The ADRB3 Trp64Arg variant and BMI: a meta-analysis of 44 833 individuals.

BACKGROUND: The beta-3 adrenergic receptor gene (ADRB3) is part of the adrenergic system, which is known to play a key role in energy metabolism. The association between the Trp64Arg variant in the ADRB3 and body mass index (BMI) has been widely examined, but previous studies have been small and results have been inconsistent. METHODS: We assessed the association between the ADRB3 Trp64Arg variant and BMI in a large UK population-based cohort of 4854 middle-aged men and women. We also performed a meta-analysis of 97 studies, involving 44 833 individuals, to place our findings in context. RESULTS: Although we found no significant difference in BMI (0.20 kg/m(2), P=0.40) between the Trp64Trp homozygotes and Arg64 allele carriers in our UK population-based cohort, the meta-analysis showed significant association between the Arg64Trp variant and BMI, with Arg64-allele carriers having a 0.24 kg/m(2) (P=0.0002) higher BMI compared with noncarriers. However, we also found substantial heterogeneity among the studies (P=2.2 x 10(-14)). The difference in East Asians (0.31 kg/m(2), P=0.001) was 3.9 times larger than that in Europeans in whom no significant association was observed (0.08 kg/m(2), P=0.36). This was consistent with the chronological cumulative decrease in the effect size, which decreased steadily in Europeans and reached nonsignificance after 11 studies in 1996. In East Asians, the cumulative effect size decreased after the first reports, but reached a steady state at a significant effect size of 0.24 kg/m(2) in 2000. Although the funnel plot indicated no apparent publication bias, smaller studies tended to report greater differences in BMI, compared with larger studies. CONCLUSIONS: Collectively, these data suggest that the Trp64Arg ADRB3 genetic variant might be associated with BMI in East Asians, but not Europeans. More generally, our study shows the importance of meta-analyses in the field of genetic association studies for common traits. Each genetic variant makes only a small contribution to variation in BMI, and large sample sizes are needed to reliably assess and interpret gene-phenotype associations.

Advancement of magma fragmentation by inhomogeneous bubble distribution.

Decompression times reported in previous studies suggest that thoroughly brittle fragmentation is unlikely in actual explosive volcanic eruptions. What occurs in practice is brittle-like fragmentation, which is defined as the solid-like fracture of a material whose bulk rheological properties are close to those of a fluid. Through laboratory experiments and numerical simulation, the link between the inhomogeneous structure of bubbles and the development of cracks that may lead to brittle-like fragmentation was clearly demonstrated here. A rapid decompression test was conducted to simulate the fragmentation of a specimen whose pore morphology was revealed by X-ray microtomography. The dynamic response during decompression was observed by high-speed photography. Large variation was observed in the responses of the specimens even among specimens with equal bulk rheological properties. The stress fields of the specimens under decompression computed by finite element analysis shows that the presence of satellite bubbles beneath a large bubble induced the stress concentration. On the basis of the obtained results, a new mechanism for brittle-like fragmentation is proposed. In the proposed scenario, the second nucleation of bubbles near the fragmentation surface is an essential process for the advancement of fragmentation in an upward magma flow in a volcanic conduit.

5-HT2A promoter polymorphism is not associated with anorexia nervosa in Japanese patients.

Genetic factors have been implicated in playing a significant role in susceptibility to anorexia nervosa (AN). Among many candidate genes for AN, an association with the A allele of the -1438G/A polymorphism in the promoter region of the 5-HT2A receptor has been reported. However, these findings are controversial and all patients studied to date have been Caucasian. This study was designed to determine whether this association is reproducible in Japanese subjects. This case-control study of a cohort of 75 female Japanese AN sufferers and 127 normal female control subjects revealed no significant association between the 5-HT2A promoter polymorphism and AN. Thus, at least for Japanese subjects, the A-allele of the -1438G/A polymorphism in the promoter region of the 5-HT2A receptor gene does not contribute to a predisposition to AN.

Analysis of tumor necrosis factor-alpha gene promoter polymorphisms in anorexia nervosa.

