Therapeutic potential of AAV-FL-Klotho in obesity: Impact on weight loss and lipid metabolism in mice.

Klotho, an anti-aging protein, has gained attention for its protective effects against various diseases, including metabolic disorders, through recombinant Klotho administration. However, the potential of Klotho as a target for gene therapy requires further exploration, as it remains relatively understudied in the context of metabolic disorders. In this study, we demonstrate that AAV-full length(FL)-Klotho administration induces weight loss in mice and provides protection against high-fat diet (HFD)-induced obesity and hepatic steatosis, concurrently reducing the weights of white adipose tissue and liver. AAV-FL-Klotho administration also enhanced thermogenic gene expression in brown adipose tissue (BAT) and improved the morphology of interscapular BAT. The weight loss effect of AAV-FL-Klotho was found to be, at least in part, mediated by UCP1-dependent thermogenesis in brown adipocytes, potentially influenced by hepatokines secreted from AAV-FL-Klotho-transduced hepatocytes. These findings suggest that AAV-FL-Klotho is an attractive candidate for gene therapy to combat obesity. Nevertheless, unbiased experiments have also revealed disturbances in lipid metabolism due to AAV-FL-Klotho, as evidenced by the emergence of lipomas and increased expression of hepatic lipogenic proteins.

A pontomesencephalic PACAPergic pathway underlying panic-like behavioral and somatic symptoms in mice.

Panic disorder is characterized by uncontrollable fear accompanied by somatic symptoms that distinguish it from other anxiety disorders. Neural mechanisms underlying these unique symptoms are not completely understood. Here, we report that the pituitary adenylate cyclase-activating polypeptide (PACAP)-expressing neurons in the lateral parabrachial nucleus projecting to the dorsal raphe are crucial for panic-like behavioral and physiological alterations. These neurons are activated by panicogenic stimuli but inhibited in conditioned fear and anxiogenic conditions. Activating these neurons elicits strong defensive behaviors and rapid cardiorespiratory increase without creating aversive memory, whereas inhibiting them attenuates panic-associated symptoms. Chemogenetic or pharmacological inhibition of downstream PACAP receptor-expressing dorsal raphe neurons abolishes panic-like symptoms. The pontomesencephalic PACAPergic pathway is therefore a likely mediator of panicogenesis, and may be a promising therapeutic target for treating panic disorder.

NG ,NG -Dimethylarginine Dimethylaminohydrolase 1 Expression Is Dispensable for Cold- or Diet-Induced Thermogenesis.

The strategy to activate thermogenic adipocytes has therapeutic potential to overcome obesity as they dissipate surplus energy as heat through various mechanisms. NG,NG-dimethylarginine dimethylaminohydrolases (DDAHs) are enzymes involved in the nitric oxide-protein kinase G signaling axis which increases thermogenic gene expression. However, the role of DDAHs in thermogenic adipocytes has not been elucidated. The adipocyte-specific Ddah1 knockout mice are generated by crossing Ddah1fl/fl mice with adiponectin Cre recombinase mice. Adipocyte-specific DDAH1 overexpressing mice are generated using adeno-associated virus-double-floxed inverse open reading frame (AAV-DIO) system. These mice are analyzed under basal, cold exposure, or high-fat diet (HFD) conditions. Primary inguinal white adipose tissue cells from adipocyte-specific Ddah1 knockout mice expressed comparable amounts of Ucp1 mRNA. Adipocyte-specific DDAH1 overexpressing mice do not exhibit enhanced activation of thermogenic adipocytes. In addition, when these mice are exposed to cold environment or fed an HFD, their body temperature/weight and thermogenesis-related gene and protein expressions are unchanged. These findings indicate that DDAH1 does not play a role in either cold- or diet-induced thermogenesis. Therefore, adipocyte targeting DDAH1 gene therapy for the treatment of obesity is unlikely to be effective.

Genome-wide CRISPR/Cas9 knockout screening to mitigate cell growth inhibition induced by histone deacetylase inhibitors in recombinant CHO cells.

