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2.
Int J Obes (Lond) ; 34(2): 302-8, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19901952

RESUMO

BACKGROUND: Although several studies have shown a positive association between evidence of anti-adenovirus 36 (Ad-36) antibodies (Ad-36 exposure) and (1) obesity and (2) serum cholesterol in animals, there is limited research demonstrating this association in humans. There is also limited research on transmission, presentation and demographics of Ad-36 infection. DESIGN: (1) Body mass (body mass index (BMI)), (2) fasting serum cholesterol and triglyceride levels and (3) demographic characteristics were compared between Ad-36 seropositive and seronegative groups. The majority of subjects were matched as cases versus controls on a number of demographic variables. SUBJECTS: A total of 150 obese and 150 lean active-duty military personnel were studied. MEASUREMENTS: Subjects completed a questionnaire regarding demographic and behavioral characteristics. Subject serum samples were tested by serum neutralization assay for the presence of anti-Ad-36 antibodies. RESULTS: In all, 34% of obese and 39% of lean subjects had Ad-36 exposure, an insignificant difference. Serum cholesterol and triglyceride levels were significantly higher among the obese subjects than among the lean, but there were no associations between serum cholesterol and triglyceride levels and Ad-36 exposure. Positive associations were found between Ad-36 exposure and age, race and gender. CONCLUSION: The study stands in contrast to previous work that has shown a positive relationship between Ad-36 exposure and (1) obesity, and (2) levels of serum cholesterol and triglycerides. In this study there was no association in either case. Unanticipated relationships between Ad-36 exposure and age, race and gender were found, and this is the first time that such a link between Ad-36 exposure and demographics has been found.


Assuntos
Infecções por Adenovirus Humanos/sangue , Anticorpos Antivirais/sangue , Colesterol/sangue , Militares , Obesidade/sangue , Triglicerídeos/sangue , Infecções por Adenovirus Humanos/etnologia , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/imunologia , Adolescente , Adulto , Índice de Massa Corporal , Estudos de Casos e Controles , Jejum/sangue , Feminino , Humanos , Masculino , Obesidade/virologia , Inquéritos e Questionários , Estados Unidos/epidemiologia , Adulto Jovem
3.
Br J Health Psychol ; 25(3): 405-427, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32304286

RESUMO

Objectives Health literacy encompasses a broad skill set linked to patients' self-management ability and the complexity of their health care environments. Self-management in the context of multimorbidity is particularly challenging, placing patients at risk of poor clinical outcomes. This study aimed to explore the prognostic associations between health literacy domains, depression, and 12-month health care utilization and mortality in patients with diabetes and end-stage renal disease (DM-ESRD). Design Observational study. Methods Patients with DM-ESRD undergoing haemodialysis were recruited. Information on all-cause hospitalization/admission and mortality of participants was recorded. Negative binomial and Cox regressions were used to model risk factors for hospitalization and mortality. Results A total 221 participants [median age: 59 years, 61.6% men, 54.8% Chinese] were recruited. Differences in health literacy were found as a function of age, ethnicity, relationship status, and education. After adjusting for demographic and clinical factors, the HLQ domain Actively Managing My Health remained independently associated with lower rates of hospitalization (incidence rate ratio (IRR) = 0.674, 95% CI [0.490, 0.925], p = .02) and mortality (hazard ratio = 0.382, 95% CI [0.160, 0.848], p = .02). Cumulative hospitalization days were associated with employment status (IRR = 2.242, 95% CI [1.223, 4.113], p = .009), albumin (IRR = 0.918, 95% CI [0.854, 0.988], p = .02), HbA1c (IRR = 1.183, 95% CI [1.028, 1.360], p = .02), comorbidity burden (IRR = 1.137, 95% CI [1.003, 1.289], p = .04), and depression (IRR = 1.059, 95% CI [1.003, 1.118], p = .04) but no health literacy domains. Conclusions Health literacy skills related to Actively Managing My Health predict hospitalization and mortality independently of other risk factors. The HLQ provides an assessement of novel health literacy parameters which offer new insights into patients' status and behaviours and may strengthen interventions to improve clinical services, and patient outcomes in DM-ESRD. Statement of contribution What is already known on this subject? Patients with diabetes (DM) comprise the fastest growing segment of patients with end-stage renal disease (ESRD). Health literacy (HL) is pivotal for managing the complex treatment guidelines for DM-ESRD. Most prior work on HL focused on functional HL and shown significant associations with mortality and hospitalization. Limited research has investigated wider HL skills in relation to clinical outcomes. What does this study add? Supporting patients in Actively Managing my health liteacy skills is critical in decreasing probability of hospitalization and morbidity. The presence of symptoms of depression is associated with longer hospitalization period.


