Genetic diversity of Lactobacillus delbrueckii isolated from raw milk in Hokkaido, Japan.

Lactic acid bacteria (LAB) play important roles in acid production and flavor formation in fermented dairy products. Lactic acid bacteria strains with distinct characteristics confer unique features to products. Diverse LAB have been identified in raw milk and traditional fermented milk prepared from raw milk. However, little is known about LAB in raw milk in Japan. To preserve diverse LAB as potential starters or probiotics for future use, we have isolated and identified various kinds of LAB from raw milk produced in Japan. In this study, we focused on Lactobacillus delbrueckii, one of the most important species in the dairy industry. We identified L. delbrueckii subspecies isolated from raw milk in Hokkaido, Japan, by analyzing intraspecific diversity using 4 distinct methods, hsp60 cluster analysis, multilocus sequence analysis, core-genome analysis, and whole-genome analysis based on average nucleotide identity. The subspecies distribution and a new dominant subset of L. delbrueckii from raw milk in Japan were revealed. The discovery of new strains with different genotypes is important for understanding the geographic distribution and characteristics of the bacteria and further their use as a microbial resource with the potential to express unconventional flavors and functionalities. The strains identified in this study may have practical applications in the development of fermented dairy products.

Diet-induced alteration of fatty acid synthase in prostate cancer progression.

Fatty acid synthase (FASN) is a cytosolic metabolic enzyme that catalyzes de novo fatty acid synthesis. A high-fat diet (HFD) is attributed to prostate cancer (PCa) progression, but the role FASN on HFD-mediated PCa progression remains unclear. We investigated the role of FASN on PCa progression in LNCaP xenograft mice fed with HFD or low-fat diet (LFD), in PCa cells, and in clinical PCa. The HFD promoted tumour growth and FASN expression in the LNCaP xenograft mice. HFD resulted in AKT and extracellular signal-regulated kinase (ERK) activation and 5' adenosine monophosphate-activated protein kinase (AMPK) inactivation. Serum FASN levels were significantly lower in the HFD group (P=0.026) and correlated inversely with tumour volume (P=0.022). Extracellular FASN release was enhanced in the PCa cells with phosphatidylinositol 3-kinase (PI3K)/mitogen-activated protein kinase (MAPK) inhibition and AMPK signalling activation. FASN inhibition resulted in decrease of PCa cell proliferation through PI3K/MAPK downregulation and AMPK activation. Furthermore, AMPK activation was associated with FASN downregulation and PI3K/MAPK inactivation. Clinically, high FASN expression was significantly associated with high Gleason scores and advanced pathological T stage. Moreover, FASN expression was markedly decreased in the PCa response to androgen deprivation therapy and chemotherapy. HFD modulates FASN expression, which may be an important mechanism in HFD-associated PCa progression. Furthermore, a critical stimulatory loop exists between FASN and the PI3K/MAPK system, whereas AMPK signalling was associated with suppression. These may offer appropriate targets for chemoprevention and cancer therapy in HFD-induced PCa.

Utilization of native oxygen in Eu(RE)-doped GaN for enabling device compatibility in optoelectronic applications.

The detrimental influence of oxygen on the performance and reliability of V/III nitride based devices is well known. However, the influence of oxygen on the nature of the incorporation of other co-dopants, such as rare earth ions, has been largely overlooked in GaN. Here, we report the first comprehensive study of the critical role that oxygen has on Eu in GaN, as well as atomic scale observation of diffusion and local concentration of both atoms in the crystal lattice. We find that oxygen plays an integral role in the location, stability, and local defect structure around the Eu ions that were doped into the GaN host. Although the availability of oxygen is essential for these properties, it renders the material incompatible with GaN-based devices. However, the utilization of the normally occurring oxygen in GaN is promoted through structural manipulation, reducing its concentration by 2 orders of magnitude, while maintaining both the material quality and the favorable optical properties of the Eu ions. These findings open the way for full integration of RE dopants for optoelectronic functionalities in the existing GaN platform.

Coping strategies and risk of cardiovascular disease incidence and mortality: the Japan Public Health Center-based prospective Study.

