Molecular and morphological analyses revealed a cryptic species of dojo loach Misgurnus anguillicaudatus (Cypriniformes: Cobitidae) in Japan.

Although it has been reported that populations of the Japanese dojo loach Misgurnus anguillicaudatus (Cypriniformes: Cobitidae) belong to two distinct mitochondrial (mt)DNA (Type I and Type II), the taxonomic status of the species remains unresolved. To address this question, nuclear DNA and morphological analyses were performed on M. anguillicaudatus population in the Nakaikemi Wetland, where Type I and Type II lineages are sympatric. Results suggest the existence of a cryptic species (Type I) within the Japanese dojo loach.

Weight change after 20 years of age and the incidence of dyslipidemia: a cohort study of Japanese male workers.

BACKGROUND: While heavier weight is known to increase the incidence of dyslipidemia, limited data are available on the relationship between weight gain and its development. METHODS: A total of 2647 males were categorized into the following four groups according to the difference between their self-reported weight at 20 years of age and their measured weight in 1994-95: a loss of ≥5% (decrease), loss of <5% or gain of <5% (no change), gain of ≥5 to <15% (increase) and gain of ≥15% (sizable increase). They were followed up until their 2002-03 health examination. Using the 'no change' group as reference, the multivariable-adjusted odds ratio (adjusted for age, body mass index at 20 years of age, physical activity, smoking and alcohol intake) and 95% confidence interval (95% CI) for the incidence of dyslipidemia were determined using logistic regression models. RESULTS: A total of 1342 participants developed dyslipidemia during the follow-up period. The 'increase' and 'sizable increase' groups had odds ratios for the incidence of dyslipidemia of 1.97 (95% CI, 1.59-2.45) and 2.68 (2.15-3.34), respectively, demonstrating that there was a significant dose-response association between weight gain since 20 years of age and the incidence of dyslipidemia (P < 0.001 for trend). CONCLUSION: These results suggest that dyslipidemia could be prevented by avoiding weight gain in adulthood.

Effect of replacing corn with whole-grain paddy rice and brown rice in broiler diets on growth performance and intestinal morphology.

The present study was conducted to investigate replacing corn with whole-grain paddy rice (WPR) and whole-grain brown rice (BR) in broiler chicken diets and its effect on growth performance and histological structures of the intestinal villi. Marshall Chunky male chicks (14 days old) were divided into five groups with four replicates of four chicks each. In the dietary treatments, corn in the basal diet was replaced with WPR and BR. The chickens received five experimental diets consisting of corn, WPR and BR in ratios of 100:0:0 (Control), 50:0:50 (50Corn + 50BR), 50:25:25 (50Corn + 25WPR + 25BR), 25:50:25 (25Corn + 50WPR + 25BR) and 0:50:50 (50WPR + 50BR) respectively. Feed and water were provided ad libitum for 35 day. No significant differences were found in feed intake, body weight gain and feed efficiency among the treatment groups. The relative weights of the gizzard in the 50Corn + 25WPR + 25BR, 25Corn + 50WPR + 25BR and 50WPR + 50BR groups were significantly higher than that of the Control and 50Corn + 50BR groups (p < 0.05). The gizzard pH of the experimental groups was lower than those of Control (p < 0.05). The ileal crypt of birds on the Control diets was deeper (p < 0.05) than those observed in the experimental birds. Moreover, the ileal villus height: crypt depth ratio increased (p < 0.05) in the 50WPR + 50BR group (p < 0.05) compared with the Control group. No specific changes were observed in the epithelial cells on the duodenal apical surface among the groups except that the villus of the 25Corn + 50WPR + 25BR group had cell clusters. The jejunal and ileal villus apical surface of the experimental groups showed similar morphology to the Control group. These findings suggest that WPR and BR can totally replace corn in broiler diets without negatively affecting growth performance.

Effects of whole-grain paddy rice replacement with or without enzyme addition on broiler performance and intestinal morphology.

