Association between frailty and cognitive status among ambulating Korean elderly: An observational study.

We aimed to determine the association between frailty and cognitive status of the elderly population in Korea. We examined data from 9920 elders who participated in the 2020 Survey of Living Conditions and Welfare Needs of Korean Older Persons. Frailty was assessed using the Korean version of the Fatigue, Resistance, Ambulation, Illnesses, and Weight Loss scale. The Korean mini-mental status examination was used to test cognitive function. Several logistic regression analysis was performed, with correction for several confounding variables (socioeconomic, health behavior, psychological characteristics, and functional status), to evaluate the relationship between frailty and cognitive state. Of the elderly population in Korea, 1451 (14.6%) were frail and 5977 (60.3%) were pre-frail. Compared to the non-frail group (20.3%), cognitive impairment was considerably higher in the pre-frail (33.1%) and frail (39.8%) groups. When compared to the non-frail group, cognitive impairment was substantially linked to a higher risk of frailty after adjustment (pre-frail odds ratio [OR]: 1.66, 95% confidence interval [CI]: 1.47-1.88; frail OR: 2.00, 95% CI: 1.68-2.37). When cognitive impairment and frailty subcomponents were present, there was a higher likelihood of severe resistance (OR: 1.89; 95% CI: 1.70-2.11) and ambulation (OR: 1.46, 95% CI: 1.32-1.63) issues. Frailty is associated with cognitive impairment.

Effects of stress on burnout among infection control nurses during the COVID-19 pandemic: the mediating effects of social support and self-efficacy.

BACKGROUND: This study investigated the mediating effects of self-efficacy and social support on the relationship between stress and burnout among infection control nurses (ICNs) during an emerging infectious disease pandemic. METHODS: The study participants encompassed 210 ICNs with at least six months' experience in an infection control unit at a general hospital in South Korea during the COVID-19 pandemic. Data were analyzed using independent t-tests or one-way analysis of variance (ANOVA), while descriptive statistics were performed using SPSS/WIN 26.0 software. Hayes's PROCESS macro 4.2 software was used to verify the significance of the indirect effects of the mediators. RESULTS: Stress had a significant positive effect on burnout (ß = 0.80, p < .001), accounting for 73% of the variance. Self-efficacy (ß = - 0.26, p < .001) and social support (ß = - 0.11, p = .034) had a significant negative effect on burnout, accounting for 78% of the variance. Stress was lower when self-efficacy and social support were entered into the model (ß = 0.80 → 0.59), indicating that self-efficacy and social support mediated the relationship between stress and burnout. CONCLUSION: This study is significant in that it confirms the effects of self-efficacy and social support on the relationship between stress and burnout among ICNs. The results highlight the importance of establishing organizational support systems and developing and implementing programs for enhancing self-efficacy in order to reduce burnout among ICNs.

Enhanced Electrical Properties and Stability in IGZO TFTs via Low-Temperature Activation with MgOx Layer.

We propose the introduction of a magnesium oxide (MgOx) layer to reduce the temperature required for the activation of indium gallium zinc oxide (IGZO) thin films. By incorporating the MgOx layer between the IGZO channel layer and the gate insulator layer, the required activation temperature is lowered from 300 to 200 °C while enhancing the electrical performance of the IGZO thin-film transistor (TFT). Specifically, the field effect mobility is improved from 6.40 to 16.12 cm2/(V s), the on/off current ratio is enhanced from 1.62 × 109 to 7.16 × 109, and subthreshold swing is enhanced from 0.48 to 0.46 V/decade. Furthermore, IGZO TFTs with the MgOx layer exhibit enhancements in threshold voltage (VTH) shift compared to TFTs without the MgOx layer under positive bias stress (VGS = 20 V and VDS = 0.1 V for 10,000 s) and negative bias stress (VGS = -20 V and VDS = 0.1 V for 10,000 s): the VTH shifts are decreased from 2.40 to 1.72 V and from 0.56 to 0.53 V, respectively. These enhancements are verified through various analyses and are attributed to the diffusion of Mg atoms into the IGZO front channel during the low-temperature activation process, which results in the formation of Mg-doped IGZO between the MgOx and IGZO channel layers.

