Cacao Procyanidins-Induced Lifespan Extension in Caenorhabditis elegans in a Nervous System and CaMKII-Dependent Manner.

Procyanidins are gaining attention due to their potential health benefits. We found that cacao liquor procyanidin (CLPr) from Theobroma cacao seeds increased the lifespan of Caenorhabditis elegans, a representative model organism for aging studies. The genetic dependence of the lifespan-extending effect of CLPr was consistent with that of blueberry procyanidin, which is dependent on unc-43, osr-1, sek-1, and mev-1, but not on daf-16, sir-2.1, or skn-1. The lifespan-extending effect of CLPr was inhibited by neuron-specific RNA interference (RNAi) targeting unc-43 and pmk-1, and in worms with loss-of-function mutations in the odr-3, odr-1, or tax-4 genes, which are essential in sensory neurons, including AWC neurons. It was also inhibited in worms in which AWC neurons or AIB interneurons had been eliminated, and in worms with loss-of-function mutations in eat-4 or glr-1, which are responsible for glutamatergic synaptic transmission. These results suggest that the lifespan-extending effect of CLPr is dependent on the nervous system. In addition, it also requires unc-43 and pmk-1 expression in nonneuronal cells, as demonstrated by the experiments with RNAi in wild-type worms, the neuronal cells of which are not affected by systemic RNAi. The osr-1 gene is expressed in hypodermal and intestinal cells and regulates the response to osmotic stress along with unc-43/calcium/calmodulin-dependent protein kinase II and the p38 mitogen-activated protein kinase pathway. Consistent with this, CLPr improved osmotic stress tolerance in an unc-43- and pmk-1-dependent manner, and it was also dependent on AWC neurons. The lifespan-extending and osmotic-tolerance-improving activities were attributed to procyanidins with a tetrameric or higher-order oligomeric structure.

Expression of DEP Domain-Containing 1B in Canine Lymphoma and Other Types of Canine Tumours.

Canine lymphoma is the most common haematological malignancy in dogs and is typically treated with multidrug chemotherapy. Most cases are at risk of relapse after several courses of chemotherapy and the oncogenic mechanism remains unknown. This study was aimed at identifying genes expressed in canine lymphoma by cDNA microarray. We found elevated expression of Dishevelled, EGL-10 and pleckstrin (DEP) domain-containing 1B (DEPDC1B) in canine lymphoma cells compared with cells and tissues from healthy dogs. Canine DEPDC1B protein was detected in 13 of 41 lymphoma specimens by immunohistochemistry, but was not detected in lymph nodes from normal dogs. Immunoreactive DEPDC1B protein was also detected in several other types of canine tumour. This is the first report documenting the association of DEPDC1B with canine cancer and the results suggest that DEPDC1B might serve as a potential marker or therapeutic target for canine malignancies.

Establishment of rat anti-canine DEP domain containing 1B (DEPDC1B) monoclonal antibodies.

DEP domain-containing 1B (DEPDC1B) is involved in the regulation of cell de-adhesion and actin cytoskeleton activity during the G2/M transition of the cell cycle, and its overexpression has been proven to be associated with cancer progression in several human cancers. Canine DEPDC1B was identified as a gene that was overexpressed in canine lymphoma tissues in our previous study. However, in dogs, the protein expression of DEPDC1B remains to be determined due to the lack of a specific monoclonal antibody. Here, we developed rat monoclonal antibodies against canine DEPDC1B and characterized their applicability for immunodetection assays. Our findings demonstrated that these antibodies are functional and can be important tools to investigate the precise role of DEPDC1B in canine tumors.

Electrical Conductivity, pH, Minerals, and Sensory Evaluation of Airag (Fermented Mare's Milk).

