RESUMEN
BACKGROUND: Most published studies on seafarer health have focused on patterns of mortality, injury and communicable diseases. Little information is available regarding lifestyle-related cardio-metabolic disease in maritime populations. AIMS: To describe health characteristics of a population of US inland waterway merchant marine captains and pilots. METHODS: A cross-sectional study of the health characteristics of mariners required to complete the United States Coast Guard physical assessment at a regional medical centre from 2003-10. Variables collected included self-reported smoking status, body mass index, fasting lipids, glucose and triglyceride levels, blood pressure and treadmill time and maximal oxygen uptake as measured using the Bruce Protocol. Major medical conditions related to lifestyle and risk for metabolic syndrome were also assessed. RESULTS: There were 388 participants. The study population had high prevalence of obesity (61%), smoking (41%), high triglycerides (42%), low HDL cholesterol (47%), high blood pressure (42%), high fasting glucose (22%) and three or more features of the metabolic syndrome (39%). CONCLUSIONS: This population exhibited a high prevalence of chronic disease risk factors and could potentially benefit from health promotion programmes aimed at improving health and fitness.
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Enfermedades Cardiovasculares/epidemiología , Enfermedades Metabólicas/epidemiología , Medicina Naval/estadística & datos numéricos , Enfermedades Profesionales/epidemiología , Adulto , Anciano , Glucemia/metabolismo , Presión Sanguínea/fisiología , Índice de Masa Corporal , Enfermedades Cardiovasculares/sangre , Enfermedades Cardiovasculares/fisiopatología , Estudios Transversales , Estado de Salud , Humanos , Hiperlipidemias/epidemiología , Hipertensión/epidemiología , Estilo de Vida , Metabolismo de los Lípidos/fisiología , Masculino , Enfermedades Metabólicas/sangre , Enfermedades Metabólicas/fisiopatología , Persona de Mediana Edad , Obesidad/epidemiología , Enfermedades Profesionales/sangre , Enfermedades Profesionales/fisiopatología , Prevalencia , Factores de Riesgo , Fumar/epidemiología , Estados Unidos/epidemiología , Evaluación de Capacidad de Trabajo , Adulto JovenRESUMEN
AIM: We examined the association of fasting plasma glucose (FPG), 2-h plasma glucose (2hPG) and HbA1c with retinopathy and microalbuminuria using both deciles of glycaemia and change point models, to validate current diagnostic criteria for diabetes and to identify therapeutic thresholds for glycaemic control. METHODS: The Australian Diabetes Obesity and Lifestyle study (AusDiab), conducted in 1999-2000, included adults aged > or =25 years from 42 randomly selected areas of Australia. Retinopathy and albuminuria were assessed in participants identified as having diabetes (based on self report and oral glucose tolerance test), impaired fasting glucose, impaired glucose tolerance and in a random sample with normal glucose tolerance. Data were available for 2,182 participants with retinal photographs and 2,389 with urinary albumin/creatinine results. RESULTS: The prevalence of retinopathy in the first 8 deciles of FPG and HbA1c and the first 9 deciles of 2hPG were 7.2, 6.6, and 6.3%, respectively and showed no variation with increasing glucose or HbA1c. Above these levels, the prevalence rose markedly to 18.6% in the top 2 deciles of FPG, 21.3% in the top 2 deciles of HbA1c and 10.9% in the top decile of 2hPG. The thresholds for increasing prevalence of retinopathy were 7.1 mmol/l for FPG, 6.1% for HbA1c and 13.1 mmol/l for 2hPG. The prevalence of microalbuminuria rose gradually across deciles of each glycaemic measure. Thresholds were less clear than for retinopathy, but were seen at a FPG of 7.2 mmol/l and HbA1c of 6.1%, with no evidence of a threshold effect for 2hPG. CONCLUSIONS: The prevalence of retinopathy rose dramatically in the highest deciles of each glycaemic measure, while for microalbuminuria the increase of prevalence was more gradual. The FPG values corresponded well with the WHO diagnostic cut-point for diabetes, however the 2hPG value did not. HbA1c thresholds were similar for both retinopathy and microalbuminuria and compared well to values shown in other studies. These results support current targets for FPG and HbA1c in preventing microvascular complications.
