Embryo splitting can increase the quantity but not the quality of blastocysts

AbstractObjectiveIn this study, we investigated the developmental potential of single blastomeres that were obtained from 4-cell mice embryos that were split during the blastocyst stage.Materials and MethodsImprinting Control Region (ICR) mice (age: 6–8 weeks), were superovulated and mated with a single fertile male of the same strain. We obtained 2-cell embryos that were then cultured in 4 groups (×4) with Human tubal fluid (HTF) supplemented with 12% fetal bovine serum. When these embryos reached the 4-cell stage, their zonae pellucidae were removed and every single blastomere was isolated by repeated pipetting with Ca/Mg2+-free medium. The isolated blastomeres (study group) and the intact embryos (control group) were then cultured to determine the blastocyst formation rate and quality.ResultsWe collected a total of 936 embryos from 524 morphologically intact, top-grade embryos in the 4-cell stage from 80 stimulated mice. We used 356 of these embryos to isolate the blastomeres. The remaining 168 embryos were cultured as controls. A total of 1312 single blastomeres were obtained and cultured in vitro. Among these, 620 blastocysts were harvested from the original embryos compared with 136 blastocysts that were harvested from the control group. The overall blastocyst formation rate was 174.2% (620 blastocysts from 356 embryos) for the study group compared with 81.5% (136 blastocysts from 168 embryos) for the control group. The study group was 43.3% (268 of 620) top-grade blastocysts compared with 91% (152 of 168) of the control group. Taken together, the percentage of top-grade blastocysts obtained per original embryo in the split group was 75.4% (174.2%×43.3%) compared with 74.2% (81.5%×91%) for the control group.ConclusionsEmbryo splitting can increase the number of blastocysts. However, the percentage of available top-grade blastocysts is the same compared with nonsplit embryos. Embryo splitting may not be a cost-effective technique for the generation of high-quality mouse blastocysts

Symbiodinium spp. associated with scleractinian corals from Dongsha Atoll (Pratas), Taiwan, in the South China Seal

Dongsha Atoll (also known as Pratas) in Taiwan is the northernmost atoll in the South China Sea and a designated marine national park since 2007. The marine park's scope of protection covers the bio-resources of its waters in addition to uplands, so it is important to have data logging information and analyses of marine flora and fauna, including their physiology, ecology, and genetics. As part of this effort, we investigated Symbiodinium associations in scleractinian corals from Dongsha Atoll through surveys carried out at two depth ranges (shallow, 1-5 m; and deep, 10-15 m) in 2009 and during a bleaching event in 2010. Symbiodinium composition was assessed using restriction fragment length polymorphism (RFLP) of 28S nuclear large subunit ribosomal DNA (nlsrDNA). Our results showed that the 796 coral samples from seven families and 20 genera collected in 2009 and 132 coral samples from seven families and 12 genera collected in 2010 were associated with Symbiodinium C, D and C+D. Occurrence of clade D in shallow water (24.5%) was higher compared to deep (14.9%). Due to a bleaching event in 2010, up to 80010 of coral species associated with Symbiodinium C underwent moderate to severe bleaching. Using the fine resolution technique of denaturing gradient gel electrophoresis (DGGE) of internal transcribed spacer 2 (ITS2) in 175 randomly selected coral samples, from 2009 and 2010, eight Symbiodinium C types and two Symbiodinium D types were detected. This study is the first baseline survey on Symbiodinium associations in the corals of Dongsha Atoll in the South China Sea, and it shows the dominance of Symbiodinium Glade C in the population

Ethical preferences for influencing superiors: A 41-society study

With a 41-society sample of 9990 managers and professionals, we used hierarchical linear modeling to investigate the impact of both macro-level and micro-level predictors on subordinate influence ethics. While we found that both macro-level and micro-level predictors contributed to the model definition, we also found global agreement for a subordinate influence ethics hierarchy. Thus our findings provide evidence that developing a global model of subordinate ethics is possible, and should be based upon multiple criteria and multilevel variables