Elevated plasma tumor necrosis factor-alpha (TNFalpha) levels and enhanced spontaneous TNFalpha release from peripheral blood mononuclear cells in patients with anorexia nervosa (AN) have been reported. TNFalpha activates the hypothalamic-pituitary-adrenal axis and reduces food intake, which is characteristic of eating disorders. Recently, three novel polymorphisms in the 5'-flanking region of the TNFalpha gene were reported at positions -1031 (T --> C substitution), -863 (C --> A) and -857 (C --> T). Differences in these alleles are reportedly related to altered TNFalpha-transcriptional promoter activity. Therefore, we performed a case-control association analysis to determine whether any of those three polymorphisms in the TNFalpha promoter region were involved in a predisposition to AN. The results of our analysis of a cohort of 79 female Japanese AN sufferers and 127 normal female control subjects provide no support for the hypothesis that -1031T/C, -863 C/A and -857C/T polymorphisms in the TNFalpha gene promoter region influence the susceptibility to AN.

Mapping of cyclosporin A binding sites in cyclophilin A by using synthetic peptides.

In order to map cyclosporin A (CsA) binding sites of cyclophilin (CyP), we synthesized the complete set of overlapping 157 octapeptides corresponding to human CyP A using the multi-pin peptide synthesis system. The pin-coupled synthetic octapeptides were examined in terms of binding ability to CsA by a modification of the enzyme-linked immunosorbent assay. Significant binding of CsA was detected with 35 synthetic N alpha-acetylated octapeptides possessing the N-terminal amino acids corresponding to the residues in positions 24-26, 42-44, 69-73, 75, 76, 89-91, 102, 116, 124-131, 144-151 and 152 in human CyP A, respectively. Other eight octapeptides showed moderate CsA binding activity. The distinct binding of octapeptides covering the C-terminal region of the CyP A was particularly significant. These data are to be compared with the information provided by X-ray and NMR studies on the CsA binding sites and furnish thus a test of the reported method. The present study also gave added insight into the CsA interaction sites of CyP.

Effects of compounds related to beta-hydroxyglutamic acid (BHGA) on identifiable giant neurones of an African giant snail (Achatina fulica Férussac).

The present study aimed to further elucidate the pharmacological features, with respect to sensitivity to L-BHGA agonists, of the receptors sensitive to beta-hydroxy-L-glutamic acid (L-BHGA) in five Achatina giant neurones: PON (periodically oscillating neurone), d-RPLN (dorsal-right parietal large neurone), VIN (visceral intermittently firing neurone), RAPN (right anterior pallial neurone) and v-RCDN (ventral-right cerebral distinct neurone). Of these neurones, d-RPLN and RAPN were depolarized by L-BHGA, while PON, VIN and v-RCDN were inhibited. Threo-beta-hydroxy-DL-aspartic acid markedly depolarized d-RPLN and RAPN (effective potency quotient (EPQ) in relation to the more effective L-BHGA isomer: 1 for d-RPLN and 0.3 for RAPN). This compound produced only slight inhibitory effects on PON, VIN and v-RCDN with EPQs calculated to be less than 0.03, less than 0.03 and 0.03, respectively. On the other hand, erythro-beta-hydroxy-DL-aspartic acid at 10(-3) M was almost ineffective, except on v-RCDN where it elicited some slight inhibitory effects (EPQ: 0.01). L-Aspartic and D-aspartic acid at 10(-3) M, also had almost no effect except for slight effects of D-aspartic acid on d-RPLN (EPQ: 0.1). N-Methyl-L- and N-methyl-D-aspartic acid were slightly effective only on v-RCDN (EPQ: less than 0.01 and 0.01, respectively). The other compounds, including beta-hydroxypyrroglutamic acid (cyclic BHGA) and proline derivatives, were almost ineffective at 10(-3) M; very weak effects were occasionally observed on some neurones.(ABSTRACT TRUNCATED AT 250 WORDS)

Change in solubility of insulin in therapeutic mixtures of insulin preparations: HPLC analysis.

We systematically and quantitatively observed the changes in solubility when short- and intermediate-acting insulin preparations were mixed together, using high-performance liquid chromatography (HPLC) and centrifugation. We tested 30 combinations of 16 kinds of commercial insulin preparations, including six short-acting and eight intermediate-acting preparations. The soluble insulin component in the short-acting preparations examined was in all cases insolubilized to various extents after mixing. Particularly remarkable was the complete precipitation of the insulin from Actrapid Human when it was mixed with Monotard Human (1:1). This sort of drastic change in insulin solubility after mixture was reflected in the changes in blood glucose level after the subcutaneous injection of the mixture to rabbits. The information provided by pharmaceutical companies on absorption kinetics and action profiles of short-acting insulin preparations may no longer be valid when mixtures with longer-acting preparations are used.

Demonstration of two distinct insulin-binding components in solubilized human placental membranes by radioimmunoassay using synthetic peptide and anti-synthetic peptide antibody.