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Albuminuria/complicaciones , Diabetes Mellitus/diagnóstico , Retinopatía Diabética/complicaciones , Anciano , Anciano de 80 o más Años , Albuminuria/sangre , Australia , Glucemia/análisis , Estudios Transversales , Complicaciones de la Diabetes/sangre , Complicaciones de la Diabetes/diagnóstico , Diabetes Mellitus/sangre , Retinopatía Diabética/sangre , Femenino , Hemoglobina Glucada/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Factores de Riesgo , Sensibilidad y Especificidad , Organización Mundial de la SaludRESUMEN
We investigated the epidemiology of systemic lupus erythematosus (SLE) in the Marshfield Epidemiologic Study Area (MESA), a defined rural region where nearly all residents obtain their health care from a large clinic system. Computerized medical records were searched to identify MESA residents diagnosed with SLE from 1991 through 2001. Medical records were manually reviewed for all selected patients to identify cases of SLE using the 1982 revised American College of Rheumatology criteria. Patients with > or = 4 criteria were classified as definite SLE. Age- and gender-specific SLE incidence rates (1991-2001), the population prevalence rate of SLE on 31 December 2001 and survival rates were calculated. We identified 117 MESA residents with definite SLE. The average age-adjusted incidence of definite SLE was 5.1 per 100 000 per year (95% CI: 3.6, 6.6) and the age-adjusted population prevalence was 78.5 per 100 000 (95% CI: 59.0, 98.0). The mean age at diagnosis among the 44 incident cases was 51.7 years (range: 14-90 years). Positive anti-nuclear antibody (ANA), hematologic abnormalities, arthritis and renal disease were common at diagnosis. Five- and 10-year survival rates were 88% and 76%, respectively. Epidemiologic characteristics of SLE in this rural Caucasian population are generally similar to those reported by other US studies. One notable difference is a relatively high incidence of SLE in older adults.
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Lupus Eritematoso Sistémico/epidemiología , Población Rural/estadística & datos numéricos , Población Blanca/estadística & datos numéricos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Lupus Eritematoso Sistémico/diagnóstico , Masculino , Persona de Mediana Edad , Wisconsin/epidemiologíaRESUMEN
The fetal environment may be a contributing factor in the etiology of some adult diseases. This study examined whether birth weight, birth length and gestational age are associated with the subsequent development of systemic lupus erythematosus (SLE). The Marshfield Clinic Lupus Registry was searched to identify patients who were born at Saint Joseph's Hospital in Marshfield, Wisconsin, USA. Birth data on each case and five age-, sex-, and race-matched controls were recorded from medical and delivery room register records. Perinatal data were obtained for 23 cases and 115 controls. The unadjusted mean birth weight was similar for cases (3407 +/- 581 g) and controls (3422 +/- 514 g). Birth length was not different between groups. Birth weight adjusted for gestational age, analysed by conditional logistic regression, was not statistically significantly different between groups. We concluded that birth weight and length were similar among SLE cases and controls, suggesting that these perinatal characteristics are not associated with subsequent SLE.
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Peso al Nacer , Lupus Eritematoso Sistémico/etiología , Puntaje de Apgar , Estatura , Estudios de Casos y Controles , Femenino , Edad Gestacional , Humanos , Recién Nacido , Modelos Logísticos , Masculino , Factores de RiesgoRESUMEN
PURPOSE: To evaluate trends in health service delivery for cataract surgery at the Royal Victorian Eye and Ear Hospital, Melbourne, Australia. METHODS: A retrospective study was conducted of computerized hospital admissions for cataract surgery from July 1994 through June 1999. Main outcome measures included the annual number of cataract surgeries, waiting time, percentage of patients hospitalized overnight, length of overnight hospitalization, type of cataract surgery, surgery duration and total time in the operating theatre. RESULTS: Although the annual number of patients undergoing cataract surgery increased from 3395 to 4796 over the 5-year study interval, there was a 13-day reduction in mean waiting time for public patients, a 30% reduction in overnight hospitalizations and a half-day reduction in length of hospitalization for all patients. There was also an increasing trend in use of phacoemulsification cataract extraction. CONCLUSIONS: The health-care delivery for cataract surgery appears to have improved during the past 5 years at the Royal Victorian Eye and Ear Hospital.
