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Habitual exercise training is recommended to young people for their health promotion, but adherence may be influenced by atmospheric temperature (T a ) if performed outdoors. We compared the adherence to and the effects of a home-based interval walking training (IWT) program on sedentary female college students between winter and summer. For summer training over 176 days, 48 subjects (18-22 years old) were randomly divided into two groups: the control group (CNTsummer, n = 24), which maintained a sedentary lifestyle as before, and the IWT group (IWTsummer, n = 24), which performed IWT while energy expenditure was monitored by accelerometry. For winter training over 133 days, another group of 47 subjects (18-24 years old) was randomly divided into CNTwinter (n = 24) and IWTwinter (n = 23), as in summer. The peak T a per day was 26 ± 6 °C (SD) (range of 9-35 °C) in summer, much higher than 7 ± 5 °C (range of - 3-20 °C) in winter (P < 0.001). During a ~ 50-day vacation period, participants walked 2.1 ± 0.3 (SE) days/week in IWTsummer, less than 4.2 ± 0.3 days/week in IWTwinter (P < 0.001), with half of the energy expenditure/week for fast walking during the winter vacation (P < 0.02), whereas both IWT groups walked ~ 2 days/week during a school period (P > 0.8). After training, the peak aerobic capacity and knee flexion force increased in IWTwinter (P < 0.01) but not in CNTwinter (P > 0.3). Conversely, these parameters decreased in the summer groups. Thus, the adherence to and effects of IWT on sedentary female college students in Japan decreased in summer at least partially due to a high T a .
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Cooperación del Paciente , Estaciones del Año , Caminata/fisiología , Adulto , Terapia por Ejercicio , Femenino , Humanos , Japón , Conducta Sedentaria , Estudiantes , Universidades , Adulto JovenRESUMEN
Patients with diabetes mellitus are believed to be suspected to be immunocompromized hosts. Many reports have pointed out that diabetic patients are susceptible to certain infections such as surgical site infections, malignant otitis externa, mucormycosis, and necrotizing fasciitis. But their etiology seems to be non-uniform, heterogenous and individualized. Above all, obesity-related infections are also increasing accompanied with the recent rising incidence of obesity. Further studies should be addressed about the relationships between infections and diabetes which include the factors of body mass index, life style, degree of diabetes complications, and poor glycemic control duration. They could live a normal life the same as healthy subjects if good glycemic control is achieved without hypoglycemia.
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Complicaciones de la Diabetes , Infecciones/etiología , Diabetes Mellitus , Humanos , Control de Infecciones , Infecciones/patología , Necrosis , Obesidad/complicaciones , Factores de Riesgo , VacunaciónRESUMEN
It has recently been reported that expression of heme oxygenase-1 (HO-1) plays a protective role against many diseases. Furthermore, n-3 polyunsaturated fatty acids (PUFAs) were shown to induce HO-1 expression in several cells in vitro, and in a few cases also in vivo. However, very few reports have demonstrated that n-3 PUFAs induce HO-1 in vivo. In this study, we examined the effect of fish-oil dietary supplementation on the distribution of fatty acids and their peroxidative metabolites and on the expression of HO-1 in multiple tissues (liver, kidney, heart, lung, spleen, intestine, skeletal muscle, white adipose, brown adipose, brain, aorta, and plasma) of C57BL/6 mice. Mice were divided into 4 groups, and fed a control, safflower-oil, and fish-oil diet for 3 weeks. One group was fed a fish-oil diet for just 1 week. The concentration of fatty acids, 4-hydroxy hexenal (4-HHE), and 4-hydroxy nonenal (4-HNE), and the expression of HO-1 mRNA were measured in the same tissues. We found that the concentration of 4-HHE (a product of n-3 PUFAs peroxidation) and expression of HO-1 mRNA were significantly increased after fish-oil treatment in most tissues. In addition, these increases were paralleled by an increase in the level of docosahexaenoic acid (DHA) but not eicosapentaenoic acid (EPA) in each tissue. These results are consistent with our previous results showing that DHA induces HO-1 expression through 4-HHE in vascular endothelial cells. In conclusion, we hypothesize that the HO-1-mediated protective effect of the fish oil diet may be through production of 4-HHE from DHA but not EPA in various tissues.