AIMS: Coping strategies may be significantly associated with health outcomes. This is the first study to investigate the association between baseline coping strategies and cardiovascular disease (CVD) incidence and mortality in a general population cohort. METHODS AND RESULTS: The Japan Public Health Center-based prospective Study asked questions on coping in its third follow-up survey (2000-04). Analyses on CVD incidence and mortality included 57 017 subjects aged 50-79 without a history of CVD and who provided complete answers on approach- and avoidance-oriented coping behaviours and strategies. Cox regression models, adjusted for confounders, were used to determine hazard ratios (HRs) according to coping style. Mean follow-up time was 7.9 years for incidence and 8.0 years for mortality.The premorbid use of an approach-oriented coping strategy was inversely associated with incidence of stroke (HR = 0.85; 95% CI, 0.73-1.00) and CVD mortality (HR = 0.74; 95% CI, 0.55-0.99). Stroke subtype analyses revealed an inverse association between the approach-oriented coping strategy and incidence of ischaemic stroke (HR = 0.79; 95% CI, 0.64-0.98) and a positive association between the combined coping strategy and incidence of intra-parenchymal haemorrhage (HR = 2.03; 95% CI, 1.01-4.10). Utilizing an avoidance coping strategy was associated with increased mortality from ischaemic heart disease (IHD) only in hypertensive individuals (HR = 3.46; 95% CI, 1.07-11.18). The coping behaviours fantasizing and positive reappraisal were associated with increased risk of CVD incidence (HR = 1.24; 95% CI, 1.03-1.50) and reduced risk of IHD mortality (HR = 0.63; 95% CI, 0.40-0.99), respectively. CONCLUSION: An approach-oriented coping strategy, i.e. proactively dealing with sources of stress, may be associated with significantly reduced stroke incidence and CVD mortality in a Japanese population-based cohort.

Detection of methicillin-resistant Staphylococcus pseudintermedius ST169 and novel ST354 SCCmec II-III isolates related to the worldwide ST71 clone.

The recent appearance of methicillin-resistant Staphylococcus pseudintermedius (MRSP) is a concern for both veterinary and human healthcare. MRSP clonal lineages with sequence type (ST) 71-spa t02-staphylococcal cassette chromosome mec (SCCmec) II-III and ST68-spa t06-SCCmec V have spread throughout Europe and North America, respectively. The current study compared the molecular characteristics of 43 MRSP isolates from dogs in Japan with those of MRSP from previous reports using multilocus sequence typing based on seven housekeeping genes, SCCmec typing, and detection of antimicrobial resistance genes. Three related clonal lineages, ST71, ST169, and the newly registered ST354, were observed in SCCmec II-III isolates from Japan, despite MRSP SCCmec II-III isolates being thought to belong to a single clonal lineage. The majority of SCCmec II-III isolates belonging to ST169 (9/11) and ST354 (3/3), but not ST71 (0/11), harboured tetM. Four STs were observed for the SCCmec V isolates; however, neither ST68 nor related STs were found in the Japanese MRSP isolates. In conclusion, MRSP SCCmec II-III isolates from Japan belonged to ST71 and related STs (ST169 and ST354). A variety of MRSP SCCmec V clones, including some novel clones, were identified.

Cognitive and affective impairments of a novel SCA/MND crossroad mutation Asidan.

BACKGROUND: A variety of hereditary spinocerebellar ataxia (SCA) develops a broad spectrum of both ataxia and non-ataxia symptoms. Cognitive and affective changes are one such non-ataxia symptoms, but have been described only in hereditary SCAs with exonic CAG gene expansion. METHODS: We newly found intronic hexanucleotide GGCCTG gene expansion in NOP56 gene as the causative mutation (=SCA36) in nine unrelated Japanese familial SCA originating from Asida river area in the western part of Japan, thus nicknamed Asidan for this mutation. These patients show unique clinical balance of cerebellar ataxia and motor neuron disease (MND), locating on the crossroad of these two diseases. In the nine families, 14 patients were clinically examined and genetically confirmed to Asidan. In the present study, we examined cognitive and affective analyses on 12 patients (seven men and five women) who agreed to join the examination with average age at onset of 53.1 ± 3.2 years, average duration of 12.1 ± 5.2 years, and current average age at 65.1 ± 6.2 years. RESULTS: The 12 Asidan patients demonstrated a significant decrease in their frontal executive functions measured by frontal assessment battery (FAB) and Montreal cognitive assessment (MoCA) compared with age- and gender-matched controls, whilst mini-mental state examination (MMSE) and Hasegawa dementia score-revised (HDS-R) were within normal range. The decline of frontal executive function was related to their disease duration and scale for the assessment and rating of ataxias (SARA). They also demonstrated mild depression and apathy. Single-photon emission tomography (SPECT) analysis showed that these Asidan patients showed decline of regional cerebral blood flow (rCBF) in a particular areas of cerebral cortices such as Brodmann areas 24 and 44-46. CONCLUSION: These data suggest that the patients with Asidan mutation show unique cognitive and affective characteristics different from other hereditary SCAs with exonal CAG expansion or MND.