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Extracción de Catarata/tendencias , Atención a la Salud/tendencias , Servicios de Salud/tendencias , Anciano , Femenino , Investigación sobre Servicios de Salud , Hospitalización/tendencias , Hospitales Especializados , Humanos , Masculino , Oftalmología , Estudios Retrospectivos , Factores de Tiempo , Victoria , Listas de EsperaRESUMEN
PURPOSE: To determine the efficacy and safety of topical autologous serum as a treatment of dry eye patients. METHODS: A 2-month, prospective, single-masked, placebo-controlled study was conducted in patients with bilateral severe dry eye. One eye was randomized to receive the patient's own serum as a tear substitute, and the fellow eye received unpreserved normal saline solution as a placebo. Subjective symptoms and clinical parameters of dry eye including conjunctival impression cytology were assessed at baseline and 1 week, 1 month, and 2 months after treatment. RESULTS: Twelve dry eye patients were enrolled. Both subjective symptoms (discomfort, foreign-body sensation, dryness, and photophobia), objective signs (fluorescein and rose bengal staining and conjunctival impression cytology) improved significantly in treated eyes compared with baseline. Control eyes also had improvement in symptoms, signs, and rose bengal staining compared with baseline. Neither Schirmer test results nor tear break-up time improved in either group. The means score of all parameters were improved in both groups, and the results of conjunctival impression cytology were better in treated eyes; however, these results are not significantly different. There were no serious adverse effects observed in this study. CONCLUSIONS: There was a trend toward improvement in symptoms and signs of dry eye including cytologic changes after application of autologous serum in severe dry eye patients. However, this trend was not statistically significant. A larger scale study is warranted.
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Sangre , Síndromes de Ojo Seco/terapia , Administración Tópica , Adulto , Anciano , Síndromes de Ojo Seco/fisiopatología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Seguridad , Método Simple Ciego , Resultado del TratamientoRESUMEN
The aim of this study was to identify changes in cartilage intermediate layer protein/nucleotide pyrophosphohydrolase (CILP/NTPPH) expression in articular cartilage during aging. Adult (3-4 years old) and young (7-10 days old) porcine articular hyaline cartilage and fibrocartilage were studied by Northern blot analysis, in situ hybridization, and immunohistochemistry using a complementary DNA (cDNA) probe encoding porcine CILP/NTPPH and antibody to a synthetic peptide corresponding to a CILP/NTPPH sequence. Northern blot analysis of chondrocytes showed lower expression of CILP/NTPPH messenger RNA (mRNA) in young cartilage than in adult cartilage. In adult cartilage, extracellular matrix from the surface to the middeep zone was immunoreactive for CILP/NTPPH, especially in the pericellular matrix surrounding the middeep zone chondrocytes. In young cartilage, chondrocytes were moderately immunoreactive for CILP/NTPPH throughout all zones except the calcified zone. The matrix of young cartilage was negative except in the superficial zone. In young cartilage, CILP/NTPPH mRNA expression was undetectable. In adult cartilage, chondrocytes showed strong mRNA expression for CILP/NTPPH throughout middeep zones. Protein and mRNA signals were not detectable below the tidemark. CILP/NTPPH secretion into matrix around chondrocytes increases with aging. In this extracellular site it may generate inorganic pyrophosphate and contribute to age-related calcium pyrophosphate dihydrate crystal deposition disease.
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Envejecimiento/metabolismo , Condrocitos/enzimología , Proteínas de la Matriz Extracelular/metabolismo , Pirofosfatasas/metabolismo , Animales , Northern Blotting/métodos , Cartílago Articular/citología , Cartílago Articular/enzimología , Proteínas de la Matriz Extracelular/genética , Expresión Génica , Hialina , Pirofosfatasas/genética , PorcinosRESUMEN
PURPOSE: To evaluate the safety and efficacy of laser in situ keratomileusis (LASIK) compared to photorefractive keratectomy (PRK) for the correction of low or moderate myopia (-0.50 to -6.00 D) at 6 months after surgery. METHODS: The study population comprised a non-randomized consecutive series of 622 eyes of 392 patients who were treated with the Nidek EC-5000 excimer laser. LASIK was performed using the ACS Chiron microkeratome on 314 eyes and surface PRK on 308 eyes. All patients were treated using a standard protocol, then assessed at 1, 3, and 6 months postoperatively. RESULTS: Forty-four percent of the LASIK group and 67% of the PRK group attended their 6-month examination. Eighty percent of patients (111 eyes) after LASIK and 65% (136 eyes) after PRK had an uncorrected visual acuity of 20/20 or better. Spherical equivalent refraction was within +/-0.50 D of intended refraction in 78% (109 eyes) for LASIK and 82% (170 eyes) for PRK. Loss of two more lines of best spectacle-corrected visual acuity at 6 months occurred in 1.4% (2 eyes) of the LASIK group and 1.0% (2 eyes) of the PRK group. CONCLUSION: At 1 month follow-up, the percentage of eyes that achieved 20/20 uncorrected visual acuity was greater in the LASIK group than in the PRK group. At 6 months, visual and refractive outcomes of LASIK and PRK were similar. Although flap related complications occurred only after LASIK, the overall risk of loss of best spectacle-corrected visual acuity was not significantly greater than for PRK.