Traditional airag (fermented mare's milk) is a sour, slightly alcoholic drink handmade by Mongolian nomads. As airag is not heated after production, the fermentation continues to proceed and the taste changes rapidly. The objective of this study was to investigate the association of the sensory taste evaluation of airag with some properties-electrical conductivity (EC), pH and concentrations of macro minerals (calcium (Ca), phosphorous (P), sulfur (S), magnesium (Mg), potassium (K), and sodium (Na))-of airag. We held an airag contest in Mogod county, one of the most famous airag production areas, in order to collect samples of airag for the analysis of airag properties and to conduct an airag taste evaluation by Mongolian people. The results of the statistical analysis indicated that the EC-value was related to the evaluation score of airag. Except for EC, no statistically significant relationship between the taste score and the other properties was found in this study. It was concluded that the EC-value would be a simple measurement indicator for evaluating the quality of airag on site.

C-reactive protein and incident hypertension in a worksite population of Japanese men.

Inflammation plays a key role in the pathogenesis of cardiovascular diseases via the development of atherosclerosis. Here, we evaluated the impact of serum C-reactive protein (CRP) and the white blood cell (WBC) count on the risk of hypertension in middle-aged Japanese men at a work site. We evaluated a total of 2991 Japanese male workers without hypertension who ranged in age from 18 to 64 years (mean age 40.4 ± 0.2 years) at a worksite in 2010. The hazard ratio (HR) for incident hypertension was estimated according to quartile levels of serum high-sensitivity CRP (hs-CRP) or WBC count. These men were followed up for 5 years from 2010 to 2015. During the follow-up period, 579 (19.4%) subjects developed hypertension. In a multivariable analysis, the risk of incident hypertension was significantly increased with higher hs-CRP levels: HR 1.00 (reference) for the lowest quartile, 1.39 (1.04-1.85) for the 2nd quartile, 1.46 (1.08-1.98) for the 3rd quartile, and 1.57 (1.17-2.11) for the highest quartile. In contrast, the WBC count was not associated with a greater risk of incident hypertension after multivariable adjustment. These findings suggest that higher levels of serum hs-CRP, but not the WBC count, are associated with the future incidence of hypertension in middle-aged Japanese men.

Impact of obstructive sleep apnea on long-term blood pressure variability in Japanese men: a cross-sectional study of a work-site population.

Blood pressure variability (BPV) has been shown to be associated with cardiovascular diseases. The effects on long-term BPV of obstructive sleep apnea (OSA) are not yet known. We evaluated a total of 1653 Japanese male workers (18-69 years) at a work site to diagnose OSA, and we divided them into three groups: non-OSA (apnea-hypopnea index (AHI): < 5, n = 1414), mild-to-moderate OSA (5 ≤ AHI < 30: n = 131) and severe OSA (AHI ≥ 30: n = 108). The standard deviation and coefficient of variation of the subjects' BPV were calculated by using their annual blood pressure measurements at routine physical examinations from 2012 to 2015 (four measurements). The multivariable-adjusted BPV of systolic blood pressure (SBP) was significantly higher in the severe-OSA group compared to the non-OSA group. A multiple regression analysis also revealed that OSA was positively associated with BPV of SBP. We focused on the mild-to-moderate OSA group to evaluate the association of OSA treatment with BPV, because most of the severe-OSA subjects were being treated with continuous positive airway pressure or an oral appliance. The BPV of both systolic and diastolic blood pressure was significantly decreased in the treated subjects. These findings suggest that OSA is associated with increases in long-term BPV which was improved by the treatment of OSA in Japanese men of a work-site population.

Impact of serum uric acid on incident hypertension in a worksite population of Japanese men.