f Electron contribution to the change of electronic structure in CeRu2Si2 with temperature: a Compton scattering study.

High resolution Compton profiles have been measured in the single crystal of CeRu(2)Si(2) above and below the Kondo temperature to elucidate the change of the Ce-4f electron from localized to itinerant states. Two-dimensional electron occupation number densities projected on the first Brillouin zone, which are obtained after a series of analyses, clearly specify the difference between itinerant and localized states. The contribution of Ce-4f electrons to the electronic structure is discussed by contrast with a band calculation.

Role of oxygen electrons in the metal-insulator transition in the magnetoresistive oxide La2-2xSr1+2xMn2O7 probed by compton scattering.

We have studied the [100]-[110] anisotropy of the Compton profile in the bilayer manganite. Quantitative agreement is found between theory and experiment with respect to the anisotropy in the two metallic phases (i.e., the low temperature ferromagnetic and the colossal magnetoresistant phase under a magnetic field of 7 T). Robust signatures of the metal-insulator transition are identified in the momentum density for the paramagnetic phase above the Curie temperature. We interpret our results as providing direct evidence for the transition from the metalliclike to the admixed ionic-covalent bonding accompanying the magnetic transition. The number of electrons involved in this phase transition is estimated. Our study demonstrates the sensitivity of the Compton scattering technique for identifying the number and type of electrons involved in the metal-insulator transition.

Autosomal dominant moyamoya disease maps to chromosome 17q25.3.

BACKGROUND: Moyamoya disease (MMD) is an idiopathic steno-occlusive cerebrovascular disease that represents an important cause of stroke. However, etiology of the disease has remained largely unknown. METHODS: We previously showed that the inheritance pattern of MMD is autosomal dominant with incomplete penetrance. Here, we report the genome-wide parametric linkage analysis for MMD in 15 extended Japanese families. We conducted linkage analyses under two diagnostic classifications: narrow and broad. Affected member-only analysis was applied due to incomplete and age-dependent penetrance of the disease. RESULTS: Under both classifications, significant evidence of linkage was only observed on chromosome 17q25.3, with maximum multipoint logarithm of odds (lod) scores of 6.57 (under the narrow classification) and 8.07 (under the broad classification) at D17S704. Haplotype analysis revealed segregation of a disease haplotype in all families but one, and informative crossovers enabled mapping of the MMD locus to a 3.5-Mb region between D17S1806 and the telomere of 17q, encompassing 94 annotated genes. CONCLUSIONS: Our data suggest that there is a major gene locus for autosomal dominant moyamoya disease on chromosome 17q25.3.

Smoking status, sports participation and mortality from coronary heart disease.

BACKGROUND: Since smoking and exercise have opposite effects on coronary risk factors, the hypothesis was proposed that smoking might weaken the protective effect of exercise on prevention of coronary heart disease. OBJECTIVE: To determine the effect of smoking on the relationship between sports participation and mortality from coronary heart disease. DESIGN: Population-based prospective cohort study in Japan. PARTICIPANTS: A total of 76 832 Japanese men and women, aged 40-79 years with no history of stroke, coronary heart disease, or cancer, completed a self-administered questionnaire between 1988 and 1990. MAIN OUTCOME MEASURES: Systematic mortality surveillance was carried out through 2003, and 638 deaths from coronary heart disease (496 myocardial infarction) were identified. RESULTS: People who reported the longest time in sports participation (>or=5 hours/week) had an approximately 50-80% lower age-adjusted risk of mortality from coronary heart disease compared with those in the second lowest category (1-2 hours/week) among never and ex-smokers, but no association was found among current smokers. Adjustment for known risk factors and exclusion of subjects who died within 2 years of the baseline inquiry did not substantially alter these associations. The multivariable hazard ratios (95% confidence interval) of coronary heart disease for the >or=5 hours/week versus 1-2 hours/week of sports participation were 0.44 (0.23 to 0.86) among never smokers, 0.18 (0.05 to 0.60) among ex-smokers, and 0.82 (0.47 to 1.40) among current smokers. Similar associations were found for men and women. CONCLUSIONS: Smoking may reduce the beneficial effect of sports participation for reduction of fatal coronary heart disease.