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Córnea/cirugía , Queratomileusis por Láser In Situ/métodos , Miopía/cirugía , Queratectomía Fotorrefractiva/métodos , Adulto , Córnea/fisiopatología , Estudios de Seguimiento , Humanos , Láseres de Excímeros , Miopía/fisiopatología , Complicaciones Posoperatorias , Refracción Ocular , Reoperación , Resultado del Tratamiento , Agudeza VisualRESUMEN
PURPOSE: To compare the incidence of corneal perforation in eyes treated with topical fluoroquinolone or fortified antibiotics for microbial keratitis. METHODS: A retrospective medical record review of patients hospitalized for bacterial keratitis from January 1991 through November 1999. RESULTS: Two hundred seventy-seven cases of bacterial keratitis were identified. The incidence of corneal perforations was significantly higher in fluoroquinolone-treated eyes (18 out of 142, 12.7%) compared with eyes treated with fortified antibiotics (1 out of 135, 0.7%) (chi-square = 13.6, degrees of freedom (df) = 2, P <.001). Within the fluoroquinolone-treated group, all corneal perforations occurred in those eyes treated with ofloxacin (18 out of 125, 14.4%). Corneal perforations that occurred in the ofloxacin-treated eyes were not associated with any particular microbial isolate. CONCLUSION: Our data suggest an increased risk of corneal perforation after fluoroquinolone treatment for bacterial keratitis compared with treatment with fortified antibiotics. Further studies are warranted to verify this association and establish possible mechanisms by which topical fluoroquinolones may alter corneal collagen or keratocyte function.
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Antiinfecciosos/efectos adversos , Córnea/efectos de los fármacos , Enfermedades de la Córnea/inducido químicamente , Úlcera de la Córnea/tratamiento farmacológico , Infecciones Bacterianas del Ojo/tratamiento farmacológico , Ofloxacino/efectos adversos , Bacterias/aislamiento & purificación , Córnea/microbiología , Úlcera de la Córnea/microbiología , Infecciones Bacterianas del Ojo/microbiología , Humanos , Incidencia , Estudios Retrospectivos , Rotura EspontáneaRESUMEN
In 1996, a questionnaire was distributed to 4000 Australian commercial pilots for the quantification of symptoms of dry eye. The 1246 respondents ranged in age from 18-71 years (mean age 40.4 years), and 98.2% were men. Symptoms of dry eye during flight were reported by 901 (72.3%, 95% CI = 69.7 74.8) while only 67 (5.4%, 95% CI = 4.2-6.8) reported dry eye independent of flight. In univariate analyses, the following factors were associated with dry eye in flight: age, use of distance glasses, smoking, average weekly number of flight hours, type of aircraft usually flown, and normal cruising altitude. In a multiple logistic regression model, only the class of aeroplane (large jets vs. propeller-driven planes) (OR = 1.75, 95% CI = 1.34-2.28) and estimated number of flight hours per week (OR = 1.58, 95% CI = 1.34-1.86) were related to dry eye symptoms. In conclusion, self-reported symptoms of dry eye are common in Australian pilots and are associated with aeroplane class and flying time.
