Possible association of oestrogen and Cryba4 with masticatory muscle tendon-aponeurosis hyperplasia.

OBJECTIVE: Masticatory muscle tendon-aponeurosis hyperplasia, which is associated with limited mouth opening, progresses very slowly from adolescence. The prevalence rates of this disease are higher among women than among men, suggesting oestrogen involvement. As parafunctional habits are frequently observed, mechanical stress is likely involved in the pathogenesis and advancement of this disease. To elucidate the pathological condition, we examined the effect of oestrogen on tenocyte function and the relationship between mechanical stress and crystallin beta A4 (Cryba4), using murine TT-D6 tenocytes. MATERIALS AND METHODS: Cell proliferation assays, RT-PCR, real-time RT-PCR, Western blot analysis and mechanical loading experiments were performed. RESULTS: The physiological dose of oestrogen increased the levels of scleraxis and tenomodulin in TT-D6 tenocytes. In contrast, forced expression of Cryba4 inhibited scleraxis expression in these cells. Surprisingly, oestrogen significantly promoted cell differentiation in the Cryba4-overexpressing TT-D6 tenocytes. Moreover, tensile force induced Cryba4 expression in these tendon cells. CONCLUSION: Oestrogen and Cryba4 may be associated with the progression of masticatory muscle tendon-aponeurosis hyperplasia.

Predictive performance of a genetic risk score using 11 susceptibility alleles for the incidence of Type 2 diabetes in a general Japanese population: a nested case-control study.

AIMS: To assess the predictive ability of a genetic risk score for the incidence of Type 2 diabetes in a general Japanese population. METHODS: This prospective case-control study, nested within a Japan Public Health Centre-based prospective study, included 466 participants with incident Type 2 diabetes over a 5-year period (cases) and 1361 control participants, as well as 1463 participants with existing diabetes and 1463 control participants. Eleven susceptibility single nucleotide polymorphisms, identified through genome-wide association studies and replicated in Japanese populations, were analysed. RESULTS: Most single nucleotide polymorphism loci showed directionally consistent associations with diabetes. From the combined samples, one single nucleotide polymorphism (rs2206734 at CDKAL1) reached a genome-wide significance level (odds ratio 1.28, 95% CI 1.18-1.40; P = 1.8 × 10-8 ). Three single nucleotide polymorphisms (rs2206734 in CDKAL1, rs2383208 in CDKN2A/B, and rs2237892 in KCNQ1) were nominally significantly associated with incident diabetes. Compared with the lowest quintile of the total number of risk alleles, the highest quintile had a higher odds of incident diabetes (odds ratio 2.34, 95% CI 1.59-3.46) after adjusting for conventional risk factors such as age, sex and BMI. The addition to the conventional risk factor-based model of a genetic risk score using the 11 single nucleotide polymorphisms significantly improved predictive performance; the c-statistic increased by 0.021, net reclassification improved by 6.2%, and integrated discrimination improved by 0.003. CONCLUSIONS: Our prospective findings suggest that the addition of a genetic risk score may provide modest but significant incremental predictive performance beyond that of the conventional risk factor-based model without biochemical markers.

Cytotoxicity of Natural Killer Cells Activated Through NKG2D Contributes to the Development of Bronchiolitis Obliterans in a Murine Heterotopic Tracheal Transplant Model.

Bronchiolitis obliterans after lung transplantation is a major cause of postoperative mortality in which T cell-mediated immunity is known to play an important role. However, the exact contribution of natural killer (NK) cells, which have functions similar to CD8+ T cells, has not been defined. Here, we assessed the role of NK cells in murine bronchiolitis obliterans through heterotopic tracheal transplantations and found a greater percentage of NK cells in allografts than in isografts. Depletion of NK cells using an anti-NK1.1 antibody attenuated bronchiolitis obliterans in transplant recipients compared with controls. In terms of NK cell effector functions, an improvement in bronchiolitis obliterans was observed in perforin-KO recipient mice compared to wild type (WT). Furthermore, we found upregulation of NKG2D-ligand in allografts and demonstrated the significance of this using grafts expressing Rae-1, a murine NKG2D-ligand, which induced severe bronchiolitis obliterans in WT and Rag-1 KO recipients. This effect was ameliorated by injection of anti-NKG2D blocking antibody. Together, these results suggest that cytotoxicity resulting from activation of NK cells through NKG2D leads to the development of murine bronchiolitis obliterans.

