Successful Long-Term Preservation of Rat Sperm by Freeze-Drying

Background: Freeze-drying sperm has been developed as a new preservation method where liquid nitrogen is no longer necessary. An advantage of freeze-drying sperm is that it can be stored at 4uC and transported at room temperature. Although the successful freeze-drying of sperm has been reported in a number of animals, the possibility of long-term preservation using this method has not yet been studied. Methodology/Principal Findings: Offspring were obtained from oocytes fertilized with rat epididymal sperm freeze-dried using a solution containing 10 mM Tris and 1 mM EDTA adjusted to pH 8.0. Tolerance of testicular sperm to freeze-drying was increased by pre-treatment with diamide. Offspring with normal fertility were obtained from oocytes fertilized with freeze-dried epididymal sperm stored at 4uC for 5 years. Conclusions and Significance: Sperm with –SS – cross-linking in the thiol-disulfide of their protamine were highly tolerant to freeze-drying, and the fertility of freeze-dried sperm was maintained for 5 years without deterioration. This is the first report to demonstrate the successful freeze-drying of sperm using a new and simple method for long-term preservation

三重県における地域別食生活実態の公衆栄養学的考察 : 第17報 昭和60年,三重県阿山郡大山田村東小学校学童の家庭における栄養摂取状態(自然科学編)

三重県における学童の健康をめぐる食生活の今日的な課題とその栄養指導対策を得る目的で,昭和53年より平坦部市街化地域の鈴鹿市庄野小学校および山間部の阿山郡大山田村東小学校を対象として,毎年継続実施してきたが,今回は後者の本県において循環器疾患およびガン死亡率が高い地域であり,かつ,上野保健所管内のうちでも,さらに高率村である阿山郡大山田村東小学校学童のみを対象としてその家庭食事(朝食・夕食および間食)の栄養摂取状態について調査を行なった。(1)栄養素別摂取量の各栄養素別標準量に対する有意差を検討した結果は,男子においては,総たん白質および動物性たん白質の摂取量が99%の信頼度で標準量を上回っていた。女子においては,動物性たん白質の摂取量が99%の信頼度で標準量を上回っていたが,植物性たん白質の摂取量のみは99%の信頼度で標準量を下回っていた。カルシウムについては女子のみが95%の信頼度で標準量を下回っていた。ビタミンについてはビタミンB_2が女子のみ99%の信頼度で標準量を下回っていた。(2)栄養摂取量の標準量に対する充足率について男子と女子との間の有意差をうると,エネルギー,総たん白質,動物性たん白質およびビタミンB_1の各栄養素に有意差がみられとくにビタミンB_1において,女子は男子のそれよりも摂取不足が著しいことが注目された。なお,カルシウム,ビタミンB_2および繊維は男子・女子ともに摂取不足がみられた。(3)性別,各栄養素別摂取量に対する未満児を個人別にみた場合は,前述のごとく充足していた栄養素においても個人差が大きく,その摂取量の未満児が,かなり認められた。(4)PFC比の平均値については,男子・女子とも適正な範囲と見なされ,かつ,男子と女子との間には有意差はみられなかった。(5)殻物エネルギー比と動物性たん白質比との相関関係は男子・女子ともに99%の信頼度で認められた。動物性たん白質比と植物性脂肪/動物性脂肪との間の相関関係は男子において95%の信頼度で認められたが,女子においてはその相関関係は認められなかった。(6)植物性脂肪/動物性脂肪については,男子0.80,女子0.98であり,やや不適正な値を示したが,本調査では一価不飽和脂肪酸含量の多い魚油を考慮した場合は適正な値であった。魚油を含むその植物性脂肪/動物性脂肪についてもその変動係数は大きく,1.0以下の者が男子43.8%,女子30.0%の存在は否定しえず,脂肪の質についての特別な指導が必要と考えられる。(7)食塩摂取過剰と粗繊維の摂取不足傾向がみられた。(8)孤食学童・家族全員食別学童の栄養摂取量において,孤食学童にビタミンAの不足が95%信頼度で認められた。ついで,カルシウムおよびビタミンB_2においても摂取不足の傾向は認められた。Studies have been conducted continuously each year since 1978 at the Shono Elementary School in Suzuka city and the Higashi Elementary School in Ohyamada-mura, Ayama-gun, for the purposes of determining the elements of present-day diets as they relate to the health of students and to determine nutritional guidelines. The present study, however, focussed only on the students at Higashi Elementary School in Ayama-gun, which is located in a region which has a high incidence of cardiovascular disease and a high cancer-related death rate, even throughout the jurisdiction of the Ueno Health Center, and encompassed the conditions of nutrient intake of at-home diets (breakfast, dinner and between meal snacks). 1) The results of investigations into significant differences in the consumed and standard amounts of each nutrient showed, with 99% reliability, that the consumption of total and animal proteins by boys exceeded the standard levels. While the consumption of animal proteins for girls was shown with 99% reliability to be above standard levels, the consumption of vegetable proteins was below standard levels. The intake of calcium was shown, with 95% reliability, to be below the standard level only for girls, while consumption of vitamin B_2 was also shown to be below the standard level only for girls. 2) In examining differences between boys and girls in meeting standard levels of consumed nutrients, significant differences were found in the consumption of each of the energy用roducing nutrients such as total proteins, animal proteins and vitamin B_1. An inadequacy of vitamin B_1 consumption in particular, was much worse for girls than for boys. Insufficient intake of calcium and vitamin B_2 were also observed. 3) In examining individual differences of sex and amounts of each consumed nutrient for pupils who had below standard levels consumption, it was observed that great individual differences occured even in nutrients with adequate consumption levels. 4) The mean value in the PFC ratio for boys and girls fell withim the appropriate region, and no significant differences were abserved between them. 5) A correlation between the ratio of energy from cereals and the ratio of animal proteins was observed in both and girls with 99% reliability. While a correlation between the animal protein ratio and vegetable fats/animal fats was observed with 95% reliability for boys, no such correlation was observed for girls. 6) In regard to the ratio of vegetable and animal fats, values of 0.80 for boys and 0.98 for girls were determined and while these are somewhat inappropriate values, they are appropriate for this study when considered in light of the many mono・unsaturated fatty acids found in fish oils. The coefficient of variation is large for the vegetable fat/animal fat ratio which includes fish oils. The existence of pupils for whom this coefficient was less than 1.0 is undeniable at 43.8% of the boys and 30.0% of the girls. It is beliened that particular guidance is required concerning fatty substances in the diet. 7) A tendency for excessive salt intake and inadequate intake of rough fiver were observed. 8) In differentiating nutrient intake between students who ate alone and those who ate with their families, an insufficiency of vitamin A intake was observed, with 95% reliability, for students who ate alone. A tendency was also noticed towards inadequate calcium and vitamin B_2 intake

