Cell-free extract from porcine induced pluripotent stem cells can affect porcine somatic cell nuclear reprogramming.

Pretreatment of somatic cells with undifferentiated cell extracts, such as embryonic stem cells and mammalian oocytes, is an attractive alternative method for reprogramming control. The properties of induced pluripotent stem cells (iPSCs) are similar to those of embryonic stem cells; however, no studies have reported somatic cell nuclear reprogramming using iPSC extracts. Therefore, this study aimed to evaluate the effects of porcine iPSC extracts treatment on porcine ear fibroblasts and early development of porcine cloned embryos produced from porcine ear skin fibroblasts pretreated with the porcine iPSC extracts. The Chariot(TM) reagent system was used to deliver the iPSC extracts into cultured porcine ear skin fibroblasts. The iPSC extracts-treated cells (iPSC-treated cells) were cultured for 3 days and used for analyzing histone modification and somatic cell nuclear transfer. Compared to the results for nontreated cells, the trimethylation status of histone H3 lysine residue 9 (H3K9) in the iPSC-treated cells significantly decreased. The expression of Jmjd2b, the H3K9 trimethylation-specific demethylase gene, significantly increased in the iPSC-treated cells; conversely, the expression of the proapoptotic genes, Bax and p53, significantly decreased. When the iPSC-treated cells were transferred into enucleated porcine oocytes, no differences were observed in blastocyst development and total cell number in blastocysts compared with the results for control cells. However, H3K9 trimethylation of pronuclear-stage-cloned embryos significantly decreased in the iPSC-treated cells. Additionally, Bax and p53 gene expression in the blastocysts was significantly lower in iPSC-treated cells than in control cells. To our knowledge, this study is the first to show that an extracts of porcine iPSCs can affect histone modification and gene expression in porcine ear skin fibroblasts and cloned embryos.

Distinct TLR-mediated pathways regulate house dust mite-induced allergic disease in the upper and lower airways.

BACKGROUND: Allergic rhinitis (AR) and asthma are 2 entities of allergic airway diseases that frequently occur together, which is referred to as united airways. In contrast to this general concept, we hypothesized that innate immunity of the upper and lower airways is respectively distinctive, because the immunologic conditions of the nasal and lung mucosa as well as the functions of the immune cells within their epithelia are different. OBJECTIVE: We wanted to identify distinctive mechanisms of innate immunity in the nose and lung mucosa, which are responsible for house dust mite (HDM)-induced AR and allergic asthma (AA), respectively. METHODS: We constructed a mouse model of AR or AA induced by sensitization and consequent provocation with HDM extracts. RESULTS: HDM-derived ß-glucans, rather than LPS, were proven to be essential to activating innate immunity in the nasal mucosa and triggering AR, which depended on Toll-like receptor 2 (TLR2), but not on TLR4; however, the LPS/TLR4 signaling axis, rather than ß-glucans/TLR2, was critical to HDM-induced AA. These differences were attributed to the specific role of ß-glucans and LPS in inducing the surface expression of TLR2 and TLR4 and their translocation to lipid rafts in nasal and bronchial epithelial cells, respectively. We also showed that dual oxidase 2-generated reactive oxygen species mediate both ß-glucan-induced TLR2 activation and LPS-induced TLR4 activation. CONCLUSIONS: We describe a novel finding of distinctive innate immunity of the nose and lungs, respectively, which trigger AR and AA, by showing the critical role of HDM-induced TLR activation via dual oxidase 2-mediated reactive oxygen species.

Reciprocal longitudinal effects among Korean young adolescent' negative peer relationships, social withdrawal, and smartphone dependence.

The research should consider the complex dynamics of social interaction to better understand smartphone dependence among adolescents. In social situations, adolescents can choose to interact with their peers or use their smartphones, and smartphone dependence can drive adolescents far away from interaction with their peers. Adolescents, conversely, may use smartphones to avoid peer interaction because they have experienced social withdrawal or negative peer relationships. Previous research has not discussed enough what precedes or follows in longitudinal relationships. This study aims to examine the reciprocal longitudinal effects between adolescents' negative peer relationships, social withdrawal, and smartphone dependence. The analysis used longitudinal panel data collected yearly between 2018 and 2020 from the Korean Children and Youth Panel Survey (KCYPS) sampled from adolescents in the Republic of Korea and employed autoregressive cross-lagged modeling on 2230 participants. The reciprocal longitudinal relationships differed depending on the sub-factors of smartphone dependence. For instance, the sub-factor of smartphone dependence, including virtual life orientation and withdrawal, had reciprocal longitudinal relationships with negative peer relationships. The other sub-factor of smartphone dependence, including daily life disturbance and tolerance, influenced negative peer relationships consistently, but the converse path did not. In addition, social withdrawal longitudinally affected negative peer relationships the following year. However, the reciprocal longitudinal relationships between social withdrawal and smartphone dependence were not significant in any path in this research. This research provides practical implications for intervention to reduce smartphone dependence among young adolescents.