Inheritance pattern of familial moyamoya disease: autosomal dominant mode and genomic imprinting.

BACKGROUND: Although the aetiology of moyamoya disease (MMD) has not been fully clarified, genetic analysis of familial MMD (F-MMD) has considerable potential to disclose it. OBJECTIVE: To determine the inheritance pattern and clinical characteristics of F-MMD to enable precise genetic analyses of the disease. METHODS: 15 highly aggregated Japanese families (52 patients; 38 women and 14 men) with three or more affected members were examined. The difference in categories of age at onset (child onset, adult onset and asymptomatic) between paternal and maternal transmission was compared by chi2 statistics. RESULTS: In all families there had been three or more generations without consanguinity, and all types of transmission, including father-to-son, were observed. Among a total of 135 offspring of affected people, 59 (43.7%) were patients with MMD or obligatory carriers. Affected mothers were more likely to produce late-onset (adult-onset or asymptomatic) female offspring (p = 0.007). CONCLUSIONS: The mode of inheritance of F-MMD is autosomal dominant with incomplete penetrance. Thus, in future genetic studies on F-MMD, parametric linkage analyses using large families with an autosomal dominant mode of inheritance are recommended. Genomic imprinting may be associated with the disease.

Parental longevity and mortality amongst Japanese men and women: the JACC Study.

OBJECTIVES: To examine whether the risk of mortality varies according to parents' age at death. DESIGN AND SUBJECTS: A large prospective study in Japanese men and women from 45 communities across Japan. A total of 51 485 men and women aged 40-79 years completed self-administered questionnaires at baseline and followed up for 9.6 years. RESULTS: The risk of mortality from stroke, cardiovascular disease, and all causes was 20-30% lower in men and women with fathers who died at age > or = 80 years, compared with those with fathers whose age at death was <60 years. A similar reduction was found when the age at death of mothers was > or = 85 years compared with <65 years. Furthermore, the risk reduction was more evident amongst persons with both parents being long-lived parents compared with those with being short-lived parents, especially for death from cardiovascular disease. CONCLUSIONS: Our findings indicate that parental longevity could be a predictor for reduced risk of mortality from stroke, cardiovascular disease, and all causes for both Japanese men and women.

KNT-artificial neural network model for flux prediction of ultrafiltration membrane producing drinking water.

This paper describes the prediction of flux behavior in an ultrafiltration (UF) membrane system using a Kalman neuro training (KNT) network model. The experimental data was obtained from operating a pilot plant of hollow fiber UF membrane with groundwater for 7 months. The network was trained using operating conditions such as inlet pressure, filtration duration, and feed water quality parameters including turbidity, temperature and UV254. Pre-processing of raw data allowed the normalized input data to be used in sigmoid activation functions. A neural network architecture was structured by modifying the number of hidden layers, neurons and learning iterations. The structure of KNT-neural network with 3 layers and 5 neurons allowed a good prediction of permeate flux by 0.997 of correlation coefficient during the learning phase. Also the validity of the designed model was evaluated with other experimental data not used during the training phase and nonlinear flux behavior was accurately estimated with 0.999 of correlation coefficient and a lower error of prediction in the testing phase. This good flux prediction can provide preliminary criteria in membrane design and set up the proper cleaning cycle in membrane operation. The KNT-artificial neural network is also expected to predict the variation of transmembrane pressure during filtration cycles and can be applied to automation and control of full scale treatment plants.

Gender-related difference, geographical variation and time trend in dietary cadmium intake in Japan.

