Structural and functional glycosphingolipidomics by glycoblotting with aminooxy-functionalized gold nanoparticle

Glycosphingolipids (GSLs) synthesized in Golgi apparatus by sequential transfer of sugar residues to a ceramide lipid anchor are ubiquitously distributing on vertebrate plasma membranes. Standardized method allowing for high throughput structural profiling and functional characterization of living cell surface GSLs is of growing importance because they function as crucial signal transduction molecules in various processes of dynamic cellular recognitions. However, methods are not available for amplification of GSLs, while the genomic scale PCR amplification permits large-scale mammalian proteomic analysis.　Here we communicate such an approach to a novel "omics", namely glycosphingolipidomics based on the glycoblotting method. The method, which involves selective ozonolysis of the C-C double bond in ceramide moiety and subsequent enrichment of generated GSL-aldehydes by chemical ligation using aminooxy-functionalized gold nanoparticle (aoGNP) should be of widespread utility for identifying and characterizing whole GSLs present in the living cell surfaces. The present protocol using glycoblotting permitted MALDI-TOFMS-based high throughput structural profiling of mouse brain gangliosides such as GM1, GD1a/GD1b, and GT1b for adult or GD3 in case for embryonic mouse. When mouse melanoma B16 cells were subjected to this protocol, it was demonstrated that gangliosides enriched from the plasma membranes are only GM3 bearing microheteogeneity in the structure of N-acyl chain. Surface plasmon resonance analysis revealed that aoGNP displaying whole GSLs blotted from mouse B16 melanoma cell surfaces can be used directly for monitoring specific interaction with self-assembled monolayer (SAM) of Gg3Cer (gangliotriaosylceramide). Our results indicate that GSL-selective enrichment onto aoGNP from living cell surfaces allows for rapid reconstruction of plasma membrane models mimicking intact GSL-microdomain feasible for further structural and functional characterization

Distribution of Air Temperature in Tajimi City in Summer

To research the distribution of air temperature in the city of Tajimi, Gifu prefecture, a total of 15 thermometers were placed at the schools and parks in the city of Tajimi, and the city of Kasugai, Aichi prefecture in August 2010. From the distribution of monthly average air temperature, air temperature at the center of Tajimi city was higher than the suburbs. Also, the days of daily minimum air temperature more than or equal to 25°C and daily maximum air temperature more than or equal to 35°C at the elementary school near the center of Tajimi city was more than those at other schools. This tendency appeared more clearly on the days of daily minimum air temperature more than or equal to 25°C. Also, the air temperature near the center of the city was higher than that ofthe suburbs in the early morning. Thus, it was indicated that the air temperature was hard to decrease as the bottom of the basin. From these results, the influence of urbanization to the formation of the daily minimum temperature in Tajimi city was indicated

Five isoforms of the phosphatidylinositol 3-kinase regulatory subunit exhibit different associations with receptor tyrosine kinases and their tyrosine phosphorylations

AbstractThere are five isoforms of the regulatory subunit for the heterodimeric type of phosphatidylinositol 3-kinase. These five regulatory subunit isoforms were overexpressed using an adenovirus transfection system, and their own tyrosine phosphorylations and associations with various tyrosine kinase receptors were investigated. When overexpressed in CHO-PDGFR cells, the associations of these regulatory subunit isoforms with the platelet-derived growth factor receptor were similar. However, when overexpressed in CHO-IR cells, p55γ exhibited a significantly lower ability to bind with IRS-1 upon insulin stimulation, as compared with other regulatory subunit isoforms. Furthermore, p55α and p55γ were found to be tyrosine-phosphorylated. Finally, interestingly, when overexpressed in CHO-EGFR cells or A431 cells and stimulated with epidermal growth factor (EGF), phosphorylated EGF receptor was detected in p85α, p85β and p50α immunoprecipitates, but not in p55α and p55γ immunoprecipitates. In addition, EGF-induced tyrosine phosphorylation was observed in p85α, p85β, p55α and p55γ, but not in p50α, immunoprecipitates. Thus, each regulatory subunit exhibits specific responses regarding both the association with tyrosine-phosphorylated substrates and its own tyrosine phosphorylation. These results suggest that each isoform possesses specific roles in signal transduction, based on its individual tyrosine kinase receptor

Observational Study and Numerical Prediction Experiments on Wet-Bulb Globe Temperature in Tajimi, Gifu Prefecture: Consideration of Uncertainty with a Physics Parameterization Scheme and Horizontal Resolution of the Weather Research and Forecasting Model