Association between Circadian Rhythm-Disturbing Factors and Metabolic Syndrome in Korean Adults: Korea National Health and Nutrition Examination Survey (2016-2020).

Background: Circadian misalignment is associated with metabolic syndrome. This study aimed to examine the association between circadian rhythm-disturbing factors and metabolic syndrome. Methods: We used data from the 7th and 8th Korea National Health and Nutrition Examination Survey conducted between 2016 and 2020, which surveyed 16,253 individuals. Circadian rhythm-disturbing factors were defined as follows: sleep duration outside the reference group (6-8 hours), irregular breakfast, shift work, and physical inactivity. The adjusted odds ratio (aOR) for metabolic syndrome was calculated based on the number of circadian rhythm-disturbing factors present in adults over the age of 19 years. Results: Among a total of 16,253 participants (mean age 48.2±15 years), metabolic syndrome was found in 5,237 participants (29.3 %). The participants were classified into three categories based on the number of circadian rhythm-disturbing factors as follows: 2,627 (15.6%) did not have any factors, 6,406 (38.13%) had one factor, and 7,220 (46.3%) had two or more factors. Participants with a single circadian rhythm-disturbing factor were 21% more likely to have metabolic syndrome (aOR, 1.21; 95% confidence interval [CI], 1.08-1.36), and participants with two or more factors were 27% more likely to have metabolic syndrome (aOR, 1.27; 95% CI, 1.12-1.43). Conclusion: Circadian rhythm-disturbing factors were significantly associated with the prevalence of metabolic syndrome in Korean adults. This finding has potential clinical implications for maintaining circadian rhythms by avoiding certain factors to prevent metabolic syndrome. Further studies are required to confirm these findings.

Interaction between healthcare providers and parents of children or adolescents with epilepsy: A qualitative systematic review and meta-synthesis.

AIMS: This study aimed to explore the interactions between healthcare providers and parents of children or adolescents with epilepsy. A qualitative systematic review based on the theory-generating meta-synthesis research approach proposed by Finfgeld-Connett (2018) was applied. MATERIALS AND METHODS: We searched for empirical qualitative studies in five electronic databases (PubMed, Embase, CINAHL, Cochrane Library, and Web of Science), from January 1, 2003 to February 9, 2023. Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were used to guide the selection process, and two researchers independently assessed the methodological quality of the articles using the Critical Appraisal Skills Programme Qualitative Studies Checklist. RESULTS: Of the 4,768 studies initially imported for screening, 27 studies were reviewed and synthesized. Only one qualitative study directly focused on the interactions between parents and healthcare providers, but various studies mentioning such interaction as themes or sub-themes of other phenomena allowed us to draw out common attributes. Defining attribute, "journey through the three stages of interaction," were derived as follows: Stage 1: trust vs. mistrust; Stage 2: autonomy vs. doubt; Stage 3: adaptation. The antecedents included encounters with healthcare providers and parent empowerment. A patient-centered approach was found to be the consequence. CONCLUSION: It is important for parents of children or adolescents with epilepsy to empower themselves and increase their interactions. Considering the stage of interaction, healthcare providers and researchers should explore strategies to promote effective communication. Further research is required to develop strategies aimed at supporting parents and healthcare providers to achieve the tasks at each stage and maintain Stage 3, "adaptation."

177Lu Anti-Angiogenic Radioimmunotherapy Targeting ATP Synthase in Gastric Cancer Model.