A cluster randomized trial on the effect of a multifaceted intervention improved the technical quality of diabetes care by primary care physicians: The Japan Diabetes Outcome Intervention Trial-2 (J-DOIT2).

AIMS: To evaluate the effect of multifaceted interventions using the Achievable Benchmark of Care (ABC) method for improving the technical quality of diabetes care in primary care settings. METHODS: We conducted a 1-year cluster randomized controlled trial in 22 regions divided into an intervention group (IG) or control group (CG). Physicians in the IG received a monthly report of their care quality, with the top 10% quality of diabetes care scores for all physicians being the achievable benchmark. The change in quality-of-care scores between the IG and CG during follow-up was analysed using a generalized linear model considering clustering. RESULTS: A total of 2199 patients were included. Their mean (sd) age was 56.5 ± 5.9 years and the mean (sd) HbA1c level was 56.4 ± 13.3 mmol/mol (7.4 ± 1.2%). The quality-of-care score in the CG changed from 50.2%-point at baseline to 51%-point at 12 months, whereas the IG score changed from 49.9%-point to 69.6%-point, with statistically significant differences between the two groups during follow-up [the effect of intervention was 19.0%-point (95% confidence interval 16.7%- to 21.3%-point; P < 0.001)]. CONCLUSIONS: Multifaceted intervention, measuring quality-of-care indicators and providing feedback regarding the quality of diabetes care to physicians with ABC, was effective for improving the technical quality of care in patients with Type 2 diabetes in primary care settings. ( TRIAL REGISTRATION: umin.ac.jp/ctr as UMIN000002186).

Short sleep duration in association with CT-scanned abdominal fat areas: the Hitachi Health Study.

OBJECTIVE: To examine the relationship between short sleep duration and body mass index (BMI), waist circumference (WC), visceral fat area (VFA) and subcutaneous fat area (SFA) among a working population in Japan. DESIGN: Health-center-based, cross-sectional study. SUBJECTS: The study subjects included 5400 men and 642 women aged 30 to 75 years who underwent an abdominal computed tomography (CT) scanning examination in a comprehensive health checkup. MEASUREMENTS: Height and weight were measured, and BMI was calculated. WC, VFA and SFA were measured using a CT scanner. Sleep duration was self-reported. Analysis of covariance was used to estimate adjusted means of BMI, WC, VFA and SFA across categories of sleep duration with adjustments for potential confounders. Trend of the association was assessed using multiple linear regression analysis. RESULTS: In men, the mean values of BMI, WC and SFA decreased with increasing sleep duration after adjustment for age, physical activity, smoking and drinking (P-value for trend <0.001). Additional adjustment for physical illnesses did not attenuate the explanatory power of the models (P-value for trend <0.001). In addition, the association between sleep duration and SFA did not change after controlling for VFA (P-value for trend <0.001). The mean values of SFA for subjects sleeping '<5 h', '5 to <6 h', '6 to <7 h' and '7 h' per day were 145.8±67.4 cm(2), 138.7±61.5 cm(2), 134.7±60.4 cm(2) and 132.5±49.2 cm(2), respectively. Sleep duration was not appreciably associated with VFA. In women, no significant association was detected in any models. CONCLUSION: Shorter sleep duration is associated with higher BMI, WC and SFA in men. Further research is needed to explicate the biological mechanisms behind these relationships and to see whether interventions addressing inadequate sleep could treat or prevent obesity by taking gender differences into consideration.

Depletion of homeodomain-interacting protein kinase 3 impairs insulin secretion and glucose tolerance in mice.