Matrix-assisted laser desorption ionization-time of flight mass spectrometry identification of peptide citrullination site using Br signature.

Enzymatic conversion of arginine to citrulline by peptidyl arginine deiminase is associated with peptide presentation and development of autoimmunity in rheumatoid arthritis. In order to facilitate identification of the citrullination site, citrulline residue was modified using 4-bromophenyl glyoxal, and 194Da mass increase and incorporation of the Br signature were confirmed by MALDI-TOF MS. Using this approach, we identified four and five citrullination sites of bovine serum albumin and bovine fibrinogen, respectively. MALDI-TOF/TOF MS was used to unambiguously identify two citrullination sites from bovine fibrinogen.

Enhancing spatial resolution in Fourier transform infrared spectral image via machine learning algorithms.

Owing to the intrinsic signal noise in the characterization of chemical structures through Fourier transform infrared (FT-IR) spectroscopy, the determination of the signal-to-noise ratio (SNR) depends on the level of the concentration of the chemical structures. In situations characterized by limited concentrations of chemical structures, the traditional approach involves mitigating the resulting low SNR by superimposing repetitive measurements. In this study, we achieved comparable high-quality results to data scanned 64 times and superimposed by employing machine learning algorithms such as the principal component analysis and non-negative matrix factorization, which perform the dimensionality reduction, on FT-IR spectral image data that was only scanned once. Furthermore, the spatial resolution of the mapping images correlated to each chemical structure was enhanced by applying both the machine learning algorithms and the Gaussian fitting simultaneously. Significantly, our investigation demonstrated that the spatial resolution of the mapping images acquired through relative intensity is further improved by employing dimensionality reduction techniques. Collectively, our findings imply that by optimizing research data through noise reduction enhancing spatial resolution using the machine learning algorithms, research processes can be more efficient, for instance by reducing redundant physical measurements.

Higher food literacy scores are associated with healthier diet quality in children and adolescents: the development and validation of a two-dimensional food literacy measurement tool for children and adolescents.

BACKGROUND/OBJECTIVES: Most child and adolescent food literacy measurement tools focus on nutrition and food safety. However, the importance of aspects related to the food system such as food distribution and food waste and their effects on environmental sustainability is growing. We therefore developed and validated a two-dimensional tool for children (8-12 years old) and adolescents (13-18 years old) that can comprehensively measure food literacy. The association of food literacy with diet quality and self-reported health was assessed. SUBJECTS/METHODS: First, we developed a food literacy conceptual framework that contains food system and literacy dimensions through a literature review, focus group interviews, and expert review. After a face validity study, we conducted the main survey (n = 200) to validate the questionnaire. Construct validity and reliability were assessed using exploratory factor analysis (EFA), confirmatory factor analysis (CFA), and Cronbach's alpha. RESULTS: As a result of the Delphi study, content validity was confirmed for the remaining 30 items after two items were excluded (content validity ratio = 0.86). Eleven items were excluded from the EFA results, while the CFA results indicated appropriate fit indices for the proposed model (comparative fit index = 0.904, root mean square error of approximation = 0.068). The final food literacy questionnaire consisted of 19 questions and comprised 5 factors: production, distribution, selection, preparation and cooking, and intake. Food literacy was positively associated with diet quality, as assessed by the Nutrition Quotient score, in both children and adolescents and with self-reported health in adolescents. CONCLUSIONS: Based on this integrated conceptual framework, a two-dimensional 19-item food literacy measurement tool was developed and verified for practical use to improve the diet quality and food-related environmental sustainability awareness of children and adolescents.

Development and Validation of a Food Literacy Assessment Tool for Community-Dwelling Elderly People.

Food literacy refers to the knowledge, skills, and attitudes required for individuals to choose foods that promote health. As the rate of diet-related diseases increases, food literacy is becoming more important. However, there are no tools available to evaluate food literacy among the Korean elderly. We derived 547 questions from a literature review and, after three rounds of Delphi surveys, selected 33 preliminary questions. We calculated the content validity ratio of the questions and applied a face validity procedure. We then selected 32 questions, assessed their validity, and distributed them as a questionnaire to 205 elderly people. We then conducted exploratory factor analysis (EFA) to determine the validity of the questionnaire and used an internal consistency index (Cronbach's α coefficient) to determine reliability. Based on the factor analysis, 13 questions were selected, distributed among three factors, and evaluated using the Kaiser-Meyer-Olkin (KMO) and Bartlett sphericity tests. The factor analysis showed that KMO was 0.872, which is a highly acceptable score, and the Bartlett sphericity test was χ2 = 1,374.69 at p = 0.00. The food literacy questionnaire developed in this study will likely be helpful for improving the healthcare of elderly people.