This study investigated a novel radioimmunotherapy strategy for targeting tumor angiogenesis. We developed a radiopharmaceutical complex by labeling an anti-adenosine triphosphate synthase (ATPS) monoclonal antibody (mAb) with the radioisotope 177Lu using DOTA as a chelating agent. 177Lu-DOTA-ATPS mAb demonstrated high labeling efficiency (99.0%) and stability in serum. MKN-45 cancer cells exhibited the highest cellular uptake, which could be specifically blocked by unlabeled ATPS mAb. In mice, 177Lu-DOTA-ATPS mAb accumulated significantly in tumors, with a tumor uptake of 16.0 ± 1.5%ID/g on day 7. 177Lu-DOTA-ATPS mAb treatment significantly reduced the viability of MKN-45 cells in a dose-dependent manner. In a xenograft tumor model, this radioimmunotherapy strategy led to substantial tumor growth inhibition (82.8%). Furthermore, combining 177Lu-DOTA-ATPS mAb with sunitinib, an anti-angiogenic drug, enhanced the therapeutic efficacy of sunitinib in the mouse model. Our study successfully developed 177Lu-DOTA-ATPS mAb, a radioimmunotherapy agent targeting tumor blood vessels. This approach demonstrates significant promise for inhibiting tumor growth, both as a single therapy and in combination with other anti-cancer drugs.

Genetic modifiers of somatic expansion and clinical phenotypes in Huntington's disease reveal shared and tissue-specific effects.

Huntington's disease (HD), due to expansion of a CAG repeat in HTT , is representative of a growing number of disorders involving somatically unstable short tandem repeats. We find that overlapping and distinct genetic modifiers of clinical landmarks and somatic expansion in blood DNA reveal an underlying complexity and cell-type specificity to the mismatch repair-related processes that influence disease timing. Differential capture of non-DNA-repair gene modifiers by multiple measures of cognitive and motor dysfunction argues additionally for cell-type specificity of pathogenic processes. Beyond trans modifiers, differential effects are also illustrated at HTT by a 5'-UTR variant that promotes somatic expansion in blood without influencing clinical HD, while, even after correcting for uninterrupted CAG length, a synonymous sequence change at the end of the CAG repeat dramatically hastens onset of motor signs without increasing somatic expansion. Our findings are directly relevant to therapeutic suppression of somatic expansion in HD and related disorders and provide a route to define the individual neuronal cell types that contribute to different HD clinical phenotypes.

Psychometric Properties of the Fall Risk Perception Questionnaire-Short Version for Inpatients in Acute Care Hospitals.

PURPOSE: Patients' perception of fall risk is a promising new indicator for fall prevention. Therefore, a fall risk perception questionnaire that can be used rapidly and repeatedly in acute care settings is required. This study aimed to develop a short version of the fall risk perception questionnaire (Short-FRPQ) for inpatients. METHODS: For the psychometric measurements, 246 inpatients were recruited from an acute care hospital. The construct (using confirmatory factor analysis and discriminant validity of each item), convergent, and known-group validities were tested to determine the validity of the Short-FRPQ. McDonald's omega coefficient was used to examine the internal consistency of reliability. RESULTS: In the confirmatory factor analysis, the fit indices of the Short-FRPQ, comprising 14 items and three factors, appeared to be satisfactory. The Short-FRPQ had a significantly positive correlation with the original scale, the Korean Falls Efficacy Scale-International, and the Morse Fall Scale. The risk of falls group, assessed using the Morse Fall Scale, had a higher score on the Short-FRPQ. McDonald's omega coefficient was .90. CONCLUSION: The Short-FRPQ presents good reliability and validity. As patient participation is essential in fall interventions, evaluating the fall risk perception of inpatients quickly and repeatedly using scales of acceptable validity and reliability is necessary.

[Support Needs for Health Promotion of Community-Dwelling People with Disabilities: Perspectives of Operators Managing Disability Supportive Housing].

PURPOSE: Recent studies have focused on policies aimed at supporting the independence of individuals with disabilities in communities. As part of this initiative, supportive housing, integrated care, and residential spaces offer tailored services based on individual needs and autonomy. The attitudes and knowledge of the administrators supporting supportive housing residents regarding health management can influence the well-being of individuals with disabilities. Therefore, this study aimed to explore the challenges faced by supporting housing workers in enhancing the self-management skills of individuals with disabilities. METHODS: In this qualitative study, focus group interviews were conducted in August 2023 with nine administrators working to support housing in Seoul. Qualitative content analysis was used to analyze the interview data. RESULTS: The needs and challenges in enhancing the self-management skills of individuals with disabilities were as follows: (1) the complexity of health management challenges, (2) bidirectional strategies for strengthening health management capabilities, and (3) support for systematic health management. Additionally, eight subthemes were derived. CONCLUSION: By investigating the difficulties experienced and identifying the necessary support requirements for supportive housing workers, this study seeks to uncover insights and identifies areas for improvement and strategies for health management. This study acknowledges the educational and institutional support necessary to improve the health and quality of life of individuals with disabilities residing in supportive housing.