AIMS/HYPOTHESIS: Insufficient insulin secretion and reduced pancreatic beta cell mass are hallmarks of type 2 diabetes. Here, we focused on a family of serine-threonine kinases known as homeodomain-interacting protein kinases (HIPKs). HIPKs are implicated in the modulation of Wnt signalling, which plays a crucial role in transcriptional activity, and in pancreas development and maintenance. The aim of the present study was to characterise the role of HIPKs in glucose metabolism. METHODS: We used RNA interference to characterise the role of HIPKs in regulating insulin secretion and transcription activity. We conducted RT-PCR and western blot analyses to analyse the expression and abundance of HIPK genes and proteins in the islets of high-fat diet-fed mice. Glucose-induced insulin secretion and beta cell proliferation were measured in islets from Hipk3 ( -/- ) mice, which have impaired glucose tolerance owing to an insulin secretion deficiency. The abundance of pancreatic duodenal homeobox (PDX)-1 and glycogen synthase kinase (GSK)-3ß phosphorylation in Hipk3 ( -/- ) islets was determined by immunohistology and western blot analyses. RESULTS: We found that HIPKs regulate insulin secretion and transcription activity. Hipk3 expression was most significantly increased in the islets of high-fat diet-fed mice. Furthermore, glucose-induced insulin secretion and beta cell proliferation were decreased in the islets of Hipk3 ( -/- ) mice. Levels of PDX1 and GSK-3ß phosphorylation were significantly decreased in Hipk3 ( -/- ) islets. CONCLUSIONS/INTERPRETATION: Depletion of HIPK3 impairs insulin secretion and glucose tolerance. Decreased levels of HIPK3 may play a substantial role in the pathogenesis of type 2 diabetes.

Haemoglobin A1c cut-off point to identify a high risk group of future diabetes: results from the Omiya MA Cohort Study.

AIMS: Using the HbA(1c) level to define diabetes has several advantages and these advantages also apply to define a high-risk group. However, the risk of diabetes increases as HbA(1c) increases and a certain degree of arbitrariness in the cut-off for the high risk group is unavoidable. The aim of this study was to determine the HbA(1c) cut-off for defining a high-risk group that corresponds to the fasting plasma glucose cut-off by comparing the risk of diabetes against the fasting plasma glucose and HbA(1c) levels in the Japanese population. METHODS: A retrospective cohort study was conducted using data from annual health examinations performed in Omiya city. A total of 11,271 subjects between the ages of 40 and 79 years without diabetes at baseline were followed for up to 7 years. According to the new diagnostic criteria, diabetes was defined as an fasting plasma glucose level ≥ 7 mmol/l or an HbA(1c) level ≥ 48 mmol/mol (≥ 6.5%) or a self-report. The HbA(1c) cut-off corresponding to the fasting plasma glucose cut-off was determined using the incidence, hazard ratio, and a receiver operating characteristic analysis. RESULTS: Eight hundred and sixty subjects developed diabetes. The incidence, hazard ratio, and receiver operating characteristic analysis all indicated that an HbA(1c) cut-off of 39 mmol/mol (5.7%) corresponded to an fasting plasma glucose level of 5.6 mmol/l. CONCLUSIONS: Our results suggested that the HbA(1c) cut-off for high-risk of diabetes should be 39 mmol/mol (5.7%), consistent with the American Diabetes Association recommendation. Further research is needed to determine whether our results are applicable to other populations.

Borg proteins control septin organization and are negatively regulated by Cdc42.

The Cdc42 GTPase binds to numerous effector proteins that control cell polarity, cytoskeletal remodelling and vesicle transport. In many cases the signalling pathways downstream of these effectors are not known. Here we show that the Cdc42 effectors Borg1 to Borg3 bind to septin GTPases. Endogenous septin Cdc10 and Borg3 proteins can be immunoprecipitated together by an anti-Borg3 antibody. The ectopic expression of Borgs disrupts normal septin organization. Cdc42 negatively regulates this effect and inhibits the binding of Borg3 to septins. Borgs are therefore the first known regulators of mammalian septin organization and provide an unexpected link between the septin and Cdc42 GTPases.

Adiponectin stimulates glucose utilization and fatty-acid oxidation by activating AMP-activated protein kinase.

Adiponectin (Ad) is a hormone secreted by adipocytes that regulates energy homeostasis and glucose and lipid metabolism. However, the signaling pathways that mediate the metabolic effects of Ad remain poorly identified. Here we show that phosphorylation and activation of the 5'-AMP-activated protein kinase (AMPK) are stimulated with globular and full-length Ad in skeletal muscle and only with full-length Ad in the liver. In parallel with its activation of AMPK, Ad stimulates phosphorylation of acetyl coenzyme A carboxylase (ACC), fatty-acid oxidation, glucose uptake and lactate production in myocytes, phosphorylation of ACC and reduction of molecules involved in gluconeogenesis in the liver, and reduction of glucose levels in vivo. Blocking AMPK activation by dominant-negative mutant inhibits each of these effects, indicating that stimulation of glucose utilization and fatty-acid oxidation by Ad occurs through activation of AMPK. Our data may provide a novel paradigm that an adipocyte-derived antidiabetic hormone, Ad, activates AMPK, thereby directly regulating glucose metabolism and insulin sensitivity in vitro and in vivo.

