Combination of restriction endonuclease digestion with the ΔΔCt method in real-time PCR to monitor etoxazole resistance allele frequency in the two-spotted spider mite.

Monitoring resistance allele frequency at the early stage of resistance development is important for the successful acaricide resistance management. Etoxazole is a mite growth inhibitor to which resistance is conferred by an amino acid substitution in the chitin synthase 1 (CHS1; I1017F) in T. urticae. If the susceptible allele can be specifically digested by restriction endonuclease, the ΔΔCt method using real-time PCR for genomic DNA (RED-ΔΔCt method) may be available for monitoring the resistance allele frequency. We tested whether the etoxazole resistance allele frequency in a pooled sample was accurately measured by the RED-ΔΔCt method and validated whether the resistance variant frequency was correlated with etoxazole resistance phenotype in a bioassay. Finally, we performed a pilot test using field populations. Strong linearity of the measures by the RED-ΔΔCt method with practical resistance allele frequencies; resistance allele frequency in the range between 0.5% to at least 0.75% was strictly represented. The strong linear relationship between hatchability of haploid male eggs after the etoxazole treatments (phenotype) and resistance allele frequencies in their mothers provided direct evidence that I1017F is a primary resistance factor to etoxazole in the strains used for experiments. The pilot test revealed a significant correlation between egg hatchability (including both diploid female eggs and haploid male eggs) and estimators in field populations. Consequently, we concluded that the RED-ΔΔCt method is a powerful tool for monitoring a resistance allele in a pooled sample.

A Patient with Rett Syndrome Maintained Motor Function by Periodic Rehabilitation Therapy and Proactive Daily Activities.

BACKGROUND: The motor functions of Rett syndrome patients degrade during the course of the disease. Some patients, however, are able to maintain their motor functions by undertaking exercise programs. CASE: A 2-year-old girl was diagnosed with Rett syndrome after the identification of a mutation in the gene encoding the methyl-CpG-binding protein. The patient started weekly rehabilitation therapy; however, over time, this reduced to monthly sessions, and subsequently, because of worsening epileptic seizures, even fewer sessions were possible. When the patient was 12 years old, intensive rehabilitation therapy was undertaken for 1 month; therapy involved 80 min of in-hospital therapy on weekdays and walking exercises at home at the weekend. An accelerometer (Actiwatch 2, standard type, Philips Respironics) indicated that more than 60 min of daily training at an intensity of 3 metabolic equivalents (METs) or more was achieved at weekends. The patient took the 10-m walk test, and the average time reduced from 18.6 to 13.5 s and the number of steps reduced from 32 to 23 between the start and finish of the first 1-month intensive training regime. After being discharged from the hospital, the patient maintained walking exercises at school during weekdays and performed more than 50 min/day of activity at at least 3 METs at home on weekends, in addition to a weekly home-visit rehabilitation therapy and the annual in-hospital 1-month rehabilitation therapy. Four years later, the patent's average time and number of steps required to walk 10 m remained lower (12.7 s, 24 steps) than those recorded at the first evaluation. DISCUSSION: Periodic rehabilitation therapy and proactive walking exercises at more than 3 METs for a duration of up to 50 min/day were instrumental in maintaining the motor functions of a Rett syndrome patient.

LDL cholesterol and the development of stroke subtypes and coronary heart disease in a general Japanese population: the Hisayama study.

BACKGROUND AND PURPOSE: Although the relation between serum LDL cholesterol level and coronary heart disease (CHD) is well established, its relation with stroke subtypes is less clear. METHODS: A total of 2351 inhabitants age >or=40 years in a Japanese community were followed up for 19 years. RESULTS: During follow-up, 271 subjects developed stroke and 144 developed CHD. Whereas the age- and sex-adjusted incidences of CHD significantly increased with increasing LDL cholesterol levels (P for trend <0.001), the associations between LDL cholesterol level and the incidences of ischemic or hemorrhagic stroke were not significant. The age- and sex-adjusted incidences of atherothrombotic infarctions (ATIs) and lacunar infarctions (LIs) significantly increased with increasing LDL cholesterol level (P for trend=0.03 for ATIs and=0.02 for LIs), but no such association was observed for cardioembolic infarction. After multivariate adjustment, the positive associations of LDL cholesterol level with the risks of ATI and CHD remained significant (P for trend=0.02 for ATIs and=0.03 for CHD), whereas the association with LIs was not significant. The risk of ATI significantly increased in the fourth quartile of LDL cholesterol compared with the first quartile (multivariate-adjusted hazard ratio=2.84; 95% CI, 1.17 to 6.93). The multivariate-adjusted risks for developing nonembolic infarction (ATIs and LIs) and CHD were significantly elevated in the groups with elevated LDL cholesterol values with and without the metabolic syndrome. CONCLUSIONS: Our findings suggest that an elevated LDL cholesterol level is a significant risk factor for developing ATI as well as CHD, and these associations are independent of the metabolic syndrome.