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Aldehídos/metabolismo , Ácidos Grasos Omega-3/metabolismo , Hemo-Oxigenasa 1/biosíntesis , Especificidad de Órganos , Aldehídos/sangre , Animales , Ácido Araquidónico/sangre , Ácidos Docosahexaenoicos/sangre , Ácido Eicosapentaenoico/sangre , Inducción Enzimática , Ácidos Grasos Omega-3/sangre , Hemo-Oxigenasa 1/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Oxidación-Reducción , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
There is no exercise training regimen broadly available in the field to increase physical fitness and prevent lifestyle-related diseases in middle-aged and older people. We have developed interval walking training (IWT) repeating five or more sets of 3 min fast walking at 70% peak aerobic capacity for walking (w ) per day with intervening 3 min slow walking at 40% w , for 4 days week(1), for 5 months. Moreover, to determine w in individuals and also to measure their energy expenditure even while incline walking, we have developed a portable calorimeter. Further, to instruct subjects on IWT even if they live remotely from the trainers, we have developed e-Health Promotion System. This transfers individual energy expenditure during IWT stored on the meter to a central server through the internet; it sends back the achievement to individuals along with advice generated automatically by the sever according to a database on 4000 subjects. Where we found that 5 months of IWT increased physical fitness and improved the indices of lifestyle-related diseases by 10-20% on average. Since our system is run at low cost with fewer staff for more subjects, it enables us to develop exercise prescriptions appropriate for individuals.
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Ejercicio Físico/fisiología , Aptitud Física/fisiología , Envejecimiento , Umbral Anaerobio/fisiología , Calorimetría , Métodos Epidemiológicos , Promoción de la Salud , Humanos , Población , Caminata/fisiologíaRESUMEN
The co-occurrence of resistance to thyroid hormone beta (RTHß) and myotonic dystrophy type 1 (DM1) was observed in a Japanese family. Two mutations, P453A and C36Y, were identified in the thyroid hormone receptor beta (THRB) gene. Whereas family members with THRBP453A exhibited RTHß, two members with THRBC36Y but without THRBP453A had normal thyroid function. Two members, one with RTHß and the other without, had a triplet expansion in the dystrophia myotonia protein kinase gene, a hallmark of DM1. The member with both RTHß and DM1 developed atrial fibrillation at the age of 16 years, suggesting a synergistic impact on the heart.
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Mutación , Distrofia Miotónica/genética , Proteína Quinasa de Distrofia Miotónica/genética , Receptores beta de Hormona Tiroidea/genética , Síndrome de Resistencia a Hormonas Tiroideas/genética , Repeticiones de Trinucleótidos , Adolescente , Adulto , Fibrilación Atrial/etiología , Aleteo Atrial/etiología , Femenino , Predisposición Genética a la Enfermedad , Herencia , Humanos , Masculino , Persona de Mediana Edad , Distrofia Miotónica/complicaciones , Distrofia Miotónica/diagnóstico , Linaje , Fenotipo , Síndrome de Resistencia a Hormonas Tiroideas/complicaciones , Síndrome de Resistencia a Hormonas Tiroideas/diagnósticoRESUMEN
