Na+/K+ ATPase α1 and ß3 subunits are localized to the basolateral membrane of trophectoderm cells in human blastocysts.

STUDY QUESTION: Is there a relation between specific Na+/K+ ATPase isoform expression and localization in human blastocysts and the developmental behavior of the embryo? SUMMARY ANSWER: Na+/K+ ATPase α1, ß1 and ß3 are the main isoforms expressed in human blastocysts and no association was found between the expression level of their respective mRNAs and the rate of blastocyst expansion. WHAT IS KNOWN ALREADY: In mouse embryos, Na+/K+ ATPase α1 and ß1 are expressed in the basolateral membrane of trophectoderm (TE) cells and are believed to be involved in blastocoel formation (cavitation). STUDY DESIGN, SIZE, DURATION: A total of 20 surplus embryos from 11 patients who underwent IVF and embryo transfer at a university hospital between 2009 and 2018 were analyzed. PARTICIPANTS/MATERIALS, SETTING, METHODS: After freezing and thawing Day 5 human blastocysts, their developmental behavior was observed for 24 h using time-lapse imaging, and the expression of Na+/K+ ATPase isoforms was examined using quantitative RT-PCR (RT-qPCR). The expressed isoforms were then localized in blastocysts using fluorescent immunostaining. MAIN RESULTS AND THE ROLE OF CHANCE: RT-qPCR results demonstrated the expression of Na+/K+ ATPase α1, ß1 and ß3 isoforms in human blastocysts. Isoforms α1 and ß3 were localized to the basolateral membrane of TE cells, and ß1 was localized between TE cells. A high level of ß3 mRNA expression correlated with easier hatching (P = 0.0261). LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: The expression of mRNA and the localization of proteins of interest were verified, but we have not been able to perform functional analysis. WIDER IMPLICATIONS OF THE FINDINGS: Of the various Na+/K+ ATPase isoforms, expression levels of the α1, ß1 and ß3 mRNAs were clearly higher than other isoforms in human blastocysts. Since α1 and ß3 were localized to the basolateral membrane via fluorescent immunostaining, we believe that these subunits contribute to the dilation of the blastocoel. The ß1 isoform is localized between TE cells and may be involved in tight junction formation, as previously reported in mouse embryos. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the JSPS KAKENHI (https://www.jsps.go.jp/english/index.html), grant number 17K11215. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors have no conflicts of interest.

Enhanced Material Assimilation in a Toroidal Plasma Using Mixed H_{2}+Ne Pellet Injection and Implications to ITER.

The ablation and assimilation of cryogenic pure H_{2} and mixed H_{2}+Ne pellets, which are foreseen to be used by the ITER tokamak for mitigating thermal and electromagnetic loads of major disruptions, are observed by spatially and temporally resolved measurements. It is experimentally demonstrated that a small fraction (here ≈5%) of neon added to hydrogenic pellets enhances the core density assimilation with reduced outward transport for the low magnetic-field side injection. This is consistent with theoretical expectations that line radiation increased by doped neon in dense plasmoids suppresses the plasmoid pressure and reduces the E[over →]×B[over →] transport of the ablated material.

PolyTag: A peptide tag that affords scaffold-less covalent protein assembly catalyzed by microbial transglutaminase.

Assembling proteins in close vicinity to each other provides an opportunity to gain unique function because collaborative and even synergistic functionalities can be expected in an assembled form. There have been a variety of strategies to synthesize functional protein assemblies but site-specific covalent assembly of monomeric protein units without impairing their intrinsic function remains challenging. Herein we report a powerful strategy to design protein assemblies by using microbial transglutaminase (MTG). A serendipitous discovery of self-crosslinking of enhanced green fluorescent protein (EGFP) fused with StrepTag I at the C-terminus revealed that EGFP was assembled through the crosslinking of the Lys (K) residue in the C-terminus of EGFP and the Gln (Q) residue in StrepTag I (AWRHPQFGG). Site-directed mutagenesis of the residues next to the K and Q yielded EGFP assemblies with higher molecular weights. An optimized peptide tag comprised of both K and Q residues (HKRWRHYQRGG) enabled the assembly of different types of proteins of interest (POI) when it was fused to either the N- or C-terminus. The peptide tag that enabled the self-polymerization of the functional POI without a scaffold was designated as a 'PolyTag'.