[Emergency anterior approach for decortication with right pneumonectomy in a patient with chronic expanding hematoma].

The patient was a 68 year-old woman who had a history of treatment of pulmonary tuberculosis 35 years ago. She has experienced dyspnea and hemosputa since several years ago and has been followed up as having chronic empyema. She was admitted to our hospital due to recent exacerbation of symptoms. X-ray films and computed tomography scans of the chest showed the right thoracic cavity to be totally filled with a mass and the shift of mediastinum to the left side. After several days from admission, she needed mechanical ventilation support due to dyspnea exacerbation. Emergency decortication with right pneumonectomy through median sternotomy with anterolateral incision was performed. Postoperative course was uneventful. Pathlogical diagnosis was chronic expanding hematoma.

[Management of surgical approach for intractable secondary spontaneous pneumothorax].

Secondary spontaneous pneumothorax (SSP) such as lymphangioleiomyomatosis (LAM), bronchiolitis obliterans (BO) is intractable or repeated the recurrence of pneumothorax. The most effective chemical pleurodesis for intractable pneumothorax is talc poudrage and so on that is associated with a reduction in the rate of pneumothorax recurrence. However, severe and broad pleural adhesion due to the pleural interventional procedures sometimes cause serious bleeding when the patients undergo lung transplantation. We must be considered for new approaches to these intractable secondary pneumothoraces which replaced traditional conservative and surgical approaches. We had proposed new 2 approaches of total pleural covering (TPC) and awake surgical intervention (ASI) for intractable pneumothorax. We applied the TPC modified with coverage of air leak points with polyglycolic acid (PGA) sheet to 5 patients with intractable bilateral pneumothorax to reduce the risk of excessive bleeding by chemical pleurodesis in lung transplantation. The bilateral pneumothorax was well controlled, and no recurrence has been observed. TPC is reliable procedure for management intractable bilateral SSP. For 12 high-risk patients with other underling pulmonary diseases on general poor conditions, a surgical intervention was performed in awake condition. The air leaks were stopped in 11 cases except for 1 case. The recurrence of pneumothorax after surgery was 2 cases. ASI for intractable secondary pneumothorax can be applicable to selected patients with deteriorated general condition.

Distal Vessel Imaging via Intra-arterial Flat Panel Detector CTA during Mechanical Thrombectomy.

BACKGROUND AND PURPOSE: Obtaining information on invisible vasculature distal to the occlusion site helps to deploy a stent retriever safely during mechanical thrombectomy for large-vessel occlusion. It is essential to reduce the amount of contrast used for detecting the vessels distal to the occlusion site because acute ischemic stroke patients tend to have chronic kidney disease and patients with severe chronic kidney disease are at an increased risk of contrast-associated acute kidney injury. We assessed whether vessels distal to the occlusion site during acute ischemic stroke with large-vessel occlusion could be visualized on angiographic images using flat panel detector CT acquired following intra-arterial diluted contrast injection, compared with MRA findings. MATERIALS AND METHODS: Between May 2019 and January 2020, we enrolled 28 consecutive patients with large-vessel occlusions of the anterior circulation eligible for mechanical thrombectomy following MR imaging. The patients underwent CBV imaging using flat panel detector CT with an intra-arterial diluted contrast injection instead of intravenous injection. Flat panel detector CT angiographic images reconstructed from the same dataset were evaluated for image quality, collateral status of the MCA territory, and visualization of the vessels distal to the occlusion site. These findings were compared with MRA findings. RESULTS: Twenty-two patients were retrospectively examined. Flat panel detector CT angiographic image quality in 20 patients (91%) was excellent or good. The distal portion of the occluded vessel segment was visualized in 14 patients (70%), while the proximal portion of the segment adjacent to the occluded vessel in 3 (15%) was visualized. No visualization was observed in only 1 patient (5%) with no collateral supply. Flat panel detector CT angiographic images were shown to evaluate vessels distal to the occlusion site more accurately than MRA. CONCLUSIONS: In acute ischemic stroke with large-vessel occlusion, flat panel detector CT angiographic images could successfully visualize vessels distal to the occlusion site with a small amount of contrast material.