OBJECTIVE: To examine whether high-intensity interval walking training increased thigh muscle strength and peak aerobic capacity and reduced blood pressure more than moderate-intensity continuous walking training. PARTICIPANTS AND METHODS: From May 18, 2004, to October 15, 2004 (5-month study period), 60 men and 186 women with a mean +/- SD age of 63 +/- 6 years were randomly divided into 3 groups: no walking training, moderate-intensity continuous walking training, and high-intensity interval walking training. Participants in the moderate-intensity continuous walking training group were instructed to walk at approximately 50% of their peak aerobic capacity for walking, using a pedometer to verify that they took 8000 steps or more per day for 4 or more days per week. Those in the high-intensity interval walking training group, who were monitored by accelerometry, were instructed to repeat 5 or more sets of 3-minute low-intensity walking at 40% of peak aerobic capacity for walking followed by a 3-minute high-intensity walking above 70% of peak aerobic capacity for walking per day for 4 or more days per week. Isometric knee extension and flexion forces, peak aerobic capacity for cycling, and peak aerobic capacity for walking were all measured both before and after training. RESULTS: The targets were met by 9 of 25 men and 37 of 59 women in the no walking training group, by 8 of 16 men and 43 of 59 women in the moderate-intensity continuous walking training group, and by 11 of 19 men and 31 of 68 women in the high-intensity interval walking training group. In the high-intensity interval walking training group, isometric knee extension increased by 13%, isometric knee flexion by 17%, peak aerobic capacity for cycling by 8%, and peak aerobic capacity for walking by 9% (all, P<.001), all of which were significantly greater than the increases observed in the moderate-intensity continuous walking training group (all, P<.01). Moreover, the reduction in resting systolic blood pressure was higher for the high-intensity interval walking training group (P=.01). CONCLUSION: High-intensity interval walking may protect against age-associated increases in blood pressure and decreases in thigh muscle strength and peak aerobic capacity.
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Presión Sanguínea , Aptitud Física , Caminata , Anciano , Índice de Masa Corporal , Metabolismo Energético , Femenino , Frecuencia Cardíaca , Humanos , Masculino , Persona de Mediana Edad , Fuerza MuscularRESUMEN
BACKGROUND & AIMS: A fiber-rich diet has a cardioprotective effect, but the mechanism for this remains unclear. We hypothesized that a fiber-rich diet with brown rice improves endothelial function in patients with type 2 diabetes mellitus. METHODS: Twenty-eight patients with type 2 diabetes mellitus at a single general hospital in Japan were randomly assigned to a brown rice (n = 14) or white rice (n = 14) diet and were followed for 8 weeks. The primary outcome was changes in endothelial function determined from flow debt repayment by reactive hyperemia using strain-gauge plethysmography in the fasting state. Secondary outcomes were changes in HbA1c, postprandial glucose excursions, and markers of oxidative stress and inflammation. The area under the curve for glucose after ingesting 250 kcal of assigned rice was compared between baseline (T0) and at the end of the intervention (T1) to estimate glucose excursions in each group. RESULTS: Improvement in endothelial function, assessed by fasting flow debt repayment (20.4% vs. -5.8%, p = 0.004), was significantly greater in the brown rice diet group than the white rice diet group, although the between-group difference in change of fiber intake was small (5.6 g/day vs. -1.2 g/day, p<0.0001). Changes in total, HDL-, and LDL-cholesterol, and urine 8-isoprostane levels did not differ between the two groups. The high-sensitivity C-reactive protein level tended to improve in the brown rice diet group compared with the white rice diet group (0.01 µg/L vs. -0.04 µg/L, p = 0.063). The area under the curve for glucose was subtly but consistently lower in the brown rice diet group (T0: 21.4 mmol/L*h vs. 24.0 mmol/L*h, p = 0.043, T1: 20.4 mmol/L*h vs. 23.3 mmol/L*h, p = 0.046) without changes in HbA1c. CONCLUSIONS: Intervention with a fiber-rich diet with brown rice effectively improved endothelial function, without changes in HbA1c levels, possibly through reducing glucose excursions.
