Involvement of regulatory elements on corticotropin-releasing factor gene promoter in hypothalamic 4B cells.

INTRODUCTION: Corticotropin-releasing factor (CRF) plays a central role in controlling the hypothalamic-pituitary-adrenal (HPA) axis during stressful periods. CRF is synthesized and secreted in the hypothalamic paraventricular nucleus (PVN) in response to stress, and stimulates ACTH in the pituitary corticotrophs. ACTH stimulates the release of glucocorticoids from the adrenal glands, and glucocorticoids sequentially inhibit hypothalamic PVN production of CRF and pituitary production of ACTH. The effects of glucocorticoids on CRF gene regulation, however, are possibly tissue-specific since glucocorticoids stimulate CRF gene expression in the placenta and the bed nucleus of the stria terminalis, while they inhibit it in the hypothalamus. METHODS AND RESULTS: In a hypothalamic cell line, 4B, we found that forskolin-stimulated CRF gene transcription was mediated by a functional cAMP-response element (CRE), which included -220 to -233 bp on the CRF 5'-promoter region. Protein kinase A, protein kinase C, and p38 mitogen-activated protein kinase pathways contributed to forskolin-induced transcriptional activity of CRF in hypothalamic 4B cells. Glucocorticoid-dependent repression of cAMP-stimulated transcriptional activity of CRF was localized to promoter sequences between -278 and -233 bp, which included a glucocorticoid regulatory element and a serum response element. CONCLUSION: Taken together, these findings indicate that the regulatory elements, including CRE, negative glucocorticoid regulatory element, and a serum response element on the promoter, contribute to the regulation of CRF gene transcription in hypothalamic 4B cells.

A case of thyrotropin-producing pituitary adenoma, accompanied by an increase in anti-thyrotropin receptor antibody after tumor resection.

We describe a rare, but interesting, case of TSH-producing adenoma (TSHoma), accompanied by increases in both anti-TSH receptor antibody (TRAb) and thyroid-stimulating antibody (TSAb) after tumor resection. A 21-yr-old woman was referred to our department for further evaluation of pituitary tumor. In a nearby hospital, she had been diagnosed as having pituitary tumor. Her serum free T4, free T3, and TSH levels were all elevated concomitantly. On the basis of a diagnosis of pituitary adenoma with TSH production, transsphenoidal resection of the pituitary adenoma was performed. Two weeks after the operation, the blood concentrations of TSH were undetectable, whereas both TRAb and TSAb levels were elevated. TSAb levels gradually increased further from 2 weeks to 3 months after the operation, accompanied by an increase in TSH and free T4 levels. TSH is an important hormone in maintaining physiology and regulating immunomodulators in thyrocytes, as it can influence a variety of immune-regulating cytokine-like activities and inhibit expressions of Fas antigen, intracellular adhesion molecule-1, and class II trans-activator. Changes in TSH would modulate the immune circumstances in the thyroid, and then induce TRAb and TSAb. Autoimmune parameters with thyroid function should be observed carefully when managing patients with TSHoma.

Gene analysis of the calcium channel 1 subunit and clinical studies for two patients with hypokalemic periodic paralysis.

Hypokalemic periodic paralysis (HypoPP) is a skeletal muscle disorder in which episodic attacks of muscle weakness occur; they are associated with decreased serum potassium (K+) levels. Recent molecular approaches have clarified that the condition is caused by mutations in the skeletal muscle voltage-gated calcium channel 1 subunit (CACNA1S). We describe two unrelated patients with HypoPP, followed by their relevant clinical studies and gene analysis. Clinical studies included an oral glucose tolerance test (OGTT), food-loading and insulin tolerance tests (ITT). For Case 1, serum K+ levels were extremely decreased following insulin tolerance testing compared with levels for controls. These results support the hypothesis that no efflux of K+ ion occurs in patients because of low activity of adenosine triphosphate (ATP)-sensitive K+ channel (KATP) channels. Mutational analysis of the CACNA1S gene showed a duplicate insertion of 14 base pairs (bp) from 52 to 65 in intron 26, present in the heterozygous state in both patients. No other mutations were detected in the CACNA1S gene, the muscle sodium channel gene (SCN4A) or the voltage-gated K+ channel gene (KCN3) of either patient. Further analysis showed that this duplicate insertion of 14 bp in intron 26 of the CACNA1S gene was found in 23.7% of healthy subjects. K+ dynamics studies are useful for confirming this syndrome, while further gene analysis for various ion channels using amplification and direct sequencing are required to evaluate the molecular basis of the disorder in the individual patient.

Urocortin III, a brain neuropeptide of the corticotropin-releasing hormone family: modulation by stress and attenuation of some anxiety-like behaviours.