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Medicina Aeroespacial/estadística & datos numéricos , Aeronaves , Síndromes de Ojo Seco/epidemiología , Adolescente , Adulto , Anciano , Australia/epidemiología , Femenino , Encuestas Epidemiológicas , Humanos , Masculino , Persona de Mediana Edad , Prevalencia , Encuestas y CuestionariosRESUMEN
PURPOSE: Cataract surgery is increasing in Australia and represents a significant burden on limited health resources. This study examines the frequency and outcomes of cataract surgery for patients who were hospitalized overnight compared with those treated as day surgery cases. METHODS: Medical records of 671 consecutive admissions for cataract surgery at the Royal Victorian Eye and Ear Hospital were reviewed. Data analysed included demographic features, insurance status, length of hospitalization, ophthalmic conditions, medical conditions, social problems and planned surgical technique. Ophthalmic and anaesthetic complications, active ophthalmic and medical interventions were also studied. RESULTS: Of the 671 patient admissions for cataract during the study period, 226 (33.4%) were hospitalized overnight. Factors significantly associated with overnight hospitalization in univariate analyses include older age, female sex, country residence, Veterans' Affairs insurance, monocular vision status, pre-existing ischaemic heart disease, pre-existing asthma/chronic obstructive lung disease, absence of carer, transportation problems, planned extra-capsular cataract extraction technique, ophthalmic complications and active ophthalmic and/or medical interventions. After adjusting for possible confounding factors using backwards stepwise multivariate logistic regression models all except pre-existing ischaemic heart disease and ophthalmic complications were significantly associated with overnight admission for cataract surgery. In total, 14 cases (2.1%) needed active ophthalmic and/or medical interventions, 13 overnight cases and one day case. CONCLUSIONS: These data suggest that many patients who are hospitalized overnight for cataract surgery could be safely treated as day cases. Such a shift in the pattern of care for cataract surgery could provide a significant potential for health care savings.
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Atención Ambulatoria/estadística & datos numéricos , Extracción de Catarata/estadística & datos numéricos , Hospitalización/estadística & datos numéricos , Hospitales Especializados/estadística & datos numéricos , Oftalmología , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Evaluación de Resultado en la Atención de Salud , Pacientes Ambulatorios/estadística & datos numéricos , Admisión del Paciente/estadística & datos numéricos , Estudios Retrospectivos , VictoriaRESUMEN
OBJECTIVE: To evaluate patterns of self-management, healthcare utilisation and screening for major complications among Tasmanians with insulin-treated diabetes. MAIN OUTCOME MEASURES: Frequency of self-monitoring of blood glucose, health care utilisation and screening for diabetic complications. DESIGN AND SETTING: A questionnaire survey of 1517 people listed on the Tasmanian Diabetes Register in 1995-1997. RESULTS: Response rate was 79.5%. Self-monitoring of blood glucose was reported by 98% of respondents, daily self-monitoring by 74%. About 41% of respondents were being managed jointly by GPs and diabetes specialists, 29% solely by GPs and 25% solely by diabetes specialists. Over 96% visited the doctor treating their diabetes more than once a year, but 21% reported they had never visited a diabetes educator and 43% reported they had never visited a dietitian. Most respondents aged > or = 25 years (90%) reported having an eye examination within the past two years, almost all by an eye specialist. Blood pressure was commonly assessed, but most adults indicated that the doctor treating their diabetes did not routinely examine their feet. Nearly 19% of respondents smoked cigarettes. CONCLUSIONS: Some aspects of diabetes self-care and medical care have improved in Tasmania since the 1984 survey (eg, frequency of self-monitoring of blood glucose rose from 50% to 98%). However, our findings suggest that further improvements are needed to increase daily self-monitoring of blood glucose, attendance at diabetes educator and dietitian services, and foot examinations by doctors. Additional efforts are also needed to lower the prevalence of smoking.
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Diabetes Mellitus Tipo 1/prevención & control , Servicios de Salud/estadística & datos numéricos , Aceptación de la Atención de Salud/estadística & datos numéricos , Autocuidado/métodos , Adolescente , Adulto , Anciano , Automonitorización de la Glucosa Sanguínea , Diabetes Mellitus Tipo 1/complicaciones , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Diabetes Mellitus Tipo 1/metabolismo , Medicina Familiar y Comunitaria , Femenino , Encuestas de Atención de la Salud , Humanos , Masculino , Tamizaje Masivo/estadística & datos numéricos , Medicina , Persona de Mediana Edad , Especialización , Encuestas y Cuestionarios , TasmaniaAsunto(s)
Diabetes Mellitus/epidemiología , Hemoglobina Glucada/análisis , Adulto , Anciano , Anciano de 80 o más Años , Diabetes Mellitus/sangre , Diabetes Mellitus/diagnóstico , Femenino , Humanos , Masculino , Tamizaje Masivo , Persona de Mediana Edad , Prevalencia , Valores de Referencia , Salud Rural , Victoria/epidemiologíaRESUMEN
OBJECTIVE: Quantification of serum nucleotide pyrophosphohydrolase (NTPPHase) activity in healthy subjects and in patients with various rheumatic diseases or with quad/hemiplegia, hemodialysis, or renal transplant. METHODS: Colorimetric assay of enzyme activity in serum. RESULTS: Serum NTPPHase activity in 85 healthy subjects was independent of age or sex and was highly reproducible in each individual. The biologic and methodologic coefficients of variation were nearly identical. Elevated enzyme levels were found in sera from patients with osteoarthritis/spondylosis, calcium pyrophosphate dihydrate (CPPD) crystal deposition, scleroderma, fibromyalgia, or hemodialysis. Renal transplant patients receiving cyclosporine had the highest enzyme activity of any group, whereas transplant patients not taking this drug had normal levels. Histograms of values in all groups showed a normal distribution. CONCLUSION: Serum NTPPHase activity levels were significantly elevated in patients with degenerative arthritis whether or not CPPD crystals were present, in patients with either scleroderma or fibromyalgia, and in patients receiving hemodialysis therapy or taking cyclosporine.