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Diabetes Mellitus Tipo 2/fisiopatología , Fibras de la Dieta , Endotelio Vascular/fisiopatología , Oryza , Anciano , Femenino , Humanos , Japón , Masculino , Persona de Mediana EdadRESUMEN
Osteoblasts and adipocytes are believed to share a common progenitor. Peroxisome proliferator-activated receptor gamma (PPARgamma) plays a key role in the switching of these two cell lineages. Here, we demonstrated the differentiation of ES cells into an osteoblastic lineage using siRNA against PPARgamma without the addition of any osteogenic factors. We found that PPARgamma-siRNA downregulated the expression of aP2 mRNA and lipid accumulation, whereas it upregulated the expression of osteocalcin and calcium deposition. These results suggested that ES cells were directed into an osteoblastic lineage. Therefore, transient suppression using PPARgamma-siRNA may be a novel tool to induce differentiation of ES cells into osteoblasts.
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Diferenciación Celular/fisiología , Linaje de la Célula/fisiología , Embrión de Mamíferos/fisiología , Osteoblastos/fisiología , PPAR gamma/metabolismo , Células Madre/fisiología , Adipocitos/citología , Adipocitos/fisiología , Animales , Calcificación Fisiológica/efectos de los fármacos , Calcificación Fisiológica/fisiología , Calcio/metabolismo , Diferenciación Celular/efectos de los fármacos , Línea Celular , Linaje de la Célula/efectos de los fármacos , Embrión de Mamíferos/citología , Ratones , Osteoblastos/citología , Osteopontina , PPAR gamma/antagonistas & inhibidores , PPAR gamma/genética , ARN Mensajero/biosíntesis , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/farmacología , Sialoglicoproteínas/biosíntesis , Células Madre/citología , Factores de Transcripción/biosíntesis , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/fisiologíaRESUMEN
Production of functional adipocytes is important in adipocyte research and regenerative medicine. In this paper, we describe the differentiation of monkey embryonic stem (ES) cells into insulin-responsive adipocytes. Treatment of embryoid body (EB) outgrowth with adipogenic hormones induced the expression of adipocyte-specific genes, such as PPARgamma, C/EBPalpha, aP2, insulin receptor, and GLUT4. Expression of adipocytokines, leptin and adiponectin, was also detected. Furthermore, translocation of GLUT4 was observed by insulin stimulation in differentiated adipocytes. These results suggested that monkey ES cells can be a useful tool for studying adipogenesis in primate.
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Adipocitos/citología , Adipogénesis , Diferenciación Celular , Células Madre/citología , Adipocitos/química , Adipocitos/efectos de los fármacos , Adiponectina/análisis , Adiponectina/genética , Animales , Proteína alfa Potenciadora de Unión a CCAAT/análisis , Proteína alfa Potenciadora de Unión a CCAAT/genética , Células Cultivadas , Citocinas/análisis , Citocinas/genética , Regulación del Desarrollo de la Expresión Génica , Transportador de Glucosa de Tipo 4/análisis , Transportador de Glucosa de Tipo 4/genética , Inmunohistoquímica , Insulina/farmacología , Leptina/análisis , Leptina/genética , Macaca fascicularis , PPAR gamma/análisis , PPAR gamma/genética , Receptor de Insulina/análisis , Receptor de Insulina/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Madre/química , Células Madre/efectos de los fármacos , Translocación Genética/efectos de los fármacosRESUMEN
AIMS: We assessed the association between smoking status and mild cognitive impairment (MCI) in Japanese diabetic patients. METHODS: This cross-sectional study included 323 diabetic patients, aged 40-79 years, who were referred to an outpatient diabetic clinic between January and July 2013 at Shiga University of Medical Science Hospital (Otsu, Japan). Cognitive function was assessed using the mini-mental state examination (MMSE), and patients were classified into two categories: normal cognitive function (MMSE score ≥27) and MCI (MMSE score 22-26). Logistic regression analysis was used to estimate the multivariable-adjusted odds ratios (ORs) and 95 % confidence intervals (CIs) for MCI in current smokers and ex-smokers compared with never-smokers. RESULTS: Of the 323 patients, 55 (17.0 %), 134 (41.5 %), and 134 (41.5 %) were current smokers, ex-smokers, and never-smokers, respectively. Of these, 68 (21.0 %) patients had MCI. After adjusting for age, sex, systolic blood pressure, body mass index, high-density lipoprotein cholesterol, estimated glomerular filtration rate, hemoglobin A1c, insulin therapy, sulfonylurea, history of coronary heart disease, exercise habit, drinking status, and education, the OR for MCI was 3.62 (95 % CI 1.26-10.40) in current smokers compared with never-smokers. In addition, the multivariable-adjusted ORs for MCI were 3.02 (95 % CI 0.64-14.32) in current smokers <30.0 pack-years and 4.90 (95 % CI 1.32-18.16) in current smokers ≥30.0 pack-years, compared with never-smokers (p for trend = 0.017). CONCLUSIONS: Current smoking, especially current smoking for which cumulative lifetime exposure was high, was associated with MCI, as assessed using the MMSE in Japanese diabetic patients.