Cbl-b enhances Runx2 protein stability and augments osteocalcin promoter activity in osteoblastic cell lines.

Cbl-b is a member of Cbl family of E3 ubiquitin (Ub) ligase. Besides the important role in ubiquitination process, other members of Cbl family have been suggested to show non-ubiquitination-related function in regulation of osteoblastic differentiation. However, the role of Cbl-b in regulation of osteoblastic function has not been known yet. To elucidate the role of Cbl-b in regulation of osteoblastic function, we examined its effects on Runx2, a master gene of osteoblastic differentiation. We co-expressed Cbl-b and Runx2 in osteoblastic cell lines and tested their effects on osteocalcin promoter activity together with the expression of Runx2 and its downstream genes. Luciferase assay demonstrated that Cbl-b synergistically enhances osteocalcin promoter activity in conjunction with the effect on Runx2. Co-transfection of Cbl-b and Runx2 further upregulated Runx2 protein levels without any alteration in Runx2 mRNA expression. The upregulation of Runx2 protein by Cbl-b was inhibited by the treatment with lactacystin, a specific inhibitor of the 26S proteasome. These results indicated that Cbl-b would control Runx2 protein levels at the post-translational event. Moreover, the upregulation of downstream genes of Runx2 such as osteocalcin and alkaline phosphatase mRNA was also observed. These data propose the involvement of Cbl-b in the regulation of osteoblast-related genes expression.

Enhancement of osteoclastic bone resorption and suppression of osteoblastic bone formation in response to reduced mechanical stress do not occur in the absence of osteopontin.

Reduced mechanical stress to bone in bedridden patients and astronauts leads to bone loss and increase in fracture risk which is one of the major medical and health issues in modern aging society and space medicine. However, no molecule involved in the mechanisms underlying this phenomenon has been identified to date. Osteopontin (OPN) is one of the major noncollagenous proteins in bone matrix, but its function in mediating physical-force effects on bone in vivo has not been known. To investigate the possible requirement for OPN in the transduction of mechanical signaling in bone metabolism in vivo, we examined the effect of unloading on the bones of OPN(-/-) mice using a tail suspension model. In contrast to the tail suspension-induced bone loss in wild-type mice, OPN(-/-) mice did not lose bone. Elevation of urinary deoxypyridinoline levels due to unloading was observed in wild-type but not in OPN(-/-) mice. Analysis of the mechanisms of OPN deficiency-dependent reduction in bone on the cellular basis resulted in two unexpected findings. First, osteoclasts, which were increased by unloading in wild-type mice, were not increased by tail suspension in OPN(-/-) mice. Second, measures of osteoblastic bone formation, which were decreased in wild-type mice by unloading, were not altered in OPN(-/-) mice. These observations indicate that the presence of OPN is a prerequisite for the activation of osteoclastic bone resorption and for the reduction in osteoblastic bone formation in unloaded mice. Thus, OPN is a molecule required for the bone loss induced by mechanical stress that regulates the functions of osteoblasts and osteoclasts.

Mutants in the ADP-ribosyltransferase cleft of cholera toxin lack diarrheagenicity but retain adjuvanticity.

Cholera toxin (CT), the most commonly used mucosal adjuvant in experimental animals, is unsuitable for humans because of potent diarrhea-inducing properties. We have constructed two CT-A subunit mutants, e.g., serine-->phenylalanine at position 61 (S61F), and glutamic acid-->lysine at 112 (E112K) by site-directed mutagenesis. Neither mutant CT (mCT), in contrast to native CT (nCT), induced adenosine diphosphate-ribosylation, cyclic adenosine monophosphate formation, or fluid accumulation in ligated mouse ileal loops. Both mCTs retained adjuvant properties, since mice given ovalbumin (OVA) subcutaneously with mCTs or nCT, but not OVA alone developed high-titered serum anti-OVA immunoglobulin G (IgG) antibodies (Abs) which were largely of IgG1 and IgG2b subclasses. Although nCT induced brisk IgE Ab responses, both mCTs elicited lower anti-OVA IgE Abs. OVA-specific CD4+ T cells were induced by nCT and by mCTs, and quantitative analysis of secreted cytokines and mRNA revealed a T helper cell 2 (Th2)-type response. These results now show that the toxic properties of CT can be separated from adjuvanticity, and the mCTs induce Ab responses via a Th2 cell pathway.