To investigate the effect of replacing maize with whole-grain paddy rice (WPR) in broiler chicken diets, with or without enzyme addition, on growth performance and histological structures of the intestinal villi, 14-d-old Marshall Chunky male chicks were divided into 4 groups with 4 replicates of 4 chicks each. The experimental diets containing different concentrations of WPR were as follows: (1) 0 g/kg (Control); (2) 141.5 g/kg, grower, and 125.0 g/kg, finisher (25WPR); (3) 283.0 g/kg, grower, and 250.0 g/kg, finisher (50WPR); (4) 283.0 g/kg, grower, and 250.0 g/kg, finisher, and enzyme supplementation (50WPR + enzyme). All diets were formulated to be isocaloric and isonitrogenous and provided ad libitum for 35 d. There were no differences among the diets on the growth performance and digestive organ size. The villus height and cell mitosis number of all intestinal segments did not change in any treatment. The ileal villus area, duodenal cell area, duodenal and jejunal goblet cell number in the 50WPR group increased significantly relative to the control but not when enzyme was included. In the scanning electron microscope results, all experimental groups showed clear protuberant cells and cell clusters on the villus apical surface of the duodenum. In the jejunum, cell clusters and areas having cells with no microvilli were frequently found in both the 50WPR and 50WPR + enzyme groups. In conclusion, broilers fed on diets replacing maize with WPR showed hypertrophied villi of duodenum and ileum and epithelial cells in duodenum and jejunum, especially in the 50WPR group, without negatively affecting growth performance. These findings suggest that WPR can replace maize up to a level of 50% (283.0 g/kg, starter, and 250.0 g/kg, finisher) in broiler diets without enzyme supplementation. However, further studies are needed to improve our knowledge of the influence of WPR on higher numbers of birds.

Dose-Response Relationships between Diet Quality and Mortality among Frail and Non-Frail Older Adults: A Population-Based Kyoto-Kameoka Prospective Cohort Study.

OBJECTIVES: Although better diet quality is inversely associated with mortality risk, the association between diet quality and mortality remains unclear in frail and non-frail older adults. Thus, we aimed to examine this association in older Japanese adults. DESIGN: A prospective cohort study. SETTING AND PARTICIPANTS: We used the data of 8,051 Japanese older adults aged ≥65 years in the Kyoto-Kameoka study. MESUREMENTS: Dietary intake was estimated using a validated food frequency questionnaire. Diet quality was evaluated by calculating the adherence scores to the Japanese Food Guide Spinning Top (range, 0 [worst] to 80 [best]), which were stratified into quartiles. Frailty status was assessed using the validated self-administered Kihon Checklist (KCL) and the Fried phenotype (FP) model. Survival data were collected between February 15, 2012 and November 30, 2016. Statistical analysis was performed using the multivariate Cox proportional hazard analysis and the spline model. RESULTS: During the median 4.75-year follow-up (36,552 person-years), we recorded 661 deaths. After adjusting for confounders, compared with the bottom adherence score quartile, the top quartile was associated with lower hazard ratio (HR) of mortality in frailty (HR, 0.73; 95% confidence interval [CI], 0.54-1.00) and non-frailty, as defined by the KCL (HR, 0.72; 95% CI, 0.52-1.01). In the spline model, regardless of frailty status defined by the KCL and FP model, adherence score showed a strongly dose-dependent inverse association with mortality up to approximately 55 points; however, no significant differences were observed thereafter. This association was similar to the results obtained in individuals with physical, cognitive, and depression as domains of KCL in the spline model. CONCLUSIONS: Our findings demonstrate an L-shaped association between diet quality and mortality in both frail and non-frail individuals. This study may provide important knowledge for improving poor diet quality in older individuals with frailty or domains of frailty.

Long-term outcomes of hemostatic therapy for variceal bleeding and the challenge pending in the post-direct-acting antivirals era.

