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

Prevalence of tuberculosis in small pulmonary nodules obtained by video-assisted thoracoscopic surgery

SummaryThe prevalence of tuberculosis in small solitary lesions of the lung obtained by video-assisted thoracoscopic surgery (VATS) is still unclear. Of 103 lung lesions resected by VATS in 98 patients (47 men, 51 women), 19 were identified macroscopically as inflammatory changes, 78 were neoplastic, and 6 were undefined. Presumptive diagnosis based on microscopic analysis of fresh specimen smears treated with Papanicolaou stain was performed in 19 lesions. Of these, 11 lesions had epithelioid cells, granulomas with caseous necrosis and Langerhans-type giant cells. The 6 undefined lesions were non-inflammatory benign changes. Isolation and identification of tuberculosis were based on microscopic findings of fresh material smears and sections of fixed specimens stained with Ziehl–Neelsen's dye, cultivation using egg-based Ogawa medium, and in situ hybridization between polymerase chain reaction (PCR) products of each of the 11 lesions and specific DNA sequences for Mycobacterium tuberculosis, M. avium, and M. intracellulare. Of these 11 lesions, M. tuberculosis was confirmed in one (0.96%) by PCR and M. avium was confirmed in four by culture and PCR. Of the 78 malignant lesions, final pathologies were primary lung cancer (n=59, 70.2%) and pulmonary metastatic cancer (n=19, 22.6%). The most frequent primary malignant cancer was adenocarcinoma, which was found in 19 men and 28 women in the present study. Eight lesions in 8 men were squamous cell carcinomas. The results of the present study suggested that even though the prevalence of lung tuberculosis is low, attention should be paid to the presence of M. tuberculosis in specimens obtained by VATS

スリランカにおける出生過程の分析

本稿はスリランカのDHS93のデータを用いて、30-49才女子の出生行動のパターンから教育水準間の出生数格差の変動から考察をしている。PB_aの放物線は教育水準の上昇に伴ってPB_aの高さが低く、右にシフトしている。このシフトは有配偶率（MR_a）の増加パターンからみられる教育水準の上昇による結婚年令が遅れたためであり、CNB_aの格差の発生となることを明らかにした。さらにPB_mの分布からは出生行動の期間が教育水準の上昇に伴い短縮される傾向が明確に示され、さらに40-49才女子に比べ全ての教育水準において30-39才女子のほうが短縮されていた。結婚年数を時間軸にしたとき１年目の値が際立って高く、その後の右下がりの傾きは教育水準が高いほど大きく、これがCNB_aの格差を発生させていることを明らかにしている。このような分析によりスリランカの出生数の低下は結婚年令の上昇による出生行動期間のシフトが大きな影響を与えていることが明らかになった。Total fertility rate (TFR) in Sri Lanka was 2.3 in 1993. The higher mother's education level was, the lower mean Children ever born (CEB) were. This paper considers differences of births between 4 educational levels by conducting two analyses: birth probability by mother's age (PB_a) and period of marriage (PB_m). PBa curves draw inverse—U—shaped and shift rightward according as educational level goes up. PB_m curves draw downward sloping from 1 year and drop rapidly according as educational level goes up. The result of comparisons between educational levels is that education contributes to delay age of marriage, shorten reproductive period and lower birth probability. Younger cohort who is 30's years' females shows these trends clearly. In Sri Lanka, lower births can be explained by change in both reproductive period and its behavior

A Quantitative Analysis of Endogenous Target Fertility in Sri Lanka

Even though the economic model of fertility seems theoretically plausible, application of the model to empirical data often gives us ambiguous impression about its validity. One source of the ambiguity that is examined by an empirical analysis in this paper comes from simultaneity of demand for and supply of children. Fertility differentials are explained by differences in educational attainment, work experience prior to marriage, possession of durable goods, job categories of the husband and the residual of the age at marriage. Differentials in parity progression are largely explained by the age of marriage or the previous childbirth. The effect of socio-economic variables is parity dependent in supply of children. Differentials in spacing of childbirths are explained by the expected demand and unexpected additional demand for children and the age difference from the mean age at marriage or the previous childbirth

Diet-induced obesity in zebrafish shares common pathophysiological pathways with mammalian obesity