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AIMS: This study assessed the association between attentional function and postural instability in older Japanese patients with diabetes. METHODS: This cross-sectional study included 168 older patients with diabetes who were referred to an outpatient diabetic clinic between June and July 2013. The Trail Making Test-A (TMT-A) was used to evaluate attentional function. Posturography was used to evaluate postural sway. Indices of postural sway were the total length and the enveloped area. Analysis of covariance was used to estimate the multivariable-adjusted means of indices of postural sway according to tertile of TMT-A. RESULTS: After adjustment for age, sex, regular exercise, diabetic retinopathy, bilateral numbness and/or paresthesia in the feet, hemoglobin A1c level, quadriceps strength, and Mini-Mental State Examination score, patients with lower attentional function had higher postural sway length (tertile 3 vs. tertile 1, p = 0.010) and enveloped area (tertile 3 vs. tertile 1, p = 0.030) levels than those with higher attentional function. CONCLUSIONS: Among older patients with diabetes who did not have dementia, patients with lower attentional function may have more postural instability than those with higher attentional function.
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AIM: This study assessed the association between symptoms of bilateral numbness and/or paresthesia in the feet and postural instability in Japanese patients with diabetes. METHODS: This cross-sectional study included 303 patients with diabetes, aged 40-88 years, who were referred to an outpatient diabetic clinic between January and July 2013 at Shiga University of Medical Science Hospital. A posturography test was used to evaluate postural sway in patients. Indices of postural sway were the total length and the enveloped area. Analysis of covariance was used to estimate the multivariable-adjusted means of indices of postural sway according to the presence or absence of symptoms of bilateral numbness and/or paresthesia in the feet. RESULTS: Of 303 patients, 35 (11.6 %) had symptoms of bilateral numbness and/or paresthesia in the feet. After adjustment for age, sex, diabetic retinopathy, regular exercise, body mass index, hemoglobin A1c level, and quadriceps' strength, patients with symptoms had higher levels of postural sway length and an enveloped area in the posturography test than those without symptoms. In addition, we observed similar results when we analyzed 234 patients aged ≥60 years. CONCLUSIONS: Our findings suggest that patients who had symptoms of bilateral numbness and/or paresthesia in the feet may have more postural instability than those without symptoms.
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We herein report a case involving a woman with type 1 diabetes and a history of metal allergy who developed a local delayed-type (type IV) allergy to zinc-containing insulin. She had been treated by continuous subcutaneous insulin infusion, but her glycemic control was poor, and she developed diabetic ketoacidosis. Her plasma insulin concentration was unexpectedly low during use of insulin lispro, but it was recovered by changing from the zinc-containing insulin lispro to the zinc-free insulin glulisine. Intradermal tests showed no reactions to various insulins except for zinc chloride. A skin biopsy at the injection site of insulin lispro showed invasion of lymphocytes, neutrophils, and eosinophils, but a skin biopsy at the injection site of insulin glulisine showed invasion of only lymphocytes. A drug lymphocyte stimulation test against polaprezinc, an antiulcer drug containing zinc, was positive. Therefore, we diagnosed the patient with local delayed allergy to zinc-containing insulin. Insulin allergy should be considered as a possible cause of poor glycemic control and diabetic ketoacidosis in patients with type 1 diabetes.
