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Abstract Background Spina bifida (SB) is a chronic condition requiring lifelong self-management, underscoring the need to establish a dedicated cohort for longitudinal monitoring of health outcomes. The purpose of this study was to describe the development and initial implementation of a single-center prospective cohort study of children with SB and their parents living in South Korea and to describe demographics, clinical outcomes, psychosocial characteristics, and family data for this cohort. Methods This cohort was established through expert panel formation, identification of health indicators based on the Life Course Model for Spina Bifida, creation of a cohort database system, and quality control planning. Participants, children aged 4–12 years with SB and their parents, were recruited from a large SB clinic at a tertiary hospital in South Korea. Two approaches were used to collect data: (1) diagnosis and clinical outcomes (e.g., lesion level, surgical history, laboratory results) were collected from the electronic medical record by pediatric nurse practitioners and (2) demographics, psychosocial characteristics and family data were collected from online self-reported questionnaires completed by children with SB and their parents if the child with SB was aged 7–12 years and by only the parents if the child with SB was aged 4–6 years. Results Between September 2022 and September 2023, 162 children (mean age 7.6 ± 2.6 years) and their parents participated, with 35.8% (4–6 years), 29% (7–9 years), and 35.2% (10–12 years). Diagnoses included lipomyelomeningocele (51.2%), myelomeningocele (27.2%), and tethered cord syndrome (20.4%). Clean intermittent catheterization was used by 38.3% and enemas by 22.2%. Moreover, 30.9% experienced urinary incontinence and 26.5% experienced fecal incontinence. Ambulation assistive devices were used by 14.8%, and ventriculoperitoneal shunts by only 4.3%. Conclusions This cohort provides a comprehensive understanding of demographics, and clinical and psychosocial outcomes for children with SB in South Korea. The dataset offers opportunities for data-driven, life-course tailored interventions to meet the specific needs of this population and their families.

Abstract A germinal disc located on the egg yolk surface drives genetic modification. Windowed and surrogate eggshell incubation methods have been developed, but these exhibit limited abilities to generate transgenic chickens. In the present study, we investigated the frequency of observing the germinal disc according to the preincubation positioning direction and time and found that it depended on those conditions, but only a few chicks (2.8–5.6%) hatched using the windowed method. Then, we attempted to improve surrogate method via one- or two-step procedures. All eggs, including surrogates that were 10 g heavier than the donor eggs, were obtained from a poultry flock of the same age. With the one-step surrogate method, where the donor egg was transferred only once through a 3.5 cm hole on the point end, at the first day of preincubation, into the surrogate egg, the survival rate at day 4 was 30.8%, and the hatching rate was 11.8%. With the two-step surrogate method (transfer was on the 1st and 4th day of incubation), the survival rate at day 4 was improved to 90.7%, and a hatching rate of 70.0% was achieved. Therefore, this method can be effective for in ovo artificial incubation.

Chicken embryonic stem (ES) cells have great potential and provide a powerful tool to investigate embryonic development and to manipulate genetic modification in a genome. However, very limited studies are available on the functional characterization and robust expansion of chicken ES cells compared to other species. Here, we have developed a method to generate chicken embryonic stem cell-like cells under pluripotent culture conditions. The chicken embryonic stem cell-like cells were cultivated long-term over several passages of culture without loss of pluripotency in vitro and had the specific expression of key stem cell markers. Furthermore, they showed severe changes in morphology and a significant reduction in pluripotent genes after siRNA-mediated NANOG knockdown. Collectively, these results demonstrate the efficient generation of chicken embryonic stem cell-like cells from EGK stage X blastoderm-derived singularized cells and will facilitate their potential use for various purposes, such as biobanking genetic materials and understanding stemness in the fields of animal biotechnology.

