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

Measurements of Gamma-Ray Production Cross Sections of Nb and Cu for 15-MeV Neutrons

Gamma-ray production cross sections of Nb and Cu are measured for 14.9 -MeV neutrons. The accuracy of the experimental results is confirmed by comparing those with other experipental data obtained by similar energy neutrons. Hence, the data given in this paper are useful in the evaluation of secondary gamma-ray production data for fusion use. Based on the measured data, some comments are given to the evaluated data in the ENDF/B-Ⅳ file : (1) the gamma-ray spectrum of Cu data is not adequate, (2) the gamma-ray yield data of the file for Nb is overestimated

Domination number of annulus triangulations

An {\em annulus triangulation} $G$ is a 2-connected plane graph with two disjoint faces $f_1$ and $f_2$ such that every face other than $f_1$ and $f_2$ are triangular, and that every vertex of $G$ is contained in the boundary cycle of $f_1$ or $f_2$. In this paper, we prove that every annulus triangulation $G$ with $t$ vertices of degree 2 has a dominating set with cardinality at most $\lfloor \frac{|V(G)|+t+1}{4} \rfloor$ if $G$ is not isomorphic to the octahedron. In particular, this bound is best possible

Proliferation of Luteal Steroidogenic Cells in Cattle

The rapid growth of the corpus luteum (CL) after ovulation is believed to be mainly due to an increase in the size of luteal cells (hypertrophy) rather than an increase in their number. However, the relationship between luteal growth and the proliferation of luteal steroidogenic cells (LSCs) is not fully understood. One goal of the present study was to determine whether LSCs proliferate during CL growth. A second goal was to determine whether luteinizing hormone (LH), which is known have roles in the proliferation and differentiation of follicular cells, also affects the proliferation of LSCs. Ki-67 (a cell proliferation marker) was expressed during the early, developing and mid luteal stages and some Ki-67-positive cells co-expressed HSD3B (a steroidogenic marker). DNA content in LSCs isolated from the developing CL increased much more rapidly (indicating rapid growth) than did DNA content in LSCs isolated from the mid CL. The cell cycle-progressive genes CCND2 (cyclin D2) and CCNE1 (cyclin E1) mRNA were expressed more strongly in the small luteal cells than in the large luteal cells. LH decreased the rate of increase of DNA in LSCs isolated from the mid luteal stage but not in LSCs from the developing stage. LH suppressed CCND2 expression in LSCs from the mid luteal stage but not from the developing luteal stage. Furthermore, LH receptor (LHCGR) mRNA expression was higher at the mid luteal stage than at the developing luteal stage. The overall results suggest that the growth of the bovine CL is due to not only hypertrophy of LSCs but also an increase in their number, and that the proliferative ability of luteal steroidogenic cells decreases between the developing and mid luteal stages

Decomposition Process of Woody Debris and Fungal Community Structure

We examined the relationship between the community structure of wood-decaying fungi, detected by high-throughput sequencing, and the decomposition rate using 13 years of data from a forest dynamics plot. For molecular analysis and wood density measurements, drill dust samples were collected from logs and stumps of Fagus and Quercus in the plot. Regression using a negative exponential model between wood density and time since death revealed that the decomposition rate of Fagus was greater than that of Quercus. The residual between the expected value obtained from the regression curve and the observed wood density was used as a decomposition rate index. Principal component analysis showed that the fungal community compositions of both Fagus and Quercus changed with time since death. Principal component analysis axis scores were used as an index of fungal community composition. A structural equation model for each wood genus was used to assess the effect of fungal community structure traits on the decomposition rate and how the fungal community structure was determined by the traits of coarse woody debris. Results of the structural equation model suggested that the decomposition rate of Fagus was affected by two fungal community composition components: one that was affected by time since death and another that was not affected by the traits of coarse woody debris. In contrast, the decomposition rate of Quercus was not affected by coarse woody debris traits or fungal community structure. These findings suggest that, in the case of Fagus coarse woody debris, the fungal community structure is related to the decomposition process of its host substrate. Because fungal community structure is affected partly by the decay stage and wood density of its substrate, these factors influence each other. Further research on interactive effects is needed to improve our understanding of the relationship between fungal community structure and the woody debris decomposition process

Farming strategy of African smallholder farmers in transition from traditional to alternative agriculture : the case of the Nupe in central Nigeria