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The objective of this study was to explore predictors, including social factors, lifestyle factors, and factors relevant to glycemic control and treatment, for mild and severe hypoglycemia in insulin-treated Japanese diabetic patients. This study included 123 insulin-treated diabetic patients who were referred to the diabetes clinic between January and July 2013 at Shiga University of Medical Science Hospital. After a survey examining the various factors, patients were followed for 6 months. During the follow-up period, blood glucose was self-monitored. Mild hypoglycemia was defined as blood glucose level 50-69 mg/dl, and severe hypoglycemia was defined as blood glucose level ≤49 mg/dl. Multinomial logistic regression was used to estimate the adjusted odds ratio (OR) and 95% confidence interval (CI) of each factor for mild and severe hypoglycemia. During the 6-month follow-up period, 41 (33.3%) patients experienced mild hypoglycemia, and 20 (16.3%) experienced severe hypoglycemia. In multivariable-adjusted analyses, assistance from family members at the time of the insulin injection [presence/absence, OR (95% CI): 0.39 (0.16-0.97)] and drinking [current drinker/non- and ex-drinker, OR (95% CI): 4.89 (1.68-14.25)] affected mild hypoglycemia. Assistance from family members at the time of insulin injection [presence/absence, OR (95% CI): 0.19 (0.05-0.75)] and intensive insulin therapy [yes/no, OR (95% CI): 3.61 (1.06-12.26)] affected severe hypoglycemia. In conclusion, our findings suggest that not only a factor relevant to glycemic control and treatment (intensive insulin therapy) but also a social factor (assistance from family members) and a lifestyle factor (current drinking) were predictors for mild or severe hypoglycemia in Japanese insulin-treated diabetic patients.
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Diabetes Mellitus/tratamiento farmacológico , Hipoglucemia/inducido químicamente , Insulina/efectos adversos , Anciano , Consumo de Bebidas Alcohólicas/sangre , Antropometría , Automonitorización de la Glucosa Sanguínea , Cuidadores , Diabetes Mellitus/sangre , Quimioterapia Combinada , Escolaridad , Familia , Femenino , Estudios de Seguimiento , Hábitos , Encuestas de Atención de la Salud , Humanos , Hipoglucemia/epidemiología , Hipoglucemiantes/efectos adversos , Hipoglucemiantes/uso terapéutico , Insulina/administración & dosificación , Insulina/uso terapéutico , Japón , Estilo de Vida , Masculino , Persona de Mediana Edad , Factores de Riesgo , Factores Socioeconómicos , Encuestas y CuestionariosRESUMEN
BACKGROUND: To assess dyslipidemia, measurement of low-density lipoprotein cholesterol via either Friedewald equation (LDL-F) or direct assay (LDL-D), and non-high-density lipoprotein cholesterol (non-HDL-C) are recommended with some guidelines showing preference to direct over calculated measurements. However, direct comparisons of their respective associations with cardiovascular disease (CVD) risk are currently unavailable. OBJECTIVE: In this study, we evaluated the clinical effectiveness of LDL-F and non-HDL-C vs LDL-D and their associations with CVD. METHODS: This retrospective cohort study comprised apparently healthy Japanese individuals who underwent an annual health check-up between 2005 and 2007 and completed a 5-year follow-up visit. The incidence of CVD, including coronary and cerebrovascular diseases, during a 5-year follow-up period was evaluated using multivariate logistic regression. RESULTS: At baseline, 26,739 participants (mean age, 47 years; 49.0% men) were enrolled, and 292 (1.09%) incidents of CVD were identified at follow-up. Baseline LDL-F, LDL-D, and non-HDL-C were all significantly associated with CVD, although the effect appeared higher for LDL-F, particularly for coronary heart disease. Increased risks of CVD were observed for high LDL-F (≥130 mg/dL), despite being categorized into the lower LDL category based on LDL-D (odds ratio [OR], 1.85; 95% confidence interval [CI], 1.19-2.87) and non-HDL-C (OR, 1.75; 95% CI, 1.22-2.52). Without high LDL-F, no CVD associations were found for high LDL-D (P = .62) or non-HDL-C (P = .93). CONCLUSION: Despite growing availability of direct assays and increasing evidence of non-HDL-C utility, the Friedewald equation may offer better clinical utility for CVD prevention, especially in the screening of apparently healthy individuals.
