USA.


Assuntos
Elefantes , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose/veterinária , Animais , Incidência , Prevalência , Especificidade da Espécie , Fatores de Tempo , Tuberculose/epidemiologia , Tuberculose/microbiologia , Estados Unidos/epidemiologia
13.
Virology ; 401(1): 49-60, 2010 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-20206959

RESUMO

The vaccinia H5 protein has been implicated in several steps of virus replication including DNA synthesis, postreplicative gene transcription, and virion morphogenesis. Our recent mapping of mutants in the consolidated Condit-Dales collection identified a temperature-sensitive vaccinia mutant in the H5R gene (Dts57). We demonstrate here that Dts57 has a DNA negative phenotype, strongly suggesting a direct role for H5 in DNA replication. We used a temperature shift protocol to determine the impact of H5 temperature sensitivity on postreplicative gene expression and observed changes in the pattern of postreplicative viral mRNA metabolism consistent with a role of H5 in postreplicative transcription. Finally, using a rifampicin release temperature shift protocol, we show that H5 is involved in multiple steps of virion morphogenesis. These data demonstrate directly that H5 plays roles in DNA replication, transcription and morphogenesis in vivo.


Assuntos
Proteínas de Ligação a DNA/fisiologia , Vaccinia virus/fisiologia , Proteínas Virais/fisiologia , Replicação do DNA , DNA Viral/biossíntese , Humanos , Transcrição Gênica , Vírion/fisiologia , Replicação Viral
14.
Virology ; 375(1): 213-22, 2008 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-18314155

RESUMO

Complementation analysis of the combined Condit/Dales collection of vaccinia virus temperature-sensitive mutants has been reported (Lackner, C.A., D'Costa, S.M., Buck, C., Condit, R.C., 2003. Complementation analysis of the Dales collection of vaccinia virus temperature-sensitive mutants. Virology 305, 240-259), however not all complementation groups have previously been assigned to single genes on the viral genome. We have used marker rescue to map at least one representative of each complementation group to a unique viral gene. The final combined collection contains 124 temperature-sensitive mutants affecting 38 viral genes, plus five double mutants.


Assuntos
Mapeamento Cromossômico , Mutação , Vaccinia virus/genética , Animais , Linhagem Celular , Chlorocebus aethiops , Genes Virais , Teste de Complementação Genética , Temperatura Alta , Análise de Sequência de DNA , Ensaio de Placa Viral
15.
Virology ; 366(1): 62-72, 2007 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-17499330

RESUMO

The vaccinia virus temperature-sensitive mutations Cts6 and Cts9 were mapped by marker rescue and DNA sequencing to the A28 gene. Cts6 and Cts9 contain an identical 2-bp deletion truncating the A28 protein and removing the fourth conserved cysteine near the C-terminus. Cts9 mutant virions produced at 40 degrees C were non-infectious and unable to cause cytopathic effect. However, the mutant A28 protein localized to purified mature virions (MV) at 31 degrees C and 40 degrees C. MV of Cts9 produced at 40 degrees C bound to cells but did not enter cells. Low pH treatment of Cts9-infected cells at 18 h p.i. failed to produce fusion from within at 40 degrees C, but gave fusion at 31 degrees C. Adsorption of Cts9 mutant virions to cells followed by low pH treatment showed a defect in fusion from without. The Cts9 phenotype suggests that the A28 protein is involved in both virus entry and cell-cell fusion, and supports the linkage between the two processes.