It is worthwhile to understand farming strategies of smallholder farmers in sub-Saharan Africa, especially those of farmers who are in transition from traditional to alternative agriculture in terms of adoption of innovative technologies. In a case study of inland valleys in central Nigeria, we investigated the farming strategy of Nupe farmers who have a long-term tradition of wet rice cultivation and indigenous methods of land preparation for soil, water and weed management. In this region, a new method of land preparation has recently been introduced along with a recommendation to use improved seeds and chemical fertilizers. Our findings reveal that Nupe farmers directly sow traditional seeds and apply a marginal amount of fertilizer to paddy plots prepared by labor-saving methods on drought-prone hydromorphic valley fringes and flood-susceptible valley bottoms, whereas they preferentially transplanted improved seedlings and applied a relatively large quantity of fertilizer to paddy fields prepared by a labor-intensive and mechanized method on a valley position where they can access to optimum water condition (less risky against the drought and flood)

世界各地の日本語学習者の文法学習・語彙学習についてのビリーフ : ノンネイティブ日本語教師・日本人大学生・日本人教師と比較して

名古屋外国語大学Nagoya University of Foreign Studies本研究では,日本語学習者の文法学習と語彙学習に対するビリーフについて,世界各地の学習者を対象とした先行研究結果を取り出して地域的特徴を考察した。さらに,先行研究で明らかになっているノンネイティブ日本語教師のビリーフの傾向,日本人大学生や日本人教師のビリーフ調査の結果とも比較した。その結果,文法学習も語彙学習も大切だというビリーフに学生が強く賛成し,現地のノンネイティブ日本語教師と同じ傾向を示す地域(南アジア,東南アジア),そのようなビリーフに学生は賛成するが,それほど強く賛成するわけではなく,現地のノンネイティブ日本語教師の傾向とも近い地域(西欧,大洋州),前2者の中間程度の強さで学生がビリーフに賛成し,教師のビリーフとはやや異なる傾向の地域(中南米,東南アジア・東アジアや大洋州の一部)など,地域による違いが見られた。さらに,日本人の結果を見てみると,日本人大学生や教師歴がごく短い日本人教師は,文法学習も語彙学習も大切ではないというビリーフを持ち,世界各地の学習者とは異なる傾向を示す。一方,経験豊富な教師は世界各地の学習者と同じような傾向であることも分かった。最後に,こういった傾向を把握したうえで,文法・語彙のシラバス・教材作成と普及を行う必要があることを指摘した。This study compares the results of previous researches on beliefs about grammar and vocabulary learning, which were obtained from surveys of Japanese learners abroad, and analyses the regional learner belief characteristics. These results were also compared to the beliefs of non-native Japanese teachers, Japanese university students, and native Japanese teachers. The following characteristics were identified. Learners in South Asia and Southeast Asia strongly agree with the belief that grammar and vocabulary learning are important to language learning. However, those in Western Europe and Oceania weakly agree with this belief. These tendencies are in accordance with non-native teachers\u27 beliefs in these areas. Learners in Central and South America, and some in Southeast Asia, East Asia, and Oceania moderately agree with the abovementioned belief; this tendency is different from the non-native teachers\u27 beliefs in these areas. These results were then compared with beliefs of Japanese university students and native Japanese teachers. Japanese university students and Japanese teachers with little teaching experience were found to disagree with learners\u27 beliefs. On the other, beliefs of Japanese teachers with long term teaching experiences were found to have similar tendency with learners\u27 beliefs. The results of this study should be used to develop syllabi and learning materials for teaching Japanese grammar and vocabulary

Inactivation of Vibrio vulnificus hemolysin through mutation of the N- or C-terminus of the lectin-like domain

Vibrio vulnificus is an etiological agent causing serious systemic infections in the immunocompromised humans or cultured eels. This species commonly produces a hemolytic toxin consisting of the cytolysin domain and the lectin-like domain. For hemolysis, the lectin-like domain specifically binds to cholesterol in the erythrocyte membrane, and to form a hollow oligomer, the toxin is subsequently assembled on the membrane. The cytolysin domain is essential for the process to form the oligomer. Three-dimensional structure model revealed that two domains connected linearly and the C-terminus was located near to the joint of the domains. Insertion of amino acid residues between two domains was found to cause inactivation of the toxin. In the C-terminus, deletion, substitution or addition of an amino acid residue also elicited reduction of the activity. However, the cholesterol-binding ability was not affected by the mutations. These results suggest that mutation of the C- or N-terminus of the lectin-like domain may result in blockage of the toxin assembly