Abstract Background Transition to adulthood to live independently while self-managing health and daily life without parental help is crucial for young adults with chronic conditions. Despite its importance as a precursor to effectively managing lifelong conditions, little is known about the experiences of young adults with spina bifida (SB) in transition to adulthood in Asian countries. This study aimed to explore the experiences of Korean young adults with SB to identify the facilitators or barriers to the transition from adolescence to adulthood from their perspectives. Methods This study used a qualitative descriptive design. The data were collected in South Korea through three focus group interviews with 16 young adults with SB, aged 19–26, from August to November 2020. We conducted a qualitative content analysis using a conventional approach to identify the factors that facilitated and hindered the participants’ transition to adulthood. Results Two themes emerged as facilitators and barriers to the transition to adulthood. a) Facilitators: understanding and acceptance of SB, acquiring self-management skills, parenting styles encouraging autonomy, parents’ emotional support, school teachers’ thoughtful consideration, and participation in self-help groups. b) Barriers: overprotective parenting style, experience of being bullied by peers, damaged self-concept, concealing one’s chronic condition from others, and the lack of privacy in school restrooms. Conclusions Korean young adults with SB shared their experiences of struggling to properly manage their chronic conditions on their own, particularly concerning regular bladder emptying, during the transition from adolescence to adulthood. To facilitate the transition to adulthood, education on the SB and self-management for adolescents with SB and on parenting styles for their parents are important. To eliminate barriers to the transition to adulthood, improving negative perceptions of disability among students and teachers and making school restrooms CIC-friendly are needed.

Recently, we reported the robust in vitro three-dimensional (3D) expansion of intestinal organoids derived from adult bovine (> 24 months) samples. The present study aimed to establish an in vitro 3D system for the cultivation of intestinal organoids derived from growing cattle (12 months old) for practical use as a potential alternative to in vivo systems for various purposes. However, very few studies on the functional characterization and 3D expansion of adult stem cells from livestock species compared to those from other species are available. In this study, intestinal crypts, including intestinal stem cells, from the small intestines (ileum and jejunum) of growing cattle were isolated and long-term 3D cultures were successfully established using a scaffold-based method. Furthermore, we generated an apical-out intestinal organoid derived from growing cattle. Interestingly, intestinal organoids derived from the ileum, but not the jejunum, could be expanded without losing the ability to recapitulate crypts, and these organoids specifically expressed several specific markers of intestinal stem cells and the intestinal epithelium. Furthermore, these organoids exhibited key functionality with regard to high permeability for compounds up to 4 kDa in size (e.g., fluorescein isothiocyanate [FITC]-dextran), indicating that apical-out intestinal organoids are better than other models. Collectively, these results indicate the establishment of growing cattle-derived intestinal organoids and subsequent generation of apical-out intestinal organoids. These organoids may be valuable tools and potential alternatives to in vivo systems for examining host-pathogen interactions involving epithelial cells, such as enteric virus infection and nutrient absorption, and may be used for various purposes.

ABSTRACT: To date, many transgenic (TG) chicken lines have been developed, but few studies have performed a comparative analysis of their mortality, growth, and egg productivity. Previously, we reported the production of 3D8 scFv TG chickens showing antiviral activity. Here, we performed a biometric characterization of TG offspring female chickens. We selected 40 TG and 40 non-TG offspring female chicks among newly hatched chicks produced via artificial insemination of semen from heterotypic 3D8 scFv males into wild-type female chickens. Serum was collected at 14 wk of age, and serum concentrations of biochemical parameters, cytokines, and sex hormones were analyzed. Mortality and growth were monitored daily from 1 to 34 wk, egg productivity was monitored daily from 20 to 34 wk, and the weekly average values were used for analyses. Some serum parameters and cytokines were significantly different between non-TG and TG offspring female chickens. The levels of phosphorus (PHOS), total protein (TP), albumin (ALB), globulin (GLOB), and alanine aminotransferase (ALT) were significantly higher in non-TG chickens (P < 0.05). The levels of alkaline phosphatase (ALP) and gamma-glutamyltransferase (GGT) were significantly higher in TG chickens (P < 0.05). The levels of insulin growth factor-1 (IGF-1), interferon-gamma (INF-γ), interleukin-4 (IL-4), and IL-8 were significantly lower in TG chickens (P < 0.05). Despite these differences, the mortality rates, body weight, egg production rates, and egg weight were not significantly different in the experimental groups of non-TG and TG offspring female chickens (P > 0.05). In conclusion, ubiquitous expression of the 3D8 scFv gene in TG offspring female chickens does not affect some biometric characteristics, including mortality, growth, and egg productivity.

