Current status and legal/ethical problems in the research use of the tissues of aborted human fetuses in Japan

To date, there is no law regulating the research use of human aborted fetuses in Japan. The aim was to review the current status with historical background and legal/ethical problems limiting the research use of the tissues of aborted human fetuses. We reviewed literature via PubMed, Web of Science, Scopus, Japana Centra Revuo Medicina and CiNii, reports from various committees and research groups from Ministry of Health, Labour and Welfare (MHLW), and domestic books. Aborted human fetal tissues used for research purposes were first documented in the 1920s. The first guideline, the Peel Code was released in 1972. Since then, in Western countries, the research use of aborted fetuses has been less restricted compared with that of embryos, due to the following guidelines outlined by expert groups. Currently, aborted fetal tissues are commercially available for research purposes in the United States. In Japan, only four indications are presented in “a public statement permitting research use of deceased fetuses' and ‘neonates’ organs, etc.” (1987). In the 2000s, expert committees of the MHLW concluded that research use of human aborted fetuses should be discontinued, and that comprehensive rules and independent regulations should be implemented. This issue has not been discussed in the Japanese legislature since 2003. Establishment of laws and guidelines for this issue is insufficient not only in Japan but also in other countries. It is important to secure transparency for making laws and guidelines and in obtaining public understanding

KINEMATIC ANALYSIS ON THE PUNT KICK IN FOOTBALL GOALKEEPER

The present study aimed to investigate punt kicks by football goalkeepers on the basis of differences in the effort of the kick. Twelve experienced goalkeepers participated in the study. The participants were made aware that for the maximum distance trial (the 100% trial), they should send the ball as far as possible, and for the 80% and 60% trials they should have a more controlled approach. Punt kicks were divided into three phases: the phase of release of the ball from the left hand (BR), the phase of pivot foot ground-contact (LFC), and the phase of the ball impact (IMP). Right hip and knee joint angles were calculated. In comparison to lower intensity punt kicks, the higher effort punt kicks involved increasing the hip joint extension angle for the right foot during the backswing and lowering the knee joint angle of the right leg at the start of the forward swing, thereby producing forward swing velocity for the right foot

KINEMATIC ANALYSIS OF BASEBALL BATTING MOTION WHEN BATTING PITCHES WITH VARYING VELOCITIES

The purpose of this study was to identify effect of moving the center of gravity of the body and rotating the torso when batting pitches with varying velocity. The subjects were 10 experienced university baseball player. The subjects batted toward the center field, both fastballs and slowballs, aimed near the center of the strike zone from a pitching machine. Data were collected using a three dimensional automatic motion analysis system (Vicon MX). The rotation angle of the torso and displacement of the center of gravity were computed. Due to differences in the shoulder rotation from the latter half of stepping leg touchdown through impact, we believe that the motion is adapted to pitch differences starting from the latter half of stepping leg touchdown. Comparing the shift in the center of gravity of the body during fastballs and slowballs, the forward motion and downward sinking of the center of gravity were significantly larger for slowballs

INFLUENCE OF THE BALL SPEED ON THE DISPLACEMENT OF THE CENTER OF GRAVITY DURING BASEBALL BATTING MOTION

The purpose of this study was to investigate the modification in batting motion with different pitching speeds focusing on differences in batting technical level. The subjects were 10 experienced university baseball player. The subjects batted toward the center field, both fastballs and slowballs, aimed near the center of the strike zone from a pitching machine. Data were collected using a three dimensional automatic motion analysis system (Vicon MX). The displacement of the center of gravity(CG) were computed. Significant differences were seen due to difference in pitching speed in unskilled player. Conversely, in skilled player, no significant difference was found in the movement of CG due to the difference in pitching speed. It was revealed that it was not preferable for movement of the CG to fluctuate by difference in pitching speeds

Kinematic analysis of punt kick in football goalkeepers based on the level of kick effort

In the present study, we aimed to investigate the differences in punt kicks by football goalkeepers based on the level of effort required. Twelve experienced goalkeepers participated in the study. The participants were instructed to kick the ball as far as possible in the maximum distance trial (100% trial) and to have a more controlled approach for the 80% and 60% trials. Each punt kick was divided into three events: release of the ball from the left hand (BR), pivot foot ground-contact (LFC), and ball impact (IMP). Right lower limb joint velocity, right hip and knee joint angles, flight distance, ball velocity, and kick angle were calculated. The 80% and 100% trials yielded almost the same velocity for each part of the right leg; however, in the 60% trial, the level of kicking effort was managed by adjusting the velocity of the right ankle joint, starting from BR, in addition to adjustment of the velocity of the right knee joint at LFC. Compared to punt kicks with a lower level of effort, the punt kicks with a higher level of effort involved an increase in the hip joint extension angle for the right leg during the backswing and the lowering of the knee joint angle of the right leg at the start of the forward swing, thus producing forward swing velocity for the right foot

Visualization of the spatial positioning of the SNRPN, UBE3A, and GABRB3 genes in the normal human nucleus by three-color 3D fluorescence in situ hybridization