OBJECTIVE: Higher levels of serum uric acid are associated with an increased risk of cardiovascular diseases, which may be confounded by comorbidities. We investigated the effects of serum uric acid on the risk of hypertension in Japanese men at a worksite. METHODS: We evaluated a total of 2335 Japanese male workers without hypertension who ranged in age from 18 to 64 years at a worksite in 2009. These men were followed for 6 years from 2009 to 2015. RESULTS: During the follow-up period, 380 individuals developed hypertension. The odds ratio for the incident hypertension was estimated according to quartiles of serum uric acid levels of 5.1 or less, 5.2-5.8, 5.9-6.6, and at least 6.7 mg/dl. The multivariable-adjusted risk of incident hypertension was significantly higher in the highest serum uric acid quartile than in the lowest: odds ratio 1.00 (reference) for the lowest quartile, 1.34 (0.91-1.97) for the second quartile, 1.42 (0.97-2.06) for the third quartile, and 1.65 (1.14-2.40) for the highest quartile. In stratified analyses, the association between serum uric acid and incident hypertension was significant in the patients of aged below 45 years and without comorbidities, namely diabetes and low levels of high-density lipoprotein-cholesterol. CONCLUSIONS: Serum uric acid levels were associated with the future incidence of hypertension, and the association was observed in the younger individuals, those without diabetes, and those with preserved high-density lipoprotein cholesterol levels in a worksite population of Japanese men.

Impact of obstructive sleep apnea on blood pressure and cardiovascular risk factors in Japanese men: A cross-sectional study in work-site group.

It has been shown that obstructive sleep apnea (OSA) is related to hypertension and cardiovascular disease; however, the prevalence of OSA in general population and the impact of it on blood pressure especially in Japan has not been well determined. We have conducted a screening test for OSA from 2003 to 2011. In addition, a cross-sectional analysis was performed in 2012 to determine the association of OSA and cardiovascular risk factors in Japanese men (18-69 years of age; mean age, 44.4 ± 0.2). The study group consisted of 2208 male employees, and OSA was evaluated by using the 4% oxygen desaturation index and apnea-hypopnea index (AHI). The prevalence of mild-to-moderate (5≤AHI<30) and severe (AHI≥30) OSA in the studied subjects were 7.1%, and 6.1%, respectively. Among the 135 severe OSA subjects, 105 (77.8%) had been treated with continuous positive airway pressure. Both systolic and diastolic blood pressures (DBP) were significantly increased in the subjects with severe OSA compared with those without OSA. These associations in DBP remained observed after adjustment for age, body mass index (BMI), estimated glomerular filtration rate, HbA1c, current alcohol intake, current smoking habits, and OSA treatment. DBP in severe OSA subjects were significantly increased in 1807 subjects who were not treated for hypertension or OSA. However, the levels of blood pressures were not decreased by OSA treatment. These results suggest that the prevalence of OSA is relatively high in middle-aged Japanese men and that blood pressures were elevated in the subjects with severe OSA.

Movements of dams milked for fermented horse milk production in Mongolia.

Airag, (Fermented horse milk) is a traditional milk product in Mongolia. Herders separate foals from their dams and tie them at a milking site during the daytime to produce airag. To evaluate the effects of horse management on the movement of dams, we tracked three dams in a herd in camp 1 during summer and camp 2 during autumn of 2013 and analyzed their movements during the milking (daytime) and non-milking (nighttime) periods in an area famous for its high-quality airag. Dams were gathered every 1.7 ± 0.0 h between 07.46 and 15.47 hours at the milking sites and milked 4.6 ± 0.2 times/day during the study period (86 days). Daily cumulative and maximum linear distances from the milking sites were longer (P < 0.01) during the non-milking period than during the milking period. Daily home ranges were 91 and 26 times greater during the non-milking period (P < 0.001) in camps 1 and 2, respectively. The greater range during the non-milking period would reflect the spatial distributions of water, salt and forage. The dams initially used similar areas and gradually shifted their daily home ranges after several days. This shift suggests that the dams grazed farther afield as forage availability declined around the milking site. For better airag production and sustainable pasture use, our results provide insights useful for evaluating the effects of milking management on vegetation and soil in those pastures, for selecting the appropriate milking times and frequency, and for choosing the right timing to shift milking sites.

Effect of losartan on serum uric acid in hypertension treated with a diuretic: the COMFORT study.