Background and study aims: This study evaluated the longterm outcomes of mainly endoscopic hemostatic therapy for gastrointestinal variceal bleeding and of the transition of hemostatic therapy. Patients and methods: Among 1,163 patients treated for gastrointestinal varices between April 2006 and June 2020, a total of 125 patients who underwent emergency hemostatic therapy were enrolled. Survival rates and secondary evaluation points were analyzed. Additionally, patients were classified into two groups: the previous and latter term. Patients' background, therapeutic method, and treatment results were compared between the groups. Results: 94.4% had cirrhosis. The average Child-Pugh score was 8.90. Successful primary hemostasis rate was 98.4%, and 5.6% died within 2 weeks, all with a Child-Pugh score ≥9. The respective 1- and 5-year survival rates for Child-Pugh grade A/B were 81.3% and 55.4%, while those for Child-Pugh grade C were 58.1% and 17.8%. Child-Pugh grade C or hepatocellular carcinoma was significantly associated with poor prognosis. In total, 21.6% experienced variceal re-bleeding; 62.9% of these cases were triggered by continued alcohol consumption. There was no significant difference in survival between patients with and without variceal re-bleeding and in post-treatment survival between the previous and latter terms. In the latter term, the number of cases caused by continued alcohol consumption significantly increased. Conclusions: Multidisciplinary treatment and continuation of proper management after hemostatic therapy for variceal bleeding are crucial. Continued alcohol consumption leads to variceal bleeding and re-bleeding; its proper management, including alcohol abstinence, is one of the major challenges left in the post-directacting antivirals era.

Impact of surgical menopause on lipid and bone metabolism.

OBJECTIVE: To clarify the effects of ovariectomy on lipid and bone metabolism. METHODS: This study was a prospective study with a longitudinal design within 1 year after surgery. Sixty-two premenopausal women were recruited and divided into two groups: group M (preservation of ovary, n=27) and group BSO (bilateral ovariectomy, n=35). Serum lipid levels, urinary N-telopeptide of type I collagen (NTx) and bone mineral density (BMD) were measured. We also examined the number of postoperative episodes requiring pharmacological intervention. RESULTS: There was a significant elevation in the level of low density lipoprotein cholesterol in group BSO from 6 to 12 months compared with the baseline level; the level did not change in group M. The NTx level significantly increased from 6 to 12 months, and the BMD was significantly decreased by as much as 6.7% at 12 months in group BSO; these variables did not change in group M. The effect of lipid and bone metabolism in group BSO was observed when the ages of the two groups were matched. Carbohydrate metabolism and arterial stiffness, measured by pulse wave velocity, did not change throughout the study period in either group. No subjects in group M required medication expect for two patients whose ovarian function was diminished by postoperative radiation and by natural menopause. Eleven women received medication in group BSO: nine for climacteric disorders using hormone therapy (25.7%), and two for dyslipidemia using statins (5.7%). CONCLUSIONS: Bilateral ovariectomy seems to cause dyslipidemia and serious loss of bone mineral density within only 1 year, and patients who lose ovarian function may require careful medical care.

Phosphotyrosine-modified proteins are concentrated at the membranes of epithelial and endothelial cells during tissue development in chick embryos.

We have used high affinity polyclonal antibodies specific for phosphotyrosine (PTyr) residues to examine the localization in various chick embryonic tissues in situ of PTyr-modified proteins by immunocytochemical methods. During the period from 9 to 21 d of development, most tissues exhibit elevated levels of PTyr-modified proteins as determined by immunoblotting experiments of tissue extracts with the anti-PTyr antibodies (Maher, P. A., and E. B. Pasquale. 1988. J. Cell Biol. 106:1747-1755). By immunofluorescence labeling of semithin frozen sections, the highest concentrations of PTyr immunolabeling in all of the embryonic tissues examined were localized to the membranes of the epithelial and endothelial cells with other cells showing no detectable labeling. These results were confirmed by immunoelectron microscopic labeling, which showed particularly high concentrations of PTyr-modified proteins close to the membranes at the apical junctions. The corresponding adult tissues showed no labeling. It is proposed that these results reflect the molecular basis for the functional plasticity of epithelial and endothelial cell junctions during embryonic development.

Identification of glutamic acid decarboxylase gene and distribution of GABAergic nervous system in the planarian Dugesia japonica.

