Fertilization failure and oocyte activation

Recent reports have suggested that when unfertilized oocytes with a sperma-tozoon after intracytoplasmic sperm injection (ICSI) are properly activated, the activated oocytes develop normaly similar to fertilized oocytes. However, human oocytes do not easily respond to universal activators of mammalian oocytes like ethanol or calcium ionophore A23187, which induce a calcium transient in ooplasm. Puromycin activates human oocytes at a rate of 90%, but more than two thirds of the parthenogenones possess 2 haploid pronuclei or 1 diploid pronucleus without extrusion of the second polar body. Therefore, the activation method which produces one pronucleus with extrusion of the second polar body in oocytes without a spermatozoon is necessary for producing embryos with normal karyotypes. Recently, we found the oocyte activation method which produced parthenogenones displaying one pronucleus with extrusion of the second polar body. Using our method (a combination of calcium ionophore A23187 and puromycin), the activation rate was approximately 90% and the proportion of parthenogenones dis-playing one pronucleus with extrusion of the second polar body was approximately 80% in human aged and mouse young oocytes. When human unfertilized oocytes following ICSI were activated by this method, two pronuclei were formed with extrusion of the second polar body in 30% of the oocytes. Four cleaved parthenogenones (or embryos) showed normal karyotypes. However, the cytotoxic, teratogenetic and mutagenetic activity of Ca ionophore and puromycin should be approved prior to the clinical adaptation of the method

Testing Orem’s self-care deficit theory : dependent care agency for duration of breast-feeding

Aims : The purpose of this study was to test of Orem’s theory of self-care deficit. Specifically, limiting factors influencing the duration of breast-feeding were analyzed with Orem model to clarify the ability needed by a mother for continuing breast-feeding. Background : The mean incidence of mothers with breast-feeding, mixtrophic breast-feeding or bottled milk is 52.6%, 43.2%, and 4.8%, respectively until one month after birth. However, the incidence of mothers with breast-feeding decreases to 46.2% just one month after birth. Although midwives instruct the importance of breast-feeding and promote breast-feeding by using various kinds of empirical care, there is not the standardized care with a high effect for continuation of breastfeeding. Method : descriptive study. Design Methodology : One hundred thirty-three mothers who delivered their babies in the University Hospital were included in this research. The hospital experienced around 400 birth in a year. Sixty-nine mothers of them fed a baby with breast milk (the breast-feeding group). The other mothers fed a baby with bottled milk or with bottled milk and breast milk (the mixed group). Results : According to Orem model, comments obtained by interviews were categorized, and we found 3 factors impairing continuation of breast-feeding, “crying of a child”, “inappropriate advice” and “previous experience”. The limiting factors were related to each other, and inexperience of a life with a baby, the time-consuming life without composure, and unskilled responses to the shortage of milk enhanced restriction of adaptability caused by a difference between an imagined baby and a real baby. Conclusions:This study support’s Orem’s model. Factors influencing the duration of breast-feeding were clarifyed by Orem’s limitaitions factors

Relationship between mammary gland structures during pregnancy and breast-feeding

Study Purpose Diagnostic indices for prediction of breast milk production and assessment of care necessary for mammary gland structure during the pregnancy period were investigated. Methods Development of mammary gland during the pregnancy period was examined via ultrasound methodology. Each subject underwent three measurements once per trimester. Quantity of post-partum breast milk production was also measured ; furthermore, the relationship between mammary gland image and production quantity was evaluated. Results and conclusion As previously reported, mammary gland reaches a certain level of development during the course of pregnancy. Four distinct types of developmental pattern were observed. Mammary gland exhibited remarkable development up to the 15th week of pregnancy in comparison with non-pregnant adult females in type 1. In type 2, mammary gland displayed significant development from the 16th to the 35th week of pregnancy. In types 3 and 4, mammary gland tissue development, which was apparent from 35 weeks, was characterized by gland thickness of > 20 mm and < 20 mm, respectively. Among these four types, type 1 mammary gland demonstrated the largest level of breast milk production. Therefore, we suggest that understanding with respect to mammary gland structure during pregnancy contributes toward individual support in breast-feeding following childbirth

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

ANATOMICAL STUDIES OF CORPUS PARACLOACALIS VASCULARIS IN COCKS

In the corpus paracloacalis vascularis of cocks the authors distinguished 5 different tissues : the capsule, the trabecula, the capillary cord, the peripheral lymphatic space, and the internal lymphatic space. In the peripheral and internal lymphatic spaces almost no argyrophilic fibers were found. This research work was performed with the following observation in mind, "the lymphatic spaces of the corpus paracloacalis vascularis were not the ordinary lymphatic sinuses, but a plexus of lymphatic capillaries"

FINE STRUCTURES OF CORPUS PARACLOACALIS VASCULARIS IN COCKS

The corpus paracloacalis vascularis in cocks was observed under scanning (SEM) and transmission electron microscopes (TEM). Under the SEM it was clearly demonstrated that the organs consisted of the capsule, trabeculae, capillary cords and lymphatic spaces. The capillary cords were arranged in anastomosing thick columns or sheets partitioned by lymphatic spaces under the SEM. The fine structures of the lymphatic spaces perfectly corresponded with the ordinary lymphatic capillaries under the TEM. The blood capillaries in the capillary cords consisted of a layer of endothelial cells and comparatively numerous pericytes, but some characteristic features were noted ; the endothelial cells were surrounded by a thicker, denser basement membrane and included numerous dilated rough-surfaced endoplasmic reticulums and unusual large pinocytotic vesicles, 300～500 mμ in size, which were often opened on the inner and outer surfaces of the endothelium

THE LYMPHATIC SYSTEM OF THE CORPUS PARACLOACALIS VASCULARIS AND THE SECOND FOLD IN THE MALE DOMESTIC FOWL

The lymphatic system of the corpus paracloacalis vascularis and the second fold of the cloaca in the male domestic fowl was investigated by means of the Latex-cast specimens. The lymphatic system of the corpus paracloacalis vascularis (CV) was a complex pear-shaped lymphatic spongy body with a stalk. The lymphatics of the second fold of both sides were connected and formed an arch-shaped, spongy body. The body was divided into 4 parts : the connecting part ; the phallus part ; the round fold part ; and the lymph fold part. The spongy body of the CV was closely connected to that of the second fold ; it was suggested that both bodies, as a system, play a role in the erection of the copulatory organ. The writers recommend the term "Corpus paraclocalis vascularis-Second fold lymphatic system" for these lymphatic structures