Following its discovery 20 years ago, corticotropin-releasing hormone (CRH) has been postulated to mediate both hormonal and behavioural responses to stressors. Here, we characterize and describe a behavioural role for the murine gene, UcnIII, which encodes a recently discovered CRH-related neuropeptide, urocortin III. We found that mouse UcnIII is expressed predominantly in regions of the brain known to be involved in stress-related behaviours, and its expression in the hypothalamus increases following restraint. In addition, we found that intracerebroventricular administration of mUcnIII stimulates behaviours that are associated with reduced anxiety, including exploration of an open field and decreased latency to enter the lit compartment of a dark-light chamber, but has no effect on the elevated-plus maze. Finally, we found that mUcnIII does not exert any effects on the hormonal stress response. Based upon our findings, UcnIII may be an endogenous brain neuropeptide that is modulated by stress and stimulates behaviours associated with reduced anxiety. In this capacity, UcnIII may attenuate stress-related behaviours, which may be useful both to help cope with stressful situations as well as to avoid pathology associated with excessive reaction to stressors.

Psychosocial correlates of depressive symptoms among Japanese high school students.

PURPOSE: To determine the psychosocial factors associated with the presence and persistence of depressive symptoms among high school students in Okinawa, Japan. METHODS: The study sample was 3202 students from 12 public senior high schools. Students completed self-administered questionnaires from October through December 1997. We measured depressive symptomatology using the Center for Epidemiologic Studies Depression Scale. We asked students to report whether they had depressive symptoms at any time in the immediate past week, and whether those symptoms persisted for 5-7 days. The psychosocial variables examined were life stressors, perceived social support, health practices, self-esteem, and Locus of Control. The relationship between the psychosocial factors and depressive symptoms was examined using hierarchical multiple regression analyses. RESULTS: After controlling for the effects of demographic and other psychosocial variables, presence of depressive symptoms was positively associated with life stressors in the domains of friends, family, and teachers. Similarly, persistence of depressive symptoms was also positively associated with life stressors in the domains of friends and teachers. Presence and persistence of depressive symptoms were negatively associated with positive health practices, more social support, high self-esteem, and internal Locus of Control. CONCLUSIONS: The psychosocial variables associated with presence and persistence of depressive symptoms were remarkably consistent. Life stressors might be risk factors; on the contrary, positive health practices, perceived social support, high self-esteem, and internal Locus of Control might be protective factors of depressive symptoms among Japanese adolescents.

Patterns of health-risk behavior among Japanese high school students.

This study explored patterns of health-risk behaviors among Japanese high school students and examined if a cluster and an accumulation of health-risk behaviors existed. Self-administered questionnaires were employed in 1999 using a sample of 1,466 students (male 50.5%, female 49.5%) in grades 10 through 12 at seven public senior high schools in Okinawa, Japan. Health-risk behaviors studied included cigarette smoking, alcohol use, thinner use, nonuse of seat belts, suicide ideation, sexual intercourse, weight loss practices, and physical inactivity. Among male and female students, cigarette smoking, alcohol use, and sexual intercourse clustered. Accumulation of these risk behaviors also occurred because the observed proportion was greater than the expected proportion assuming independent occurrence. Vocational high school students and upper graders were strongly associated with accumulation of health risk behaviors. These findings identify a high-risk target group among Japanese adolescents and suggest that preventive intervention strategies should take into consideration the cluster and accumulation of health-risk behaviors.

[Factors related to ADL of stroke patients three months after discharge].

Focusing on the ADL of stroke patients, 94 hemiplegia cases in rehabilitation units of acute care general hospitals were studied to identify factors affecting ADL, measured using the Barthel Index, 3 months after discharge. The main results were as follows: 1. The 8 independent variables: age, gender, length of hospitalization, degree of paralysis of lower extremities at discharge, motivation toward rehabilitation, sight, Barthel scores at discharge, and the expression of one's intentions emerged as the 8 reliable predictive factors (R = .904, R2 = .817). 2. It may be deduced that in older patients in particular, a status of amyotrophy has a significant influence on ADL after discharge. The 94 stroke patients were divided into two age groups-44 patients aged 64 years or younger, and 50 patients aged 65 years or older. Barthel scores at 3 months after discharge were analyzed by multiple regression analysis in both groups. The result was that ADL, after discharge, of the aged 65 years or older group, was more affected by their status at discharge, than was it with the 64 years or younger group (64 years or younger; R = .871, R2 = .758, 65 years or older; R = .934, R2 = .873). 3. For patients in the 64 years or younger group, Barthel scores at 3 months after discharge were significantly related to age, length of hospitalization, sight, Barthel scores at discharge and expression of one's intentions. For patients in the 65 years or older group, the results were significantly related to sight, Barthel scores at discharge, and motivation for rehabilitation.

[The relationship of chewing ability to nutrient and food intakes in the community elderly].

