Pharmacogenetic Approach to Tramadol Use in the Arab Population.

Tramdol is one of most popular opioids used for postoperative analgesia worldwide. Among Arabic countries, there are reports that its dosage is not appropriate due to cultural background. To provide theoretical background of the proper usage of tramadol, this study analyzed the association between several genetic polymorphisms (CYP2D6/OPRM1) and the effect of tramadol. A total of 39 patients who took tramadol for postoperative analgesia were recruited, samples were obtained, and their DNA was extracted for polymerase chain reaction products analysis followed by allelic variations of CYP2D6 and OPRM A118G determination. Numerical pain scales were measured before and 1 h after taking tramadol. The effect of tramadol was defined by the difference between these scales. We concluded that CYP2D6 and OPRM1 A118G single nucleotide polymorphisms may serve as crucial determinants in predicting tramadol efficacy and susceptibility to post-surgical pain. Further validation of personalized prescription practices based on these genetic polymorphisms could provide valuable insights for the development of clinical guidelines tailored to post-surgical tramadol use in the Arabic population.

Seroprevalence of anti-diphtheria toxoid antibody and implications for vaccination policy in Vietnam's South-central coast: a cross-sectional study.

BACKGROUND: Diphtheria is a re-emerging infectious disease and public health concern worldwide and in Vietnam with increasing cases in recent years. This study aimed to assess the anti-diphtheria toxoid antibodies status in Khanh Hoa Province and identify factors contributing to the vaccination policy in the south-central coast of Vietnam. METHODS: This was a cross-sectional study to evaluate the seroprevalence of anti-diphtheria toxoid antibodies among 1,195 participants, aged 5 - 40 years in Khanh Hoa Province, Vietnam. Immunoglobulin G antibody levels against diphtheria were detected using a commercial anti-diphtheria toxoid enzyme-linked immunosorbent assay (SERION ELISA classic Diphtheria Immunoglobulin G) and were categorized following the World Health Organization guidelines. RESULTS: The mean anti-diphtheria toxoid antibody levels were 0.07 IU/ml (95% Confidence Interval: 0.07-0.08). Anti-diphtheria toxoid antibody levels were found to be associated with age and history of diphtheria vaccination. The 5-15 years age group had the highest levels (0.09 IU/ml), while the older age group had the lowest antibody level (p < 0.001). Individuals who received three doses (adjusted Odds ratio: 2.34, 95%CI: 1.35 - 4.07) or 4+ doses (adjusted Odds ratio: 2.45, 95%CI: 1.29 - 4.64) had a higher antibody level compared to those who received only one dose regardless of age. CONCLUSION: It is crucial to promote routine vaccination coverage to over 95% for children under one year of age with three primary doses of the diphtheria-containing vaccine, including additional doses at 18 months and 7 years of age. Booster doses should be promoted and administered to adolescents and adults every 10 years.

Enantioselective Michael addition of 3-hydroxy-2-pyridone to nitroolefins using cinchona-derived bifunctional organocatalysts.

Despite the extensive use of N-heteroarenes in pharmaceuticals and natural products, efficient methods for selective alkylation at the C-4 position of 2-pyridone are scarce. We developed an enantioselective Michael addition of 3-hydroxy-2-pyridone to nitroolefins at the C-4 position using cinchona-derived bifunctional squaramide organocatalysts, achieving up to 95% yield and >99% ee. This selectivity is driven by the bifunctional organocatalysts' hydrogen bonding interactions with 3-hydroxy-2-pyridone and nitroolefins under mild conditions. This method demonstrates the Michael reaction's versatility with various nitroolefins, providing a sustainable approach for synthesizing chiral N-heteroarenes with high enantioselectivity and regioselectivity under environmentally friendly conditions.

Whole genome and reverse protein phase array landscapes of patient derived osteosarcoma xenograft models.