Recent progress has been made to establish intestinal organoids for an in vitro model as a potential alternative to an in vivo system in animals. We previously reported a reliable method for the isolation of intestinal crypts from the small intestine and robust three-dimensional (3D) expansion of intestinal organoids (basal-out) in adult bovines. The present study aimed to establish next-generation intestinal organoids for practical applications in disease modeling-based host-pathogen interactions and feed efficiency measurements. In this study, we developed a rapid and convenient method for the efficient generation of intestinal organoids through the modulation of the Wnt signaling pathway and continuous apical-out intestinal organoids. Remarkably, the intestinal epithelium only takes 3-4 days to undergo CHIR (1µM) treatment as a Wnt activator, which is much shorter than that required for spontaneous differentiation (7 days). Subsequently, we successfully established an apical-out bovine intestinal organoid culture system through suspension culture without Matrigel matrix, indicating an apical-out membrane on the surface. Collectively, these results demonstrate the efficient generation and next-generation of bovine intestinal organoids and will facilitate their potential use for various purposes, such as disease modeling, in the field of animal biotechnology.

ABSTRACT: NANOG, as a transcription factor, plays a key role in maintaining pluripotency in higher vertebrates. Thus, NANOG gene expression is a critical index for the transition from somatic cells to the pluripotent stage. Here, we established chicken knock-in DF1 cells in which the red fluorescent protein (RFP) gene was specifically inserted into the transcriptional start site of the NANOG gene through the CRISPR‒Cas9 (clustered regularly interspaced short palindromic repeat-CRISPR associated protein 9) technical platform. Subsequently, 4 transcription factors (Pou5f3, Sox2, Nanog, and Lin28A) were introduced into the NANOG-RFP DF1 cells, and finally, the induced pluripotent cells were established and examined by endogenous NANOG promoter-controlled RFP gene expression. The development of induced pluripotent stem cells (iPSCs) in avians would be useful for practical applications in the field of avian biotechnology, including biobanking genetic materials and restoring endangered species. In this study, a reporter cell line system was established to efficiently identify the induced pluripotent stage, and it will facilitate potential use for various purposes in the field of avian experimental models.

Objective Transgenic hens hold a great promise to produce various valuable proteins. Through virus transduction into stage X embryo, the transgene expression under the control of constructed chicken ovalbumin promoters has been successfully achieved. However, a validation system that can evaluate differently developed ovalbumin promoters in in vitro, remains to be developed. Methods In the present study, chicken oviduct epithelial cells (cOECs) were isolated from oviduct tissue and shortly cultured with keratinocyte complete medium supplemented with chicken serum. The isolated cells were characterized with immunofluorescence, western blot, and flow cytometry using oviduct-specific marker. Chicken mutated ovalbumin promoter (Mut-4.4-kb-pOV) was validated in these cells using luciferase reporter analysis. Results The isolated cOECs revealed that the oviduct-specific marker, ovalbumin protein, was clearly detected by immunofluorescence, western blot, and flow cytometry analysis revealed that approximately 79.40% of the cells contained this protein. Also, luciferase reporter analysis showed that the constructed Mut-4.4-kb-pOV exhibited 7.1-fold (p

To evaluate the effectiveness of acupuncture treatment for shoulder impingement syndrome (SIS) a literature review was conducted of randomized controlled trials (RCTs) where acupuncture was used as an intervention for patients diagnosed with SIS. Relevant clinical studies (N = 181) were retrieved from several databases based on the inclusion/exclusion criteria, and the interventions and results were analyzed. Six RCTs were selected to review based on the inclusion and exclusion criteria. In all 6 studies, the acupuncture treatment group showed significant positive changes in indicators evaluating pain, disability, and quality of life. A significant decrease in the evaluation indicators (Shoulder Pain and Disability Index, numeric rating scale, Visual Analogue Scale, Constant Murley Shoulder Assessment Score, patient’s global assessment, and doctor’s global assessment) and a significant increase in the questionnaire scores (UCLA, AL-score, EuroQol 5 Dimension Self-Report Questionnaire, and disabilities of the arm, shoulder, and hand) were observed. In addition, 1 study showed similar improvements in pain and quality of life measures in the acupuncture group and corticosteroid injection group. No major side effects were reported. Acupuncture may be an effective and safe treatment for SIS however, further RCTs are required.