The purpose of the present study is to investigate the relationship of chewing ability to nutrient and food intakes in the community elderly. Subjects consisted of 145 elderly residents (56 males, 89 females) ranging in age from 65 to 79 years, living at home in a village of Okinawa Prefecture. Chewing ability was divided into two categories: being able to chew ordinary foods was defined as "good", and being able to chew only soft foods as "bad". To determine quantitative food intakes 24-hour dietary records were utilized which were confirmed by nutritionists during interviews. Good chewing ability was found in 82.1% for males and 83.1% for females with no statistically significant difference concerning age or sex. Teeth status was significantly related to chewing ability. For both males and females, protein intake was lower in the group with bad chewing ability than in the good group. Only in males intakes of total energy, fats, Ca, and Fe were lower in the group with bad chewing ability. Concerning food items, intakes of dark green and yellow vegetables and fat in males, and intake of rice in females, were lower in the group with bad chewing ability.

[Risk factors for falls among the elderly living in a Japanese rural community].

The purpose of the present study is to explore risk factors for falls among the elderly in a Japanese rural community. Subjects comprised 658 elderly persons aged 65 and over living in a rural village of Akita Prefecture (response rate; 96.1%). In 1988, the baseline medical health examinations and an interview survey were performed and repeated at the follow-up survey one year later. Results obtained were as follows; 1) The rate of falls at the follow-up survey was 17.3% in men, 15.6% in women. There was no difference in rate of falls between sexes. The rate of falls in the old-old did not significantly differ from that in the young-old in both sexes. 2) Chi square test or Cochran-Armitage test was performed by sex to examine the relationship between the factors investigated at the baseline survey and occurrence of falls at the follow-up survey. Significant factors in men were income, spouse, need for assistance in daily living, history of stroke, fall experience within one year preceding the baseline survey, ECG abnormalities, standing time with eyes open, grip strength, TMIG (Tokyo Metropolitan Institute of Gerontology) index of competence. In women, mobility, need for assistance in daily living, admission to a hospital within one year preceding the baseline survey, fall experience within one year preceding the baseline survey, standing time with eyes open, grip strength, TMIG index of competence were significant. 3) Using the significant variables in univariate analysis, multivariate logistic regression analysis controlling for age was performed by sex. Significant risk factors for falls in men were fall experience within one year preceding the baseline survey and ECG abnormalities, while for women fall experience within one year preceding the baseline survey was significant. The present longitudinal study revealed that falls experience within one year preceding the baseline survey was the highest risk factor for falls in the elderly residents in the community. This indicates that even a simple interview on fall experience can identify high risk elderly.

[Chewing ability in relation to physical health status].

The purpose of the present study is to explore relationship of chewing ability to physical health status in the elderly. Subjects were 722 community elderly adults aged 65 years and over (participation rate 88.6%). They were divided into 2 groups according to chewing ability as rated by a questionnaire: good and bad. Results obtained were as follows; 1. Those who had good ability of chewing accounted for 86% of the subjects. The chewing ability was determined by dental status. Those with more natural teeth were better able to chew. 2. Multiple regression analyses controlled by age and sex showed that good chewing ability had significantly positive effect on weight, body mass and length of time for standing on one leg with eyes open.

Diagnosis of a case of Gitelman's syndrome based on renal clearance studies and gene analysis of a novel mutation of the thiazide-sensitive Na-Cl cotransporter.

Gitelman's syndrome is a recessively inherited renal tubular disorder characterized by low plasma potassium and magnesium levels, reduced calcium excretion, metabolic alkalosis, and increased plasma renin activity and plasma aldosterone concentration with normal blood pressure levels. A 23-yr-old man was referred to our department for further evaluation of hypokalemia. The patient also had hypomagnesemia and markedly reduced urinary calcium excretion. Renal clearance studies and gene analysis of the thiazide-sensitive Na-Cl cotransporter (TSC) were performed in the patient. In response to an iv injection of furosemide, chloride clearance (CCl) increased markedly, while distal fractional chloride reabsorption CH2O/(CH2O+CCl) was considerably reduced. In contrast, thiazide ingestion had no significant effects on these parameters. The patient had compound heterozygous mutations in the alleles encoding the TSC gene, one of which has not been formerly reported. Renal clearance studies and TSC gene analysis by amplification and direct sequencing are useful diagnostic tools for confirming a diagnosis of Gitelman's syndrome.

Gender differences in the association between psychosocial factors and depressive symptoms in Japanese junior high school students.