Histone deacetylase inhibitors (iHDACs) have been extensively studied as enhancers of therapeutic protein production in recombinant Chinese hamster ovary (CHO) (rCHO) cell cultures. However, the addition of iHDACs reduces the viable cell concentration (VCC) in rCHO cell cultures, thereby reducing their potential to enhance therapeutic protein production. To mitigate the negative effects of iHDACs on VCC, screening using a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-based single-gene knockout (KO) library in rCHO cells was performed in the presence of CI994, a member of iHDACs, and 10 potential KO genes that enhanced the VCC of CI994-treated rCHO cells were identified. Among these, Bcor was validated as a promising KO target that improved VCC without negatively affecting the specific productivity in the presence of CI994. Bcor KO increased the VCC and therapeutic protein concentrations in both batch and fed-batch cultures in the presence of CI994. Taken together, these findings highlight the potential of the whole-genome CRISPR/Cas9-based single-gene KO cell library to identify KO target genes for the development of iHDAC-resistant rCHO cells for enhanced therapeutic protein production.

Do expressive writing interventions have positive effects on Koreans?: a meta-analysis.

Introduction: Expressive Writing (EW) is an intervention that focuses on individuals' writing down their thoughts and feelings about trauma or stressful events. Meta-analyses on EW studies have confirmed that EW has a positive effect. However, the heterogeneity of studies is high, so many studies have investigated boundary conditions and moderators. One of these moderators is the cultural difference in emotional suppression. Since EW focuses on the expression of suppressed thoughts and emotions, its effect might be slightly different for people in Asian cultures who show a high tendency to suppress their emotions. This study attempted to confirm the effect size of the EW interventions in Korea and examine whether these studies have different effect size from those based on Western cultures. Method: A total of 29 studies published in Korea until 2021 were analyzed. The effect size was calculated using the "dmetar," "meta," and "metafor" packages of the statistical program R 4.0.4. Results: The results were as follows. First, the effect size of EW intervention was 0.16, and we found that studies in the Korean context showed no significant difference from studies based on western meta-analysis. Second, the moderating variables that influenced the EW intervention were the writing type, the number of sessions, the time per session, and the measurement time. Discussion: The results of this study suggest that EW interventions benefit Koreans. And it is at least harmless and has a positive effect considering the efficiency and conciseness of interventions. Furthermore, the finding shows that EW interventions can be helpful even in the general population without apparent psychological problems. By considering moderators, we could structure more effective form of EW interventions for Koreans.

Comparative analysis of infertility healthcare utilization before and after insurance coverage of assisted reproductive technology: A cross-sectional study using National Patient Sample data.

This study aims to analyze the types and cost of infertility care provided in a clinical setting to examine the changes of healthcare utilization for infertility after the 2017 launch of assisted reproductive technology (ART) health insurance coverage in South Korea. Health Insurance Review Assessment-National Patient Sample data from 2016 and 2018 were analyzed comparatively. Data related to receiving medical service under the International Classification of Diseases 10th revision code N97 (female infertility) or N46 (male infertility) at least once were analyzed, including patients' characteristics and healthcare utilization (type of healthcare facility and treatment approach). Between 2016 and 2018, the percentage of patients aged 30-34 receiving infertility care dropped; the percentages of patients in older age groups increased. The number of female patients remained comparable, whereas the number of male patients increased by 23%. Average visits per patient increased by about 1 day from 2016 to 2018. Total annual infertility care claim cost increased from $665,391.05 to $3,214,219.48; the per-patient annual cost increased from $114.76 to $522.38. The number of claims and cost of treatment and surgery increased markedly, as did the number of claims and cost of gonadotropins. With its focus on health insurance coverage of ART and results demonstrating increases in medical services, medications, cost, and patient utilization, this study reveals the significant effects of national health policies on the treatment, cost, and management of infertility.

AAV-mediated skeletal muscle specific irisin expression does not contribute to weight loss in mice.