Isotope Effect on Energy Confinement Time and Thermal Transport in Neutral-Beam-Heated Stellarator-Heliotron Plasmas.

The isotope effect on energy confinement time and thermal transport has been investigated for plasmas confined by a stellarator-heliotron magnetic field. This is the first detailed assessment of an isotope effect in a stellarator heliotron. Hydrogen and deuterium plasmas heated by neutral beam injection on the Large Helical Device have exhibited no significant dependence on the isotope mass in thermal energy confinement time, which is not consistent with the simple gyro-Bohm model. A comparison of thermal diffusivity for dimensionally similar hydrogen and deuterium plasmas in terms of the gyroradius, collisionality, and thermal pressure has clearly shown robust confinement improvement in deuterium to compensate for the unfavorable mass dependence predicted by the gyro-Bohm model.

The motion and distribution of the gas in the central 300 pc of the galaxy as revealed by spectra of H3.

Spectroscopy of absorption lines of H3+ in the central molecular zone (CMZ) of the Galaxy show that a previously largely unknown component of the interstellar medium there, warm (T∼200 K) and diffuse (n ≲ 102 cm-3) gas, makes up a large fraction of the volume of the CMZ, and that this gas is moving radially outward from the centre. These discoveries upend the generally accepted understanding that the interstellar environment of the CMZ comprises almost entirely an ultra-hot plasma and dense molecular clouds. The radial momentum associated with the diffuse gas in the CMZ exceeds that of the ejecta of thousands of core-collapse supernovae and implies some extraordinary past activity in the centre, possibly associated with the supermassive black hole, Sgr A*. We speculate that within approximately 106 years, gravity could halt the expansion of the diffuse gas and that contraction towards the centre could then commence. This article is part of a discussion meeting issue 'Advances in hydrogen molecular ions: H3+, H5+ and beyond'.

Predictive performance of a genetic risk score using 11 susceptibility alleles for the incidence of Type 2 diabetes in a general Japanese population: a nested case-control study.

AIMS: To assess the predictive ability of a genetic risk score for the incidence of Type 2 diabetes in a general Japanese population. METHODS: This prospective case-control study, nested within a Japan Public Health Centre-based prospective study, included 466 participants with incident Type 2 diabetes over a 5-year period (cases) and 1361 control participants, as well as 1463 participants with existing diabetes and 1463 control participants. Eleven susceptibility single nucleotide polymorphisms, identified through genome-wide association studies and replicated in Japanese populations, were analysed. RESULTS: Most single nucleotide polymorphism loci showed directionally consistent associations with diabetes. From the combined samples, one single nucleotide polymorphism (rs2206734 at CDKAL1) reached a genome-wide significance level (odds ratio 1.28, 95% CI 1.18-1.40; P = 1.8 × 10-8 ). Three single nucleotide polymorphisms (rs2206734 in CDKAL1, rs2383208 in CDKN2A/B, and rs2237892 in KCNQ1) were nominally significantly associated with incident diabetes. Compared with the lowest quintile of the total number of risk alleles, the highest quintile had a higher odds of incident diabetes (odds ratio 2.34, 95% CI 1.59-3.46) after adjusting for conventional risk factors such as age, sex and BMI. The addition to the conventional risk factor-based model of a genetic risk score using the 11 single nucleotide polymorphisms significantly improved predictive performance; the c-statistic increased by 0.021, net reclassification improved by 6.2%, and integrated discrimination improved by 0.003. CONCLUSIONS: Our prospective findings suggest that the addition of a genetic risk score may provide modest but significant incremental predictive performance beyond that of the conventional risk factor-based model without biochemical markers.

