Klotho is a protein that plays different functions in female fertility. We have previously reported that klotho protein supplementation during in vitro maturation improves porcine embryo development, while klotho knockout for somatic cell cloning completely blocks full-term pregnancy in vivo. However, the effects of the microinjection of klotho protein or klotho knockdown dual vector in porcine embryos at different time points and the specific molecular mechanisms remain largely unknown. In this study, we injected the preassembled cas9 + sgRNA dual vector, for klotho knockdown, into the cytoplasm of the germinal vesicle stage of oocytes and into porcine embryos after 6-h parthenogenetic activation. Similarly, the klotho protein was inserted into the cytoplasm of germinal vesicle stage oocytes and porcine embryos after 6-h parthenogenetic activation. Compared with the controls, the microinjection of klotho dual vector markedly decreased the blastocyst formation rates in germinal vesicle stage oocytes and activated embryos. However, the efficiency of blastocyst formation when klotho protein was inserted before in vitro maturation was significantly higher than that after klotho protein insertion into parthenogenetically activated embryos. These results indicated that klotho knockdown may impair embryo development into blastocyst irrespective of injection timing. In addition, klotho protein injection timing in pig embryos may be an important factor for regulating embryo development.

Radiocarbon (14C) in natural samples undergoes changes due to variations in atmospheric CO2 resulting from anthropogenic activities. To analyze the variation of the 14C ratio in atmospheric CO2, deciduous tree leaves were collected in Gyeongju, a popular tourist city in South Korea. Leaf samples were collected from Prunus subg. Cerasus trees at five different sampling points throughout the city over 3 years (2018, 2020, and 2021). The 14C data of the samples were categorized into three groups (downtown, rural, and tourist sites) and analyzed for variations among the different years. The 14C ratio at downtown sites was stable after 2018, the rural site ratio increased between 2018 and 2020 and then decreased between 2020 and 2021, and the tourist site ratio increased after 2018. We theorize that the increased 14C ratio at the tourist site was caused by a decrease in tourism after 2018.

Nosocomial transmission of COVID-19 among immunocompromised hosts can have a serious impact on COVID-19 severity, underlying disease progression and SARS-CoV-2 transmission to other patients and healthcare workers within hospitals. We experienced a nosocomial outbreak of COVID-19 in the setting of a daycare unit for paediatric and young adult cancer patients. Between 9 and 18 November 2020, 473 individuals (181 patients, 247 caregivers/siblings and 45 staff members) were exposed to the index case, who was a nursing staff. Among them, three patients and four caregivers were infected. Two 5-year-old cancer patients with COVID-19 were not severely ill, but a 25-year-old cancer patient showed prolonged shedding of SARS-CoV-2 RNA for at least 12 weeks, which probably infected his mother at home approximately 7–8 weeks after the initial diagnosis. Except for this case, no secondary transmission was observed from the confirmed cases in either the hospital or the community. To conclude, in the day care setting of immunocompromised children and young adults, the rate of in-hospital transmission of SARS-CoV-2 was 1.6% when applying the stringent policy of infection prevention and control, including universal mask application and rapid and extensive contact investigation. Severely immunocompromised children/young adults with COVID-19 would have to be carefully managed after the mandatory isolation period while keeping the possibility of prolonged shedding of live virus in mind.

Oxidative stress causes several diseases and dysfunctions in cells, including oocytes. Clearly, oxidative stress influences oocyte quality during in vitro maturation and fertilization. Here we tested the ability of coenzyme Q10 (CoQ10) to reduce reactive oxygen species (ROS) and improve mouse oocyte quality during in vitro culture. Treatment with 50 μM CoQ10 efficiently reduced ROS levels in oocytes cultured in vitro. The fertilizable form of an oocyte usually contains a cortical granule-free domain (CGFD). CoQ10 enhanced the ratio of CGFD–oocytes from 35% to 45%. However, the hardening of the zona pellucida in oocytes was not affected by CoQ10 treatment. The in vitro maturation capacity of oocytes, which was determined by the first polar body extrusion, was enhanced from 48.9% to 75.7% by the addition of CoQ10 to the culture medium. During the parthenogenesis process, the number of two-cell embryos was increased by CoQ10 from 43.5% to 67.3%. Additionally, treatment with CoQ10 increased the expression of Bcl2 and Sirt1 in cumulus cells. These results suggested that CoQ10 had a positive effect on ROS reduction, maturation rate and two-cell embryo formation in mouse oocyte culture.