As part of research on the actual conditions of the thermal environment surrounding the city of Tajimi, Gifu prefecture, and the city of Kasugai, Aichi prefecture, the surface air temperature and wet-bulb globe temperature (WBGT) were investigated by observation at these locations on clear-sky days in August 2010. Numerical prediction experiments on the WBGT were performed to confirm the utility of the Weather Research and Forecasting (WRF) model. Sensitivity experiments utilizing physics parameterization schemes and horizontal resolution of the WRF model were conducted to confirm the predicted WBGT. The resultsshowed that the maximum sensitivity with the parameterization scheme was 8.4°C for the daytime average, and especially, the simple thermal diffusion surface (SLAB) scheme caused an over/estimation of 6.8°C. On the other hand, the maximum sensitivity with horizontal resolution was 0.5°C, which is much less than that with the parameterization scheme

The Tokyo Oldest Old Survey on Total Health (TOOTH): A longitudinal cohort study of multidimensional components of health and well-being

<p>Abstract</p> <p>Background</p> <p>With the rapid worldwide increase in the oldest old population, considerable concern has arisen about the social and economic burden of diseases and disability in this age group. Understanding of multidimensional structure of health and its life-course trajectory is an essential prerequisite for effective health care delivery. Therefore, we organized an interdisciplinary research team consisting of geriatricians, dentists, psychologists, sociologists, and epidemiologists to conduct a longitudinal observational study.</p> <p>Methods/Design</p> <p>For the Tokyo Oldest Old Survey on Total Health (TOOTH) study, a random sample of inhabitants of the city of Tokyo, aged 85 years or older, was drawn from the basic city registry. The baseline comprehensive assessment consists of an in-home interview, a self-administered questionnaire, and a medical/dental examination. To perform a wide variety of biomedical measurements, including carotid ultrasonography and a detailed dental examination, participants were invited to our study center at Keio University Hospital. For those who were not able to visit the study center, we provided the option of a home-based examination, in which participants were simultaneously visited by a geriatrician and a dentist. Of 2875 eligible individuals, a total of 1152 people were recruited, of which 542 completed both the in-home interview and the medical/dental examination, with 442 completed the in-home interview only, and another 168 completed self or proxy-administered data collection only. Carotid ultrasonography was completed in 458 subjects, which was 99.6% of the clinic visitors (n = 460). Masticatory assessment using a colour-changeable chewing gum was completed in 421 subjects, a 91.5% of the clinic visitors.</p> <p>Discussion</p> <p>Our results demonstrated the feasibility of a new comprehensive study that incorporated non-invasive measurements of subclinical diseases and a detailed dental examination aiming at community-dwelling individuals aged 85 years or older. The bimodal recruitment strategy is critically important to capture a broad range of health profiles among the oldest old. Results form the TOOTH study will help develop new models of health promotion, which are expected to contribute to an improvement in lifelong health and well-being.</p> <p>Trial Registration</p> <p>This study has been registered in the UMIN-Clinical Trial Registry (CTR), ID: UMIN000001842.</p

Effects of a high-sodium diet on renal tubule Ca2+ transporter and claudin expression in Wistar-Kyoto rats

Background: Urinary Ca^{2+} excretion increases with dietary NaCl. NaCl-induced calciuria may be associated with hypertension, urinary stone formation and osteoporosis, but its mechanism and long-term effects are not fully understood. This study examined alterations in the expressions of renal Ca^{2+} transporters, channels and claudins upon salt loading to better understand the mechanism of salt-induced urinary Ca^{2+} loss. Methods: Eight-week old Wistar-Kyoto rats were fed either 0.3% or 8% NaCl diet for 8 weeks. Renal cortical expressions of Na+/Ca2+ exchanger 1 (NCX1), Ca^{2+} pump (PCMA1b), Ca^{2+} channel (TRPV5), calbindin-D28k, and claudins (CLDN-2, -7, -8, -16 and −19) were analyzed by quantitative PCR, western blot and/or immunohistochemistry. Results: Fractional excretion of Ca^{2+} increased 6.0 fold with high-salt diet. Renal cortical claudin-2 protein decreased by approximately 20% with decreased immunological staining on tissue sections. Claudin-16 and −19 expressions were not altered. Renal cortical TRPV5, calbindin-D28k and NCX1 expressions increased 1.6, 1.5 and 1.2 fold, respectively. Conclusions: Chronic high-salt diet decreased claudin-2 protein and increased renal TRPV5, calbindin-D28k, and NCX1. Salt loading is known to reduce the proximal tubular reabsorption of both Na+ and Ca^{2+}. The reduction in claudin-2 protein expression may be partly responsible for the reduced Ca^{2+} reabsorption in this segment. The concerted upregulation of more distal Ca^{2+}-transporting molecules may be a physiological response to curtail the loss of Ca^{2+}, although the magnitude of compensation does not seem adequate to bring the urinary Ca^{2+} excretion down to that of the normal-diet group

Avaliação de um Treinamento de Habilidades Sociais (THS) com universitários e recém-formados