Eight-week stress management exercise program improves mental health and autonomic nervous system of adults exposed to chronic stress.

BACKGROUND: This study investigated a stress management exercise program's effect on mental health, muscle pain, and autonomic nervous system activity in adults exposed to chronic stress. METHODS: Thirty mothers between 40 to 59 years old raising children with disabilities were equally allocated into the yoga (YG) or control group (CG). The YG participated in 16 prop-assisted yoga sessions twice weekly for eight weeks, focusing on relaxing and strengthening neck, shoulder, back, and waist stress-related muscles. The Beck Depression Inventory, State-Trait Anxiety Inventory, Parenting Stress Scale, Stress Response Inventory, and Brief Resilience Scale assessed mental health variable changes. Visual Analog Scales (VAS) were used to self-report neck, shoulders, back, and waist pain and assess stress, anxiety, relaxation, and confidence. We also measured pain, psychological VAS, and Heart Rate Variability (HRV) immediately before and after each yoga session. RESULTS: The YG exhibited significant reductions in muscle pain, depression, stress response, fatigue, and frustration, whereas the CG expressed no change. Prop-assisted yoga immediately improved pain and psychological VAS. HRV analyses revealed instantly reduced heart rates and an R-R interval and standard deviation normal to normal (SDNN) increase. CONCLUSIONS: Prop-assisted yoga may serve as a viable, safe, and efficient alternative therapeutic modality for managing chronic stress-related conditions.

Collaborative project-based learning in global health: Enhancing competencies and skills for undergraduate nursing students.

BACKGROUND: Despite the importance of collaboration and communication in global health, existing educational approaches often rely on traditional one-way instruction from instructor to student. Therefore, this study aimed to evaluate the effectiveness of a newly developed undergraduate curriculum on global health in enhancing nursing students' competencies in global health and communication, problem-solving, and self-directed learning skills. METHODS: A 15-week course "Global Health and Nursing" was designed for undergraduate nursing students, and a collaborative project-based learning method was used. Study participants were undergraduate nursing students enrolled in the course. The study was a multi-method study and included quantitative and qualitative components. It employed a one-group pretest-posttest design to quantitatively assess the impact of the curriculum. Additionally, student experiences with the learning process were qualitatively explored through a focus group interview. A total of 28 students participated in this study, and 5 of them participated in the focus group interview. RESULTS: The collaborative project-based learning method significantly improved global health competency (t = - 10.646, df = 22, p < 0.001), with a large effect size. It also improved communication skills (t = - 2.649, df = 22, p = 0.015), problem-solving skills (t = - 3.453, df = 22, p = 0.002), and self-directed learning skills (t = - 2.375, df = 22, p = 0.027). Three themes were found through the focus group interview: (a) Promoting global health competency; (b) Fostering life skills through collaborative projects; and (c) Recommendations for future classes. The focus group interview indicated that overall, the study participants were satisfied with the collaborative project-based method for global health education. CONCLUSIONS: This study confirms that project-based learning significantly boosts the competencies and skills of students, recommending its broader adoption in nursing education. Nursing instructors should consider adopting this teaching approach for global health education at the undergraduate level. Future studies may employ a longitudinal design to assess the prolonged effects of the collaborative project-based learning approach, particularly focusing on the long-term retention of skills and the broader applicability of this model across different educational settings.

The relationship between dementia caregivers and quality of life in South Korean populations.