A peptide corresponding to the 957-980 sequence of human placental insulin receptor precursor (HIRP) was synthesized and antisera were produced against the synthetic peptide. Anti-synthetic HIRP(957-980) serum HIR-27 was proved to cross-react with HIRP-related proteins in solubilized human placental membranes. A radioimmunoassay developed with the antiserum and synthetic peptide HIRP(957-980) enabled us to separate, in combination with gel filtration, two insulin-binding components in solubilized human placental membranes which conceivably correspond to the alpha 2 beta 2 and alpha beta structures of the placental insulin receptor. The two components were shown to be distinct in insulin-binding behavior depending on conditions of pH and ionic strength in the binding assay.

Enhancement effects of barbital on the teratogenicity of aminopyrine.

Aminopyrine and its compound with barbital have been used in humans as analgesics and antipyretics. A compound, pyrabital (2 molecules of aminopyrine and 1 molecule of barbital) given daily on Days 9, 10 and 11 of gestation produced significant yields of fetal deaths and malformations in ICR/Jcl mice. Most malformations induced were ruptured omphaloceles (eventration of the abdominal viscera), which were associated with malrotation of the intestine, cleft palates, and tail anomalies, finger and toe anomalies. Aminopyrine also induced significant yields of fetal deaths and malformations. However, the incidence of fetal deaths and malformations induced by a dose of pyrabital was significantly higher than that by an equivalent dose of aminopyrine which was contained in pyrabital. When aminopyrine (0.21 mg/g) and barbital (0.09 mg/g) were given in 2 separate injections to pregnant mice, teratogenicity was approximately equal to that by the equivalent dose of pyrabital (0.3 mg/g). Consequently, potent teratogenicity of pyrabital is not caused by the compound, but only by the coexistence of barbital and aminopyrine. Such enhancement effects of barbital may be due to the induction of enzymes responsible for transforming aminopyrine to teratogenic forms, because pretreatment with barbital and phenobarbital similarly enhanced embryotoxicity of aminopyrine.

Cytogenotoxicities of sublimed urethane gas to the mouse embryo.

Urethane (ethyl carbamate) which has long been used for commonly used drugs and has proven to be useful in the formation of products in every-day use, is volatile, and small amounts sublime spontaneously. Pregnant ICR mice were maintained in the vinyl chamber (45 liter) which was ventilated 4 times per hour. To inhale urethane gas, air was passed first through a glass bottle containing 500 g of crystalline urethane and then into the vinyl chamber. Concentration of the sublimed urethane gas in the chamber was 1.28 +/- 0.08 mg/l, and sublimed urethane gas produced significantly high incidence of chromosomal aberrations in the cells of whole embryo, when mice inhaled it for 48 h from day 9 to day 11 of pregnancy. High and significant incidence of chromosomal aberrations (36.0%) was detected in the embryo 3 h after urethane gas inhalation, but decreased to 5.3% at 24 h after exposure and showed no significant differences from controls after 48 h, while the incidence in bone marrow cells from the adult (pregnant) mice was lower (21.5%) at 3 h after exposure but a significant increase remained until 72 h after exposure. A majority of chromosomal aberrations was chromatid types. As a consequence of cellular damages by urethane gas inhalation during pregnancy, significantly high incidence of fetal deaths and congenital malformations (cleft palate, polydactyly, tail anomaly etc.) was induced in the offspring. Thus, we must be aware of the risk of volatile chemicals, because it is difficult to perceive and avoid hazardous exposure via respiration.

Gel chromatographic analysis of cyclosporin and its metabolites in human blood compartments.

Gel chromatography combined with specific and non-specific cyclosporin radioimmunoassays was adopted for quantitative analysis of cyclosporin and metabolites in free and protein-bound forms in blood compartments of kidney transplant patients. The analytical method was proved to be useful for the purpose, although plasma protein-bound forms of neither cyclosporin nor metabolites could be quantitated in the system. The present study also provided, by gel chromatographic analysis, additional examples to prove that concentrations of cyclosporin metabolites in blood compartments may not be deduced or inferred simply from those of cyclosporin.

In-vivo distribution and erythrocyte binding characteristics of cyclosporin in renal transplant patients.