Lithium-ion batteries (LIBs) are widely used, and extensive research has been conducted on new electrode materials. Silicon has been widely studied as an anode material. However, Si has a problem which is the large volume expansion. This study improved the electrochemical properties of Si thin films by employing a Ti-Nb-Zr SMA as a replacement candidate. The electrodes were prepared using two-step DC magnetron sputtering, followed by heat treatment to improve the adhesion between the Si thin film and the Ti-Nb-Zr film of the electrodes. X-ray diffraction confirmed that the SMA changed from the β (BCC) phase structure to the α (HCP) crystalline phase because of the volume changes arising from the Li-Si reactions during the lithiation/delithiation process. Additionally, the heat treatment significantly improved the adhesion between the active materials and substrate, improving the internal charge transfer bond and the martensite properties of the alloy allows it to work in the full sense. This study highlights a high-capacity, long cycle life, and free-standing electrode improvement by combining the shape-memory behavior recrystallization mechanism of heat-treated superelastic passive Ti-Nb-Zr for Si anode materials.

This study examined the potential benefits of male specific-pathogen-free (SPF) White Leghorn embryos in cellular agriculture for sustainable and ethical poultry meat production—addressing traditional farming challenges, including disease outbreaks of Salmonella and Avian influenza. We isolated myogenic precursor cells (MPCs) from the thigh muscles (Musculus femoris) of 12.5-day-old embryos from 10 SPF White Leghorns that tested negative for Salmonella. We randomly selected MPCs from three males and three females, isolated them using a modified pre-plating (pp) method, and compared their in vitro development. After 1 h (pp1) and 2 h (pp2) of incubation, they were transferred to a new dish to remove fast-adhering cells and cultured (pp3). Isolated MPCs had a 69% positive reaction to Pax7. During proliferation, no differences were observed in PAX7, MYF5, or MYOD expression between the male and female MPCs. However, after five days of differentiation, the expression of late myogenic factors—MYOG and MYF6—significantly increased in all MPCs. Notably, MYOG expression was 1.9 times higher in female than in male MPCs. This impacted MYMK’s expression pattern. Despite this, the myotube fusion index did not differ between the sexes. Muscle cells from male SPF-laying chicken embryos are promising for developing clean animal-cell-derived protein sources via resource recycling.

Spinel LiMn2O4 (LMO) is a state-of-the-art cathode material for Li-ion batteries. However, the operating voltage and battery life of spinel LMO needs to be improved for application in various modern technologies. Modifying the composition of the spinel LMO material alters its electronic structure, thereby increasing its operating voltage. Additionally, modifying the microstructure of the spinel LMO by controlling the size and distribution of the particles can improve its electrochemical properties. In this study, we elucidate the sol-gel synthesis mechanisms of two common types of sol-gels (modified and unmodified metal complexes)—chelate gel and organic polymeric gel—and investigate their structural and morphological properties and electrochemical performances. This study highlights that uniform distribution of cations during sol-gel formation is important for the growth of LMO crystals. Furthermore, a homogeneous multicomponent sol-gel, necessary to ensure that no conflicting morphologies and structures would degrade the electrochemical performances, can be obtained when the sol-gel has a polymer-like structure and uniformly bound ions; this can be achieved by using additional multifunctional reagents, namely cross-linkers.

ABSTRACT: A surrogate eggshell incubation system is a well-defined method to apply to avian genetic modification. In this study, we tried to investigate whether the egg weight differences between donor and surrogate eggs have an effect on donor viability. The groups were divided by egg weight differences between the donor and surrogate eggs into 4 in each system. The viability at d 4 was evaluated at the end of System II, the embryos alive were transferred into the second surrogate eggshells, and the viability at d 5, 6 was evaluated at early phase of System III. Then, the viability of System III was evaluated at different incubation period: d 6–12, d 13–18, d 19–21, and hatching rate was evaluated at d 22. Although the effect of egg weight differences between the donor and surrogate eggs was not observed, a specific group in System III showed higher survival and hatching rate than other group (P > 0.05).