The experiments reported in this research paper aimed to determine the effect of supplementing different forms of L-methionine (L-Met) and acetate on protein synthesis in immortalized bovine mammary epithelial cell line (MAC-T cells). Treatments were Control, L-Met, conjugated L-Met and acetate (CMA), and non-conjugated L-Met and Acetate (NMA). Protein synthesis mechanism was determined by omics method. NMA group had the highest protein content in the media and CSN2 mRNA expression levels (P < 0.05). The number of upregulated and downregulated proteins observed were 39 and 77 in L-Met group, 62 and 80 in CMA group and 50 and 81 in NMA group from 448 proteins, respectively (P < 0.05). L-Met, NMA and CMA treatments stimulated pathways related to protein and energy metabolism (P < 0.05). Metabolomic analysis also revealed that L-Met, CMA and NMA treatments resulted in increases of several metabolites (P < 0.05). In conclusion, NMA treatment increased protein concentration and expression level of CSN2 mRNA in MAC-T cells compared to control as well as L-Met and CMA treatments through increased expression of milk protein synthesis-related genes and production of the proteins and metabolites involved in energy and protein synthesis pathways.

Allicin (AL) regulates the cellular redox, proliferation, viability, and cell cycle of different cells against extracellular-derived stress. This study investigated the effects of allicin treatment on porcine oocyte maturation and developmental competence. Porcine oocytes were cultured in medium supplemented with 0 (control), 0.01, 0.1, 1, 10 or 100 μM AL, respectively, during in vitro maturation (IVM). The rate of polar body emission was higher in the 0.1 AL-treated group (74.5% ± 2.3%) than in the control (68.0% ± 2.6%) (P < 0.1). After parthenogenetic activation, the rates of cleavage and blastocyst formation were significantly higher in the 0.1 AL-treated group than in the control (P < 0.05). The reactive oxygen species level at metaphase II did not significantly differ among all groups. In matured oocytes, the expression of both BAK and CASP3, and BIRC5 was significantly lower and higher, respectively, in the 0.1 AL-treated group than in the control. Similarly, the expression of BMP15 and CCNB1, and the activity of phospho-p44/42 mitogen-activated protein kinase (MAPK), significantly increased. These results indicate that supplementation of oocyte maturation medium with allicin during IVM improves the maturation of oocytes and the subsequent developmental competence of porcine oocytes.

Feasibility of multiwavelength Raman spectroscopy was studied as a potentialin-line monitoring technique for grain size distribution in channel poly-Si usedin three dimensional stacked NAND (3D NAND) Flash memory devices. Variouschannel poly-Si materials in 3D-NAND Flash memory devices, converted fromchemical vapor deposition (CVD) grown a-Si, were characterized usingnon-contact, multiwavelength Raman spectroscopy and high resolutioncross-sectional transmission electron microscopy (HRXTEM). The Ramancharacterization results were compared with HRXTEM images. The correlationbetween the grain size distribution characterized by multiwavelength Ramanspectroscopy and “on current” (ION) of 3D NANDFlash memory devices was investigated. Good correlation between these techniqueswas seen. Multiwavelength Raman spectroscopy is very promising as anon-destructive in-line monitoring technique for grain size distribution inchannel poly-Si used in 3D NAND Flash memory devices.