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Condrocalcinosis/sangre , Fibromialgia/sangre , Osteoartritis/sangre , Pirofosfatasas/sangre , Esclerodermia Sistémica/sangre , Condrocalcinosis/enzimología , Ciclosporina/uso terapéutico , Femenino , Fibromialgia/enzimología , Humanos , Trasplante de Riñón , Masculino , Osteoartritis/enzimología , Cuidados Posoperatorios , Valores de Referencia , Diálisis Renal , Esclerodermia Sistémica/enzimologíaRESUMEN
We investigated the association of serum antibodies to glutamic acid decarboxylase (GADab) with early start of insulin treatment (< or = 1 year from diagnosis, or < or = 2 years from diagnosis) using data from a representative sample of 374 adult-onset insulin-treated individuals from the Tasmanian Diabetes Register. Furthermore, we examined whether this association was stronger than the phenotypic characteristics (age at diagnosis, sex, family history of diabetes, level of obesity, duration of diabetes) often used for diabetes classification. In this cohort, 35.9% of males and 38.5% of females were GADab positive. Within the first year from diagnosis, 78.4% of GADab positive people compared to 44.0% of GADab negative people (p < 0.001) had started insulin treatment. Univariate associations with insulin treatment < or = 1 year from diagnosis included GADab positivity, no family history of diabetes, lower BMI for men, and GADab positivity and lower BMI for women. In multivariate models, significant associations with insulin treatment < or = 1 year from diagnosis included a family history of diabetes (OR = 0.47, 95% CI = 0.23-0.95) and GADab positivity (OR = 2.19, 95% CI = 1.01-4.73) for men, but only GADab positivity (OR = 7.53, 95% CI = 3.09-18.30) for women. Age at diagnosis was not associated with insulin treatment < or = 1 year or < or = 2 years from diagnosis for either sex. These findings indicate that a positive GADab test result is strongly associated with start of insulin treatment within 1 or 2 years from diagnosis, more so than characteristics such as level of obesity and age at diagnosis.
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Autoanticuerpos/sangre , Diabetes Mellitus Tipo 1/inmunología , Diabetes Mellitus Tipo 2/inmunología , Glutamato Descarboxilasa/inmunología , Adulto , Edad de Inicio , Anciano , Estudios de Cohortes , Diabetes Mellitus/sangre , Diabetes Mellitus/inmunología , Diabetes Mellitus Tipo 1/sangre , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Diabetes Mellitus Tipo 2/sangre , Femenino , Humanos , Hipoglucemiantes/uso terapéutico , Insulina/uso terapéutico , Masculino , Persona de Mediana Edad , Análisis Multivariante , Obesidad , Oportunidad Relativa , Fenotipo , Análisis de Regresión , Caracteres Sexuales , Tasmania , Factores de TiempoRESUMEN
The porcine 127-kDa nucleotide pyrophosphohydrolase (NTPPHase) had been previously purified from the conditioned culture media of porcine articular cartilage. Protein sequencing of an internal 61-kDa proteolytic fragment of NTPPHase (61-kDa NTPPHase) determined the 26 N-terminal amino acids. This sequence was used to amplify a DNA fragment, which was used as a probe to clone the gene encoding the 61-kDa NTPPHase from a porcine chondrocyte cDNA library. DNA sequence analysis showed the cDNA insert to be 2509 bp, corresponding to a predicted open reading frame (ORF) encoding 599 amino acids. The 26 N-terminal amino acids of the 61-kDa NTPPHase were located within the ORF immediately downstream of a putative protease recognition region, RRKRR. This is consistent with this cDNA insert representing an internal proteolytic fragment of the full length 127-kDa NTPPHase. BLAST and FASTA analysis confirmed that the deduced amino acid sequence of 61-kDa NTPPHase was unique and did not possess a high degree of homology to sequence in the non-redundant protein and nucleotide databases. Proteins that possess limited homology (< 