The pharmacokinetic parameters of cyclosporin, a potent immunosuppressive agent, show large intra- and inter-individual variability, possibly because of the different analytical methods used. A recently developed cyclosporin-specific radioimmunoassay has been used to study the in-vivo distribution and binding characteristics of cyclosporin in whole blood, plasma and erythrocytes of fifteen renal transplant patients. The profiles of cyclosporin concentration-time curves after an oral dose of cyclosporin had either one peak (ten patients, group A) or two (five patients, group B). Essentially no difference was observed between the two groups in the relationship between equilibrium cyclosporin concentrations in erythrocyte and plasma as a function of whole-blood concentration. The equilibrium in-vivo cyclosporin concentrations in erythrocyte and plasma were, however, markedly lower than those previously observed under in-vitro conditions. The ratio of cyclosporin concentration in erythrocytes (CE) to that in plasma (CP) changed with time, in inverse proportion to the change in cyclosporin concentration in blood, over the range 0.63-2.80 in individual patients with an average of 1.36 +/- 0.07 (mean +/- s.e.m.) for group A and 1.42 +/- 0.23 for group B. The apparent cyclosporin binding affinity (Kd) to erythrocytes under in-vivo conditions averaged 452.2 +/- 47.6 nM (543.5 +/- 57.2 ng mL-1) for group A and 419.4 +/- 41.2 nM (504.1 +/- 49.5 ng mL-1) for group B, whereas apparent cyclosporin binding capacity (Bmax) of the blood cell averaged 0.83 +/- 0.07 nmol mL-1 for group A and 0.78 +/- 0.07 nmol mL-1 for group B. Significantly reduced average Kd (262.7 +/- 40.2 nM or 315.8 +/- 48.9 ng mL-1, P < 0.01) and Bmax (0.56 +/- 0.08 nmol mL-1, P < 0.05) values were observed during the period after Tmax (4-12 h after the drug ingestion) in group A patients. Apparent Kd and Bmax, determined by a nonlinear regression technique, were 131.6 +/- 29.4 and 1088.0 +/- 114.7 nM (158.2 +/- 35.4 and 1307.8 +/- 137.9 ng mL-1) and 0.178 +/- 0.024 and 0.814 +/- 0.078 nmol mL-1, respectively, during the 4-12 h period in group A patients. These findings reveal distinct differences in in-vivo distribution of cyclosporin and the binding characteristics of the compound to erythrocytes from those previously observed under in-vitro conditions. The significantly lower Kd of cyclosporin binding to erythrocytes during the elimination phase suggests a potential effect of cyclosporin-containing erythrocytes or of cyclosporin contained in erythrocytes during cyclosporin treatment.

Interrater agreement on the Coping Inventory for Stressful Situations (CISS).

The Coping Inventory for Stressful Situations (CISS) is a 48-item self-report inventory designed to measure three basic coping styles: Task Oriented, Emotion Oriented, and Avoidance Oriented coping. The psychometric properties of this inventory are promising, but CISS scores have not yet been shown to reflect behavioral variation in response to stress. This study was designed as a first step toward this end by examining the relationship between self- and peer-report on the CISS. One hundred and sixty-three pairs of friends completed the CISS, a peer form of the CISS, and a friendship questionnaire. Positive but modest correlations were found for each construct. Higher correlations were obtained when comparing scores across forms completed by the same informant, indicating that examinees believe their friends cope as they do themselves. Actual friend similarity was apparent only on Avoidance Oriented coping. Neither depth of relationship nor item observability moderated peer-self agreement.

Melatonin secretion in normal pressure hydrocephalus after cerebral aneurysm rupture--investigation before and after ventriculoperitoneal shunt.

Melatonin (MLT) secretion was examined in six normal pressure hydrocephalus (NPH) patients before and after ventriculoperitoneal (VP) shunt surgery. Ten healthy subjects were used as controls. Venous blood samples were taken daily at 2 p.m., 8 p.m., 2 a.m., and 8 a.m. Radioimmunoassay of MLT used a new specific antiserum and separation method achieving low cross-reactivity and high-efficiency MLT separation. Plasma levels in the control group at 2 p.m. and 2 a.m. were significantly different, showing diurnal rhythm (DR). The patients' MLT levels before VP shunt were significantly lower than control levels and the DR was absent. Postoperatively, the values were significantly different from preoperative values only at 2 a.m., but the DR reappeared. Thus, in NPH, VP shunt surgery improved the melatonin DR, probably through normalization of the dilated third ventricle.

[Melatonin secretion in intracranial tumor].