Abstract Background The 3D8 single chain variable fragment (scFv) is a mini-antibody sequence that exhibits independent nuclease activity against all types of nucleic acids. In this research, crossing a 3D8 scFv G1 transgenic rooster with wild-type hens produced 3D8 scFv G2 transgenic chickens to evaluate suppression of viral transmission. Result The transgenic chickens were identified using genomic PCR and immunohistochemistry. To evaluate Newcastle disease virus (NDV) protection conferred by 3D8 scFv expression, transgenic, non-transgenic, and specific pathogen-free (SPF) chickens were challenged with virulent NDV by direct injection or aerosol exposure. The three groups of chickens showed no significant differences (p

Ultrasmall platinum nanoparticles (Pt-NPs) grafted with three types of hydrophilic and biocompatible polymers, i.e., poly(acrylic acid), poly(acrylic acid-co-maleic acid), and poly(methyl vinyl ether-alt-maleic acid) were synthesized using a one-pot polyol method. Their physicochemical and X-ray attenuation properties were characterized. All polymer-coated Pt-NPs had an average particle diameter (davg) of 2.0 nm. Polymers grafted onto Pt-NP surfaces exhibited excellent colloidal stability (i.e., no precipitation after synthesis for >1.5 years) and low cellular toxicity. The X-ray attenuation power of the polymer-coated Pt-NPs in aqueous media was stronger than that of the commercial iodine contrast agent Ultravist at the same atomic concentration and considerably stronger at the same number density, confirming their potential as computed tomography contrast agents.

ABSTRACT: Virus injection into EGK-X embryos is a well-defined approach in avian transgenesis. This system uses a chicken ovalbumin gene promoter to induce transgene expression in the chicken oviduct. Although a reconstructed chicken ovalbumin promoter that links an ovalbumin promoter and estrogen-responsive enhancer element (ERE) is useful, a large viral vector containing the ovalbumin promoter and a target gene restricts viral packaging capacity and produces low-titer virus particles. We newly developed recombinant chicken promoters by linking regulatory regions of ovalbumin and other oviduct-specific genes. Putative enhancer fragments of the genes, such as ovotransferrin (TF), ovomucin alpha subunit (OVOA), and ovalbumin-related protein X (OVALX), were placed at the 5`-flanking region of the 2.8-kb ovalbumin promoter. Basal promoter fragments of the genes, namely, pTF, lysozyme (pLYZ), and ovomucoid (pOVM), were placed at the 3`-flanking region of the 1.6-kb ovalbumin ERE. The recombinant promoters cloned into each reporter vector were evaluated using a dual luciferase assay in human and chicken somatic cells, and LMH/2A cells treated with 0-1,000 nM estrogen, and cultured primary chicken oviduct cells. The recombinant promoters with linking ovalbumin and TF, OVOA, pOVM, and pLYZ regulatory regions had 2.1- to 19.5-fold (P < 0.05) higher luciferase activity than the reconstructed ovalbumin promoter in chicken oviduct cells. Therefore, recombinant promoters may be used to efficiently drive transgene expression in transgenic chickens.

Lithium-ion batteries (LIBs) have been used in many fields, such as consumer electronics and automotive and grid storage, and its applications continue to expand. Several studies have attempted to improve the performance of LIBs. In particular, the use of high-capacity silicon and tin as anodes has been widely studied. Although anodes composed of silicone and tin have high theoretical capacities, poor electrical conductivity and considerable volume expansion of such anodes deteriorate the LIB performance. Thus, Li4Ti5O12 (LTO), a zero-strain material, has attracted much attention with high cycle stability and rate capability through improved electrical conductivity. However, LTO has the disadvantages of a low electrical conductivity (10−8 to 10−13 S cm−1) and moderate Li+ ion diffusion coefficient (10−9 to 10−16 cm2 s−1). In this study, the flexible and free-standing composite films were fabricated using only LTO and multi-walled carbon nanotube(CNT) with high electrical conductivity and ion diffusivity. The prepared LTO/CNT films showed a higher charge/discharge capacity than the theoretical capacity of the LTO electrode.

