DsRed gene expression by doxycycline in porcine fibroblasts and cloned embryos using transposon

To develop a transgenic pig, introduction of foreign genes into fibroblasts is required. In this study, Piggybac transposition was used to produce tetracycline dependent gene expressing cloned embryos. Red fluorescence proteins (DsRed) combined with tetracycline promoter flanked transposon sequences were transfected into fetal fibroblasts, and the transfected cells were used as the donor for somatic cell nuclear transfer. Induction of DsRed expression was successfully controlled by doxycycline treatment in donor fibroblasts and early stage embryos. In conclusion, this study suggested that Piggybac transposition could deliver genes into cells or embryos for developing transgenic pig.Keywords: Miniature pigs, transfection, Piggybac, somatic cell nuclear transfer (SCNT), RFPAfrican Journal of Biotechnology Vol. 12(21), pp. 3188-319

Growth, carcass traits, cecal microbial counts, and blood chemistry of meat-type quail fed diets supplemented with humic acid and black cumin seeds

Objective The present study attempted to determine safe and sufficient growth promoters in poultry feeding. Methods A total of 520 seven-day-old quail chicks were randomly allotted to eight treatment groups in a 4×2 factorial design experiment to evaluate the effect of different levels of humic acid (HA) and black cumin (BC) seed and their interactions on growth, carcass traits, gut microbes, and blood chemistry of growing quails. Quails were randomly distributed into 8 groups in a 4×2 factorial design, included 4 HA levels (0, 0.75, 1.5, and 2.25 g/kg diet) and 2 BC levels (0 or 5 g/ kg diet). Results Increasing HA level associated with a gradual increase in final weight, feed intake and body weight gain along with an improvement in feed conversion ratio. Dietary addition of 5 g BC powder/kg diet gave similar results. The highest level of HA (2.25 g/kg diet) recorded the best values of carcass weight, breast yield, intestinal length, and intestinal weight comparing with the control and other HA levels. Total viable microbial counts decreased (p<0.05) with increasing levels of HA except the intermediate level (1.5 g/kg diet). The concentration of serum cholesterol and low density lipoprotein (excluding that 0.75 g HA) decreased (p<0.05) and high density lipoprotein increased (p = 0.034) along with increasing HA level. The interaction between the 2.25 g HA×5 g gave the best results regarding most studied parameters. Conclusion These findings indicated that HA combined with BC could be used as effective growth promoters, with the recommended level being 2.25 g HA+5 g BC/kg of quail diet

Twelve-month observational study of children with cancer in 41 countries during the COVID-19 pandemic

Introduction Childhood cancer is a leading cause of death. It is unclear whether the COVID-19 pandemic has impacted childhood cancer mortality. In this study, we aimed to establish all-cause mortality rates for childhood cancers during the COVID-19 pandemic and determine the factors associated with mortality. Methods Prospective cohort study in 109 institutions in 41 countries. Inclusion criteria: children &lt;18 years who were newly diagnosed with or undergoing active treatment for acute lymphoblastic leukaemia, non-Hodgkin's lymphoma, Hodgkin lymphoma, retinoblastoma, Wilms tumour, glioma, osteosarcoma, Ewing sarcoma, rhabdomyosarcoma, medulloblastoma and neuroblastoma. Of 2327 cases, 2118 patients were included in the study. The primary outcome measure was all-cause mortality at 30 days, 90 days and 12 months. Results All-cause mortality was 3.4% (n=71/2084) at 30-day follow-up, 5.7% (n=113/1969) at 90-day follow-up and 13.0% (n=206/1581) at 12-month follow-up. The median time from diagnosis to multidisciplinary team (MDT) plan was longest in low-income countries (7 days, IQR 3-11). Multivariable analysis revealed several factors associated with 12-month mortality, including low-income (OR 6.99 (95% CI 2.49 to 19.68); p&lt;0.001), lower middle income (OR 3.32 (95% CI 1.96 to 5.61); p&lt;0.001) and upper middle income (OR 3.49 (95% CI 2.02 to 6.03); p&lt;0.001) country status and chemotherapy (OR 0.55 (95% CI 0.36 to 0.86); p=0.008) and immunotherapy (OR 0.27 (95% CI 0.08 to 0.91); p=0.035) within 30 days from MDT plan. Multivariable analysis revealed laboratory-confirmed SARS-CoV-2 infection (OR 5.33 (95% CI 1.19 to 23.84); p=0.029) was associated with 30-day mortality. Conclusions Children with cancer are more likely to die within 30 days if infected with SARS-CoV-2. However, timely treatment reduced odds of death. This report provides crucial information to balance the benefits of providing anticancer therapy against the risks of SARS-CoV-2 infection in children with cancer

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10&nbsp;years; 78.2% included were male with a median age of 37&nbsp;years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020

Sex differences in single IVF-derived bovine embryo cultured in chemically defined medium