Assuntos
Diabetes Mellitus , Letramento em Saúde , Falência Renal Crônica , Complicações do Diabetes , Feminino , Humanos , Falência Renal Crônica/complicações , Masculino , Pessoa de Meia-Idade , Aceitação pelo Paciente de Cuidados de Saúde , Pacientes , Estudos Prospectivos
4.
Science ; 155(3759): 189-91, 1967 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-17738221

RESUMO

A piston core taken off of the coast of Oregon in 358 meters of water contained an indurated calcareous layer composed partly of dolomite with a composition Ca(58.7)Mg(41.3). Dolomites of this chemical composition are typical of the supratidal environment. However, the dolomite has isotopic composition delta0(18) = 5.8 per mille, deltaC(13) = 35.1 per mille relative to the Chicago PDB-I standard. The unusual carbon isotope ratio is similar to that of calcites produced as a byproduct of bacterial breakdown of hydrocarbons.

5.
AIDS ; 15(15): F41-7, 2001 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-11600844

RESUMO

OBJECTIVE: To describe the genetic diversity of HIV-1 in South America by full genome sequencing and analysis. METHODS: Purified peripheral blood mononuclear cell DNA from HIV-infected individuals in Argentina, Uruguay and Bolivia was used to amplify full HIV-1 genomes. These were sequenced using the ABI 3100 automated sequencer and phylogenetically analysed. RESULTS: Twenty-one HIV-1 strains from three South American countries, 17 of which were pre-screened by envelope heteroduplex mobility assay (HMA), were studied. Ten out of 10 HMA subtype F and four out of seven HMA subtype B strains were actually BF recombinants upon full genome analysis. Two BF recombinants from Argentina and two from Uruguay had the same structure, representing a new circulating recombinant form termed CRF12_BF(ARMA159). Twelve other BF recombinants had structures related to CRF12 but with additional segments of subtype B; each was unique. BF recombinants were temporally and geographically widespread, found as early as 1986-1987 in vertically infected Argentinian children and in Argentina, Uruguay, and Bolivia.


Assuntos
Infecções por HIV/epidemiologia , HIV-1/classificação , HIV-1/genética , Recombinação Genética , Adulto , Feminino , Infecções por HIV/virologia , Análise Heteroduplex , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA , América do Sul/epidemiologia
6.
AIDS ; 14(12): 1785-91, 2000 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-10985316

RESUMO

OBJECTIVE: Genotype determination and risk group analysis of HIV-1 infected individuals in selected regions of South America. DESIGN: Cross-sectional convenience sampling of HIV-1-positive individuals in Peru, Ecuador, Uruguay and Paraguay from March, 1994 through September, 1998. METHODS: HIV-1-positive subjects were identified through the national AIDS surveillance program in each country. A standardized questionnaire was used to obtain demographic, clinical and risk factor data on each study subject. Viral DNA was extracted from participants' peripheral blood mononuclear cells either directly or after co-cultivation. A nested PCR was used to obtain selected fragments of the envelope genes for genotyping by the heteroduplex mobility assay (HMA). A 600 bp sequence encompassing the V3 loop was sequenced from a selection of 23 of these samples for phylogenetic analysis and confirmation of HMA genotype. RESULTS: Among the 257 successfully genotyped HIV-1-positive samples, genotype B was found in 98.3% (228/232) of those obtained from subjects in Peru, Ecuador, and Paraguay. In contrast, 56% (14/25) of the samples from Uruguay were genotype F, and the remainder were genotype B. Genotype F was detected for the first time in Peru (2/224) and Paraguay (1/4), and genotype A for the first time in Peru (1/224). Phylogenetic analysis confirmed the genotype identified by HMA in the 23 samples sequenced. There was no detectable genetic clustering of HIV-1 within the different high-risk groups or geographic locations. CONCLUSIONS: These findings verify and extend the presence of several different HIV-1 genotypes in South America.