Automated brain tissue and myelin volumetry based on quantitative MR imaging with various in-plane resolutions.

BACKGROUND AND PURPOSE: Segmented brain tissue and myelin volumes can now be automatically calculated using dedicated software (SyMRI), which is based on quantification of R1 and R2 relaxation rates and proton density. The aim of this study was to determine the validity of SyMRI brain tissue and myelin volumetry using various in-plane resolutions. METHODS: We scanned 10 healthy subjects on a 1.5T MR scanner with in-plane resolutions of 0.8, 2.0 and 3.0mm. Two scans were performed for each resolution. The acquisition time was 7-min and 24-sec for 0.8mm, 3-min and 9-sec for 2.0mm and 1-min and 56-sec for 3.0mm resolutions. The volumes of white matter (WM), gray matter (GM), cerebrospinal fluid (CSF), non-WM/GM/CSF (NoN), brain parenchymal volume (BPV), intracranial volume (ICV) and myelin were compared between in-plane resolutions. Repeatability for each resolution was then analyzed. RESULTS: No significant differences in volumes measured were found between the different in-plane resolutions, except for NoN between 0.8mm and 2.0mm and between 2.0mm and 3.0mm. The repeatability error value for the WM, GM, CSF, NoN, BPV and myelin volumes relative to ICV was 0.97%, 1.01%, 0.65%, 0.86%, 1.06% and 0.25% in 0.8mm; 1.22%, 1.36%, 0.73%, 0.37%, 1.18% and 0.35% in 2.0mm and 1.18%, 1.02%, 0.96%, 0.45%, 1.36%, and 0.28% in 3.0mm resolutions. CONCLUSION: SyMRI brain tissue and myelin volumetry with low in-plane resolution and short acquisition times is robust and has a good repeatability so could be useful for follow-up studies.

Seminoma leading to detection of testicular feminization syndrome: a case report.

The authors here report a 54-year-old (gravida 0, para 0), who claimed to have had her menarche at age 13 and menopause at 52 years. Two months prior to presentation, the subject first noticed a hard but elastic fist-sized mass in the left inguinal region that gradually grew, causing pressure-related pain. Although the external genitalia appeared female, the vagina was short and blind-ending, and no uterus or ovaries were identified on transvaginal ultrasound. Chromosome banding results (G-band method) showed 46XY. Laparoscopy revealed no traces of a vestigial uterus or ovaries; thus, based on the appearance of the external genitalia, a diagnosis of testicular feminization syndrome was made. Pathological testing of the palpable mass led to a diagnosis of seminoma with Leydig cell hyperplasia. Thus, in this case, the development of a seminoma in an undescended testis led to the detection of testicular feminization syndrome.

Retroperitoneal leiomyosarcoma: a case report.

Retroperitoneal leiomyosarcoma is a relatively rare and aggressive tumor. Because of its rarity, it is difficult to arrive at a definite diagnosis preoperatively and to design an effective strategy. Here the authors report a case of peritoneal leiomyosarcoma in which diagnosis was difficult because the clinical course resembled that of ovarian cancer. A 77-year-old woman diagnosed with ovarian cancer underwent laparotomy. The excised tumor contained a necrotic polypoid mass that histologically displayed the features of leiomyosarcoma. The patient received adjuvant chemotherapy with a combination of gemcitabine and docetaxel but died two months after surgery owing to the aggressive behavior of the tumor. Because the preoperative diagnosis in this case was ovarian cancer, arriving at a treatment strategy assuming peritoneal leiomyosarcoma was difficult. If complete surgical resection of tumor is not performed, as in the present case, the prognosis can be extremely poor.

Follow-up study of symptomatic submucous fibroids after hysteroscopic myomectomy.