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Análisis Químico de la Sangre , Enfermedades Cardiovasculares/sangre , Enfermedades Cardiovasculares/epidemiología , HDL-Colesterol/sangre , Adulto , Anciano , Enfermedades Cardiovasculares/complicaciones , Enfermedades Cardiovasculares/prevención & control , Dislipidemias/complicaciones , Femenino , Humanos , Japón/epidemiología , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Medición de RiesgoRESUMEN
Shinshu University and an NPO "JTRC" (Jukunen Taiiku Daigaku Research Center; a health promotion and support organization) provide exercise guidance focused on walking activity with the object of enhancing strength among elderly individuals. Sanyo Electric has developed an exercise meter as a tool to promote these efforts effectively. This exercise meter is equipped with a function providing estimates of energy consumption, leg muscle strength, and endurance based on triaxial acceleration data from the lumbar region. In this paper, we provide an outline of the estimation techniques and present an application of this exercise meter.
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Small interfering RNA (siRNA) has been widely used for suppressing gene expression in various organisms. Here, we describe efficient methods to suppress target genes (EGFP or Oct4) using siRNA in mouse and monkey ES cells, and differentiation. In mouse ES cells, FACS analysis revealed that EGFP expression was suppressed in 97% of transfected cells at 48 h after transfection. In addition, cells expressed Hand1 and Cdx2, which are the marker genes of trophoblast lineage by the transient suppression of Oct4. In the case of monkey ES cells, highly efficient suppression was achieved in 98% of cells at 96 h post-transfection using the Sendai virus (hemagglutinating virus of Japan, HVJ) envelope as a carrier of siRNA. These efficient transfection methods using synthetic siRNA should contribute to evaluate specific gene function in ES cells and can be used to differentiate ES cells into desired cell lineages.
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Diferenciación Celular , Silenciador del Gen , ARN Interferente Pequeño/genética , Animales , Secuencia de Bases , Células Cultivadas , Cartilla de ADN , Citometría de Flujo , Proteínas Fluorescentes Verdes/genética , Haplorrinos , RatonesRESUMEN
Like hyperglycemia, postprandial (diet-induced) hypertriglyceridemia is thought to play crucial roles in the pathogenesis of insulin resistant/metabolic syndrome. Sterol regulatory element-binding protein-1 (SREBP-1) is a key transcription factor to induce postprandial hypertriglyceridemia. We found that insulin-resistant rats fed a diet high in fructose showed an increased proteintyrosine phosphatase 1B (PTP1B) content with strong expression of SREBP-1 mRNA in the liver. To clarify the association of PTP1B with SREBP-1 gene expression, we overexpressed PTP1B in rat hepatocytes, which led to increased mRNA content and promoter activity of SREBP-1a and -1c, resulting in the increased mRNA expression of fatty-acid synthase, one of the SREBP-1-responsive lipogenic genes. Because PTP1B overexpression increased phosphatase 2A (PP2A) activity, we inhibited PP2A activity by expression of its selective inhibitor, SV40 small T antigen and found that this normalized the PTP1B-enhanced SREBP-1a and -1c mRNA expressions through activation of the Sp1 site. These results indicate that PTP1B may regulate gene expression of SREBP-1 via enhancement of PP2A activity, thus mediating hepatic lipogenesis and postprandial hypertriglyceridemia. We demonstrate here a unique serial activation of the PTP1B-PP2A axis as a novel mechanism for the regulation of gene expression in the biosynthesis of triglyceride.