Obesity, a chronic disease, significantly increases the risk of various diseases, including diabetes, cardiovascular diseases, and cancers. Exercise is crucial for weight management not only through energy expenditure by muscle activity but also through stimulating the secretion of myokines, which affect various tissues. Irisin, derived from the proteolytic processing of fibronectin type III domain-containing protein 5 (Fndc5), is a well-studied myokine with beneficial effects on metabolism. This study explored the feasibility of adeno-associated virus (AAV)-mediated Fndc5 gene therapy to treat obesity in a mouse model using the AAV-DIO system to express Fndc5 specifically in skeletal muscle, and investigated its anti-obesity effect. Although Fndc5 was specifically expressed in the muscle, no significant impact on body weight under normal chow or high-fat diets was observed, and no change in thermogenic gene expression in inguinal white adipose tissue was detected. Notably, Fndc5 transduction did affect bone metabolism, consistent with previous reports. These findings suggest that AAV-mediated Fndc5 gene therapy may not be an efficient strategy for obesity, contrary to our expectations. Further research is needed to elucidate the complex mechanisms involved in irisin's role in obesity and related disorders.

Utilization of traditional herbal medicine formulas for unexplained female infertility in Korea: a retrospective study.

BACKGROUND: This study aimed to investigate the prescription of traditional herbal medicines for the treatment of unexplained female infertility in Korea. By analyzing the relationships among the prescriptions and between the prescriptions and treatment outcomes, we aimed to confirm the utilization of standardized prescriptions and the effectiveness of this standardization. METHODS: The data were derived from the "support projects" of the local government for infertile women to receive Korean medical treatments. The presciption data of 453 participants from 2017 to 2018 were analyzed. Data preprocessing, frequency analysis, and network analysis were conducted. For network analysis, the characteristics of the herbal medicine prescriptions were used to calculate the degree centrality, closeness centrality and eigenvector centrality. Modularity clustering was also performed for cluster analysis. RESULTS: Fifty different prescriptions were used, among which only 22 were used for participants who became pregnant. The recommended standard prescriptions for support projects were used frequently. "BaeranChacksangBang" showed the highest level of in-degree centrality. Among the prescriptions for participants who became pregnant, "JogyeongJongokTang" and "BaeranChacksangBang" were the most influential nodes. "BaeranChacksangBang", "AnjeonYicheonTang", "YukLinZu" and "JogyeongJongokTang" had high closeness centrality among the prescriptions for participants who became pregnant. Clustering analysis of the prescriptions for participants who became pregnant revealed that the prescriptions showed the best modularity when divided into five groups. CONCLUSIONS: These findings depict the utilization of Korean herbal medicine in the real world and the dynamics underlying the herbal medicine prescription patterns for infertile women.

Mitigating transcriptional bottleneck using a constitutively active transcription factor, VP16-CREB, in mammalian cells.

High-level expression of recombinant proteins in mammalian cells has long been an area of interest. Inefficient transcription machinery is often an obstacle in achieving high-level expression of recombinant proteins in mammalian cells. Synthetic promoters have been developed to improve the transcription efficiency, but have achieved limited success due to the limited availability of transcription factors (TFs). Here, we present a TF-engineering approach to mitigate the transcriptional bottlenecks of recombinant proteins. This includes: (i) identification of cAMP response element binding protein (CREB) as a candidate TF by searching for TFs enriched in the cytomegalovirus (CMV) promoter-driven high-producing recombinant Chinese hamster ovary (rCHO) cell lines via transcriptome analysis, (ii) confirmation of transcriptional limitation of active CREB in rCHO cell lines, and (iii) direct activation of the transgene promoter by expressing constitutively active CREB at non-cytotoxic levels in rCHO cell lines. With the expression of constitutively active VP16-CREB, the production of therapeutic proteins, such as monoclonal antibody and etanercept, in CMV promoter-driven rCHO cell lines was increased up to 3.9-fold. VP16-CREB was also used successfully with synthetic promoters containing cAMP response elements. Taken together, this strategy to introduce constitutively active TFs into cells is a useful means of overcoming the transcriptional limitations in recombinant mammalian cells.

Cold-induced adaptive thermogenesis is impaired by exposure of Asian sand dust in mice.