Assuntos
Variação Genética , Infecções por HIV/virologia , HIV-1/genética , Sequência de Aminoácidos , Sequência de Bases , Estudos Transversais , DNA Viral/química , Feminino , Genótipo , Proteína gp120 do Envelope de HIV/química , Proteína gp120 do Envelope de HIV/genética , Infecções por HIV/epidemiologia , HIV-1/classificação , HIV-1/imunologia , Análise Heteroduplex , Humanos , Masculino , Dados de Sequência Molecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/genética , Filogenia , Reação em Cadeia da Polimerase , Fatores de Risco , Comportamento Sexual , América do Sul/epidemiologia , Inquéritos e Questionários
7.
Am J Med Genet ; 104(4): 267-76, 2001 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-11754058

RESUMO

The Cornelia de Lange syndrome (CdLS) is an autosomal dominant multisystem disorder characterized by somatic and cognitive retardation, characteristic facial features, limb abnormalities, hearing loss, and other organ system involvement. The vast majority of cases (99%) are sporadic, with rare familial occurrences having been reported. Most individuals with CdLS do not reproduce as a result of the severity of the disorder. Maternal transmission has been well documented, as have several cases of multiple-affected children being born to apparently unaffected parents. Paternal transmission has rarely been reported. A case is reported here of a father with classic features of CdLS with a similarly affected daughter. A review of the reported familial cases of CdLS is summarized.


Assuntos
Síndrome de Cornélia de Lange/genética , Genes Dominantes/genética , Adulto , Pré-Escolar , Síndrome de Cornélia de Lange/patologia , Saúde da Família , Feminino , Humanos , Masculino , Linhagem
8.
Am J Trop Med Hyg ; 64(1-2): 93-7, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11425170

RESUMO

This report describes Trocara virus, a newly recognized member of the genus Alphavirus, that has been isolated from Aedes serratus mosquitoes collected at two widely separated sites in the Amazon Basin. Biological, antigenic and genetic characteristics of the new virus are given. Results of these studies indicate that Trocara virus is the first member of a newly discovered antigenic complex within the family Togaviridae genus Alphavirus. The public health and veterinary importance of Trocara virus is still unknown.


Assuntos
Aedes/virologia , Alphavirus/genética , Alphavirus/isolamento & purificação , Alphavirus/ultraestrutura , Animais , Brasil , Testes de Fixação de Complemento , Cricetinae , Primers do DNA , Testes de Hemaglutinação , Camundongos , Microscopia Eletrônica , Peru , Reação em Cadeia da Polimerase , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa
9.
Am J Trop Med Hyg ; 63(3-4): 209-13, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11388517

RESUMO

An outbreak of delta hepatitis occurred during 1998 among the Waorani of the Amazon basin of Ecuador. Among 58 people identified with jaundice, 79% lived in four of 22 Waorani communities. Serum hepatitis B surface antigen (HBsAg) was found in the sera of 54% of the jaundiced persons, and 14% of asymptomatic persons. Ninety-five percent of 105 asymptomatic Waorani had hepatitis B core (HBc) IgG antibody, versus 98% of 51 with jaundice. These data confirm that hepatitis B virus (HBV) infection is highly endemic among the Waorani. Sixteen of 23 (70%) HBsAg carriers identified at the onset of the epidemic had serologic markers for hepatitis D virus (HDV) infection. All 16 were jaundiced, where as only two of seven (29%) with negative HDV serology were jaundiced (P = .0006). The delta cases clustered in families, 69% were children and most involved superinfection of people chronically infected with HBV. The data suggest that HDV spread rapidly by a horizontal mode of transmission other than by the sexual route.