Single embryo culture is essential for culturing embryos derived from few oocytes obtained from elite cows through ultrasonography guidance. Bovine in vitro fertilization (IVF) and individual embryo culture is a challenge as it generally leads to impaired embryo development. In this study, we explored the embryonic development and the sex ratio of IVF-derived bovine embryo cultured individually in chemically defined two-step culture medium. Total 63 cumulus-oocyte complexes were collected, in vitro matured, in vitro fertilized and the resultant fertilized oocytes were randomly cultured individually (4 trials, 15-16 oocytes each) in microdrops of 5 mu L of a chemically defined two-step culture medium. Blastocysts were counted in every trial (n=32, 50.79%) and all of them were used for both genomic DNA and total RNA extraction, cDNA synthesis and PCR using specific primers for GAPDH, GDP6, XIST and SRY genes. Results showed significant difference in expression of XIST (positive expression in 11 blastocysts) and SRY (positive expression in 21 blastocysts) mRNAs, P&lt;.05. This result supports the hypothesis of sexual dimorphism among the pre-implantation in vitro produced embryos and provides an efficient medium for single bovine embryos in vitro production.OAIID:RECH_ACHV_DSTSH_NO:T201808499RECH_ACHV_FG:RR00200001ADJUST_YN:EMP_ID:A077836CITE_RATE:0DEPT_NM:수의학과EMAIL:[email protected]_YN:NY

Relevance of multilamellar and multicompartmental vesicles in biological fluids: understanding the significance of proportional variations and disease correlation

Abstract Extracellular vesicles (EVs) have garnered significant interest in the field of biomedical science due to their potential applications in therapy and diagnosis. These vesicles participate in cell-to-cell communication and carry a diverse range of bioactive cargo molecules, such as nucleic acids, proteins, and lipids. These cargoes play essential roles in various signaling pathways, including paracrine and endocrine signaling. However, our understanding of the morphological and structural features of EVs is still limited. EVs could be unilamellar or multilamellar or even multicompartmental structures. The relative proportions of these EV subtypes in biological fluids have been associated with various human diseases; however, the mechanism remains unclear. Cryo-electron microscopy (cryo-EM) holds great promise in the field of EV characterization due to high resolution properties. Cryo-EM circumvents artifacts caused by fixation or dehydration, allows for the preservation of native conformation, and eliminates the necessity for staining procedures. In this review, we summarize the role of EVs biogenesis and pathways that might have role on their structure, and the role of cryo-EM in characterization of EVs morphology in different biological samples and integrate new knowledge of the alterations of membranous structures of EVs which could be used as biomarkers to human diseases

Comprehensive Proteomics Analysis of In Vitro Canine Oviductal Cell-Derived Extracellular Vesicles

Dogs (Canis lupus familiaris) have unique and peculiar reproductive characteristics. While the interplay between in vitro oviductal cell-derived extracellular vesicles (OC-EVs) and cumulus-oocyte complexes in dogs has begun to be elucidated, no study has yet provided extensive information on the biological content and physiological function of OC-EVs and their role in canine oocyte development. Here, we aimed to provide the first comprehensive proteomic analysis of OC-EVs. We identified 398 proteins as present in all OC-EVs samples. The functional enrichment analysis using Gene Ontology terms and an Ingenuity Pathway Analysis revealed that the identified proteins were involved in several cellular metabolic processes, including translation, synthesis, expression, and protein metabolism. Notably, the proteins were also involved in critical canonical pathways with essential functions in oocyte and embryo development, such as ERK/MAPK, EIF2, PI3K/AKT, and mTOR signaling. These data would be an important resource for studying canine reproductive physiology and establishing a successful in vitro embryo production system in dogs

Modulation of Heat-Shock Proteins Mediates Chicken Cell Survival against Thermal Stress

Heat stress is one of the most challenging environmental stresses affecting domestic animal production, particularly commercial poultry, subsequently causing severe yearly economic losses. Heat stress, a major source of oxidative stress, stimulates mitochondrial oxidative stress and cell dysfunction, leading to cell damage and apoptosis. Cell survival under stress conditions needs urgent response mechanisms and the consequent effective reinitiation of cell functions following stress mitigation. Exposure of cells to heat-stress conditions induces molecules that are ready for mediating cell death and survival signals, and for supporting the cell&rsquo;s tolerance and/or recovery from damage. Heat-shock proteins (HSPs) confer cell protection against heat stress via different mechanisms, including developing thermotolerance, modulating apoptotic and antiapoptotic signaling pathways, and regulating cellular redox conditions. These functions mainly depend on the capacity of HSPs to work as molecular chaperones and to inhibit the aggregation of non-native and misfolded proteins. This review sheds light on the key factors in heat-shock responses for protection against cell damage induced by heat stress in chicken

The Current Trends in Using Nanoparticles, Liposomes, and Exosomes for Semen Cryopreservation

Cryopreservation is an essential tool to preserve sperm cells for zootechnical management and artificial insemination purposes. Cryopreservation is associated with sperm damage via different levels of plasma membrane injury and oxidative stress. Nanoparticles are often used to defend against free radicals and oxidative stress generated through the entire process of cryopreservation. Recently, artificial or natural nanovesicles including liposomes and exosomes, respectively, have shown regenerative capabilities to repair damaged sperm during the freeze&ndash;thaw process. Exosomes possess a potential pleiotropic effect because they contain antioxidants, lipids, and other bioactive molecules regulating and repairing spermatozoa. In this review, we highlight the current strategies of using nanoparticles and nanovesicles (liposomes and exosomes) to combat the cryoinjuries associated with semen cryopreservation