The three-dimensional (3D) structure of the genome is organized non-randomly and plays a role in genomic function via epigenetic mechanisms in the eukaryotic nucleus. Here, we analyzed the spatial positioning of three target regions; the SNRPN, UBE3A, and GABRB3 genes on human chromosome 15q11.2–q12, a representative cluster of imprinted regions, in the interphase nuclei of B lymphoblastoid cell lines, peripheral blood cells, and skin fibroblasts derived from normal individuals to look for evidence of genomic organization and function. The positions of these genes were simultaneously visualized, and all inter-gene distances were calculated for each homologous chromosome in each nucleus after three-color 3D fluorescence in situ hybridization. None of the target genes were arranged linearly in most cells analyzed, and GABRB3 was positioned closer to SNRPN than UBE3A in a high proportion of cells in all cell types. This was in contrast to the genomic map in which GABRB3 was positioned closer to UBE3A than SNRPN. We compared the distances from SNRPN to UBE3A (SU) and from UBE3A to GABRB3 (UG) between alleles in each nucleus, 50 cells per subject. The results revealed that the gene-to-gene distance of one allele was longer than that of the other and that the SU ratio (longer/shorter SU distance between alleles) was larger than the UG ratio (longer/shorter UG distance between alleles). The UG distance was relatively stable between alleles; in contrast, the SU distance of one allele was obviously longer than the distance indicated by the genome size. The results therefore indicate that SNRPN, UBE3A, and GABRB3 have non-linear and non-random curved spatial positioning in the normal nucleus, with differences in the SU distance between alleles possibly representing epigenetic evidence of nuclear organization and gene expression

Predictive Factors for Successful Vaccination Against Hepatitis B Surface Antigen in Patients Who Have Undergone Orthotopic Liver Transplantation

Post-orthotopic liver transplantation (OLT) hepatitis B recurrence is well-controlled with a nucleos(t)ide analogue and hepatitis B immunoglobulin (HBIG) combination, but the high cost and the potential risk of unknown infection associated with HBIG remain unresolved issues. Low-cost recombinant hepatitis B virus (HBV) vaccine administration is a potential solution to these problems. We retrospectively analyzed the rate and predictive factors of HBV vaccine success in 49 post-OLT patients: liver cirrhosis-type B (LC-B), n=28 patients; acute liver failure-type B (ALF-B), n=8; and non-HBV-related end-stage liver disease (non-B ESLD) who received a liver from anti-hepatitis B core antibody-positive donors, n=13. A positive anti-hepatitis B surface antibody response was achieved in 29% (8/28) of the LC-B group, 88% (7/8) of the ALF-B group, and 44% (4/9) of the adult non-B ESLD group. All four non-B ESLD infants showed vaccine success. The predictive factors for a good response in LC-B were young age, marital donor, and high donor age. ALF-B and non-B ESLD infants are thus good vaccination candidates. LC-B patients with marital donors are also good candidates, perhaps because the donated liver maintains an efficient immune memory to HBV, as the donors had already been infected in adulthood and showed adequate anti-HBV immune responses

Loss of runt-related transcription factor 3 expression leads hepatocellular carcinoma cells to escape apoptosis

Background: Runt-related transcription factor 3 (RUNX3) is known as a tumor suppressor gene for gastric cancer and other cancers, this gene may be involved in the development of hepatocellular carcinoma (HCC). Methods: RUNX3 expression was analyzed by immunoblot and immunohistochemistry in HCC cells and tissues, respectively. Hep3B cells, lacking endogenous RUNX3, were introduced with RUNX3 constructs. Cell proliferation was measured using the MTT assay and apoptosis was evaluated using DAPI staining. Apoptosis signaling was assessed by immunoblot analysis. Results: RUNX3 protein expression was frequently inactivated in the HCC cell lines (91%) and tissues (90%). RUNX3 expression inhibited 90 +/- 8% of cell growth at 72 h in serum starved Hep3B cells. Forty-eight hour serum starvation-induced apoptosis and the percentage of apoptotic cells reached 31 +/- 4% and 4 +/- 1% in RUNX3-expressing Hep3B and control cells, respectively. Apoptotic activity was increased by Bim expression and caspase-3 and caspase-9 activation. Conclusion: RUNX3 expression enhanced serum starvation-induced apoptosis in HCC cell lines. RUNX3 is deleted or weakly expressed in HCC, which leads to tumorigenesis by escaping apoptosis

Systematic documentation and analysis of human genetic variation in hemoglobinopathies using the microattribution approach

We developed a series of interrelated locus-specific databases to store all published and unpublished genetic variation related to hemoglobinopathies and thalassemia and implemented microattribution to encourage submission of unpublished observations of genetic variation to these public repositories. A total of 1,941 unique genetic variants in 37 genes, encoding globins and other erythroid proteins, are currently documented in these databases, with reciprocal attribution of microcitations to data contributors. Our project provides the first example of implementing microattribution to incentivise submission of all known genetic variation in a defined system. It has demonstrably increased the reporting of human variants, leading to a comprehensive online resource for systematically describing human genetic variation in the globin genes and other genes contributing to hemoglobinopathies and thalassemias. The principles established here will serve as a model for other systems and for the analysis of other common and/or complex human genetic diseases

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