Although low-dose methotrexate (MTX) has been used widely in treatment of a variety of dermatological diseases, including multifocal primary cutaneous anaplastic large cell lymphoma (PCALCL), it has not been established for use in the treatment guidelines for solitary or localized PCALCL. Furthermore, there has been no report of long-term follow-up data in Asian patients with PCALCL treated with low-dose MTX. To investigate the effectiveness and clinical outcome of treatment with low-dose MTX, clinical and long-term follow-up data of 7 patients with solitary or localized PCALCL were analysed retrospectively. Of the 7 patients, 6 (85.7%) showed a complete response and 1 (14.3%) showed partial remission. During follow-up, mean duration of 92.1 months, 5 patients developed one or more cutaneous relapses. At the last follow-up, all of the patients with PCALCL were alive without disease. These results indicate that low-dose MTX is a highly effective and safe treatment for solitary or localized PCALCL as well as multiple relapsed lesions.

Acute vascular rejection has been known as a main barrier occurring in a xenograted tissue of alpha 1,3-galactosyltransferase knock-out (GalT KO) pig into a non-human primate (NHP). Adenosine which is a final metabolite following sequential hydrolysis of nucleotide by ecto-nucleotidases such as CD39 and CD73, act as a regulator of coagulation, and inflammation. Thus xenotransplantation of CD39 and CD73 expressing pig under the GalT KO background could lead to enhanced survival of recipient NHP. We constructed a human CD39 and CD73 expression cassette designed for endothelial cell-specific expression using porcine Icam2 promoter (pIcam2-hCD39/hCD73). We performed isolation of endothelial cells (pAEC) from aorta of 4 week-old GalT KO and membrane cofactor protein expressing pig (GalT-MCP/-MCP). We were able to verify that isolated cells were endothelial-like cells using immunofluorescence staining analysis with von Willebrand factor antibody, which is well known as an endothelial maker, and tubal formation assay. To find optimal condition for efficient transfection into pAEC, we performed transfection with GFP expression vector using four programs of nucleofection, M-003, U-023, W-023 and Y-022. We were able find that the program W-023 was optimal for pAEC with regard to viability and transfection efficiency by flow cytometry and fluorescent microscopy analyses. Finally, we were able to obtain GalT-MCP/-MCP/CD39/CD73 pAEC expressing CD39 and CD73 at levels of 33.3% and 26.8%, respectively. We suggested that pACE isolated from GalT-MCP/-MCP pig might be provided as a basic resource to understand biochemical and molecular mechanisms of the rejections and as an alternative donor cells to generate GalT-MCP/-MCP/CD39/CD73 pig expressing CD39 and CD73 at endothelial cells.

Intestinal organoids offer great promise for disease-modelling-based host–pathogen interactions and nutritional research for feed efficiency measurement in livestock and regenerative medicine for therapeutic purposes. However, very limited studies are available on the functional characterisation and three-dimensional (3D) expansion of adult stem cells in livestock species compared to other species. Intestinal crypts derived from intestinal organoids under a 3D culture system from the small intestine in adult bovine were successfully established and characterised for functionality testing, including the cellular potentials and genetic properties based on immunohistochemistry, immunocytochemistry, epithelial barrier permeability assay, QuantSeq 3′ mRNA-Seq. data and quantitative reverse transcription-polymerase chain reaction. Intestinal organoids were long-term cultivated over several passages of culture without loss of the recapitulating capacity of crypts, and they had the specific expression of several specific markers involved in intestinal stem cells, intestinal epithelium, and nutrient absorption. In addition, they showed the key functionality with regard to a high permeability for compounds of up to FITC-dextran 4 kDa, while FITC-dextran 40 kDa failed to enter the organoid lumen and revealed that the genetic properties of bovine intestinal organoids were highly similar to those of in vivo. Collectively, these results provide a reliable method for efficient isolation of intestinal crypts from the small intestine and robust 3D expansion of intestinal organoids in adult bovine and demonstrate the in vitro 3D organoids mimics the in vivo tissue topology and functionality. Finally, intestinal organoids are potential alternatives to in vivo systems and will be facilitated as the practical model to replace animal experiments for various purposes in the fields of animal biotechnology.