PURPOSE OF INVESTIGATION: This study aimed to estimate the effectiveness of hysteroscopic myomectomy for symptomatic submucous uterine fibroids and to identify prognostic factors for persistent or recurrent symptoms. MATERIALS AND METHODS: A total of 237 patients who underwent hysteroscopic myomectomy were divided into three groups according to the classification of the European Society for Gynaecological Endoscopy: Type 0 (n=116), Type I (n=97), and Type II (n=24). Medical records and videotape records of all patients were retrospectively reviewed. RESULTS: Improvement of symptoms was achieved in 100% of Types 0 and I, and 66.7% of Type II. The five-year cumulative symptom-free rates after hysteroscopic myomectomy were 96.7% ± 1.9%, 87.8% 6.7%, and 44.5% ± 12.7% in Types 0, I, and II, respectively. The mean symptom-free periods were 46.2 ± 2.6, 47.7 ± 2.7, and 24.7 ± 6.3 months in Types 0, I, and II, respectively. Logistic regression analysis showed that co-existence of other myomas and Type II were independent prognostic factors for recurrence of symptoms. CONCLUSION: Type I fibroids are a good indication for hysteroscopic myomectomy. In Type II, some patients feel that their symptoms improve, but this curative effect could be temporary.

The relation between causes and onset time of polyhydramnios.

The aim of this analysis was to investigate the onset time and significance of maximum volume of polyhydraminios and whether the tter was associated with causes. This was a retrospective cohort study between 2012 and 2014. A total number of 68 singleton pregancies were analyzed. Gestational age at onset of polyhydramnios was 30.0 ± 2.8 (25-36) weeks in maternal factor, 30.0 ± 3.5 (25- 7) weeks in fetal factor, and 32.3 ± 2.0 (27-37) weeks in idiopathic factor. Median of maximum amniotic fluid index (AFI) was gnificantly late onset in idiopathic factor. Diabetes, gestational or pre-existing, was present in all of women (ten cases) in maternal facror. Higher AFI was found to be associated with an increased frequency of prenatally detected congenital anomalies. Abnormal fetal kary- type noted in 18/45 (40%) cases of polyhydramnios. Polyhydramnios diagnosed on ultrasound requires further maternal and fetal iagnostic tests.

Histologic chorioamnionitis prevalence in patients with premature rupture membranes.

This was a retrospective cohort study between 2002 and 2011. A total number of 150 singleton pregnancies with preterm premature rupture of membranes (PROM) (before 34 weeks) were analyzed. Histological chorioamnionitis (Blanc grade III) was significantly increased over three days from onset of premature rupture of membranes. The positive relationship was strengthened (odds ratios, 3.5; 95% confidence intervals, 1.5-5.2) over three days from onset of preterm PROM. PROM is a risk factor important for histological chorioamnionitis. To avoid neonatal infection, early termination is recommended in preterm PROM patients.

Rupture risk factors of fallopian tubal pregnancy.

The present authors analyzed patients' backgrounds and pre-surgical findings to clarify the risk factors of rupture of fallopian tubal pregnancy. The surgical findings 113 cases were clearly diagnosed as fallopian tubal pregnancy with or without rupture. Twenty-six cases of fallopian tubal pregnancy were ruptured and 87 cases were not ruptured at the time of operation. The risk factors of fallopian tubal rupture were assessed by Chi-square for independence test and multiple regression analysis. Obesity (BMI over 26), prior birth history, social welfare entitlement, ultrasonography findings of fetal heart movement, and pre-surgical serum beta-hCG level more than 3,000 mIU/ml patient were significantly higher risk in fallopian tubal rupture. Fertility treatment patient were at significantly lower risk for fallopian tubal rupture. Higher beta-hCG levels, especially >3,000 mIU/ml is associated with increased risk of fallopian tubal rupture in ectopic pregnancy.

Characterization of ASKP1240, a fully human antibody targeting human CD40 with potent immunosuppressive effects.