Ultralight vector dark matter search using data from the KAGRA O3GK run

Among the various candidates for dark matter (DM), ultralight vector DM can be probed by laser interferometric gravitational wave detectors through the measurement of oscillating length changes in the arm cavities. In this context, KAGRA has a unique feature due to differing compositions of its mirrors, enhancing the signal of vector DM in the length change in the auxiliary channels. Here we present the result of a search for U(1)B−L gauge boson DM using the KAGRA data from auxiliary length channels during the first joint observation run together with GEO600. By applying our search pipeline, which takes into account the stochastic nature of ultralight DM, upper bounds on the coupling strength between the U(1)B−L gauge boson and ordinary matter are obtained for a range of DM masses. While our constraints are less stringent than those derived from previous experiments, this study demonstrates the applicability of our method to the lower-mass vector DM search, which is made difficult in this measurement by the short observation time compared to the auto-correlation time scale of DM

Study on Symbiodinium diversity in scleractinian corals of Dong-Sha Atoll

東沙環礁為南中國海中最北端且發育完整的環礁。一九九八年的聖嬰現象造成的海水升溫，使得東沙環礁內的造礁珊瑚大量白化接續死亡。欲瞭解東沙環礁造礁珊瑚與其體內共生藻(以下合稱珊瑚共生體)的生物種系多樣性與組合、不同深度的分佈、以及其與溫度變化的相關性，以為未來擬定保育策略，特別是收集面對未來氣候變遷下的持續升溫，東沙環礁珊瑚共生體與珊瑚礁抵抗力與恢復力探討的基礎資料。 本實驗分別於2009與2010年在東沙環礁內的塊礁(Patch reef)之礁頂(Reef Top)和礁底(Reef Base)，採集了七科二十一屬，共九百零三株造礁珊瑚，利用聚合酶連鎖反應(PCR)放大共生藻的細胞核核糖體核酸大次單位(nlsrDNA)基因片段後，再利用限制性片段長度多態性(RFLP)分析來評估東沙環礁內造礁珊瑚體內共生藻的多樣性。結果顯示東沙環礁造礁珊瑚體內的共生藻主要為C系群與D系群。在2009年的771個樣本中，只與C系群共生的共生體為71.34%，只與D系群共生佔19.71%，同時與C和D系群共生為8.95%，此結果與印度太平洋其他珊瑚礁共生藻多樣性皆以C系群為主相似。進一步分析塊礁礁頂與礁底共生藻分佈，結果顯示礁頂共生藻D系群的比例24.48%顯著高於礁底共生藻D系群14.88% (chi-square, p=0.007)，這可能與礁頂年均溫較礁底年均溫高有關。在2010年因反聖嬰現象引發的白化事件中，白化珊瑚中，84.78%為C系群珊瑚共生體，D系群珊瑚共生體為10.87%，而未白化珊瑚其D系群珊瑚共生體比例為62.79%。顯示與D系群共生的珊瑚宿主可能可以抵抗因高溫引起的珊瑚白化。Dong-Sha Atoll is the northernmost and well developed atoll in the South China Sea. For further study on reef-building corals and zooxanthellae (symbionts) diversity in coral hosts and their correlation with temperature fluctuation within different depths is necessary, to enact the conservation strategy in Dong-Sha Atoll, and important basic data to understand coral reef resistance and resilience to the gradual elevated seawater temperature caused by climate change. Coral samples were collected in 2009 and 2010 period in patch reefs of Dong-Sha lagoon from 2 different depths (Reef Top 1-5 meters and Reef Base 10–15 meters). A total of 903 samples belong to 7 families and 21 genera of reef-building corals were collected. Symbiodinium diversity was assessed using restriction fragment length polymorphism (RFLP) of the nuclear large subunit ribosomal DNA (nlsrDNA) of Symbiodinium DNA. In the present study, the results showed that the reef-building corals in Dong-Sha were associated with only two Symbiodinium clades: clade C and clade D. From the analysis of 771 samples collected in 2009, the Symbiodinium clade C, D and C+D proportion was 71.34%, 19.71% and 8.95% respectively. The results were similar to the pattern observed in corals from the Indo-Pacific where the Symbiodinium diversity of coral reefs is dominated by Symbiodinium clade C. Symbiodinium distribution analysis between Reef Top and Reef Base showed that clade D proportion in Reef Top (24.48%) was higher than Reef Base (14.88%)(chi-square, p=0.007). This might be due to the higher annual average seawater temperature at the Reef Top compared to Reef Base. In 2010 bleaching event caused by La Niña, the clade C proportion in bleached corals was up to 84.78% compared to clade D proportion of 10.87%. However, 62.79% of the non-bleached corals that were associated with clade D. This may support that Symbiodinium clade D can cope with coral bleaching caused by high seawater temperature