Pig has been known to be one of the most feasible animals as a bioreactor to produce pharmaceuticals in milk and as a mediator in xenotransplantation research. Previously, we generated transgenic pigs for both purposes, which were expressing Factor 8, vWF, hTPA, and hEPO in milk, along with expression of MCP at GalT gene locus (GalT-MCP/-MCP) as well as expressing MCP at GalT gene loci with CD73 expression (GalT-MCP/+/CD73). In this study, we performed comparative analyses of sperm parameters between wild type male (WT) pig and those transgenic males to examine the effects of transgenes integrated into the pigs on motility, morphology, viability, and acrosome integrity of the spermatozoa. Our results showed that the rates of actively motile spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, GalT-MCP/+/CD73, and GalT-MCP/-MCP pigs were 85.0%, 83.3%, 82.5%, 83.3%, 82.5%, 77.5%, and 78.7%, respectively. Whereas, the rates of morphologically normal spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, GalT-MCP/+/CD73, and GalT-MCP/-MCP pigs were 90.0%, 80.0%, 80.0%, 83.3%, 85.0%, 91.8%, and 80.8%, respectively. In addition, the viability in spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, GalT-MCP/+/CD73, and GalT-MCP/-MCP pigs were 93.9%, 82.4%, 89.9%, 83.9%, 87.4%, 92.8%, and 83.6%, respectively. The rates of spermatozoa with normal acrosome integrity in WT, Factor 8, vWF, hTPA, hEPO, GalT-MCP/+/CD73, and GalT-MCP/-MCP pigs were 98.1%, 98.6%, 98.6%, 98.7%, 98.1%, 99.5%, and 95.1%, respectively. There were no significant differences in motility, morphology, viability, and acrosome integrity of the spermatozoa among WT, Factor 8, vWF, hTPA, and hEPO, GalT-MCP/+/CD73, and GalT-MCP/-MCP pigs. These mean that neither random integration nor targeted integration of the transgene into chromosome of pig effect on characteristics of spermatozoa. Ultimately, the transgenic male pigs subjected in this study could apply to propagate their progenies for production of human therapeutic proteins and advancing the xenotransplantation research.

It is very difficult to get the information about semen quality analysis in transgenic pigs because of limited numbers and research facilities. Therefore, in the present study, we analyzed the semen quality of transgenic boars generated for xenotransplantation research. Briefly, the semen samples were collected from 5 homozygous α1,3-Galactosyltransferase knock-out (GalT-/-) transgenic boars and immediately transported to the laboratory. These semen samples were decupled with DPBS and conducted to analyze semen parameters by a computer-assisted semen analysis (CASA) system. The boar semen were examined all 12 parameters such as total motility (TM), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), and hyperactivated (HYP), etc. In results, among the 5 GalT-/- boars, three boars (#134, 144, and 170) showed normal range of semen parameters, but #199 and 171 boars showed abnormal ranges of semen parameters according to standard ranges of semen parameters. Unfortunately, #171 boar showed azoospermia symptom with rare sperm counts in the original semen. Conclusively, assessment of semen parameters by CASA system is useful to pre-screening of reproductively healthy boar prior to natural mating and artificial insemination for multiplication and breeding.

With the rapid development of high-performance fibers such as carbon, enhanced glass fibers in structural applications, the use of fiber-reinforced composite (FRC) materials has also increased in many areas. Liquid composite molding (LCM) is a widely used manufacturing process in composite manufacturing; however, the rapid impregnation of resin in the reinforcing fibers during processing poses a significant issue. The optimization of resin impregnation is related to tow deformations in the reinforcing fibers. The present study therefore focuses on this tow deformation. The permeability behaviors in double-scale porous media were observed under different flow rates and viscosity conditions to examine the overall tendencies of structural changes in the reinforcement. The permeability results showed hysteresis with increasing and decreasing flow rate conditions of 50–800 mm3/s, indicating structural changes in the reinforcement. The tow behaviors of the double-scale porous media with respect to the thickness and flow rate were investigated in terms of the representative indices of the minor axis (tow thickness) and major axis. The minor axis and major axis of the tow showed decreasing and increasing trends of 2–5% and 2%, respectively, with minimum and maximum values at different positions along the reinforcement, affected by the different hydrodynamic entry lengths. Finally, the deformed tow behavior was observed microscopically to examine the behavior of the tow at different flow rates.