Mutations in SOHLH1 gene associate with nonobstructive azoospermia.

In a previous study, we found SOHLH1 (spermatogenesis and oogenesis-specific basic helix-loop-helix 1) as the first testis-specific basic helix-loop-helix transcription factor essential for spermatogonial differentiation. SOHLH1 therefore represents an excellent candidate gene for testicular failure such as nonobstructive azoospermia (NOA). We analyzed whether there were mutations in the SOHLH1 gene in 96 Korean patients with NOA. The sequence analysis discovered three novel variations: one intronic variant (c.346-1G>A), and two nonsynonymous exonic variants (c.91T>C and c.529C>A) with known single nucleotide polymorphisms (SNPs), which included six intronic variants, two synonymous, and two nonsynonymous variants. We examined the consequences of mutations in SOHLH1 using in vivo and in vitro assays. Analysis of transcripts from minigenes carrying the c.346-1G>A revealed that splicing site variation leads to the partial deletion at a cryptic splicing site within exon 4. This deletion results in SOHLH1 with a truncated bHLH domain. Transient transfection assay showed that the SOHLH1 mutant with the truncated domain disrupted the transcriptional activity of KIT promoter, whereas two missense mutations harboring either p.Arg37Gln or p.Pro269Ser did not have a significant effect on its transactivation. Our findings indicate that a splice-acceptor site mutation that probably causes a nonfunctional SOHLH1 protein results in nonobstructive azoospermia by the lack of normal spermatogenesis.

Development of a Comprehensive Food Literacy Measurement Tool Integrating the Food System and Sustainability.

The concept of food literacy is evolving and expanding, increasing the need for a comprehensive measurement tool for food literacy. This study aimed to develop a validated food literacy questionnaire based on an expanded conceptual framework for food literacy. A literature review of existing frameworks and questionnaires for food literacy and focus group interviews (n = 12) were conducted to develop a conceptual framework and candidate questions. A Delphi study (n = 15) and pilot survey (n = 10) to test the preliminary questionnaire's content and face validity were conducted, which were followed by the main survey (n = 200). Construct validity and reliability were assessed using exploratory factor analysis (EFA) and Cronbach's alpha, respectively. Criterion validity was assessed by comparing food literacy scores with food knowledge scores (FN-score) and nutrient quotient scores (NQ-score). By integrating and revising the six existing conceptual frameworks and focus group interview results, we proposed a two-dimensional conceptual framework comprising a literacy dimension and a food system dimension. After reviewing 560 items and categorizing them into 18 domains (3 in the literacy dimension × 6 in the food system dimension), 32 questions were selected. As a result of the Delphi study, two items were deleted, and content validity was confirmed for the remaining 30 items (content validity ratio (CVR) = 0.92). Ten items were revised during the face validation process, and five items were excluded as a result of the EFA. The final food literacy questionnaire comprised 25 questions related to five factors: production, selection, preparation and cooking, intake, and disposal. Food literacy scores were positively associated with FN- and NQ-scores, confirming the reliability and criterion validity of the final questionnaire. The two-dimensional food literacy conceptual framework developed in this study systematically encompasses complex food literacy concepts by adding a food systems dimension (production, selection, preparation and cooking, intake, and disposal domain) to the existing literacy dimension (functional, interactive, and critical literacy domain). Based on this integrated conceptual framework, a 25-item food literacy questionnaire was developed and validated for practical use.

Implication of the Association of Fibrinogen Citrullination and Osteoclastogenesis in Bone Destruction in Rheumatoid Arthritis.