The whole blood transcriptional regulation landscape in 465 COVID-19 infected samples from Japan COVID-19 Task Force

「コロナ制圧タスクフォース」COVID-19患者由来の血液細胞における遺伝子発現の網羅的解析 --重症度に応じた遺伝子発現の変化には、ヒトゲノム配列の個人差が影響する--. 京都大学プレスリリース. 2022-08-23.Coronavirus disease 2019 (COVID-19) is a recently-emerged infectious disease that has caused millions of deaths, where comprehensive understanding of disease mechanisms is still unestablished. In particular, studies of gene expression dynamics and regulation landscape in COVID-19 infected individuals are limited. Here, we report on a thorough analysis of whole blood RNA-seq data from 465 genotyped samples from the Japan COVID-19 Task Force, including 359 severe and 106 non-severe COVID-19 cases. We discover 1169 putative causal expression quantitative trait loci (eQTLs) including 34 possible colocalizations with biobank fine-mapping results of hematopoietic traits in a Japanese population, 1549 putative causal splice QTLs (sQTLs; e.g. two independent sQTLs at TOR1AIP1), as well as biologically interpretable trans-eQTL examples (e.g., REST and STING1), all fine-mapped at single variant resolution. We perform differential gene expression analysis to elucidate 198 genes with increased expression in severe COVID-19 cases and enriched for innate immune-related functions. Finally, we evaluate the limited but non-zero effect of COVID-19 phenotype on eQTL discovery, and highlight the presence of COVID-19 severity-interaction eQTLs (ieQTLs; e.g., CLEC4C and MYBL2). Our study provides a comprehensive catalog of whole blood regulatory variants in Japanese, as well as a reference for transcriptional landscapes in response to COVID-19 infection

DOCK2 is involved in the host genetics and biology of severe COVID-19

「コロナ制圧タスクフォース」COVID-19疾患感受性遺伝子DOCK2の重症化機序を解明 --アジア最大のバイオレポジトリーでCOVID-19の治療標的を発見--. 京都大学プレスリリース. 2022-08-10.Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge. Here we conducted a genome-wide association study (GWAS) involving 2, 393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3, 289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target

三重県における地域別食生活実態の公衆栄養学的考察 : 第15報 昭和59年,三重県鈴鹿市庄野小学校および大山田村東小学校学童の栄養摂取状態とその家族とのまどい状況(自然科学編)