Polysiloxanes (PSs) have been widely utilized in the industry as lubricants, varnishes, paints, release agents, adhesives, and insulators. In addition, their applications have been expanded to include the development of new biomedical materials. To modify PS for application in therapeutic purposes, a flexible antibacterial Cu-MOF (metal–organic framework) consisting of glutarate and 1,2-bis(4-pyridyl)ethane ligands was embedded in PS via a hydrosilylation reaction of vinyl-terminated and H-terminated PSs at 25 °C. The bactericidal activities of the resulting Cu-MOF-embedded PS (PS@Cu-MOF) and the control polymer (PS) were tested against Escherichia coli, Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus. PS@Cu-MOF exhibited more than 80% bactericidal activity toward the tested bacteria at a concentration of 100 μg⋅mL−1 and exhibited a negligible cytotoxicity toward mouse embryonic fibroblasts at the same concentration. Release tests of the Cu(II) ion showed PS@Cu-MOF to be particularly stable in a phosphate-buffered saline solution. Furthermore, its physical and thermal properties, including the phase transition, rheological measurements, swelling ratio, and thermogravimetric profile loss, were similar to those of the control polymer. Moreover, the low cytotoxicity and bactericidal activities of PS@Cu-MOF render it a promising candidate for use in medicinal applications, such as in implants, skin-disease treatment, wound healing, and drug delivery.

Although piglets have been delivered by embryo transfer (ET) with in vitro produced (IVP) embryos and blastocysts, a success rate has still remained lower level. Unlike mouse, human, and bovine, it is difficult to a production of piglets by in vitro fertilization (IVF) because of an inappropriate in vitro culture (IVC) system in pig. Therefore, the present study was conducted to investigate whether minimized exposure time in IVC can improve the pregnancy and delivery rates of piglets. Immediately after IVM, the oocytes were denuded and co-incubated with freshly ejaculated boar semen for 3.5 to 4 hours at 38.5 °C under 5% CO2 in air. To avoid long-term exposure to in vitro state, we emitted IVC step after IVF. After that the presumptive zygotes were transferred into both oviducts of the surrogate on the same day or 1 day after the onset of estrus. Pregnancy was diagnosed on day 28 after ET and then was checked regularly every month by ultrasound examination. The 3 out of 4 surrogates were determined as pregnant (75%) and a total of 5 piglets (2 females and 3 males) were delivered at 118.3 ± 2.5 days of pregnancy period. In conclusion, a short-term exposure time may be an important factor in the production of IVP-derived piglets. It can be apply to the in vitro production system of transgenic pig by IVF, cloning, and pronuclear microinjection methods.

The high-pressure gaseous hydrogen (HPGH2) storage method is widely used owing to the low density of hydrogen gas at ambient temperature and atmospheric pressure. Therefore, rigorous safety analysis of the filling and discharging of compressed gas in a hydrogen tank is required to achieve reliable operational solutions for the safe storage of hydrogen. In this study, the behavior of compressed hydrogen gas in a hydrogen tank was investigated for its discharge. Numerical models for the adaptation of gas and turbulence models were examined. Gas model effects were examined to account for hydrogen gas behavior at the discharge temperature and pressure conditions. Turbulence model effects were analyzed to consider the accuracy of each model: the assessment of the turbulence models was compared in terms of the turbulence intensity. From the study of gas model effect, the Redlich–Kwong equation was found to be one of the realistic gas models of the discharging gas flow. Among the turbulence models, the shear stress transport model and Reynolds stress model predicted the compressed gas behavior more accurately, showing a lower turbulence intensity than those of the realizable and renormalization group models.