To determine psychosocial factors associated both with depressive symptoms and with gender differences in depressive symptoms among junior high school students, we conducted self-administered questionnaires using a sample of 2,660 students of 13 public junior high schools in Okinawa, Japan. Depressive symptomatology was measured by the Center for Epidemiologic Studies Depression Scale (CES-D). The psychosocial variables examined were life stresses, social support, health practices, self-esteem, and locus of control. Depressive symptoms were positively associated with life stresses and negatively associated with health practices, social support, self-esteem, and internal locus of control. In addition, female students were likely to report more depressive symptoms, life stresses, and low self-esteem and poor health practices. After controlling for the psychosocial variables differed by gender, gender differences in depressive symptoms were eliminated. In conclusion, for depressive symptoms of junior high school students, life stresses might be risk factors, but positive health practices, social support, high self-esteem, and internal locus of control might be protective factors. Gender differences in depressive symptoms could be explained by the females' elevation on these psychosocial variables.

Covariation of cigarette smoking and other health-risk behaviors among Japanese high school students: a preliminary study.

This study aims to determine if cigarette smoking is associated with engaging in other health-risk behaviors among high school students in Japan. Self-administered anonymous questionnaires were conducted in 1999 using a sample of 1,466 students (male: 50.5%, female: 49.5%) in grades 10 through 12 at seven public senior high schools in urban areas of Okinawa, Japan. Health-risk behaviors studiedal included cigarette smoking, alcohol drinking, thinner use, sexual intercourse, suicidal ideation, nonuse of seat belts, physical inactivity, and weight loss practices. In the logistic regression models, controlled for sociodemographic variables, smoking was significantly associated with all health-risk behaviors except physical inactivity. In particular, associations of alcohol drinking and sexual intercourse with smoking were strong. Among male students, statistically significant odds ratios existed for alcohol drinking, sexual intercourse, and nonuse of seat belts. Among female students, all of the odds ratios for health-risk behaviors were statistically significant, except for physical inactivity. Generally, the odds ratios of female students were higher than those of male students. In conclusion, high school students who smoked cigarettes in this study may be at higher risk for engaging in other health-risk behaviors. Particularly, alcohol drinking and sexual intercourse are more likely to co-occur with smoking. These findings suggest that smoking prevention programs should be integrated with other health-risk behaviors.

Risk factors for cigarette smoking among high school students in Okinawa, Japan.

The purpose of this study was to examine the relationships between risk factors and smoking status among high school students in Okinawa, Japan. We also examined if there is a dose-response relation between the number of risk factors and smoking status. Self-reported questionnaires including smoking status and potential risk factors were conducted using a sample of 1,029 students of a public senior high school in Okinawa. The percentage of smokers was 40.0% for males and 10.6% for females, and it was significantly higher for males than females. As a result of multivariate analysis, we identified four significant risk factors; peer smoking, attitude of peer toward subject smoking, intention to smoke at the age 20, and alcohol drinking. The number of these risk factors was linearly associated with increased percentage of smokers, and a linear trend was significant for both gender students. Additionally, magnitude of risk for smoking among females became considerably great compared with those of males as the number of risk factors increased. In conclusion, this study was the first study in Japan to indicate a significant dose-response relationship between the number of risk factors and smoking status among high school students. We also found that females with many risk factors had extremely increased vulnerability to smoking compared to male counterpart. These findings may be useful to identify high-risk students who need more intensive smoking prevention programs and to develop the content of effective interventions.

Neuropeptide Y potentiates the luteinizing hormone (LH) response to LH-releasing hormone in men.

It has been demonstrated in several animal species that neuropeptide Y (NPY) exerts a modulatory effect on luteinizing hormone (LH) secretion. However, whether NPY plays a similar role also in humans has yet to be determined. Therefore, in this study we examined the effect of human NPY on the anterior pituitary hormone secretion in 6 normal men. Intravenous bolus injection of 100 micrograms of human NPY alone did not affect the secretion of any anterior pituitary hormone or cortisol. However, when NPY (100 micrograms) was administered simultaneously with LH-releasing hormone (LHRH, 100 micrograms), a significant potentiation was observed for LHRH-induced LH secretion. Similarly, follicle-stimulating hormone (FSH) response to LHRH was slightly potentiated by the coadministration of NPY, although this effect was not statistically significant. This is the first study to demonstrate that NPY can augment the LHRH-induced LH secretion in humans.

A role for neuromedin U in stress response.

Neuromedin U (NMU) is a hypothalamic peptide that has been recently found to reduce food intake, but few is known about its other functions in the central nervous system. We here studied behavioral activities induced by an intracerebroventricular (ICV) administration of NMU in rats and mice. NMU increased gross locomotor activity, face washing behavior, and grooming. NMU-induced stress response was significantly abolished by pretreatment with an antagonist of corticotropin-releasing hormone (CRH), alpha-helical CRH (9-41) (alpha-hCRH), or anti-CRH IgG. NMU did not induce locomotor activity in CRH knockout mice. NMU that interacts anatomically and/or functionally with the CRH system is a novel physiological regulator of stress response.