The purpose of this study was to assess the relationship between quality of life and dementia caregivers. The 2019 Korean Community Health Survey participants were assessed using the Patient Health Questionnaire-9, subjective cognitive decline (SCD) and SCD-related functional limitation, and EuroQol 5-dimension (EQ-5D). Sociodemographic and psychosocial variables were evaluated and compared between participants with dementia caregivers (n = 37,614) and non-dementia caregivers (n = 140,518). The dementia caregivers group reported significantly higher rates of depression, SCD, SCD-related functional restriction, and mean EQ-5D compared to the non-dementia caregivers group (P < .001). After adjusting for multiple confoundings, the odds ratio (OR) for depression (Patient Health Questionnaire-9 ≥ 10), SCD, SCD-related functional limitation, and lowest quartile of the EQ-5D index scores in the dementia caregivers group were 1.43 (95% confidence interval [CI], 1.29-1.59), 1.30 (95% CI: 1.24-1.36), 1.26 (95% CI: 1.20-1.32), and 1.22 (95% CI: 1.16-1.29), respectively. Physical activity (OR: 1.47; 95% CI: 1.43-1.52), self-control (OR: 1.41; 95% CI: 1.35-1.47), daily activity (OR: 1.55; 95% CI: 1.50-1.60), pain (OR: 1.62; 95% CI: 1.58-1.67), and anxiety/depression (OR: 2.17; 95% CI: 2.10-2.24) were all more common among participants in the dementia caregivers group than in the non-dementia family caregivers group. Depression, SCD, and a lower quality of life are linked to dementia caregivers, especially if there is moderate to severe anxiety or depression.

Structural and biochemical analysis of the unique interactions of the Campylobacter jejuni CorA channel protein with divalent cations.

CorA is a Mg2+ channel that plays a key role in the homeostasis of intracellular Mg2+ in bacteria and archaea. CorA consists of a cytoplasmic domain and a transmembrane domain and generates a Mg2+ pathway by forming a pentamer in the cell membrane. CorA gating is regulated via negative feedback by Mg2+, which is accommodated by the pentamerization interface of the CorA cytoplasmic domain (CorACD). The Mg2+-binding sites of CorACD differ depending on the species, suggesting that the Mg2+-binding modes and Mg2+-mediated gating mechanisms of CorA vary across prokaryotes. To define the Mg2+-binding mechanism of CorA in the Campylobacter jejuni pathogen, we structurally and biochemically characterized C. jejuni CorACD (cjCorACD). cjCorACD adopts a three-layered α/ß/α structure as observed in other CorA orthologs. Interestingly, cjCorACD exhibited enhanced thermostability in the presence of Ca2+, Ni2+, Zn2+, or Mn2+ in addition to Mg2+, indicating that cjCorACD interacts with diverse divalent cations. This cjCorACD stabilization is mediated by divalent cation accommodation by negatively charged residues located at the bottom of the cjCorACD structure away from the pentamerization interface. Consistently, cjCorACD exists as a monomer irrespective of the presence of divalent cations. We concluded that cjCorACD binds divalent cations in a unique pentamerization-independent manner.

Instantaneous Thermal Energy for Swift Synthesis of Single-Atom Catalysts for Unparalleled Performance in Metal-Air Batteries and Fuel Cells.

Based on experimental and computational evidence, phthalocyanine (Pc) compounds in the form of quaternary-bound metal-nitrogen (N) atoms are the most effective catalysts for oxygen reduction reaction (ORR). However, the heat treatment process used in their synthesis may compromise the ideal structure, causing the agglomeration of transition metals. To overcome this issue, a novel method is developed for synthesizing iron (Fe) single-atom catalysts with ideal structures supported by thermally exfoliated graphene oxide (GO). This is achieved through a short heat treatment of only 2.5 min involving FePc and N, N-dimethylformamide in the presence of GO. According to the synthesis mechanism revealed by this study, carbon monoxide acts as a strong linker between the single Fe atoms and graphene. It facilitates the formation of a structure containing oxygen species between FeN4 and graphene, which provides high activity and stability for the ORR. These catalysts possess an enormous number of active sites and exhibit enhanced activity toward the alkaline ORR. They demonstrate excellent performance when applied to real electrochemical devices, such as zinc-air batteries and anion exchange membrane fuel cells. It is expected that the instantaneous heat treatment method developed in this study will aid in the development of high-performing single-atom catalysts.