UNLABELLED: The secretion of melatonin (MLT) was examined in 6 ICT patients, 2 men and 4 women, who showed marked midline shift of CT scan findings with intracranial hypertension signs (headache, nausea, vomiting). Their ages ranged from 22 to 40 years. The controls, 6 healthy men and 4 healthy women (follicular stage) were aged from 18 to 35. With the subject recumbent, blood was drawn at 14, 20, 02 and 08 hour (Lights out, 9 pm; sunrise, 6 am). After separation of plasma by centrifugation, MLT was extracted by SEP-PAK C18 cartridge and determined by radioimmunoassay (RIA). ( METHOD: The extracted MLT, the antiserum and the 3H-MLT were incubated overnight at 4 degrees C, the antibody-combined and free 3H-MLT were separated using ammonium sulfate, and the radioactivity of the precipitate containing combined 3H-MLT was measured by liquid scintillation.) Plasma MLT concentrations of controls were 42.5 +/- 7.0 (Mean +/- SEM) pg/ml at 14 hr, 50.9 +/- 8.2 pg/ml at 20 hr, 165.1 +/- 22.7 pg/ml at 02 hr, and 49.1 +/- 5.4 pg/ml at 08 hr. The value of 14 hr. significantly differed from 02 hr, (P less than 0.01) showing a diurnal rhythm. The patients' MLT levels (pg/ml) were 19.8 +/- 6.2 at 14 hr, 15.9 +/- 5.2 at 20 hr, 52.0 +/- 17.2 at 02 hr, and 20.3 +/- 3.5 at 08 hr. These values were significantly lower than in controls (P less than 0.01) except at 14 hr. (P less than 0.05), showing no diurnal rhythm except 2 cases. In conclusion, in ICT with midline shift of CT scan melatonin secretion was inhibited and diurnal rhythm disappeared in four cases (67%).

Characteristic activities of lower limbs with body weight support ratio.

In this study, we investigated a lower limbs muscle activity during body weight support treadmill training (BWSTT). Informed consent was obtained from 16 healthy men. Experimental system consists of force plate, treadmill, three-dimensional motion analysis system, electromyograph, and body weight support device. Body weight support (BWS) was set every 15% increase from 0% to 45%. Walking speed was 4.17 km/h. The measurement data were reaction forces, joint angles, joint moments and lower limbs muscle activities. The vertical reaction force shows two peaks. Two peaks decreased with increase of BWS together. Joint angles did not show significant changes with BWS. However, only the extension of hip angle was decreased with BWS. The peaks of joint moment were decreased. Decrease of ankle joint moment was greatest compared with other moment. Decrease of peaks of muscle activity by BWS was observed during stance phase, and did not almost change during swing phase.

Improvement of the maxillary bone growth suppression in the cleft palate operation with cultured dermal substitute: animal experiment and patient reports in preliminary clinical application.

Cleft palate patients often show impaired maxillary bone growth after cleft-palate-correction surgery. We attempted to investigate and elucidate the effects of using allogeneic, cultured dermal substitute (CDS) to cover an exposed, palatal bone surface in animal experiments. Fibroblasts from the abdominal skin of Wistar rats were cultured. Subsequently, the fibroblasts were seeded onto a matrix that composed of hyaluronic acid and atelo-collagen. Forty Wistar rats (3-week-old males) were assigned to one of four groups: control, open-treatment, matrix and CDS groups. The control group (n=5) received no surgical operations. In the open-treatment group (n=11), the mucosa and periosteum of the left-half of the palate were removed surgically and the bone was exposed. In the matrix group (n=11), the area of exposed bone was covered with only the matrix, excluding any cells. In the CDS group (n=10), the area of exposed bone was covered with CDS. At 9 weeks postoperatively, biopsies of the wounds were obtained. Skull preparations were made and the palatal widths were determined. The palatal widths in the CDS group were significantly wider compared to the matrix and open-treatment groups (P<0.05). However, there were no significant differences when the CDS group was compared to the control group. Haematoxylin, eosin and CD31 immunostaining confirmed a larger number of capillaries in the CDS group. This animal experiment suggested that this procedure might provide an optimum wound-healing condition, thus, reducing the maxillary bone-growth suppression. Therefore, a preliminary clinical application in three patients was performed using the autologous CDS after the pushback method.

Lambda incision for effective tissue expansion.

By using a rectangular type tissue expander, the skin is expanded as a cuboid. To cover a skin defect effectively, it is necessary to cut the expanded skin and to spread it out as a flap. We designed the lambda incision of the lateral wall of the expanded cuboid. In principle we make one lambda incision on each lateral wall (a total of 2 incisions). However, if the defect is triangular, one lambda on one side is sufficient. If it is a trapezoid, one lambda on one side and two lambdas on the opposite side are used. The lambda incision was applied in 11 cases. In almost all, the expanded skin could be spread out well and the lesions could be resected. There were no complication due to lambda incisions.