  O objetivo desta pesquisa foi avaliar a efetividade de um treinamento de habilidades sociais, através de medidas de avaliação pré e pós-teste, com universitários e/ou recém-formados que apresentavam queixas interpessoais. Participaram 15 universitários e três recém-formados que procuraram o Centro de Psicologia Aplicada de uma universidade pública. O programa contou com vinte encontros semanais em grupo, distribuídos no ano letivo, com duração de duas horas cada; foram conduzidos seis grupos de intervenção com média de três participantes cada. Como medida de avaliação foi utilizado o IHS-Del-Prette, cujos dados foram tratados através de estatística não paramétrica (Teste Wilcoxon). Os resultados indicaram aquisições importantes apontando para a efetividade do procedimento de intervenção. De todo modo, os achados precisam ser relativizados devido ao número reduzido da amostra, ao uso do relato verbal como fonte primária de dados, pela ausência de grupo controle e de avaliação de seguimento, o que poderá ser testado em pesquisas futuras. Palavras-chave: treinamento de habilidades sociais; estudantes universitários; intervenção psicológica; prevenção.El objetivo de esta investigación es evaluar la eficacia de un entrenamiento de social, con medidas del pago diario y de la después-prueba de la evaluación, con el estudiante de universidades y/o las capacidades apenas-formadas que presentaron quejas interpersonales. 15 apenas-formaron a estudiante de universidades y tres habían participado que habían mirado el centro de la psicología aplicado de una universidad pública.  El programa contó en la reunión semanal veinte en el grupo, distribuido en el año escolar, con la duración de dos horas por cada uno; seis grupos de intervención con el promedio de cinco participantes habían sido plomo cada uno. Mientras que la medida de la evaluación fue utilizada el Su-Del-Prette, que dado habían sido tratados con la distribución de la estadística libremente (prueba de Wilcoxon). Los resultados habían indicado las adquisiciones importantes que señalaban con respecto a la eficacia del procedimiento de la intervención. Toda la manera, los resultados necesita ser relativized tuvo que el número reducido de la muestra y también para la ausencia del grupo que ha controlado y evaluación de perseguir, qué podría ser probada en la investigación futura.The objective of this research was to evaluate the effectiveness of a social skills training, through pré and post-test evaluation measurements, in a group of college students and/or recent graduates who showed interpersonal complaints. Fifteen college students and three recent graduates, who came to the Center for Applied Psychology in a public university, participated in the training. The program consisted of a series of 20 weekly group-encounters over the academic year, and each session was two hours long; six intervention groups were conducted with an average of three participants in each group. The IHS-Del-Prette was used as measurement instrument, and the data were treated through a non-parametric statistical test (Wilcoxon Test). The results indicated important acquisitions which led to the effectiveness of the intervention procedure. However, the results must be analyzed considering the small sample size, the use of oral reports as the main data source and also the absence of a control or follow-up group, which can be tested in future studies. Keywords: social skills training; college students; psychological intervention; prevention

Factors causing climatologically high temperatures in a hottest city in Japan: a multi-scale analysis of Tajimi

In this study, multi-scale climatological features of extreme high temperature (EHT) events in Tajimi, the hottest cities in Japan, were investigated using observational data collected by the Japan Meteorological Agency over the past 23 years, and original data observed by the authors over the last 3 years. Results revealed the background factors that lead to climatologically high temperatures in Tajimi: the occurrence of a characteristic pressure pattern called ‘whale’: the synoptic-scale factors, and the urbanization of Tajimi: the meso-γ-scale factors. In addition, the high temperatures measured in Tajimi are affected by the foehn-like westerly airflow coming from the mountains located in the northwest/west towards the Nobi Plain where Tajimi is located at the east end: the meso-β-scale factors, and the location of the Tajimi observation site, which is within an urbanized area where the highest temperatures tend to be observed: the micro-scale factors. In contrast, statistical analysis demonstrated that the small-scale basin effects and soil dryness around Tajimi were of lesser importance than aforementioned factors, in the occurrence of EHT events in Tajimi

Infectious cerebral thromboembolism in a patient with lung cancer during long-term administration of gefitinib: a case report

Introduction　Gefitinib is a molecularly targeted drug for oral administration, a selective epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI). It is effective for patients with lung cancer who have EGFR mutation and enables long-term survival of patients with advanced disease.Case presentation　An 84-year-old Asian female with lung cancer (adenocarcinoma, cT4N3M1b stage IV PUL BRA OSS ADR) was treated by gefitinib for two years. The primary tumor was markedly reduced, and bone metastasis had almost disappeared. However, the patient became unconsciousness with general edema and died two days after falling unconsciousness. In the autopsy, it was diagnosed as infectious cerebral thromboembolism due to infectious endocarditis. The cause of death was cardiac tamponade.Conclusions　While long-term survival can be expected in patients with advanced lung cancer by molecular-targeted agents, cautious management is warranted for complications, the same as for the primary disease