Assuntos
Surtos de Doenças , Hepatite D/epidemiologia , Vírus Delta da Hepatite/imunologia , Falência Hepática/epidemiologia , Adolescente , Adulto , Criança , Pré-Escolar , Equador/epidemiologia , Etnicidade/estatística & dados numéricos , Feminino , Anticorpos Anti-Hepatite/sangue , Antígenos de Superfície da Hepatite B/sangue , Hepatite D/complicações , Vírus Delta da Hepatite/genética , Humanos , Lactente , Falência Hepática/etiologia , Masculino , Pessoa de Meia-Idade , RNA Viral/sangue
10.
Mil Med ; 165(7 Suppl 2): 52-6, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10920641

RESUMO

Military global influenza surveillance began in 1976 as an Air Force program. In 1997, the Department of Defense (DoD) Global Emerging Infections Surveillance and Response System expanded the program to include all services. Also included were local residents in areas where DoD overseas research activities operated. This new, worldwide DoD surveillance infrastructure provides valuable information and can respond quickly to outbreaks. This was demonstrated during the current influenza season when a suspected outbreak was reported in Panama. In less than 3 weeks, specimens were collected, transported, and cultured, and isolates were subtyped and sent to the Centers for Disease Control and Prevention for further studies. This influenza surveillance initiative combines viral isolation, antigenic characterization, and molecular sequencing with clinical and public health management of information. The information obtained is shared with the Centers for Disease Control and Prevention and the World Health Organization and has contributed to important decisions in influenza vaccine composition.


Assuntos
Influenza Humana/epidemiologia , Medicina Militar/organização & administração , Vigilância da População , Saúde Global , Órgãos Governamentais , Humanos , Vacinas contra Influenza , Vigilância da População/métodos , Estados Unidos
11.
Undersea Hyperb Med ; 26(4): 261-5, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-10642074

RESUMO

Divers may be exposed to intense noise underwater. Two cases of neurologic disturbances during experimental exposures to 15 min of continuous underwater sound are described. Sound exposure in the first case consisted of a warble tone with center frequency of 240 Hz and a sound pressure level of 160 dB re 1 microPa. Symptoms during exposure consisted of somnolence, lightheadedness, and an inability to concentrate. No apparent effect on hearing was noted. In the second case, a center frequency of 1,000 Hz at 181 dB was used. Lightheadedness, inability to concentrate, agitation, and head vibrations were noted during the exposure. The diver also exhibited a temporary auditory threshold shift of 19.2 dB. In both cases, overt symptoms resolved within 30 min after exposure, but both divers reported recurrent symptoms days to weeks after the exposures. Medical histories and examinations, assessment of dive profiles, and breathing gas analysis failed to support a source other than the sound exposures to account for the symptoms observed. Potential mechanisms for the described symptoms are discussed.


Assuntos
Doenças do Sistema Nervoso Central/etiologia , Mergulho/efeitos adversos , Ruído/efeitos adversos , Adulto , Doenças do Sistema Nervoso Central/fisiopatologia , Transtornos da Audição/etiologia , Transtornos da Audição/fisiopatologia , Síndrome Neurológica de Alta Pressão/etiologia , Síndrome Neurológica de Alta Pressão/fisiopatologia , Humanos , Masculino , Exame Neurológico
12.
Artigo em Inglês | MEDLINE | ID: mdl-24342130

RESUMO

The effects of an oral fish oil treatment regimen on sensorimotor, blood-brain barrier, and biochemical outcomes of traumatic brain injury (TBI) were investigated in a juvenile rat model. Seventeen-day old Long-Evans rats were given a 15mL/kg fish oil (2.01g/kg EPA, 1.34g/kg DHA) or soybean oil dose via oral gavage 30min prior to being subjected to a controlled cortical impact injury or sham surgery, followed by daily doses for seven days. Fish oil treatment resulted in less severe hindlimb deficits after TBI as assessed with the beam walk test, decreased cerebral IgG infiltration, and decreased TBI-induced expression of the Mmp9 gene one day after injury. These results indicate that fish oil improved functional outcome after TBI resulting, at least in part from decreased disruption of the blood-brain barrier through a mechanism that includes attenuation of TBI-induced expression of Mmp9.