Immune complexes containing citrullinated fibrinogen are present in the sera and synovium of rheumatoid arthritis patients and potentially contribute to synovitis. However, fibrinogen can inhibit the osteoclastogenesis of precursor cells. We investigated the direct effect of citrullinated fibrinogen on osteoclastogenesis to understand the role of citrullination on bone erosion of rheumatoid arthritis patients. We evaluated the fibrinogen citrullination sites using mass spectrometry and quantified osteoclast-related protein and gene expression levels by Western blotting, microarray, and real-time polymerase chain reaction. Differences in spectral peaks were noted between fibrinogen and citrullinated fibrinogen at five sites in α-chains, two sites in ß-chains, and one site in a γ-chain. Transcriptome changes induced by fibrinogen and citrullinated fibrinogen were identified and differentially expressed genes grouped into three distinctive modules. Fibrinogen was then citrullinated in vitro using peptidylarginine deiminase. When increasing doses of soluble fibrinogen and citrullinated fibrinogen were applied to human CD14+ monocytes, citrullination restored osteoclastogenesis-associated changes, including NF-ATc1 and ß3-integrin. Finally, citrullination rescued the number of osteoclasts by restoring fibrinogen-induced suppression of osteoclastogenesis. Taken together, the results indicate that the inhibitory function of fibrinogen on osteoclastogenesis is reversed by citrullination and suggest that citrullinated fibrinogen may contribute to erosive bone destruction in rheumatoid arthritis.

How does culture influence the degree of romantic loneliness and closeness?

A culture promoting a strong desire for romantic relationships can greatly influence feelings of romantic loneliness and of closeness. In this study, the authors hypothesized that when not in a romantic relationship, U.S. young adults experience greater degrees of romantic loneliness because of a high desire for romantic relationships, compared with Korean young adults. The authors also predicted that when in a romantic relationship, U.S. young adults experience greater closeness to their romantic partner than do Korean young adults. Results revealed that in a sample of 227 U.S. and Korean students, U.S. students reported significantly higher levels of romantic loneliness than did Koreans when not in a romantic relationship and significantly lower levels of romantic loneliness when in a stable romantic relationship. U.S. students also reported a greater degree of closeness in romantic relationships than did Korean students. The results suggest that Western cultures' strong emphasis on the importance of romantic relationships may unduly amplify individuals' levels of loneliness.

Scriptaid improves the reprogramming of donor cells and enhances canine-porcine interspecies embryo development.

Histone methylation, histone acetylation, and DNA methylation are the important factors for somatic cell nuclear transfer (SCNT). Histone deacetylase inhibitors (HDACi) and DNA methyltransferase inhibitors (DNMTi) have been used to improve cloning efficiency. In particular, scriptaid, an HDACi, has been shown to improve SCNT efficiency. However, no studies have been performed on canines. Here, we evaluated the effects of scriptaid on histone modification in canine ear fibroblasts (cEFs) and cloned canine embryos derived from cEFs. The early development of cloned canine-porcine interspecies SCNT (iSCNT) embryos was also examined. cEFs were treated with scriptaid (0, 100, 250, 500, 750, and 1000nM) in a medium for 24h. Scriptaid treatment (all concentrations) did not significantly affect cell apoptosis. Treatment with 500nM scriptaid caused a significant increase in the acetylation of H3K9, H3K14, and H4K5. cEFs treated with 500nM scriptaid showed significantly decreased Gcn5, Hat1, Hdac6, and Bcl2 and increased Oct4 and Sox2 expression levels. After SCNT with canine oocytes, H3K14 acetylation was significantly increased in the one- and two-cell cloned embryos from scriptaid-treated cEFs. In iSCNT, the percentage of embryos in the 16-cell stage was significantly higher in the scriptaid-treated group (21.6±2.44%) than in the control (7.5±2.09%). The expression levels of Oct4, Sox2, and Bcl2 were significantly increased in 16-cell iSCNT embryos, whereas that of Hdac6 was decreased. These results demonstrated that scriptaid affected the reprogramming of canine donor and cloned embryos, as well as early embryo development in canine-porcine iSCNT, by regulating reprogramming and apoptotic genes.

In vitro development of canine somatic cell nuclear transfer embryos in different culture media.

The objective of the present study was to investigate the effects of three different culture media on the development of canine somatic cell nuclear transfer (SCNT) embryos. Canine cloned embryos were cultured in modified synthetic oviductal fluid (mSOF), porcine zygote medium-3 (PZM-3), or G1/G2 sequential media. Our results showed that the G1/G2 media yielded significantly higher morula and blastocyst development in canine SCNT embryos (26.1% and 7.8%, respectively) compared to PZM-3 (8.5% and 0%or mSOF (2.3% and 0%) media. In conclusion, this study suggests that blastocysts can be produced more efficiently using G1/G2 media to culture canine SCNT embryos.

[Development of a scale to measure self-care for korean patients with chronic obstructive pulmonary disease].