It has been shown that losartan, an angiotensin II receptor blocker (ARB), reduces serum uric acid levels. However, the effects of losartan on serum uric acid levels in the patients treated with a thiazide diuretic have not been fully elucidated. We have investigated the effects of losartan compared with other ARBs on blood variables and blood pressure control in hypertensive patients treated with a thiazide diuretic using data from the COMFORT study. The present analysis included a total of 118 hypertensive subjects on combination treatment with ARBs except for losartan and a diuretic who were randomly assigned to a daily regimen of a combination pill (losartan 50 mg/hydrochlorothiazide 12.5 mg) or to continuation of two pills, an ARB except for losartan and a diuretic. Blood pressures were evaluated at 1, 3, and 6 months after randomization and changes in blood variables including serum uric acid were evaluated during 6 months treatment period. Mean follow-up blood pressure levels were not different between the combination pill (losartan treatment) group and the control (ARBs except for losartan) group. On the other hand, serum uric acid significantly decreased in the combination pill group compared with the control group (-0.44 versus + 0.10 mg/dl; p = 0.01), although hematocrit, serum creatinine, sodium and potassium were not different between the groups. These results suggest that the treatment regimen switched from a combination therapy of ARBs except for losartan and a diuretic to a combination pill (losartan/ hydrochlorothiazide) decreases serum uric acid without affecting blood pressure control.

Does a combination pill of antihypertensive drugs improve medication adherence in Japanese? A randomized controlled trial.

BACKGROUND: In order to achieve target blood pressure levels to prevent cardiovascular disease, combination therapy of antihypertensive drugs is often required, although it is thought that requiring a patient to take many different pills would reduce adherence to the medication regimen. Whether antihypertensive treatment with a single pill combining antihypertensive drugs would improve medication adherence and blood pressure control was investigated. METHODS AND RESULTS: A total of 207 hypertensive subjects were randomly assigned to a combination pill group (losartan 50mg/hydrochlorothiazide 12.5mg; n=103) or a control group (an angiotensin receptor blocker and a thiazide diuretic; n=104). Medication adherence was evaluated by pill counts at 1, 3, and 6 months after randomization. The mean adherence rates over 6 months were not different between the 2 groups: 98% in the combination pill group and 98% in the control group. Moreover, the 2 groups included similar numbers of subjects with relatively poor adherence rates (<90%) in each treatment period. The mean blood pressures over the 6-month treatment period were not different between the groups: 131/75 mmHg in the combination pill group and 130/75 mmHg in the control group (P=0.84/0.96). CONCLUSIONS: There were no appreciable effects of the combination pill of antihypertensive drugs on medication adherence or blood pressure control in Japanese patients over a 6-month period.

Salt intake in 3-year-old Japanese children.

Salt restriction is widely recommended for the prevention and management of hypertension. It is very important to reduce salt intake during childhood. This survey was conducted to investigate salt intake in Japanese children. The study subjects, totalling 1,424, comprised 3-year-old children who received health checkups conducted by a public health center. Using first-morning urine samples, urinary concentrations of sodium, potassium and creatinine were measured. Additionally, the participants' parents filled out a questionnaire regarding their children's dietary habits. Urinary sodium and potassium concentrations were 140 ± 67 and 67 ± 41 mmol l(-1), respectively. The estimated urinary sodium excretion values had a wide distribution, with a mean value of 75 ± 47 mmol per day (4.4 g per day of salt). Sodium excretion exceeded 100 and 200 mmol per day in 336 (24%) and 32 (2.2%) subjects, respectively. Urinary sodium excretion was significantly higher in children who had older siblings than in firstborn children (78 ± 49 vs. 72 ± 45 mmol per day, P < 0.05). The urinary sodium/potassium ratio was also higher in the former group (3.0 ± 2.7 vs. 2.7 ± 2.2, P < 0.01). Sodium excretion tended to be higher in children who ate daily snacks than in those who did not (76 ± 48 vs. 71 ± 44 mmol per day, P=0.07). In contrast, potassium excretion was significantly higher and the sodium/potassium ratio was lower in children who ate fruits daily than those who did not (39 ± 29 vs. 33 ± 23 mmol per day and 2.6 ± 2.0 vs. 3.2 ± 2.7, P < 0.01, respectively). These results suggest that excess salt intake occurs in a significant number of 3-year-old Japanese children. The presence of older siblings and dietary habits of eating snacks or fruits influence their sodium and potassium intake.