<p>Abstract</p> <p>Background</p> <p>Obesity is a multifactorial disorder influenced by genetic and environmental factors. Animal models of obesity are required to help us understand the signaling pathways underlying this condition. Zebrafish possess many structural and functional similarities with humans and have been used to model various human diseases, including a genetic model of obesity. The purpose of this study was to establish a zebrafish model of diet-induced obesity (DIO).</p> <p>Results</p> <p>Zebrafish were assigned into two dietary groups. One group of zebrafish was overfed with <it>Artemia </it>(60 mg dry weight/day/fish), a living prey consisting of a relatively high amount of fat. The other group of zebrafish was fed with <it>Artemia </it>sufficient to meet their energy requirements (5 mg dry weight/day/fish). Zebrafish were fed under these dietary protocols for 8 weeks. The zebrafish overfed with <it>Artemia </it>exhibited increased body mass index, which was calculated by dividing the body weight by the square of the body length, hypertriglyceridemia and hepatosteatosis, unlike the control zebrafish. Calorie restriction for 2 weeks was applied to zebrafish after the 8-week overfeeding period. The increased body weight and plasma triglyceride level were improved by calorie restriction. We also performed comparative transcriptome analysis of visceral adipose tissue from DIO zebrafish, DIO rats, DIO mice and obese humans. This analysis revealed that obese zebrafish and mammals share common pathophysiological pathways related to the coagulation cascade and lipid metabolism. Furthermore, several regulators were identified in zebrafish and mammals, including APOH, IL-6 and IL-1β in the coagulation cascade, and SREBF1, PPARα/γ, NR1H3 and LEP in lipid metabolism.</p> <p>Conclusion</p> <p>We established a zebrafish model of DIO that shared common pathophysiological pathways with mammalian obesity. The DIO zebrafish can be used to identify putative pharmacological targets and to test novel drugs for the treatment of human obesity.</p

Spinal Deformity and the Musculoskeletal Cohort Study of the General Older Population

Article信州医学雑誌 69(3) : 111-120(2021)departmental bulletin pape

Thermoluminescence of chondrules in primitive ordinary chondrites,Semarkona and Bishunpur

The spatial distribution of the induced thermoluminescence (TL) and TL glow curves of the primitive ordinary chondrites, Semarkona (LL3.0) and Bishunpur (LL3.1), were investigated over a wide range of wavelengths using a TL spatial distribution readout system. Although bulk samples of Semarkona and Bishunpur have very low TL sensitivity, individual chondrules show a wide variety of induced TL intensity and glow curve shape. Chondrules with anorthite-normative mesostases have especially high induced TL intensity, and their TL is produced at wavelengths >480nm, compared with <480nm for the sensitivity range of the usual TL measuring systems. Some of the metamorphism-dependent TL sensitivity of type 3 ordinary chondrite therefore results in changes in the spectrum of the light produced

PD-L1 expression is associated with the spontaneous regression of patients with methotrexate-associated lymphoproliferative disorders

Background Most patients with methotrexate-associated lymphoproliferative disorder (MTX-LPD) show diffuse large B-cell lymphoma (DLBCL) or classic Hodgkin lymphoma (CHL) types. Patients with MTX-LPD often have spontaneous remission after MTX discontinuation, but chemotherapeutic intervention is frequently required in patients with CHL-type MTX-LPD. In this study, we examined whether programmed cell death-ligand 1 (PD-L1) expression levels were associated with the prognosis of MTX-LPD after MTX discontinuation. Methods A total of 72 Japanese patients diagnosed with MTX-LPD were clinicopathologically analyzed, and immunohistochemical staining of PD-L1 was performed in 20 DLBCL-type and 24 CHL-type MTX-LPD cases to compare with the clinical course. Results PD-L1 was expressed in 5.0% (1/20) of patients with DLBCL-type MTX-LPD, whereas it was expressed in 66.7% (16/24) of the patients with CHL-type MTX-LPD in more than 51% of tumor cells. Most CHL-type MTX-LPD patients with high PD-L1 expression required chemotherapy owing to exacerbations or relapses after MTX discontinuation. However, no significant differences in clinicopathologic findings at diagnosis were observed between PD-L1 high- and low-expression CHL-type MTX-LPD. Conclusion PD-L1 expression was significantly higher in patients with CHL-type than DLBCL-type MTX-LPD, suggesting the need for chemotherapy in addition to MTX discontinuation in CHL-type MTX-LPD patients to achieve complete remission. No association was observed between PD-L1 expression levels and clinical findings at diagnosis, suggesting that PD-L1 expression in tumor cells influences the pathogenesis of CHL-type MTX-LPD after MTX discontinuation