Desertification and desert sandstorms caused by the worsening global warming pose increasing risks to human health. In particular, Asian sand dust (ASD) exposure has been related to an increase in mortality and hospital admissions for respiratory diseases. In this study, we investigated the effects of ASD on metabolic tissues in comparison to diesel particulate matter (DPM) that is known to cause adverse health effects. We found that larger lipid droplets were accumulated in the brown adipose tissues (BAT) of ASD-administered but not DPM-administered mice. Thermogenic gene expression was decreased in these mice as well. When ASD-administered mice were exposed to the cold, they failed to maintain their body temperature, suggesting that the ASD administration had led to impairments in cold-induced adaptive thermogenesis. However, impaired thermogenesis was not observed in DPM-administered mice. Furthermore, mice fed a high-fat diet that were chronically administered ASD demonstrated unexplained weight loss, indicating that chronic administration of ASD could be lethal in obese mice. We further identified that ASD-induced lung inflammation was not exacerbated in uncoupling protein 1 knockout mice, whose thermogenic capacity is impaired. Collectively, ASD exposure can impair cold-induced adaptive thermogenic responses in mice and increase the risk of mortality in obese mice.

Chilly climate perceived by female engineering undergraduates: an exploratory study using concept mapping.

Introduction: Women still being a minority in engineering majors, they are reported to face discriminatory treatment in a collegiate environment. "Chilly climate," referring to such a sexist environment, may have a negative impact on women's mental health, academics, and careers. But, what exactly is it that female students in engineering perceive as chilly, and how chilly is it? This study aimed to explore the chilly campus climate perceived by female undergraduate engineering students in South Korea using the concept mapping method. Methods: Semi-structured interviews were conducted with 13 participants enrolled for more than four semesters at four-year coeducational universities. After extracting 52 representative statements, the participants were asked to classify them according to content similarity and rate the influence of each statement on their perception of the chilly climate. For concept mapping analysis, multidimensional scaling analysis (ALSCAL), hierarchical cluster analysis (Ward's method), and non-hierarchical cluster analysis (K-means method) were performed. Results: Fifty-two statements were extracted under the following four clusters: (i) "Exclusion and alienation inherent in the culture (Cluster 1)," (ii) "Sexual objectification and lack of gender sensitivity (Cluster 2)," (iii) "Male-centered academic situations (Cluster 3)," and (iv) "Prejudice and generalization (Cluster 4)." A concept map was two-dimensional: an X-axis named "context dimension," with "task: academic" and "non-task: social" at both ends, and a Y-axis named "sexism dimension", having "explicit" and "implicit" at both ends. The order of higher scores in the influence rating is as follows: Cluster 2, Cluster 3, Cluster 1, and Cluster 4. Discussion: This study is significant because it conceptualizes the subjective experience of minorities in a collegiate environment and provides influence rating results for prioritized measures. The findings will be helpful in formulating educational policies, psychological counseling, and social advocacy activities. Future research should target larger populations, and cover more diverse cultures, majors, and age groups.

The relationship between faculty interactions, sense of belonging, and academic stress: a comparative study of the post-COVID-19 college life of Korean and international graduate students in South Korea.

Objective: Rapid changes in post-COVID-19 higher education have increased students' academic stress. This study focused on graduate students' academic stress in South Korea and compared the results for Korean graduate students and those for international graduate students. Method: Using the online survey results, the study verified the relationships between faculty interactions, a sense of belonging, and academic stress among Korean and international graduate students using a mediating effects analysis and a multigroup path analysis. Results: The results were as follows. First, Korean students experienced greater academic stress, faculty interactions, and a sense of belonging, but no statistically significant difference was observed. Second, a sense of belonging had a mediating effect on the relationship between faculty interactions and academic stress. Unlike in previous studies, all paths were found to be statistically significant. Faculty interactions had a negative effect on academic stress and a positive effect on a sense of belonging. A sense of belonging had a negative effect on academic stress. Third, the comparison of Korean and international graduate students showed that international students had a greater effect of faculty interactions on academic stress. Conclusion: Through these results, we explored the post-COVID-19 academic lives of Korean and international graduate students in South Korea and built grounds for effective interventions for alleviating academic stress.

A representation of students with intellectual disabilities in South Korean online newspaper articles using keyword network analysis.