The planarian Dugesia japonica has a relatively well-organized CNS that includes the brain and the ventral nerve cords, and also has high regenerative capacity derived from pluripotent stem cells present in the mesenchymal space throughout the body. Glutamic acid decarboxylase (GAD) is the enzyme that converts glutamic acid into GABA, a major inhibitory neurotransmitter. In this study, we first identified a full-length GAD gene (DjGAD, D. japonica glutamic acid decarboxylase) in the planarian D. japonica. Whole-mount in situ hybridization revealed that a few cells expressed DjGAD mRNA, and these cells were located in both the head and pharynx regions. In order to examine the distribution pattern of DjGAD protein, we generated a mouse monoclonal anti-DjGAD antibody. The distribution pattern of DjGAD protein was very similar to that of DjGAD mRNA. A neural network of DjGAD-immunopositive cells was also clearly observed. In addition, we examined the immunofluorescence during the process of regeneration of the head from the tail piece. At day 3 of regeneration, we could detect newly formed DjGAD-immunopositive neurons in the anterior region. During day 5-7 of regeneration, reconstruction of the neural network of DjGAD-immunopositive cells occurred. DjGAD-immunoreactivity was lost in DjGAD-knockdown planarians obtained by RNA interference. The amount of GABA was significantly decreased in DjGAD-knockdown planarians, which lost negative phototaxis but not locomotion activity. These results suggest that DjGAD is clearly required for GABA biosynthesis and photosensitivity in planarians, and expression of DjGAD as detected by anti-DjGAD antibody is a useful marker for GABAergic neurons.

Long-term forskolin stimulation induces AMPK activation and thereby enhances tight junction formation in human placental trophoblast BeWo cells.

BeWo cells, derived from human choriocarcinoma, have been known to respond to forskolin or cAMP analogues by differentiating into multinucleated cells- like syncytiotrophoblasts on the surfaces of chorionic villi of the human placenta. In this study, we demonstrated that long-term treatment with forskolin enhances the tight junction (TJ) formation in human placental BeWo cells. Interestingly, AMPK activation and phosphorylation of acetyl-CoA carboxylase (ACC), a molecule downstream from AMPK, were induced by long-term incubation (>12h) with forskolin, despite not being induced by acute stimulation with forskolin. In addition, co-incubation with an AMPK inhibitor, compound C, as well as overexpression of an AMPK dominant negative mutant inhibited forskolin-induced TJ formation. Thus, although the molecular mechanism underlying AMPK activation via the forskolin stimulation is unclear, the TJ formation induced by forskolin is likely to be mediated by the AMPK pathway. Taking into consideration that TJs are present in the normal human placenta, this mechanism may be important for forming the placental barrier system between the fetal and maternal circulations.

Validation of self-reported energy intake by a self-administered diet history questionnaire using the doubly labeled water method in 140 Japanese adults.

OBJECTIVE: To validate reported energy intake (rEI) with a self-administered diet history questionnaire (DHQ) against total energy expenditure (TEE) by the doubly labeled water (DLW) method. SUBJECTS: A total of 140 healthy Japanese adults (67 men and 73 women) aged 20-59 years living in four areas in Japan. METHODS: Energy intake was assessed twice with DHQ over a 1-month period before and after TEE measurement (rEI(DHQ1) and rEI(DHQ2), respectively). TEE was measured by DLW during 2 weeks (TEE(DLW)). RESULTS: Mean rEI(DHQ1) was lower than those of TEE(DLW) by 1.9+/-2.4 MJ/day (16.4%, P<0.001) for men and 0.6+/-1.9 MJ/day (6.0%, P<0.01) for women. In men and women together, 62 subjects (44%) were defined as underreporters (rEI(DHQ1)/TEE(DLW) <0.84), 58 (41%) as acceptable reporters (0.84-1.16) and 20 (14%) as over-reporters (>1.16). Pearson correlation coefficient was 0.34 for men and 0.22 for women. After adjustment for the dietary and non-dietary factors related to rEI(DHQ1)/TEE(DLW), the correlation coefficient improved to 0.42 and 0.37, respectively. CONCLUSION: The energy intake assessed with DHQ correlated low to modestly with TEE measured by DLW. In addition, DHQ underestimated energy intake at a group level. Caution is needed when energy intake was evaluated by DHQ at both individual and group levels.

Physical activity level in healthy free-living Japanese estimated by doubly labelled water method and International Physical Activity Questionnaire.