Blocking the CD40-CD154 interaction is reported to be effective for transplantation management and autoimmune disease models in rodents and nonhuman primates. However, clinical trials with anti-CD154 mAbs were halted because of high incidence of thromboembolic complications. Thus, we generated and characterized a fully human anti-CD40 mAb ASKP1240, as an alternative to anti-CD154 mAb. In vitro ASKP1240 concentration-dependently inhibited human peripheral blood mononuclear cell proliferation induced by soluble CD154. In addition, ASKP1240 did not destabilize platelet thrombi under physiological high shear conditions while mouse anti-human CD154 mAb (mu5C8) did. And ASKP1240 itself did not activate platelet and endothelial cells. In vivo administration of ASKP1240 (1 or 10 mg/kg, intravenously) to cynomolgus monkeys, weekly for 3 weeks, significantly attenuated both delayed-type hypersensitivity and specific antibody formation evoked by tetanus toxoid. The immunosuppressive effect was well correlated with the CD40 receptor saturation. Thus, these results suggest that ASKP1240 is immunosuppressive but not prothromboembolic, and as such appears to be a promising therapeutic candidate for the management of solid organ transplant rejection and autoimmune diseases therapy.

Cooling dynamics of photoexcited C6(-) and C6H(-).

We report conclusive evidence of an efficient cooling mechanism via the electronic radiative transitions of hot small molecular anions isolated in vacuum. We stored C6(-) and C6H(-) in an ion storage ring and observed laser-induced electron detachment with delays up to several milliseconds. The terminal hydrogen atom caused a drastic change in the decay profiles. The decay of photoexcited C6H(-) is slow and nonexponential, which can be explained by depletion cooling, whereas that for C6(-) occurs extremely fast, on a time scale below 0.1 ms and can only be explained by electronic radiative cooling via low-lying electronic excited states.

Self-monitoring has potential for home exercise programmes in patients with haemophilia.

Haemophiliacs who have had to keep a physically inactive lifestyle due to bleeding during childhood are likely to have little motivation for exercise. The purpose of this study is to clarify the effectiveness of the self-monitoring of home exercise for haemophiliacs. A randomized controlled trial was conducted with intervention over 8 weeks at four hospitals in Japan. Subjects included 32 male outpatients aged 26-64 years without an inhibitor who were randomly allocated to a self-monitoring group and a control group. Individual exercise guidance with physical activity for improvement of their knee functions was given to both groups. The self-monitoring materials included an activity monitor and a feedback system so that the self-monitoring group could send feedback via the Internet and cellular phone. The self-monitoring was performed by checking exercise adherence and physical activity levels, bleeding history and injection of a coagulation factor. Both groups showed significant improvements in exercise adherence (P < 0.001) and physical function such as the strength of knee extension (P < 0.001), range of knee extension (P < 0.001), range of ankle dorsiflexion (P < 0.01), a modified Functional Reach (P < 0.05) and 10 metre gait time (P < 0.01). In particular, improvements in exercise adherence (P < 0.05), self-efficacy (P < 0.05), and strength of knee extension (P < 0.05) were significant in the self-monitoring group compared with those in the control group. No increase in bleeding frequency and pain scale was noted. The self-monitoring of home exercise for haemophilic patients is useful for the improvement of exercise adherence, self-efficacy and knee extension strength.

Direct observation of internal energy distributions of C5(-).

Photon induced decay of C5(-) has been measured in an electrostatic storage ring. The time dependence of the photo-enhanced decay is close to a 1∕t decay which indicates a thermal process. The deviation from the expected power of -1 is quantitatively explained by the small heat capacity of the anion. Measurements of the photo-enhanced decay at different storage times and photon energies allow a determination of the radiative cooling rate and the energy distribution of the ions. The average energy content between 15 and 70 ms is found to vary as time to the power -0.72, and at 50 ms the ions contain an average excitation energy of 0.5 eV. The time dependent energy distribution is consistent with cooling by infrared photon emission if published oscillator strengths are reduced by a factor 2.5, in contrast to cooling of larger molecular carbon-based ions where electronic transitions cause a much stronger cooling.