Assuntos
Barreira Hematoencefálica/enzimologia , Lesões Encefálicas/enzimologia , Óleos de Peixe/farmacologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Metaloproteinase 9 da Matriz/biossíntese , Caminhada , Animais , Barreira Hematoencefálica/patologia , Lesões Encefálicas/tratamento farmacológico , Lesões Encefálicas/patologia , Ratos , Ratos Long-Evans
13.
Influenza Other Respir Viruses ; 6(5): 358-66, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22212638

RESUMO

BACKGROUND: Since 2004, the Naval Health Research Center, with San Diego and Imperial counties, has collaborated with the US Centers for Disease Control and Prevention to conduct respiratory disease surveillance in the US-Mexico border region. In 2007, the Secretariat of Health, Mexico and the Institute of Public Health of Baja California joined the collaboration. OBJECTIVES: The identification of circulating respiratory pathogens in respiratory specimens from patients with influenza-like illness (ILI). METHODS: Demographic, symptom information and respiratory swabs were collected from enrollees who met the case definition for ILI. Specimens underwent PCR testing and culture in virology and bacteriology. RESULTS: From 2004 through 2009, 1855 persons were sampled. Overall, 36% of the participants had a pathogen identified. The most frequent pathogen was influenza (25%), with those aged 6-15 years the most frequently affected. In April 2009, a young female participant from Imperial County, California, was among the first documented cases of 2009 H1N1. Additional pathogens included influenza B, adenovirus, parainfluenza virus, respiratory syncytial virus, enterovirus, herpes simplex virus, Streptococcus pneumoniae, and Streptococcus pyogenes. CONCLUSIONS: The US-Mexico border is one of the busiest in the world, with a large number of daily crossings. Due to its traffic, this area is an ideal location for surveillance sites. We identified a pathogen in 36% of the specimens tested, with influenza A the most common pathogen. A number of other viral and bacterial respiratory pathogens were identified. An understanding of the incidence of respiratory pathogens in border populations is useful for development of regional vaccination and disease prevention responses.


Assuntos
Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Viroses/epidemiologia , Viroses/virologia , Vírus/classificação , Vírus/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Bactérias/classificação , Bactérias/isolamento & purificação , Infecções Bacterianas/epidemiologia , Infecções Bacterianas/microbiologia , Técnicas Bacteriológicas , California/epidemiologia , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Infecções Respiratórias/microbiologia , Cultura de Vírus , Adulto Jovem
14.
J Clin Microbiol ; 43(4): 1768-75, 2005 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15814997

RESUMO

Viral culture isolation has been widely accepted as the "gold standard" for laboratory confirmation of viral infection; however, it requires ultralow temperature specimen storage. Storage of specimens in ethanol at room temperature could expand our ability to conduct active surveillance and retrospective screenings of viruses with rapid and inexpensive real-time PCR tests, including isolates from remote regions where freezing specimens for culture is not feasible. Molecular methods allow for rapid identification of viral pathogens without the need to maintain viability. We hypothesized that ethanol, while inactivating viruses, can preserve DNA and RNA for PCR-based methods. To evaluate the use of ethanol-stored specimens for augmenting surveillance for detection of influenza viruses A and B and adenoviruses (AdV), paired nasal swab specimens were collected from 384 recruits with febrile respiratory illness at Fort Jackson, S.C., in a 2-year study. One swab was stored at ambient temperature in 100% ethanol for up to 6 months, and the other swab was stored at -70 degrees C in viral medium. For viral detection, frozen specimens were cultured for a variety of respiratory viruses, and ethanol-fixed specimens were tested with TaqMan (TM) probe and LightCycler SYBR green (SG) melting curve assays with at least two different PCR targets for each virus. The sensitivities of the TM and SG assays on specimens stored in ethanol for 1 month were 75% and 58% for influenza A, 89% and 67% for influenza B, and 93 to 98% and 57% for AdV, respectively. Lower specificities of the real-time assays corresponded to the increased detection of PCR-positive but culture-negative specimens. Influenza virus RNA was detected as well or better after 6 months of storage in ethanol.