Blood pressure and lipid control status in Japanese hypertensive patients.

Strict blood pressure (BP) as well as lipid control is important to prevent cardiovascular events. The purpose of this study was to evaluate BP and lipid control status in hypertensive patients. Subjects were a total of 717 hypertensive patients who had been followed at National Kyushu Medical Center in FuKuoka, Japan. Goal BP was defined as < 130/85 mmHg (< 65 years) or < 140/90 mmHg (> or = 65 years). According to the Japanese guidelines, goal LDL cholesterol (LDL-C) levels were defined based on the patient category. Average BP level and the number of anti-hypertensive drugs were 133 + or - 12/74 + or - 9 mmHg and 2.1 + or - 1.1, respectively, and the LDL-C level was 119 + or - 27 mg/dl. Goal BP was achieved in 40% of the patients of < 65 years and 67% of the elderly patients. Goal LDL-C was achieved in 65% of the patients. Even in the patients taking lipid-lowering agents (n = 178), 30% failed to achieve goal LDL-C levels. In the patients who achieved BP < 130/85 mmHg, 67% also achieved goal LDL-C, whereas 61% of the patients whose BP > or = 140/90 mmHg achieved goal LDL-C. Both goal BP and LDL-C were achieved in 39% of the male and 36% of the female patients. In contrast, neither goal BP nor goal LDL-C was achieved in 16% of the male and 17% of the female patients. Results suggest that intensive intervention should be required to achieve satisfactory BP and lipid control in hypertensive patients.

Classification of the binding modes in bovine serum albumin using terminally substituted alkane analogues.

With the fluorescence probe of 8-anilino-1-naphthalenesulfonate (ANS), the binding modes of terminally substituted alkane analogues (C(n)X; X = COOH, OH, CHO, NH(3), CONH(2)) to bovine serum albumin (BSA) were investigated using a competitive binding technique. The Scatchard plot of the fluorometric titration of BSA with ANS showed that the maximum binding number of ANS, n(max), was 3.81, with the binding constant, K(bnd), of 1.42 x 10(6) mol(-1) dm(3). The binding modes of C(n)X to BSA were analyzed based on the fluorometric titration of the ANS and BSA mixture with C(n)X. C(n)COOH completely displaced the ANS bound to BSA, whereas C(n)OH and C(n)CHO displaced only about 40% of the ANS bound to BSA. In contrast, C(n)NH(2) and C(n)CONH(2) displaced very little bound ANS. By comparing these results, we classified the binding modes of C(n)X to BSA into three types. Two of them are detectable with the ANS fluorescence and the remaining one is not detectable with the fluorescence.

Isolation of yeast Kurtzmanomyces sp. I-11, novel producer of mannosylerythritol lipid.

Yeast strains were screened for producers of glycolipid-type biosurfactants from soybean oil as a sole carbon source. The structure of the glycolipid (MEL-I-11) produced by strain I-11 was analyzed. The hydrophilic sugar moiety was mannosylerythritol and the fatty acid components were C8:0 (36.4%), C12:0 (11.9%), and C14:2 (25.9%). The MEL-I-11 was identified as 6-O-acetyl-2,3-di-O-alkanoyl-beta-D-mannopyranosyl-(1-->4)-O-meso-erythritol. The strain I-11 was identified as a Kurtzmanomyces species, a novel producer of mannosylerythritol lipid.