The purpose of this study was to analyse online newspaper articles on students with intellectual disabilities (IDs) in order to identify related social phenomena to derive implications for inclusive education. Such study has traditionally practised through content analysis and/or discourse analysis manually, which is prone to subjective interpretation. Thus, this study implemented automated analysis to objectively select and interpret a big data. A total of 8,890 online newspaper articles that were published from 1990 to April 2019 were collected through automated parsing. The entire period and decade-phase based keyword and keyword network analysis were practised in order to determine how the social perceptions and related issues had changed over time. The results indicated that there was a rapid growth in scope of articles on students with IDs over the past 30 years. The attention of media gradually expanded from special education to improving quality of lives of students with IDs and their families. Moreover, online newspaper articles seemed to focus on social controversies and incidents such as sexual assaults that are related to students with IDs. Based on the results, ways to support inclusive education as well as social inclusion of students with IDs were discussed.

Pathways to post-traumatic growth in Korean female cancer patients: the mediation effects of coping strategies and resilience.

ABSTRACTBackground: Cancer is a life-threatening disease in Korean women. Female cancer patients in Korea have reported poor physical functioning and psychological distress after diagnosis. However, some patients experience post-traumatic growth (PTG), which includes positive feelings and changes.Objective: The main purpose of this study was to examine the relationships between PTG and its associated factors in Korean female cancer patients to understand how female cancer patients achieve PTG while experiencing cancer diagnosis and treatment. This study aimed to improve the quality of life of many Korean female patients with cancer.Methods: This study investigated the structural model of the paths from the disruption of core beliefs (CBD), coping strategies, and resilience to PTG in a sample of female cancer patients. In total, 164 middle-aged women diagnosed with cancer were included in the final sample.Results: First, it was determined that the proposed structural model was substantial and had high fit indices. Second, problem-solving was positively associated by the CBD routes. Third, it was also favorably expected that problem-solving would lead to resilience and resilience would lead to PTG. These findings are important for developing future interventions for Korean female cancer patients and can be considered as an important variable to improve their PTG.

The proposed structural model observed paths of how female cancer patients achieve post-traumatic growth while experiencing cancer diagnosis and treatment.Disruption of core beliefs as a factor positively associates problem-solving in Korean female cancer patients.Problem-solving and resilience as factors positively associate post-traumatic growth in Korean female cancer patients.

Development of a model of rehabilitation exercise and sports service delivery system for health promotion of people with disabilities.

People with disabilities (PWD) suffer from chronic diseases and other problems due to their low physical activity compared to people without disabilities. Physical activity and exercise can prevent chronic diseases and improve health management. However, PWD do not receive proper rehabilitation exercise and sports services immediately after dicharge from hospitals. An effective model of rehabilitation exercise and sports service delivery system has not been established in Korea. This study aimed to present such a model for health promotion, which enables PWD to participate in exercise regularly in local communities. The model is presented by dividing it into sides of consumers and suppliers by analyzing domestic and foreign rehabilitation and sports service systems and expert meetings. The system presented in this study can help promote returning to normal social life and transition into sports in daily life for PWD. In addition, it is expected to positively affect local communities through training instructors in rehabilitation exercise and sports; it can serve as a bridge between rehabilitation in medical institutes and sports in daily life.

De Novo Transcriptome Assembly and Comparative Analysis of Differentially Expressed Genes Involved in Cold Acclimation and Freezing Tolerance of the Arctic Moss Aulacomnium turgidum (Wahlenb.) Schwaegr.

Cold acclimation refers to a phenomenon in which plants become more tolerant to freezing after exposure to non-lethal low temperatures. Aulacomnium turgidum (Wahlenb.) Schwaegr is a moss found in the Arctic that can be used to study the freezing tolerance of bryophytes. To improve our understanding of the cold acclimation effect on the freezing tolerance of A. turgidum, we compared the electrolyte leakage of protonema grown at 25 °C (non-acclimation; NA) and at 4 °C (cold acclimation; CA). Freezing damage was significantly lower in CA plants frozen at -12 °C (CA-12) than in NA plants frozen at -12 °C (NA-12). During recovery at 25 °C, CA-12 demonstrated a more rapid and greater level of the maximum photochemical efficiency of photosystem II than NA-12, indicating a greater recovery capacity for CA-12 compared to NA-12. For the comparative analysis of the transcriptome between NA-12 and CA-12, six cDNA libraries were constructed in triplicate, and RNA-seq reads were assembled into 45,796 unigenes. The differential gene expression analysis showed that a significant number of AP2 transcription factor genes and pentatricopeptide repeat protein-coding genes related to abiotic stress and the sugar metabolism pathway were upregulated in CA-12. Furthermore, starch and maltose concentrations increased in CA-12, suggesting that cold acclimation increases freezing tolerance and protects photosynthetic efficiency through the accumulation of starch and maltose in A. turgidum. A de novo assembled transcriptome can be used to explore genetic sources in non-model organisms.