Reproducibility of blood pressure variation in older ambulatory and bedridden subjects.

OBJECTIVES: We investigated the influence of ambulation on the reproducibility of circadian blood pressure variation in older nursing home residents. DESIGN: Ambulatory blood pressure monitoring was performed twice in 37 older nursing home residents. SETTING: Nursing home in Japan. PARTICIPANTS: Subjects included 18 ambulatory nursing home residents who had no limitation on physical activity and 19 bedridden residents who did not participate in physical activity. MEASUREMENTS: Twenty-four-hour, daytime, and nighttime blood pressure levels and their variability. RESULTS: The 24-hour and daytime variability of systolic blood pressure (SBP) was significantly greater in ambulatory than in bedridden subjects, whereas nighttime variability was similar. Significant correlations in SBP averaged for the whole day, daytime, and nighttime were observed between the two examinations in ambulatory (r =.80-.83) and bedridden (r =.83-.91) subjects, but the variabilities of SBP for the whole day and during the daytime of the first measurement were correlated with those of the second measurement in bedridden (r =.67 and r =.47, respectively) but not in ambulatory (r =.39 and r =.28, respectively) subjects. Significant correlations were found between the nocturnal SBP changes at two occasions in both ambulatory (r =.50) and bedridden (r =.51) subjects, but the dipper versus nondipper profiles, defined as reduction in SBP of greater than 10% versus not, showed low reproducibility in ambulatory subjects; five ambulatory (28%) and one bedridden (5%) subjects showed divergent profiles between the two examinations. CONCLUSIONS: The reproducibility of blood pressure variation in nursing home residents is influenced by ambulation.

Non-high-density lipoprotein cholesterol and the development of coronary heart disease and stroke subtypes in a general Japanese population: the Hisayama Study.

BACKGROUND AND PURPOSE: It has not been fully determined whether non-high-density lipoprotein cholesterol (non-HDLC) levels are involved in vascular events, especially stroke, in general Asian populations. We evaluated the association between non-HDLC levels and the risk of type-specific cardiovascular disease in a prospective cohort study in Japan. METHODS: A total of 2452 community-dwelling Japanese subjects aged≥40 years were followed prospectively for 24 years. RESULTS: The age- and sex-adjusted incidence of coronary heart diseases (CHD) significantly increased with elevating non-HDLC levels (P for trend<0.001), but no such association was observed for ischemic and hemorrhagic strokes. With regard to ischemic stroke subtypes, the age- and sex-adjusted incidence of lacunar infarction significantly increased with elevating non-HDLC levels (P for trend<0.01), and such tendency was seen for atherothrombotic infarction (P for trend=0.098), while a significant inverse association was observed for cardioembolic infarction (P for trend=0.007). After adjustment for confounders, namely, age, sex, diabetes, body mass index, systolic blood pressure, electrocardiogram abnormalities, current drinking, current smoking, and regular exercise, the associations remained significant for CHD [adjusted hazard ratio (HR) for a 1 standard deviation of non-HDLC concentrations=1.17, 95% confidence interval (CI)=1.02 to 1.35], atherothrombotic infarction (adjusted HR=1.39, 95% CI=1.09 to 1.79), and cardioembolic infarction (adjusted HR=0.64, 95% CI=0.47 to 0.85). CONCLUSIONS: Our findings suggest that elevated non-HDLC levels are a significant risk factor for the development of atherothrombotic infarction as well as CHD but reduce the risk of cardioembolic infarction in the general Japanese population.