Nitric oxide maintains cell survival of Trichomonas vaginalis upon iron depletion

Abstract Background Iron plays a pivotal role in the pathogenesis of Trichomonas vaginalis, the causative agent of highly prevalent human trichomoniasis. T. vaginalis resides in the vaginal region, where the iron concentration is constantly changing. Hence, T. vaginalis must adapt to variations in iron availability to establish and maintain an infection. The free radical signaling molecules reactive oxygen species (ROS) and reactive nitrogen species (RNS) have been proven to participate in iron deficiency in eukaryotes. However, little is known about the roles of these molecules in iron-deficient T. vaginalis. Methods T. vaginalis cultured in iron-rich and -deficient conditions were collected for all experiments in this study. Next generation RNA sequencing was conducted to investigate the impact of iron on transcriptome of T. vaginalis. The cell viabilities were monitored after the trophozoites treated with the inhibitors of nitric oxide (NO) synthase (L-NG-monomethyl arginine, L-NMMA) and proteasome (MG132). Hydrogenosomal membrane potential was measured using JC-1 staining. Results We demonstrated that NO rather than ROS accumulates in iron-deficient T. vaginalis. The level of NO was blocked by MG132 and L-NMMA, indicating that NO production is through a proteasome and arginine dependent pathway. We found that the inhibition of proteasome activity shortened the survival of iron-deficient cells compared with untreated iron-deficient cells. Surprisingly, the addition of arginine restored both NO level and the survival of proteasome-inhibited cells, suggesting that proteasome-derived NO is crucial for cell survival under iron-limited conditions. Additionally, NO maintains the hydrogenosomal membrane potential, a determinant for cell survival, emphasizing the cytoprotective effect of NO on iron-deficient T. vaginalis. Collectively, we determined that NO produced by the proteasome prolonged the survival of iron-deficient T. vaginalis via maintenance of the hydrogenosomal functions. Conclusion The findings in this study provide a novel role of NO in adaptation to iron-deficient stress in T. vaginalis and shed light on a potential therapeutic strategy for trichomoniasis

Immune defects in active mycobacterial diseases in patients with primary immunodeficiency diseases (PIDs)

Natural human immunity to the mycobacteria group, including Mycobacterium tuberculosis, Bacille Calmette-Guérin (BCG) or nontuberculous mycobacteria (NTM), and/or Salmonella species, relies on the functional IL-12/23-IFN-γ integrity of macrophages (monocyte/dendritic cell) connecting to T lymphocyte/NK cells. Patients with severe forms of primary immunodeficiency diseases (PIDs) have more profound immune defects involving this impaired circuit in patients with severe combined immunodeficiencies (SCID) including complete DiGeorge syndrome, X-linked hyper IgM syndrome (HIGM) (CD40L mutation), CD40 deficiency, immunodeficiency with or without anhidrotic ectodermal dysplasia (NEMO and IKBA mutations), chronic granulomatous disease (CGD) and hyper IgE recurrent infection syndromes (HIES). The patients with severe PIDs have broader diverse infections rather than mycobacterial infections. In contrast, patients with an isolated inborn error of the IL-12/23-IFN-γ pathway are exclusively prone to low-virulence mycobacterial infections and nontyphoid salmonella infections, known as Mendelian susceptibility to the mycobacterial disease (MSMD) phenotype. Restricted defective molecules in the circuit, including IFN-γR1, IFN-γR2, IL-12p40, IL-12R-β1, STAT-1, NEMO, IKBA and the recently discovered CYBB responsible for autophagocytic vacuole and proteolysis, and interferon regulatory factor 8 (IRF8) for dendritic cell immunodeficiency, have been identified in around 60% of patients with the MSMD phenotype. Among all of the patients with PIDs referred for investigation since 1985, we have identified four cases with the specific defect (IFNRG1 for three and IL12RB for one), presenting as both BCG-induced diseases and NTM infections, in addition to some patients with SCID, HIGM, CGD and HIES. Furthermore, manifestations in patients with autoantibodies to IFN-γ (autoAbs-IFN-γ), which is categorized as an anticytokine autoantibody syndrome, can resemble the relatively persistent MSMD phenotype lacking BCG-induced diseases