Effects of macronutrient intake on thigh muscle mass during home-based walking training in middle-aged and older women.

We examined whether post-exercise macronutrient supplementation during a 5-month home-based interval walking training (IWT) accelerated exercise-induced increases in skeletal muscle mass and strength in healthy middle-aged and older women. Thirty-five women (41-78 years) were randomly divided into two groups: IWT alone (CNT, n = 18) or IWT plus post-exercise macronutrient (7.6 g protein, 32.5 g carbohydrate, and 4.4 g fat) supplementation (NUT, n = 17). For IWT, all subjects were instructed to repeat five or more sets of 3-min low-intensity walking at 40% peak aerobic capacity (Vo2 peak ), followed by a 3-min high-intensity walking above 70% Vo2 peak per day for 4 or more days per week. We determined Vo2 peak , thigh muscle tissue area by computer tomography, and thigh muscle strength in all subjects before and after IWT. We found that an increase in hamstring muscle tissue area was 2.8 ± 1.2% in NUT vs -1.0 ± 0.7% in CNT and that in isometric knee flexion force was 16.3 ± 3.7% in NUT vs 6.5 ± 3.0% in CNT; both were significantly higher in NUT than in CNT (both, P < 0.001). Thus, post-exercise macronutrient supplementation enhanced the increases in thigh muscle mass and strength, although partially, in home-based IWT in middle-aged and older women.

Increase in interleukin-6 immediately after wheelchair basketball games in persons with spinal cord injury: preliminary report.

STUDY DESIGN: Case series. OBJECTIVES: To investigate the effects of wheelchair basketball game on plasma interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), C-reactive protein (CRP) and blood cell counts in persons with spinal cord injury (SCI). SETTING: The 2009 Mei-shin League of Wheelchair Basketball Games held at Wakayama, Japan. PARTICIPANTS: Five wheelchair basketball players with SCI voluntarily participated in this study. INTERVENTIONS: Blood samples were taken approximately 1 h before the player warm-up for the game and immediately after the game. MAIN OUTCOME MEASURES: IL-6, TNF-α, CRP and blood cell count were measured. RESULTS: Plasma IL-6 level and number of monocytes were significantly increased after the game, compared with pre-game measurements (P<0.05). No changes were observed in other measurements. There was a significant relationship between increased IL-6 levels and accumulated play duration. CONCLUSION: The lack of change in TNF-α and CRP levels suggested that the exercise-induced rise in IL-6 was not related to exercise-induced inflammatory response. Furthermore, the associated increase in the number of monocytes did not correlate with exercise-induced IL-6 changes, negating monocytes as the source of IL-6.

Mutations in RECQL4 cause a subset of cases of Rothmund-Thomson syndrome.

Rothmund-Thomson syndrome (RTS; also known as poikiloderma congenitale) is a rare, autosomal recessive genetic disorder characterized by abnormalities in skin and skeleton, juvenile cataracts, premature ageing and a predisposition to neoplasia. Cytogenetic studies indicate that cells from affected patients show genomic instability often associated with chromosomal rearrangements causing an acquired somatic mosaicism. The gene(s) responsible for RTS remains unknown. The genes responsible for Werner and Bloom syndromes (WRN and BLM, respectively) have been identified as homologues of Escherichia coli RecQ, which encodes a DNA helicase that unwinds double-stranded DNA into single-stranded DNAs. Other eukaryotic homologues thus far identified are human RECQL, Saccharomyces cerevisiae SGS1 and Schizosaccharomyces pombe rqh1. We recently cloned two new human helicase genes, RECQL4 at 8q24.3 and RECQL5 at 17q25, which encode members of the RecQ helicase family. Here, we report that three RTS patients carried two types of compound heterozygous mutations in RECQL4. The fact that the mutated alleles were inherited from the parents in one affected family and were not found in ethnically matched controls suggests that mutation of RECQL4 at human chromosome 8q24.3 is responsible for at least some cases of RTS.