OBJECTIVE: To measure total energy expenditure (TEE) for normal healthy Japanese by the doubly labelled water (DLW), and to compare the physical activity level (PAL) among categories classified by the categories used in daily reference intake (DRI), Japan and the International Physical Activity Questionnaire (IPAQ). SUBJECTS AND METHODS: A total of 150 healthy Japanese men and women aged 20- to 59-year-old living in four districts of Japan. TEE was measured by the DLW method, and the PAL was calculated from TEE divided by basal metabolic rate. Simultaneously with TEE measurement, the PAL was assessed employing the categories used in DRI, Japan and IPAQ. RESULTS: The average TEE and PAL were 10.78+/-1.67 MJ/day and 1.72+/-0.22 for males and 8.37+/-1.30 MJ/day and 1.72+/-0.27 for females, respectively. The subjects in the highly active categories assessed by both DRI and IPAQ showed significantly higher PAL compared with less active categories. However, PALs among light and moderate categories by DRI, and insufficient and sufficiently active by IPAQ were not significantly different. CONCLUSIONS: In developed countries, highly active subjects could be assessed by a simple questionnaire. However, the questionnaire should be improved to clarify the sedentary to moderately active subjects by assessing carefully very light to moderate physical activity.

Lipophilic beta-adrenoceptor antagonist propranolol increases the hypnotic and anti-nociceptive effects of isoflurane in a swine model.

BACKGROUND: We have previously reported that landiolol, an ultra-short-acting beta1-adrenoceptor antagonist, does not alter the anaesthetic effects of isoflurane. Here, we investigated the influence of propranolol on the electroencephalographic (EEG) effects and minimum alveolar concentration (MAC) of isoflurane. METHODS: Fourteen swine [25.0 (SD 4.0) kg] were anaesthetized by isoflurane inhalation. The inhalation concentration was decreased to 0.5% and maintained for 25 min, before being returned to 2%, and maintained for a further 25 min. End-tidal isoflurane concentrations and spectral edge frequencies were recorded. Pharmacodynamic analysis was performed using a sigmoidal inhibitory maximal effect model for spectral edge frequency vs effect-site concentration. After measurement of the EEG effect, MAC was determined using the dew-claw clamp technique, in which movement in response to clamping is recorded. After completion of control measurements, a propranolol 4 mg bolus followed by an infusion (2 mg h(-1)) was started. After a 30 min stabilization period, the inhalation concentration of isoflurane was varied as in the control period and MAC was re-assessed. RESULTS: Propranolol shifted the concentration-effect relationship to the left and decreased the effect-site concentration that produced 50% of the maximal effect from 1.30 (0.18) to 1.13 (0.17)%. Propranolol also decreased isoflurane MAC from 1.91 (0.35) to 1.54 (0.32)%. CONCLUSIONS: Propranolol alters both the hypnotic and anti-nociceptive effects of isoflurane. In contrast to landiolol, lipophilic beta-adrenoceptor antagonists may increase the potency of inhalational anaesthetics.

Do the kidneys contribute to propofol elimination?

BACKGROUND: Propofol is mainly metabolized in the liver, but extrahepatic clearance may also be important since systemic propofol clearance exceeds hepatic clearance. Recent reports suggest that the kidneys contribute to propofol elimination in humans and here we investigated renal elimination of propofol in a controlled animal study. METHODS: Seventeen swine were anaesthetized with 5% isoflurane induction and 2% isoflurane maintenance. After a left subcostal incision, the left kidney and renal pedicle were exposed by an approach via the retroperitoneum and the renal vein was identified for blood sampling. Propofol was then administered via the right jugular vein at a rate of 2 mg kg(-1) h(-1). After 120 min of pseudo-steady-state infusion of propofol (Baseline 1), cardiac output (CO) was increased by continuous infusion of dobutamine for 30 min (high-CO state). Thirty minutes after stopping dobutamine (Baseline 2), CO was decreased by bolus administration of propranolol (low-CO state). Blood samples were collected simultaneously from the renal vein (direct puncture) and the femoral artery at Baseline 1, in the high-CO state, at Baseline 2, and in the low-CO state. RESULTS: There was no significant difference in propofol concentration between femoral arterial and renal venous blood in all states. The propofol concentration significantly decreased with increased CO, but renal extraction was not observed in any state. CONCLUSIONS: The kidneys are a minor site of propofol elimination in a swine model.