Novel genetically encoded tools for imaging or silencing neuropeptide release from presynaptic terminals in vivo.

Neurons produce and release neuropeptides to communicate with one another. Despite their profound impact on critical brain functions, circuit-based mechanisms of peptidergic transmission are poorly understood, primarily due to the lack of tools for monitoring and manipulating neuropeptide release in vivo. Here, we report the development of two genetically encoded tools for investigating peptidergic transmission in behaving mice: a genetically encoded large dense core vesicle (LDCV) sensor that detects the neuropeptides release presynaptically, and a genetically encoded silencer that specifically degrades neuropeptides inside the LDCV. Monitoring and silencing peptidergic and glutamatergic transmissions from presynaptic terminals using our newly developed tools and existing genetic tools, respectively, reveal that neuropeptides, not glutamate, are the primary transmitter in encoding unconditioned stimulus during Pavlovian threat learning. These results show that our sensor and silencer for peptidergic transmission are reliable tools to investigate neuropeptidergic systems in awake behaving animals.

Results From South Korean 2022 Para Report Card on Physical Activity for Children and Adolescents With Disabilities.

South Korea has developed its first Para Report Card on physical activity (PA) for children and adolescents with disabilities. Five national surveillance databases were used to evaluate PA indicators based on the benchmarks and grading rubric provided by Active Healthy Kids Global Alliance. Report card evaluation committees were invited to grade and assess the results using strengths, weaknesses, opportunities, and threats analysis. Five indicators (overall PA, D+; organized sports and PA, D-; active transportation, D-; physical fitness, D+; and government, A+) and one additional indicator (sleep, C-) were assigned a letter grade. The other five indicators were graded as incomplete. The Para Report Card revealed a significant gap between the behavioral-indicator grades (D- to D+) and the policy-indicator grade (A+), suggesting that government strategies and investment have not yet been translated into behavioral PA among children and adolescents with disabilities.

A retrospective study of factors affecting contralateral central-neck lymph node metastasis in unilateral papillary thyroid carcinoma.

BACKGROUND: In 30-40% of papillary thyroid cancer patients, central neck lymph node metastasis occurs. As a result, prophylactic central neck lymph node dissection is performed. The extent of lymph node dissection and prophylactic central neck lymph node dissection is still debatable. The incidence of central neck lymph node metastasis and related factors were investigated in this study, and also the necessity of both central neck lymph node dissections. METHODS: Between December 2017 and December 2019, 482 patients had thyroidectomy at Pusan National University Yangsan Hospital. A retrospective study of 186 patients who had a thyroidectomy with bilateral central neck lymph node dissection for unilateral thyroid carcinoma was done. RESULTS: Ipsilateral and contralateral central neck lymph node metastasis were identified in 40.9% (76/186) and 19.3% (36/186), respectively. Male (p < 0.001), tumor size >1 cm (p = 0.047), extrathyroidal extension (p = 0.002), central neck lymph node metastases >5 (p < 0.001), lateral neck lymph node metastasis (p = 0.012), and ipsilateral central neck lymph node metastasis (p < 0.001) were associated with the contralateral central neck lymph node metastasis in univariate analysis. In a multivariate analysis, extrathyroidal extension (OR, 3.664), more than 5 central neck lymph node metastases (OR, 29.667), ipsilateral central neck lymph node metastasis (OR, 3.911), and male (OR, 5.890) were related to contralateral central neck lymph node metastasis. CONCLUSION: Male, extrathyroidal extension, and ipsilateral central neck lymph node metastasis may be considered for contralateral prophylactic central neck lymph node dissection. In the future, it is thought that more research on the recurrence rate will be required.