Assuntos
Infecções por Adenovirus Humanos/diagnóstico , Etanol , Influenza Humana/diagnóstico , Nariz/virologia , Reação em Cadeia da Polimerase/métodos , Manejo de Espécimes/métodos , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/genética , Adenovírus Humanos/isolamento & purificação , DNA Viral/análise , Humanos , Vírus da Influenza A/genética , Vírus da Influenza A/isolamento & purificação , Vírus da Influenza B/genética , Vírus da Influenza B/isolamento & purificação , Influenza Humana/virologia , Militares , Vigilância da População , Valor Preditivo dos Testes , RNA Viral/análise , Sensibilidade e Especificidade
15.
J Vet Pharmacol Ther ; 9(1): 94-100, 1986 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-3701920

RESUMO

Rapid solvent extraction combined with a radioimmunoassay using a monoclonal antibody raised against a derivative of zeranol has been used to measure the residues of the anabolic agent zeranol in the edible tissues (muscle, liver, kidney and fat) of cattle treated with Ralgro. Calibration curves, both with and without, tissue extracts exhibit good parallelism. Regression analysis for the extraction of zeranol from tissues dosed with standard amounts of zeranol have correlation coefficients of 0.979, 0.991, 0.986 and 0.985 for muscle, liver, kidney and fat, respectively. The limits of decision defined as the mean value + 3 SD for the concentrations apparently observed (noise) in tissues from animals not treated with Ralgro were 278, 121, 373 and 110 ng/kg for muscle, fat, liver and kidney, respectively. In the tissues of 4 cows implanted with Ralgro (36 mg), and sampled 70 days after implanting, the highest concentration of zeranol in each tissue was 232 ng/kg (muscle), 391 ng/kg (liver), 287 ng/kg (kidney) and 293 ng/kg (fat), and residues were detected in all samples of fat (4), 3 kidney samples and 1 liver sample.


Assuntos
Análise de Alimentos , Carne , Resorcinóis/análise , Zeranol/análise , Tecido Adiposo/análise , Animais , Bovinos , Diafragma/análise , Implantes de Medicamento , Feminino , Rim/análise , Fígado/análise , Músculos/análise , Radioimunoensaio , Zeranol/administração & dosagem
16.
J Vet Pharmacol Ther ; 6(3): 173-9, 1983 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-6632074

RESUMO

A radioimmunoassay for zeranol has been validated and used to measure the concentration of zeranol in the urine of sheep and cattle treated with zeranol (Ralgro). The assay uses an antibody raised against zeranol-16-carboxy-propyl ether conjugated to human serum albumin. In sheep and cattle urine the limits of detection were approximately 2 ng/ml and 2.5 ng/ml, respectively. In two trials 13 sheep were implanted with 12 mg zeranol at the base of the ear. The mean maximum concentrations of zeranol observed in urine were 45 ng/ml (Trial I) on day 35 and 90 ng/ml (Trial II) on day 56, and had declined to 26 ng/ml 42 days after implantation (Trial I) and 11.7 ng/ml 70 days after implantation (Trial II). In four cattle implanted with 36 mg zeranol the concentrations of zeranol in urine reached a mean maximum concentration of 13.5 ng/ml 22 days after implantation and had declined to 2.9 ng/ml 69 days after implantation.


Assuntos
Bovinos/urina , Resorcinóis/urina , Ovinos/urina , Zeranol/urina , Animais , Implantes de Medicamento , Radioimunoensaio/veterinária , Fatores de Tempo , Zeranol/administração & dosagem
17.
J Biol Chem ; 260(25): 13763-70, 1985 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-2997181