Noise reduction of electron holography observations for a thin-foiled Nd-Fe-B specimen using the wavelet hidden Markov model.

In this study, we investigate the effectiveness of noise reduction in electron holography, based on the wavelet hidden Markov model (WHMM), which allows the reasonable separation of weak signals from noise. Electron holography observations from a Nd2Fe14B thin foil showed that the noise reduction method suppressed artificial phase discontinuities generated by phase retrieval. From the peak signal-to-noise ratio, it was seen that the impact of denoising was significant for observations with a narrow spacing of interference fringes, which is a key parameter for the spatial resolution of electron holography. These results provide essential information for improving the precision of electron holography studies.

Time-to-Event Genome-Wide Association Study for Incident Cardiovascular Disease in People With Type 2 Diabetes.

OBJECTIVE: To identify genetic risk factors for incident cardiovascular disease (CVD) among people with type 2 diabetes (T2D). RESEARCH DESIGN AND METHODS: We conducted a multiancestry time-to-event genome-wide association study for incident CVD among people with T2D. We also tested 204 known coronary artery disease (CAD) variants for association with incident CVD. RESULTS: Among 49,230 participants with T2D, 8,956 had incident CVD events (event rate 18.2%). We identified three novel genetic loci for incident CVD: rs147138607 (near CACNA1E/ZNF648, hazard ratio [HR] 1.23, P = 3.6 × 10-9), rs77142250 (near HS3ST1, HR 1.89, P = 9.9 × 10-9), and rs335407 (near TFB1M/NOX3, HR 1.25, P = 1.5 × 10-8). Among 204 known CAD loci, 5 were associated with incident CVD in T2D (multiple comparison-adjusted P < 0.00024, 0.05/204). A standardized polygenic score of these 204 variants was associated with incident CVD with HR 1.14 (P = 1.0 × 10-16). CONCLUSIONS: The data point to novel and known genomic regions associated with incident CVD among individuals with T2D.

Acetyl-CoA synthetase 2 contributes to a better prognosis for liver cancer by switching acetate-glucose metabolism.

Acetyl-CoA synthetase 2 (ACSS2)-dependent acetate usage has generally been associated with tumorigenesis and increased malignancy in cancers under nutrient-depleted conditions. However, the nutrient usage and metabolic characteristics of the liver differ from those of other organs; therefore, the mechanism of ACSS2-mediated acetate metabolism may also differ in liver cancer. To elucidate the underlying mechanisms of ACSS2 in liver cancer and acetate metabolism, the relationships between patient acetate uptake and metabolic characteristics and between ACSS2 and tumor malignancies were comprehensively studied in vitro, in vivo and in humans. Clinically, we initially found that ACSS2 expression was decreased in liver cancer patients. Moreover, PET-CT imaging confirmed that lower-grade cancer cells take up more 11C-acetate but less 18F-fluorodeoxyglucose (18F-FDG); however, this trend was reversed in higher-grade cancer. Among liver cancer cells, those with high ACSS2 expression avidly absorbed acetate even in a glucose-sufficient environment, whereas those with low ACSS2 expression did not, thereby showing correlations with their respective ACSS2 expression. Metabolomic isotope tracing in vitro and in vivo revealed greater acetate incorporation, greater lipid anabolic metabolism, and less malignancy in high-ACSS2 tumors. Notably, ACSS2 downregulation in liver cancer cells was associated with increased tumor occurrence in vivo. In human patient cohorts, patients in the low-ACSS2 subgroup exhibited reduced anabolism, increased glycolysis/hypoxia, and poorer prognosis. We demonstrated that acetate uptake by ACSS2 in liver cancer is independent of glucose depletion and contributes to lipid anabolic metabolism and reduced malignancy, thereby leading to a better prognosis for liver cancer patients.

Versatile human cardiac tissues engineered with perfusable heart extracellular microenvironment for biomedical applications.