三重県における年別学童期栄養の実態とその栄養指導の方法を探る目的で,昭和53年から引続き,今回も,前回と同じ平坦部の市街地域にある鈴鹿市庄野小学校および山間部の農村地域にある阿山郡大山田村東小学校における学童各17名を対象とし,その家庭食事(朝・夕食および間食)の栄養摂取状態およびその家族のまどい状況について調査した。(1)エネルギー,たん白質,動物性たん白質,ビタミンA,ビタミンB_1,ビタミンB_2およびビタミンCの栄養素別平均摂取量については,両校ともそれぞれの標準量を上回っていた。カルシウムのみは,庄野小では標準量を上回っていたが,東小ではそれを下回っていた。しかしながら,これを個人別にみたばあいは,充足していた栄養素においても,個人差が大きく,その摂取量に対する未満者が,エネルギーでは庄野小12%,東小29%,たん白質では庄野小18%,東小29%,脂肪では庄野小9%,東小29%,カルシウムでは両校とも59%,ビタミンAでは庄野小35%,東小53%,ビタミンB_1では両校とも12%,ビタミンB_2では庄野小18%,東小12%,ビタミンCは庄野小12%,東小9%も存在していることがわかった。(2)脂肪の質の面から,平均P/S比をみた結果,庄野小では0.98,東小では0.86で,厚生省勧告の1.0～2.0より低い値であった。(3)食塩の平均摂取量は,庄野小7.4g,東小4.9gで,両校との間に有意差がみられた。そして,両校ともアメリカの食塩適正目標量3.0gのそれよりも上回っていた。(4)食品群別平均摂取量の目標量に対する充足率をみると,大豆類では庄野小26%,東小6%,緑黄色野菜類では庄野小27%,東小43%,乳類では両校とも5%内外の摂取不足が認められた。(5)家族とのまどい状況では,とくに朝食で「家族全員で食べていない」と答えた学童が庄野小では47%,東小では82%も認められたことが注目された。In order to clarify annual nutritional intake in domestic diet of school children in Mie Prefecture we have examined nutiritional intake in domestic diet in week days of school children in Shono Elementary School located in a town place in Suzukashi and those lived in Higashi Elementary School of Ohyamada-mura, Ayama-gun located between mountains since 1978. (1) Average intakes of nutrients of both schools exceed standard values except that calcium intake of Higashi Elementary School is lower than the standard value. (2) Although intakes of nutrients exceed the standard values in average, they show variation depending on individuals (coefficient of variation). All nutrients are in sufficient in some children. In particular, 59% of children in both schools show calcium poverty and 35 %and 53 % of children of Shono and Higashi Elementary Schools show vitamin A poverty respectively. (3) Average P/S ratio of fat of Shono Elementary School is 0.98 while that of Higashi Elementary School is 0.86, each showing a low ratio. (4) Salt intake of Shono Elementary School is 7.4g while that of Higashi Elementary School is 4.9g. (5) With regard to family circles, 47% and 82%, of children in Shono and Higashi Elementary Schools said that they did not have their breakfast with all family members, each showing a considerably high ratio

DNA barcoding and morphological analyses revealed validity of Diadema clarki Ikeda, 1939 (Echinodermata, Echinoidea, Diadematidae)

A long-spined sea urchin Diadema-sp reported from Japanese waters was genetically distinct from all known Diadema species, but it remained undescribed. Extensive field surveys in Japan with molecular identification performed in the present study determined five phenotypes (I to V) in Diadema-sp according to the presence and/or shape of a white streak and blue iridophore lines in the naked space of the interambulacral area. All phenotypes were distinct from Diadema setosum (Leske, 1778) and Diadema savignyi (Audouin, 1829), of which a major type (I) corresponded to Diadema clarki Ikeda, 1939 that was questioned and synonymized with D. setosum by Mortensen (1940). The holotype of D. clarki has not been found, but three unlabeled dried tests of Diadema were found among Ikeda’s original collection held in the Kitakyushu Museum of Natural History and Human History, Fukuoka, Japan. A short mtDNA COI fragment (ca. 350bp) was amplified from one of the tests, and the nucleotide sequence determined (275bp) was nearly identical with that of Diadema-sp. Arrangements of the primary tubercles on the coronal plates in Diadema-sp and the museum specimen also conformed with D. clarki, indicating that Diadema-sp is identical to D. clarki and a valid species. Narrow latitudinal distribution (31°N to 35°N) of D. clarki in Japan was observed, where it co-existed with abundant D. setosum and rare D. savignyi. No D. clarki was found in the southern islands in Japan, such as Satsunan Islands to Ryukyu Islands and Ogasawara Island, where D. setosum and D. savignyi were commonly observed