Assuntos
Genes Virais , Mutação , Vaccinia virus/genética , Vaccinia virus/patogenicidade , Regiões 3' não Traduzidas , Animais , Fusão Celular , Linhagem Celular , Chlorocebus aethiops , Vírus de DNA/genética , Vírus de DNA/patogenicidade , Vírus de DNA/fisiologia , Genoma Viral , Haplorrinos , Recombinação Genética , Temperatura , Vaccinia virus/imunologia , Vaccinia virus/fisiologia , Vacinas Virais , Vírion/genética , Vírion/fisiologia
16.
Virology ; 363(2): 319-32, 2007 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-17336362

RESUMO

Treatment of wild type vaccinia virus infected cells with the anti-poxviral drug isatin-beta-thiosemicarbazone (IBT) induces the viral postreplicative transcription apparatus to synthesize longer-than-normal mRNAs through an unknown mechanism. Previous studies have shown that virus mutants resistant to or dependent on IBT affect genes involved in control of viral postreplicative transcription elongation. This study was initiated in order to identify additional viral genes involved in control of vaccinia postreplicative transcription elongation. Eight independent, spontaneous IBT resistant mutants of vaccinia virus were isolated. Marker rescue experiments mapped two mutants to gene G2R, which encodes a previously characterized postreplicative gene positive transcription elongation factor. Three mutants mapped to the largest subunit of the viral RNA polymerase, rpo147, the product of gene J6R. One mutant contained missense mutations in both G2R and A24R (rpo132, the second largest subunit of the RNA polymerase). Two mutants could not be mapped, however sequence analysis demonstrated that neither of these mutants contained mutations in previously identified IBT resistance or dependence genes. Phenotypic and biochemical analysis of the mutants suggests that they possess defects in transcription elongation that compensate for the elongation enhancing effects of IBT. The results implicate the largest subunit of the RNA polymerase (rpo147) in the control of elongation, and suggest that there exist additional gene products which mediate intermediate and late transcription elongation in vaccinia virus.


Assuntos
RNA Polimerases Dirigidas por DNA/genética , Regulação Viral da Expressão Gênica , Vaccinia virus/genética , Proteínas Virais/genética , Animais , Domínio Catalítico/genética , Linhagem Celular , RNA Polimerases Dirigidas por DNA/química , Farmacorresistência Viral , Perfilação da Expressão Gênica , Humanos , Isatina/análogos & derivados , Isatina/farmacologia , Modelos Químicos , Mutação , Fatores de Alongamento de Peptídeos , Fenótipo , RNA Mensageiro/metabolismo , RNA Viral/metabolismo , Fatores de Elongação da Transcrição/genética , Vaccinia virus/efeitos dos fármacos , Proteínas Virais/química
17.
J Biol Chem ; 279(43): 44858-71, 2004 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-15294890

RESUMO

The vaccinia virus RNA polymerase is a multi-subunit enzyme that contains eight subunits in the postreplicative form. A prior study of a virus called IBT(r90), which contains a mutation in the A24 gene encoding the RPO132 subunit of the RNA polymerase, demonstrated that the mutation results in resistance to the anti-poxvirus drug isatin-beta-thiosemicarbazone (IBT). In this study, we utilized an in vitro transcription elongation assay to determine the effect of this mutation on transcription elongation. Both wild type and IBT(r90) polymerase complexes were studied with regard to their ability to pause during elongation, their stability in a paused state, their ability to release transcripts, and their elongation rate. We have determined that the IBT(r90) complex is specifically defective in elongation compared with the WT complex, pausing longer and more frequently than the WT complex. We have built a homology model of the RPO132 subunit with the yeast pol II rpb2 subunit to propose a structural mechanism for this elongation defect.


Assuntos
RNA Polimerases Dirigidas por DNA/genética , Farmacorresistência Viral , Isatina/análogos & derivados , Isatina/farmacologia , RNA Viral , Sarcosina/análogos & derivados , Transcrição Gênica , Vaccinia virus/metabolismo , Biotinilação , Linhagem Celular Tumoral , Citoplasma/metabolismo , DNA/metabolismo , RNA Polimerases Dirigidas por DNA/metabolismo , Relação Dose-Resposta a Droga , Células HeLa , Humanos , Modelos Genéticos , Modelos Moleculares , Mutação , Plasmídeos/metabolismo , Reação em Cadeia da Polimerase , Sais/farmacologia , Sarcosina/farmacologia , Software , Fatores de Tempo , Uridina Trifosfato/química
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