PURPOSE: The objective of this study was the development and validation of a scale to measure the self-care of patients with chronic obstructive pulmonary disease(COPD) in Korea. METHOD: Self-care scale was developed based on the self-care activities patients had to carry out in order to manage their COPD. The original scale contained 34 items rated along a five-point Likert scale and was reviewed by 18 professional nurses and 10 Korean patients with COPD for content validity. Subsequently, patients with COPD were asked to complete this 23-item scale and further tests were done with the 125 useable responses. RESULT: Factor analysis identified eight factors- "maintaining a clean air way", "taking medication", "support from family", "preventing infection", "managing symptoms", "breathing exercising", and "taking in nutrition". The internal consistency of the total scale was Cronbach's alpha=0.7226. These eight factors explained 60.8% of total variance. There was correlation among Korean Self-Care Scale score, administration level, and knowledge level but there was no correlation to patients' satisfaction with medical services. CONCLUSION: The 23 item questionnaire positively identified 8 areas defined important for COPD patients. Further studies are required to see how these can be integrated into patient education.

[Attitudes of adolescents toward suicide: Q-methodological approach].

PURPOSE: The purpose of this study was to identify attitudes of adolescents toward suicide. METHODS: Q-methodology which provides a method of analyzing the subjectivity of each item was used. Thirty middle and high school students classified 37 selected statements into a normal distribution using a 9 point scale. Collected data were analyzed using the Quanl PC Program. RESULTS: Three types of attitudes toward suicide were identified. The first type (opposing suicide-moral minded) showed an attitude of opposing suicide and thinking that suicide is a sin. The second type (understanding-empathizing suicidal person) showed an attitude of understanding the situation of the adolescents who has suicidal ideation and empathizing with them. The third type (ambivalent attitude) showed an attitude of understanding the suicidal person but, at the same time, opposing suicide. CONCLUSION: Results of the study indicate that different approaches to suicide prevention programs should be developed based on the three types of suicide attitudes among adolescents.

Effects of grape pomace on the antioxidant defense system in diet-induced hypercholesterolemic rabbits.

The effects of grape seeds extract and grape peels extract prepared from grape pomace on the activity of antioxidant enzymes, degree of lipid peroxidation in serum and liver tissue were investigated in rabbits fed on high cholesterol diet. New Zealand white rabbits were divided as follows ; 1) NOR (normal group); 2) CHOL (cholesterol group); 3) GSH (cholesterol + grape seed extract group); 4) GPE (cholesterol + grape peel extract); 5) GSP (cholesterol + grape seed powder); 6) GPP (cholesterol + grape peel powder); 7) GE (cholesterol + grape seed and peel extract); 8) GP (cholesterol + grape seed and peel powder). Eight groups of rabbits were studied for 8 weeks. At the end of the experimental period, rabbits were sacrificed and the liver tissue were removed. Then, GSH, GPx, GST, CAT and MDA in the liver were measured. In liver tissues, total glutathione contents (GSH), glutathione peroxidase (GPx) and catalase (CAT) activity, which was significantly higher by grape seed extract supplementation. The level of malondialdehyde (MDA) was lower in the serum of rabbits fed grape seed extract or grape peel powder plus cholesterol than in the serum of rabbits fed cholesterol alone. It is therefore likely that grape seed extract prepared from grape pomace functioned as antioxidants in vivo, negating the effects of the oxidative stress induced by 1% cholesterol diet. The grape seed extract was found effective in converting the oxidized glutathione into reduced glutathione, and in removing H(2)O(2) that is created by oxidative stress. The grape peel powder was found to have small influence on reduced glutathione content, CAT and GPX activity, but it increased GST activity in liver tissues, resulting in promoting the combination of lipid peroxide and glutathione (GSH), and further, lowering the formation of lipid peroxide in the serum. Therefore, grape pomace (grape seed extract and grape peel powder) supplementation is considered to activate the antioxidant enzyme system and prevent damage with hypercholesterolemia.

The oocyte-specific transcription factor, Nobox, regulates the expression of Pad6, a peptidylarginine deiminase in the oocyte.

Nobox is an oocyte-specific transcriptional regulator. Nobox deficiency disrupts early folliculogenesis and the expression of oocyte-specific genes in mice. In the present study, we found that peptidylarginine deiminase 6 (Pad6) was downregulated in Nobox-null ovaries. Pad6 is preferentially expressed in oocytes and its transcript is detectable at embryonic day 16.5. In addition, we identified one Nobox DNA-binding element (NBE) within the mouse Pad6 promoter. The NBE includes a core sequence TAATTA. Sequence-specific binding of Nobox to the TAATTA motif was confirmed. Nobox overexpression augmented transcriptional activity of a luciferase reporter driven by mouse Pad6. Our findings indicate that Nobox is a critical regulator that orchestrates oocyte-specific genes such as Pad6 during folliculogenesis.