Proprotein-processing endoprotease furin controls the growth and differentiation of gastric surface mucous cells.

Gastric surface mucous cells originate from progenitor cells at the isthmus of the gastric gland, from where the cells migrate to the luminal surface. With migration they form secretory granules and express TGF alpha. We found that proprotein-processing endoprotease furin-positive cells were layered around the upper one fourth of the gastric glands of adult rats, whereas they were distributed along an outer epithelial layer in fetal rats. Because the furin-positive cell layer was localized from the upper cell proliferating zone to the less proliferating pit-cell region in the gastric gland unit, we examined the role of furin in the growth and differentiation of surface mucous cells by using the cell line, GSM06. This cell line is derived from the gastric surface mucous cells of transgenic mice harboring the temperature-sensitive simian virus 40 T antigen. At T antigen-active temperature (33 degrees C), the cells grew to confluency, whereas at T antigen-inactive temperature (39 degrees C), the cells ceased growing. At 33 degrees C, the cells exhibited a high level of furin expression with a negligible level of periodic acid Schiff (PAS)-positive materials and a low level of TGF alpha. In contrast, at 39 degrees C the cells produced a high level of PAS-positive materials, TGF alpha, and secretory granules, with a negligible level of furin expression. To further examine the role of furin, we established a GSM06 cell line introduced with either a sense or an antisense furin cDNA. The cells with sense furin expression produced fewer PAS-positive materials and a low level of TGF alpha even at 39 degrees C, whereas the cells with antisense furin expression exhibited more PAS-positive materials and TGF alpha even at 33 degrees C. When furin expression was suppressed by its antisense oligonucleotide, the cell growth was retarded with enhanced expression of the differentiated characteristics. Thus, we conclude that furin is instrumental in controlling the growth of the surface mucous cells.

A mutation in the insulin 2 gene induces diabetes with severe pancreatic beta-cell dysfunction in the Mody mouse.

The mouse autosomal dominant mutation Mody develops hyperglycemia with notable pancreatic beta-cell dysfunction. This study demonstrates that one of the alleles of the gene for insulin 2 in Mody mice encodes a protein product that substitutes tyrosine for cysteine at the seventh amino acid of the A chain in its mature form. This mutation disrupts a disulfide bond between the A and B chains and can induce a drastic conformational change of this molecule. Although there was no gross defect in the transcription from the wild-type insulin 2 allele or two alleles of insulin 1, levels of proinsulin and insulin were profoundly diminished in the beta cells of Mody mice, suggesting that the number of wild-type (pro)insulin molecules was also decreased. Electron microscopy revealed a dramatic reduction of secretory granules and a remarkably enlarged lumen of the endoplasmic reticulum. Little proinsulin was processed to insulin, but high molecular weight forms of proinsulin existed with concomitant overexpression of BiP, a molecular chaperone in the endoplasmic reticulum. Furthermore, mutant proinsulin expressed in Chinese hamster ovary cells was inefficiently secreted, and its intracellular fraction formed complexes with BiP and was eventually degraded. These findings indicate that mutant proinsulin was trapped and accumulated in the endoplasmic reticulum, which could induce beta-cell dysfunction and account for the dominant phenotype of this mutation.

Genetic cholesteryl ester transfer protein deficiency caused by two prevalent mutations as a major determinant of increased levels of high density lipoprotein cholesterol.