17%) with the 61-kDa NTTPPHase include several prokaryotic and eukaryotic ATP pyrophosphate-lyases (adenylate cyclase). Northern blot analysis of porcine chondrocyte RNA showed that the DNA encoding the 61-kDa NTPPHase hybridized to a single 4.0-kb RNA transcript. This DNA probe also hybridized to a single species of human chondrocyte RNA. Expression of a 61-kDa protein was detected by coupled in-vitro transcription/translation. Western blot analysis of this in-vitro transcription/translation reaction detected a 61-kDa protein, using an antibody raised against the peptide sequence that was originally used to clone the 61-kDa NTPPHase. These data indicate the successful in-vitro cloning and expression of the porcine chondrocyte 61-kDa NTPPHase. Future studies that utilize the gene encoding the 61-kDa NTPPHase may allow the characterization of the role of NTPPHase in calcium pyrophosphate dihydrate (CPPD) crystal deposition disease.
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Condrocitos/enzimología , Regulación Enzimológica de la Expresión Génica/genética , Pirofosfatasas/genética , Secuencia de Aminoácidos , Aminoácidos/análisis , Animales , Secuencia de Bases , Cartílago Articular/enzimología , Clonación Molecular , ADN Complementario/genética , Biblioteca de Genes , Humanos , Datos de Secuencia Molecular , Peso Molecular , Osteoartritis , Biosíntesis de Proteínas , Pirofosfatasas/química , ARN Mensajero/análisis , Análisis de Secuencia de ADN , Especificidad de la Especie , Porcinos , Transcripción GenéticaRESUMEN
OBJECTIVE: To identify the molecular forms of ectonucleotide pyrophosphohydrolase (NTPPHase) in human synovial fluid (SF). METHODS: We examined synovial fluids from 32 patients with various joint diseases [10 calcium pyrophosphate dihydrate (CPPD) deposition disease; 7 osteoarthritis (OA); 6 rheumatoid arthritis (RA); 3 after total knee arthroplasty (TKA); 6 olecranon bursa] and 3 normal joint fluids. Joint fluids were analyzed after sequential centrifugation for NTPPHase activity and by Western blot using polyclonal antibodies against 127 kDa porcine articular cartilage vesicle-associated NTPPHase and against PC-1 and 58 kDa, 2 other ecto-NTPPHases. Lysate from human synoviocytes, porcine chondrocytes, and their conditioned media were examined using antibodies to these ecto-NTPPHases. Radiographs of joints from which fluid was obtained were graded for degenerative changes 0-4 using a standard method. RESULTS: NTPPHase activity was found in all pathological and normal SF tested and correlated with the degree of radiographic degeneration (r = 0.55, p < 0.05). NTPPHase specific activity in ultracentrifugation pellets was highest in CPPD deposition disease fluids (p < 0.05). 127 kDa enzyme was found in both sedimentable and soluble fractions from CPPD, OA, TKA, and normal fluids, and was extensively degraded in all inflammatory fluids. Intact 115 kDa PC-1 was found only in the 2 CPPD fluids with the highest NTPPHase activity. 58 kDa enzyme was found in most fluids, predominantly in the soluble fraction. 127 kDa protein was identified in human synoviocyte conditioned media but not in cell lysate, while PC-1 and 58 kDa proteins were found in the cell lysate but not in the conditioned media. CONCLUSION: There was no disease specific association with any one ecto-NTPPHase. Total enzyme activity correlated with the degree of degenerative change. The specific activity of pelletable 127 kDa enzyme was higher in fluids containing CPPD crystals. All 3 ecto-NTPPHases or their presumed degradation products were detectable in some pathologic and normal fluids. A 200 kDa reactive band often accompanied reactivity to the 127 kDa enzyme. PC-1 and 127 kDa proteins were extensively degraded in inflammatory SF, while 58 kDa protein was not. The relative contribution of each of these enzymes to inorganic pyrophosphate production by human joint tissues remains unclear.