Engineered human cardiac tissues have been utilized for various biomedical applications, including drug testing, disease modeling, and regenerative medicine. However, the applications of cardiac tissues derived from human pluripotent stem cells are often limited due to their immaturity and lack of functionality. Therefore, in this study, we establish a perfusable culture system based on in vivo-like heart microenvironments to improve human cardiac tissue fabrication. The integrated culture platform of a microfluidic chip and a three-dimensional heart extracellular matrix enhances human cardiac tissue development and their structural and functional maturation. These tissues are comprised of cardiovascular lineage cells, including cardiomyocytes and cardiac fibroblasts derived from human induced pluripotent stem cells, as well as vascular endothelial cells. The resultant macroscale human cardiac tissues exhibit improved efficacy in drug testing (small molecules with various levels of arrhythmia risk), disease modeling (Long QT Syndrome and cardiac fibrosis), and regenerative therapy (myocardial infarction treatment). Therefore, our culture system can serve as a highly effective tissue-engineering platform to provide human cardiac tissues for versatile biomedical applications.

CRISPR/Cas9-mediated knockout of scarlet gene produces eye color mutants in the soybean looper, Chrysodeixis includens.

The CRISPR/Cas9 technology has greatly progressed research on non-model organisms, demonstrating successful applications in genome editing for various insects. However, its utilization in the case of the soybean looper, Chrysodeixis includens, a notable pest affecting soybean crops, has not been explored due to constraints such as limited genomic information and the embryonic microinjection technique. This study presents successful outcomes in generating heritable knockout mutants for a pigment transporter gene, scarlet, in C. includens through CRISPR/Cas9-mediated mutagenesis. The scarlet locus identified in the genome assembly of C. includens consists of 14 exons, with a coding sequence extending for 1,986 bp. Two single guide RNAs (sgRNAs) were designed to target the first exon of scarlet. Microinjection of these two sgRNAs along with the Cas9 protein into fresh embryos resulted in the successful production of variable phenotypes, particularly mutant eyes. The observed mutation rate accounted for about 16%. Genotype analysis revealed diverse indel mutations at the target site, presumably originating from double-strand breaks followed by the nonhomologous end joining repair, leading to a premature stop codon due to frame shift. Single-pair mating of the mutant moths produced G1 offspring, and the establishment of a homozygous mutant strain occurred in G2. The mutant moths exhibited lightly greenish or yellowish compound eyes in both sexes, confirming the involvement of scarlet in pigmentation in C. includens. Notably, the CRISPR/Cas9-mediated genome editing technique serves as a visible phenotypic marker, demonstrating its proof-of-concept applicability in C. includens, as other pigment transporter genes have been utilized as visible markers to establish genetic control for various insects. These results provide the first successful case that the CRISPR/Cas9 method effectively induces mutations in C. includes, an economically important soybean insect pest.

Seasonal Stability Assessment of Reference Genes for Quantitative Real-Time Polymerase Chain Reaction Normalization in Bombus terrestris.

Bumblebees (B. terrestris) play a crucial role as highly efficient biological agents in commercial pollination. Understanding the molecular mechanisms governing their adaptation to diverse seasonal environments may pave the way for effective management strategies in the future. With the burgeoning advancement in post-genetic studies focusing on B. terrestris, there is a critical need to normalize quantitative real-time PCR (qRT-PCR) data using suitable reference genes. To address this necessity, we employed RefFinder, a software-based tool, to assess the suitability of several candidate endogenous control genes, including actin (ACT), arginine kinase (AK), elongation factor 1 alpha (EF1), glyceraldehyde-3-phosphate (GAPDH), phospholipase (PLA2), and ribosomal proteins (S18, S28). These genes were evaluated for their efficacy as biological endogenous controls by examining their expression patterns across various environmental conditions corresponding to different seasons (Spring, Summer, Autumn, Winter) and tissues (ovary, fat body, thorax, head) in bumblebees. Moreover, the study investigated the significance of selecting appropriate reference genes for three key genes involved in the juvenile hormone (JH) signaling pathways: Krüppel homolog 1 (Kr-h1), methyl farnesoate epoxidase (MFE), and Vitellogenin (Vg). Our research identifies specific genes suitable for normalization in B. terrestris, thereby offering valuable insights into gene expression and functional metabolic genetics under varying seasonal conditions. This catalog of reference genes will serve as a valuable resource for future research endeavors.