Weight change and all-cause, cancer and cardiovascular disease mortality in Japanese men and women: the Japan Public Health Center-Based Prospective Study.

BACKGROUND: It is unclear whether weight change during adulthood influences subsequent mortality in Asian populations, who have a relatively lean body mass. OBJECTIVE: To assess the relation of weight change over 5 years to all-cause, cancer and cardiovascular disease mortality among Japanese men and women. DESIGN: Subjects were 36 220 men and 44 091 women aged between 45 and 75 years without a history of serious disease at baseline. Weight change was calculated as the difference of body weight between two surveys with a 5-year interval. RESULTS: During 699 963 person-years of follow-up, we identified 4232 deaths of all-cause, 1872 cancer deaths and 1021 cardiovascular deaths. The relation between weight change and all-cause mortality was reverse J-shaped. Multivariate hazard ratios (95% confidence interval) for weight loss of 5 kg or more versus weight change of less than 2.5 kg were 1.62 (1.45-1.81) in men and 1.76 (1.51-2.05) in women, whereas those for weight gain of 5 kg or more were 1.40 (1.22-1.59) in men and 1.25 (1.02-1.54) in women. These associations remained statistically significant even after the exclusion of deaths in the first 3 years of follow-up. The weight change-mortality association was pronounced in underweight persons or in nonsmoking men. The risk of cancer mortality increased in both men and women who lost weight by 5 kg or more. With regard to cardiovascular disease, mortality risk tended to increase with weight loss both in men and women, whereas its increase with weight gain was observed only in women. CONCLUSIONS: A large weight change, both loss and gain, was associated with an increased risk of mortality. Weight loss and gain may be predictors of early death in apparently healthy adult Japanese.

[Surgical approaches for superior sulcus tumor].

Surgical approach is one of the most crucial aspects in the treatment of superior sulcus tumor (SST). Posterior approach as described by Paulson and coworkers is appropriate for the resection of SST invading posterior part of the 1st rib and the vertebrae, whereas anterior approaches as described by Masaoka, Dartevelle, Grunenwald, or Rusca are suitable for resection of SST involving subclavian vessels. We present 2 cases of SST who underwent complete resection through the posterior approach and a modified hemi-clamshell approach, respectively. We also discuss the surgical approaches for SST with referring to literatures.

Calcium, vitamin D and dairy intake in relation to type 2 diabetes risk in a Japanese cohort.

AIMS/HYPOTHESIS: Calcium and vitamin D have been implicated in the development of type 2 diabetes, but epidemiological evidence is limited. We examined prospectively the relation of calcium and vitamin D intake to type 2 diabetes risk in a Japanese cohort. METHODS: Participants were 59,796 middle-aged and older men and women, who participated in the Japan Public Health Center-based Prospective Study and had no history of type 2 diabetes or other serious diseases. Dietary intake of calcium and vitamin D were estimated using a validated food frequency questionnaire. Logistic regression was used to assess the association between intake of these nutrients and self-reported newly diagnosed type 2 diabetes. RESULTS: During a 5 year follow-up, 1,114 cases of type 2 diabetes were documented. Overall, calcium intake was not associated with a significantly lower risk of type 2 diabetes; the multivariable odds ratio for the highest vs lowest quartiles was 0.93 (95% CI 0.71-1.22) in men and 0.76 (95% CI 0.56-1.03) in women. However, among participants with a higher vitamin D intake, calcium intake was inversely associated with diabetes risk; the odds ratio for the highest vs lowest intake categories was 0.62 (95% CI 0.41-0.94) in men and 0.59 (95% CI 0.38-0.91) in women. Dairy food intake was significantly associated with a lower risk of type 2 diabetes in women only. CONCLUSIONS/INTERPRETATION: Calcium and vitamin D may not be independently associated with type 2 diabetes risk. Our finding suggesting a joint action of these nutrients against type 2 diabetes warrants further investigation.