Genetic determinants of HDL cholesterol (HDL-C) levels in the general population are poorly understood. We previously described plasma cholesteryl ester transfer protein (CETP) deficiency due to an intron 14 G(+1)-to-A mutation(Int14 A) in several families with very high HDL-C levels in Japan. Subjects with HDL-C > or = 100 mg/dl (n = 130) were screened by PCR single strand conformational polymorphism analysis of the CETP gene. Two other mutations were identified by DNA sequencing or primer-mediated restriction map modification of PCR products: a novel intron 14 splice donor site mutation caused by a T insertion at position +3 from the exon14/intron14 boundary (Int14 T) and a missense mutation (Asp442 to Gly) within exon 15 (D442G). The Int14 T mutation was only found in one family. However, the D442G and Int14 A mutations were highly prevalent in subjects with HDL-C > or = 60 mg/dl, with combined allele frequencies of 9%, 12%, 21% and 43% for HDL-C 60-79, 80-99, 100-119, and > or = 120 mg/dl, respectively. Furthermore, prevalences of the D442G and Int14 A mutations were extremely high in a general sample of Japanese men (n = 236), with heterozygote frequencies of 7% and 2%, respectively. These two mutations accounted for about 10% of the total variance of HDL-C in this population. The phenotype in a genetic compound heterozygote (Int14 T and Int14 A) was similar to that of Int14 A homozygotes (no detectable CETP and markedly increased HDL-C), indicating that the Int14 T produces a null allele. In four D442G homozygotes, mean HDL-C levels (86 +/- 26 mg/dl) were lower than in Int14 A homozygotes (158 +/- 35 mg/dl), reflecting residual CETP activity in plasma. In 47 D442G heterozygotes, mean HDL-C levels were 91 +/- 23 mg/dl, similar to the level in D442G homozygotes, and significantly greater than mean HDL-C levels in Int14 A heterozygotes (69 +/- 15 mg/dl). Thus, the D442G mutation acts differently to the null mutations with weaker effects on HDL in the homozygous state and stronger effects in the heterozygotes, suggesting dominant expression of a partially defective allele. CETP deficiency, reflecting two prevalent mutations (D442G and Int14 A), is the first example of a genetic deficiency state which is sufficiently common to explain a significant fraction of the variation in HDL-C in the general population.

Interindividual variability in sleeping metabolic rate in Japanese subjects.

INTRODUCTION: Basal metabolic rate (BMR) or sleeping metabolic rate (SMR) is the largest component of total energy expenditure (EE). An accurate prediction of BMR or SMR is needed to accurately predict total EE or physical activity EE for each individual. However, large variability in BMR and SMR has been reported. OBJECTIVES: This study was designed to develop prediction equations using body size measurements for the estimation of both SMR and BMR and to compare the prediction errors with those in previous reports. METHODS: We measured body size, height, weight and body composition (fat mass and fat-free mass) from skinfold thickness in adult Japanese men (n=71) and women (n=66). SMR was determined as the sum of EE during 8 h of sleep (SMR-8h) and minimum EE during 3 consecutive hours of sleep (SMR-3h) measured using two open-circuit indirect human calorimeters. BMR was determined using a human calorimeter or a mask and Douglas bag. RESULTS: The study population ranged widely in age. The SMR/BMR ratio was 1.01+/-0.09 (range 0.82-1.42) for SMR-8h and 0.94+/-0.07 (range 0.77-1.23) for SMR-3h. The prediction equations for SMR accounted for a 3-5% larger variance with 2-3% smaller standard error of estimate (SEE) than the prediction equations for BMR. DISCUSSION: SMR can be predicted more accurately than previously reported, which indicates that SMR interindividual variability is smaller than expected, at least for Japanese subjects. The prediction equations for SMR are preferable to those for BMR because the former exhibits a smaller prediction error than the latter.

Aquaporin-2 (AQP2): its intracellular compartment and trafficking.

Aquaporins (AQPs) are membrane proteins serving in the transfer of water and small solutes across cellular membranes. At least 7 isoforms, namely AQP1, AQP2, AQP3, AQP4, AQP6, AQP7 and AQP11 are expressed in the kidney. Among them, AQP2 plays a pivotal role in the concentration of urine. AQP2 is expressed in the principal cells of the collecting ducts. It is localized in the intracellular compartment and is translocated to the cell surface upon anti-diuretic hormone stimulation. Analyses in cultured cells expressing AQP2 have provided clues to the trafficking of AQP2. AQP2 resides in the subapical vesicles, some of which are Rab11-positive, suggesting their close relationship with apical recycling endosomes. Upon stimulation with forskolin, AQP2 is translocated to the cell surface. After washout, AQP2 is endocytosed to early endosomes and then transferred to the apical storage compartment. Some of AQP2 is excreted as exosomes. Actin cytoskeleton plays important roles in the trafficking of AQP2. Analyses of molecules found in AQP2-containing vesicles will shed light on the mechanism of AQP2 translocation.