RESUMO

The native structures of protein phosphatases have not been clearly established. Several tissues contain high molecular weight enzymes which are converted to active species of Mr approximately 35,000 by denaturing treatments or partial proteolysis. We have used a monoclonal antibody directed against purified bovine cardiac Mr = 38,000 protein phosphatase to determine whether this species is the native catalytic subunit or a proteolytic product of a larger polypeptide. Monoclonal antibody was obtained from a cloned hybrid cell line produced by the fusion of Sp2 myeloma cells with spleen cells from a mouse immunized with phosphatase coupled to hemocyanin. This antibody was specific for the Mr = 38,000 phosphatase as determined by immunoblot analysis of purified enzyme or cardiac tissue extracts after native or sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A single immunoreactive protein of Mr = 38,000 was present in cardiac tissue extracts including extracts prepared from freeze-clamped rat heart rapidly denatured in hot sodium dodecyl sulfate buffer. Precipitation of cardiac extract with 80% ethanol did not alter the Mr of the phosphatase nor did it liberate new immunoreactive material not observed in the extract. Ethanol precipitation caused the dissociation of both phosphatase activity and immunoreactivity from a high Mr form to a form of Mr between 30,000 and 40,000. An immunoreactive protein of Mr = 38,000 was identified in several bovine and rat tissues as well as tissues from rabbits, mice and chickens and human HT-29 cells. From these data we conclude that the Mr = 38,000 cardiac phosphatase is a native catalytic subunit of higher molecular complexes which are dissociated by ethanol precipitation. A very similar, or identical, protein is present in several tissues and species suggesting that this catalytic subunit is a ubiquitous enzyme important in many dephosphorylation reactions.


Assuntos
Anticorpos Monoclonais/imunologia , Miocárdio/enzimologia , Fosfoproteínas Fosfatases/análise , Animais , Bovinos , Precipitação Química , Galinhas , Etanol/farmacologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Peso Molecular , Fosfoproteínas Fosfatases/imunologia , Ratos
18.
J Biol Chem ; 262(13): 6257-65, 1987 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-3032961

RESUMO

Two forms of protein phosphatase which dephosphorylate cardiac myosin or myosin light chains and the inhibitory subunit of cardiac troponin were purified from bovine cardiac muscle. The enzymes were composed of subunits of Mr = 63,000, 55,000, and 38,000 in a 1:1:1 molar ratio (PT-1) or Mr = 63,000 and 38,000 in a 1:1 molar ratio (PT-2). Native gel electrophoresis and sucrose gradient sedimentation indicated that activity toward all three substrates was due to a single enzyme species. A monoclonal antibody and polyclonal antiserum directed against an Mr = 38,000 protein phosphatase from this tissue specifically reacted with the Mr = 38,000 subunit of PT-1 and PT-2. The specificity of antibodies for the Mr = 38,000 subunit indicated that it was distinct from the other subunits. The Mr = 63,000 subunits of PT-1 and PT-2 were identical based on mobility on sodium dodecyl sulfate gels and one-dimensional peptide maps. Specificity of antiserum against the Mr = 55,000 subunit of PT-1 showed that this subunit was a distinct protein and not derived from the Mr = 63,000 subunit by proteolysis. PT-2 but not PT-1 could interact with antiserum against the Mr = 38,000 catalytic subunit in competitive immunoassays indicating that the presence of the Mr = 55,000 subunit may alter or mask antigenic site(s). Analysis of the enzymatic properties of PT-1 and PT-2 showed that PT-2 had higher activity with myosin, myosin light chains, and phosphorylase while PT-1 had higher activity with troponin. The results indicate that the presence of the Mr = 55,000 subunit may alter the enzymatic properties of the catalytic subunit.


Assuntos
Miocárdio/enzimologia , Fosfoproteínas Fosfatases/isolamento & purificação , Animais , Anticorpos Monoclonais , Sítios de Ligação , Bovinos , Eletroforese em Gel de Poliacrilamida , Técnicas de Imunoadsorção , Cinética , Substâncias Macromoleculares , Peso Molecular , Contração Miocárdica , Fosfatase de Miosina-de-Cadeia-Leve , Fosfoproteínas Fosfatases/imunologia , Especificidade por Substrato
19.
J Vet Pharmacol Ther ; 9(4): 353-8, 1986 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-3806777