OBJECTIVES: The present analysis was initiated to examine possible gender-dependency, geographic variation, and time-dependent changes in dietary intake of cadmium (Cd-F) among general populations in Cd-non-polluted areas in Japan. The role of rice as Cd-F source was also within the scope. METHODS: Two databases on Cd and nutritional analyses were re-visited. Both databases were established through collection of 24-h food duplicate portion samples from residents in areas with no known Cd pollution, and contained information on Cd and energy contents in the duplicate portion, together with daily rice consumption, the gender, the age and the location of the residence of each sample donor. The first and the second databases were established through surveys in the years around 1980 on 564 cases and around 1995 on 702 cases, respectively. The two databases were combined for evaluation by multiple regression (MRA) and other analyses. RESULTS: The analyses showed that men tended to take more Cd than women, more clearly so in the 1980 survey than in the 1995 survey. When Cd-F in the 1995 survey was compared with that in the 1980 survey, a substantial decrease was observed, e.g. by 30% (from 37.5 to 26.2 microg/day) in case of women. Cd-F values varied subject to the survey sites in a wide range (e.g. from 20 to 86 microg/day among women in the 1980 survey). In MRA with Cd-F as a dependent variable and survey sites and food intake factors (e.g. rice and energy intakes) as independent variables, the survey sites could explain more than 53% and 35% of total variation in Cd-F in the 1980 and 1995 surveys, respectively. Rice consumption was also influential to Cd-F in both surveys with partial correlation coefficients of 0.36 and 0.21, respectively, the influence being stronger in the 1980 survey than in the 1995 survey. A significant correlation was detected between the 1980 and 1995 survey results both in Cd-F and in rice consumption. CONCLUSION: Geographic and gender-related differences, and time-dependent decrease in dietary Cd intake in Japan were detected through analyses of food duplicate-based databases. The leading role of rice as dietary Cd source was also observed.

Perfluorooctane sulfonate concentrations in surface water in Japan.

Perfluorooctane sulfonate (PFOS) is a class of specialty chemicals used in a variety of applications, and has been found to be globally distributed in many living organisms including humans. Several analytical methods have been developed for determination of PFOS in environmental samples and biological matrices. However, these methods employ liquid chromatography/tandem mass spectrometry (LC/MS/MS), an instrumentation which has limited accessibility because it is expensive to use and maintain. In the present study we present the development of a robust analytical method using liquid chromatography/mass spectrometry (LC/MS) in combination with solid phase extraction. The high yield and concentration of the present method enabled us to quantify PFOS as low as 0.1 ng/L. This method was applied to the determination of PFOS in 142 surface water samples collected from various geographic locations around Japan. The geometric mean (geometric standard deviation) (ng/L) for river samples (n = 126) was 2.37 (4.13), with a median of 1.68 and a range of 0.3-157 ng/L, and for coastal sea water samples (n = 16) was 1.52 (4.14), with a median of 1.21 and a range of 0.2-25.2 ng/L. However, the concentrations in most of the samples were much lower than the values reported in the US, except for those from the Jinzu (135.0 ng/L) and Tama (157 ng/L) Rivers. Because surface waters in the Ara (13.0-38.5 ng/L), Tama (0.7-157.0 ng/L), and Yodo (0.9-27.3 ng/L) Rivers, sources of drinking water for more than eight million people, were moderately contaminated with PFOS, more work is needed to assess exposure to PFOS.

Increased expression of glutathione S-transferase in renal proximal tubules in the early stages of diabetes: a study of type-2 diabetes in the Akita mouse model.

BACKGROUND/AIM: The objective of this study was to examine whether the gene expression profile in the kidney is modified by hyperglycemia in the early stage of diabetes. METHODS: We analyzed the expression of kidney mRNAs using cDNA array membranes including 588 genes in the kidney of the Akita mouse, a model of type-2 diabetes, after exposure to hyperglycemia for a moderate length of time, but before the manifestation of diabetic glomerulosclerosis. Western blot analysis and immunohistochemical studies were performed to confirm whether the protein for the increasingly expressed mRNA was highly expressed in the kidney of the diabetic mouse. RESULTS: Two of the 10 detected mRNAs, glutathione S-transferase (GST) alpha and mu, in the kidneys from diabetic mice showed a more than twofold increased expression in comparison to those of control mice. Western blot analysis in kidney tissue extracts confirmed increases in GST alpha and mu at protein levels in the diabetic mice. Immunohistochemical studies revealed strong staining for those proteins in the proximal tubules of diabetic mice. CONCLUSION: These data collectively indicate that expression of GSTs is increased in epithelial cells in proximal tubules even at the early stage of diabetes, probably in response to oxidative stress triggered by hyperglycemia or other toxic effects of glucose.