In the present article, the intermixing and clustering of U/Nd, O, and vacancies were studied by both laboratory and synchrotron-based x-ray diffraction in U1−yNdyO2−x alloys. It was found that an increased holding time at the high experimental temperature during initial alloy preparation results in a lower disorder of the Nd distribution in the alloys. Adjustment of the oxygen concentration in the U1−yNdyO2−x alloys with different Nd concentrations was accompanied by the formation of vacancies on the oxygen sublattice and a nanocrystalline component. The lattice parameters in the U1−yNdyO2−x alloys were also found to deviate significantly from Vegard's law when the Nd concentration was high (53%) and decreased with increasing oxygen concentration. Such changes indicate the formation of large vacancy concentrations during oxygen adjustment at these high temperatures. The change in the vacancy concentration after the oxygen adjustment was estimated relative to Nd concentration and oxygen stoichiometry.

The Clostridium-related poultry disease, necrotic enteritis (NE), causes substantial economic losses on a global scale. In the present study, a mixture of two plant-derived phytonutrients, Capsicum oleoresin and turmeric oleoresin (XT), was evaluated for its effects on local and systemic immune responses using a co-infection model of experimental NE in commercial broilers. Chickens were fed from hatch with a diet supplemented with XT, or with a non-supplemented control diet, and either uninfected or orally challenged with virulent Eimeria maxima oocysts at 14 d and Clostridium perfringens at 18 d of age. Parameters of protective immunity were as follows: (1) body weight; (2) gut lesions; (3) serum levels of C. perfringens α-toxin and NE B-like (NetB) toxin; (4) serum levels of antibodies to α-toxin and NetB toxin; (5) levels of gene transcripts encoding pro-inflammatory cytokines and chemokines in the intestine and spleen. Infected chickens fed the XT-supplemented diet had increased body weight and reduced gut lesion scores compared with infected birds given the non-supplemented diet. The XT-fed group also displayed decreased serum α-toxin levels and reduced intestinal IL-8, lipopolysaccharide-induced TNF-α factor (LITAF), IL-17A and IL-17F mRNA levels, while cytokine/chemokine levels in splenocytes increased in the XT-fed group, compared with the animals fed the control diet. In conclusion, the present study documents the molecular and cellular immune changes following dietary supplementation with extracts of Capsicum and turmeric that may be relevant to protective immunity against avian NE.

Using immunostaining methodology, we traced the axonal projection of FMRFamide (Phe-Met-Arg-Phe-NH2)-like immunoreactive (LI) medial neurosecretory cells (MNCs) and lateral neurosecretory cells (LNCs) from the brain into the ventral nerve cord (VNC) and retrocerebral complex in Bombyx mori (L.) (Lepidoptera: Bombycidae). Of the seven pairs of FMRFamide-LI MNCs, one pair extended its axons from the brain pars intercerebralis into the VNC ipsilateral connective where they appeared to terminate. The axons of the remaining MNCs ran through decussation in the brain median region and contralateral nervi corporis cardiaci (NCC) I out of the brain, and eventually innervated the contralateral corpus cardiacum (CC). Axons from the single pair of FMRFamide-LI LNCs projected into the ipsilateral NCC II fused with NCC I without decussation in the brain, and finally terminated in the CC. These results suggest that transport of the FMRFamide-like neuropeptide from may be related to the modulation of functions such as gut contraction in MNCs terminating in the VNC, and regulation of production and/or secretion of specific hormones such as juvenile hormone in MNCs and LNCs terminating in the CC.

We have propsed MgO/AZO bi-layer transparent conducting oxide (TCO) for thin film solar cells. From XRD analysis, it was observed that the full width at half maximum of AZO decreased when it was grown on MgO precursor. The Hall mobility of MgO/AZO bi-layer was 17.5cm2/Vs, whereas that of AZO was 20.8cm2/Vs. These indicated that the crystallinity of AZO decreased by employing MgO precursor. However, the haze (=total diffusive transmittance/total transmittance) characteristics of highly crystalline AZO was significantly improved by MgO precursor. The average haze in the visible region increased from 14.3 to 48.2%, and that in the NIR region increased from 6.3 to 18.9%. The reflectance of microcrystalline silicon solar cell was decreased and external quantum efficiency was significantly improved by applying MgO/AZO bi-layer TCO. The efficiency of microcrystalline silicon solar cell with MgO/AZO bi-layer front TCO was 6.66%, whereas the efficiency of one with AZO single TCO was 5.19%.