Taste preferences and body weight change in Japanese adults: the JPHC Study.

OBJECTIVE: Limited data are available with regard to longitudinal changes in body weight by food taste preference. Here, we examined the associations between taste preferences and weight change in adults for a large-scale cohort study in Japan. DESIGN: Longitudinal analysis of data from a population-based cohort study, the Japan Public Health Center-based Prospective Study (JPHC Study). SUBJECTS: A total of 29,103 middle-aged men and women, who participated in a JPHC Study and returned questionnaires on lifestyle and diet, including taste preferences, at both baseline and the 10th year of follow-up. MEASUREMENTS: We assessed the relations of preferences for rich and heavy taste and a sweet taste to weight changes between the age of 20 years and baseline and those during the 10-year follow-up period. RESULTS: Preferences for rich and heavy taste and for sweet taste were significantly positively associated with weight increases between the age of 20 years and baseline (P for trend <0.001); the fully adjusted odds ratios (95% confidence interval) comparing the 'like' versus 'dislike' groups with a preference for rich and heavy taste were 1.45 (1.31-1.24) for men and 1.28 (1.16-1.41) for women, whereas that for a sweet taste preference was 1.22 (1.09-1.36) for women. As regards weight change during the 10 years of follow-up, subjects who liked the sweet taste and those who neither liked nor disliked this taste experienced a significantly greater increase than those who disliked it in both men and women. There was no such difference for rich and heavy taste. CONCLUSION: These results suggest that food taste preferences may be an important predictor of weight changes in adults. Taste preferences need to be considered when counseling patients to achieve weight control.

Identification of a DNA sequence involved in osteoblast-specific gene expression via interaction with helix-loop-helix (HLH)-type transcription factors.

To elucidate regulatory mechanism(s) underlying differentiation of osteoblasts, we examined involvement of helix-loop-helix (HLH)-type transcription factors in osteoblast-specific expression of a phenotypic marker gene which encodes osteocalcin, a major noncollagenous bone matrix protein, exclusively expressed in osteoblasts. Overexpression of a dominant negative HLH protein, Id-1, decreased the activity of the 1.1-kb osteocalcin gene promoter cotransfected into rat osteoblastic osteosarcoma ROS17/2.8 cells. Analysis of deletion mutants revealed that a 264-bp fragment of osteocalcin promoter (-198 to +66) was sufficient for the Id-1-dependent suppression. Furthermore, the activity of the same promoter fragment (-198 to +66) was enhanced when antisense Id-1 expression vector was cotransfected. This osteocalcin gene promoter region contains two sites of an E-box motif, a consensus binding site for HLH proteins, which we refer to as OCE1 (CACATG, at -102) and OCE2 (CAGCTG, at -149), respectively. Mutagenesis in OCE1 but not OCE2 led to greater than 50% reduction in transcriptional activity of the osteocalcin gene promoter. Electrophoresis mobility shift assay indicated that factors in nuclear extracts prepared from ROS17/2.8 cells bound to the 30-bp oligonucleotide probe containing the E-box motif of OCE1. This binding was competed out by OCE1 oligonucleotide but neither by OCmE1 oligonucleotide in which E-box motif was mutated nor by OCE2. The OCE1-binding activity in the nuclear extracts of ROS17/2.8 cells was reduced by 70% when bacterially expressed Id-1 protein was added to the reaction mixture, suggesting the involvement of HLH proteins in the DNA/protein complex formation. In contrast to the osteoblast-like cells, OCE1-binding activity in the nuclear extracts of C3H10T1/2 fibroblasts was very low. However, when these fibroblasts were treated with recombinant human bone morphogenetic protein-2 which induced expression of osteocalcin as well as other phenotypic markers of osteoblasts, OCE1-binding activity was increased approximately 40-fold, indicating that OCE1 would be involved in the tissue-specific expression of the osteocalcin gene. These findings indicated for the first time that osteoblast-specific gene transcription is regulated via the interaction between certain E-box binding transcription factor(s) in osteoblasts and the OCE1 sequence in the promoter region of the osteocalcin gene.