Osteosarcoma is the most common primary bone malignancy in children and young adults, and it has few treatment options. As a result, there has been little improvement in survival outcomes in the past few decades. The need for models to test novel therapies is especially great in this disease since it is both rare and does not respond to most therapies. To address this, an NCI-funded consortium has characterized and utilized a panel of patient-derived xenograft models of osteosarcoma for drug testing. The exomes, transcriptomes, and copy number landscapes of these models have been presented previously. This study now adds whole genome sequencing and reverse-phase protein array profiling data, which can be correlated with drug testing results. In addition, four additional osteosarcoma models are described for use in the research community.

Association between sleep patterns and alcohol use disorders in workers.

Alcohol use among workers that is intended to aid sleep may lead to alcohol use disorders. This study aimed to explore the association between sleep patterns and alcohol use disorders in workers. Data from the Korea National Health and Nutrition Examination Survey conducted in 2014, 2016, 2018, and 2020 were used for this study. We included only workers aged 19 years and older. The final analysis comprised 11,972 respondents (6,472 male and 5,500 female). Multiple logistic regression analysis was used to investigate the relationship between sleep patterns and alcohol use disorders. Workers with poor sleep patterns were more likely to develop alcohol use disorders compared to those with good sleep patterns (male: adjusted odds ratio [OR] 1.22, 95% confidence interval 1.07-1.39; female: adjusted OR 1.21, 95% CI 1.03-1.41). Workers with both poor sleep quality and less than seven hours of sleep had the highest odds of alcohol use disorders in both male (adjusted OR 1.73, 95% CI 1.38-2.17) and female (adjusted OR 1.44, 95% CI 1.13-1.84). Poor sleep patterns were associated with alcohol use disorders in male who work night shift (OR: 1.74, 95% CI: 1.25-2.42) and in female who worked more than 52 hours per week (adjusted OR: 1.71, 95% CI: 1.04-2.80). Customized sleep management programs should be provided to workers in sleep-deprived working environments to prevent them from developing alcohol use disorders.

Effect of physical activity on free fatty acids, insulin resistance, and blood pressure in obese older women.

PURPOSE: Obesity is characterized by a progressive increase in body fat accompanied by insulin resistance (IR) and elevated blood pressure (BP), and presents significant health risks, particularly in aged individuals. This study aimed to evaluate the effects of physical activity (PA) on free fatty acid (FFA) levels, IR, and BP in obese older women. METHODS: Twenty-three participants were randomly assigned to either the control group (CON, n = 11) or the physical activity group (PA, n = 12). The PA group was provided with a target of achieving >7,000 steps/day for 5 days each week. Body composition, FFA levels, IR, and BP were measured at pre- and post- of the 12-week intervention. RESULTS: The analysis revealed a statistically significant interaction between FFA (p < 0.01), IR (p < 0.01), and SBP (p < 0.001). FFA (p < 0.5), IR (p < 0.5), and systolic blood pressure (SBP) (p < 0.01) were significantly decreased in the PA group compared to those in the CON group, which showed no significant changes in FFA, IR, and SBP. CONCLUSION: PA significantly decreased FFA, IR, and SBP in older women with obesity. Therefore, PA is an effective intervention for the prevention and management of obesity and cardiovascular diseases in obese older women.

A quantitative analysis of fidgeting in ADHD and its relation to performance and sustained attention on a cognitive task.