RESUMO

Two trials were conducted on steers implanted with zeranol (Ralgro) to determine the edible tissue residues and the secretion pattern in faeces, urine and bile of zeranol residues throughout and beyond the recommended withdrawal period (70 days) for this drug. In the first trial there was considerable variation in the zeranol residue concentration in all edible tissues, the highest concentrations found in the liver being significantly above the control values (P less than 0.05). In the other tissues, only fat sampled 14 days after implanting was significantly above the control value (P less than 0.05). The zeranol concentration in bile samples obtained at slaughter [70 days (18), 90 days (5) and 120 days (2)] were all higher than the apparent concentration in the bile of untreated steers. The mean concentration of zeranol in the faeces and urine varied from day to day and between animals sampled on the same day following implantation. The highest mean concentrations were observed during the first 40 days following implanting, declining steadily to approach the control values 70 days after implantation. The second trial using steers prepared with bile duct re-entrant cannulae resulted in a similar pattern of zeranol excretion in bile, faeces and urine. The highest concentrations of zeranol were observed in bile and ranged from 24 to 34 micrograms/l; there was considerable variation between animals and within animals sampled on successive days. Although the concentration declined steadily, zeranol was still readily detectable 120 days after implanting.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Bile/análise , Músculos/análise , Resorcinóis/análise , Resorcinóis/metabolismo , Zeranol/análise , Zeranol/metabolismo , Tecido Adiposo/análise , Animais , Bovinos , Implantes de Medicamento , Fezes/análise , Rim/análise , Cinética , Fígado/análise , Masculino , Orquiectomia , Distribuição Tecidual , Zeranol/urina
20.
J Clin Microbiol ; 38(4): 1527-35, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10747138

RESUMO

Specific and sensitive reverse transcription-PCR (RT-PCR) assays were developed for the detection of eastern, western, and Venezuelan equine encephalitis viruses (EEE, WEE, and VEE, respectively). Tests for specificity included all known alphavirus species. The EEE-specific RT-PCR amplified a 464-bp region of the E2 gene exclusively from 10 different EEE strains from South and North America with a sensitivity of about 3,000 RNA molecules. In a subsequent nested PCR, the specificity was confirmed by the amplification of a 262-bp fragment, increasing the sensitivity of this assay to approximately 30 RNA molecules. The RT-PCR for WEE amplified a fragment of 354 bp from as few as 2,000 RNA molecules. Babanki virus, as well as Mucambo and Pixuna viruses (VEE subtypes IIIA and IV), were also amplified. However, the latter viruses showed slightly smaller fragments of about 290 and 310 bp, respectively. A subsequent seminested PCR amplified a 195-bp fragment only from the 10 tested strains of WEE from North and South America, rendering this assay virus specific and increasing its sensitivity to approximately 20 RNA molecules. Because the 12 VEE subtypes showed too much divergence in their 26S RNA nucleotide sequences to detect all of them by the use of nondegenerate primers, this assay was confined to the medically important and closely related VEE subtypes IAB, IC, ID, IE, and II. The RT-PCR-seminested PCR combination specifically amplified 342- and 194-bp fragments of the region covering the 6K gene in VEE. The sensitivity was 20 RNA molecules for subtype IAB virus and 70 RNA molecules for subtype IE virus. In addition to the subtypes mentioned above, three of the enzootic VEE (subtypes IIIB, IIIC, and IV) showed the specific amplicon in the seminested PCR. The practicability of the latter assay was tested with human sera gathered as part of the febrile illness surveillance in the Amazon River Basin of Peru near the city of Iquitos. All of the nine tested VEE-positive sera showed the expected 194-bp amplicon of the VEE-specific RT-PCR-seminested PCR.


Assuntos
Alphavirus/classificação , Alphavirus/isolamento & purificação , Encefalomielite Equina/diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Alphavirus/genética , Animais , Vírus da Encefalite Equina do Leste/classificação , Vírus da Encefalite Equina do Leste/genética , Vírus da Encefalite Equina do Leste/isolamento & purificação , Vírus da Encefalite Equina Venezuelana/classificação , Vírus da Encefalite Equina Venezuelana/genética , Vírus da Encefalite Equina Venezuelana/isolamento & purificação , Vírus da Encefalite Equina do Oeste/classificação , Vírus da Encefalite Equina do Oeste/genética , Vírus da Encefalite Equina do Oeste/isolamento & purificação , Encefalomielite Equina/veterinária , Encefalomielite Equina/virologia , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/virologia , Cavalos , Humanos , Camundongos , Reação em Cadeia da Polimerase , RNA Viral/análise , Especificidade da Espécie
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