Introduction: Attention-Deficit/Hyperactivity Disorder (ADHD) is a neurodevelopmental disorder where hyperactivity often manifests as fidgeting, a non-goal-directed motoric action. Many studies demonstrate fidgeting varies under different conditions as a self-regulating mechanism for attention and alertness during cognitively demanding tasks. Fidgeting has also been associated with reaction time variability. However, a lack of standard variables to define and quantify fidgeting can lead to discrepancies in data and interpretability issues across studies. Furthermore, little is known about fidgeting in adults with ADHD compared to youth. This study aims to design a framework to quantify meaningful fidgeting variables and to apply them to test the relation between fidgeting and performance on a cognitive task, the Flanker, in adults with ADHD. Method: Our study included 70 adult participants diagnosed with ADHD, aged 18-50 years (30.5 ± 7.2 years). Screening included a structured clinical interview, childhood, current self and current observer ratings of ADHD symptoms. Actigraphy devices were attached to the left wrist and right ankle during completion of a cognitive control, attention task (the Flanker). Laboratory testing was subsequently completed on a single day. The relation between task performance, reaction time variability and fidgeting was examined. Results and Discussion: Our analysis revealed increased fidgeting during correct trials as defined by our new variables, consistent with previous observations. Furthermore, differences in fidgeting were observed between early and later trials while the percentage of correct trials were not significantly different. This suggests a relation between the role of fidgeting and sustaining attention. Participants with low reaction time variability, that is, those with more consistent reaction times, fidgeted more during later trials. This observation supports the theory that fidgeting aids arousal and improves sustained attention. Finally, a correlation analysis using ADHD-symptom rating scales validated the relevance of the fidget variables in relation to ADHD symptom severity. These findings suggest fidgeting may be a compensatory mechanism that aids in sustained attention for those with ADHD, although alternative explanations exist. Conclusion: Our study suggests that fidgeting may aid in sustained attention during the attention-demanding, cognitive control processes for adults with ADHD, with more fidgeting observed during correct trials and among participants with lower reaction time variability. Furthermore, the newly defined fidget variables were validated through a significant correlation with ADHD rating scales. By sharing our implementation of fidget variables, we hope to standardize and encourage further quantitative research into the role of fidgeting in ADHD.

Diagnosis of ADHD using virtual reality and artificial intelligence: an exploratory study of clinical applications.

Introduction: Diagnosis of Attention Deficit/Hyperactivity Disorder (ADHD) is based on clinical evaluation of symptoms by a psychiatrist, referencing results of psychological tests. When diagnosing ADHD, the child's behavior and functionality in real-life situations are critical components. However, direct observation by a clinician is often not feasible in practice. Therefore, such information is typically gathered from primary caregivers or teachers, which can introduce subjective elements. To overcome these limitations, we developed AttnKare-D, an innovative digital diagnostic tool that could analyze children's behavioral data in Virtual Reality using Artificial Intelligence. The purpose of this study was to explore the utility and safety of AttnKare-D for clinical application. Method: A total of 21 children aged between 6 and 12 years were recruited for this study. Among them, 15 were children diagnosed with ADHD, 5 were part of a normal control group, and 1 child was excluded due to withdrawal of consent. Psychological assessments, including K-WISC, Conners CPT, K-ARS, and K-CBCL, were conducted for participants and their primary caregivers. Diagnoses of ADHD were confirmed by child and adolescent psychiatrists based on comprehensive face-to-face evaluations and results of psychological assessments. Participants underwent VR diagnostic assessment by performing various cognitive and behavioral tasks in a VR environment. Collected data were analyzed using an AI model to assess ADHD diagnosis and the severity of symptoms. Results: AttnKare-D demonstrated diagnostic performance with an AUC of 0.893 when compared to diagnoses made by child and adolescent psychiatrist, showing a sensitivity of 0.8 and a specificity of 1.0 at a cut-off score of 18.44. AttnKare-D scores showed a high correlation with K-ARS scores rated by parents and experts, although the correlation was relatively low for inattention scores. Conclusion: Results of this study suggest that AttnKare-D can be a useful tool for diagnosing ADHD in children. This approach has potential to overcome limitations of current diagnostic methods, enhancing the accuracy and objectivity of ADHD diagnoses. This study lays the groundwork for further improvement and research on diagnostic tools integrating VR and AI technologies. For future clinical applications, it is necessary to conduct clinical trials involving a sufficient number of participants to ensure reliable use.

Synthesis of Isoamyl Fatty Acid Ester, a Flavor Compound, by Immobilized Rhodococcus Cutinase.

Isoamyl fatty acid esters (IAFEs) are widely used as fruity flavor compounds in the food industry. In this study, various IAFEs were synthesized from isoamyl alcohol and various fatty acids using a cutinase enzyme (Rcut) derived from Rhodococcus bacteria. Rcut was immobilized on methacrylate divinylbenzene beads and used to synthesize isoamyl acetate, butyrate, hexanoate, octanoate, and decanoate. Among them, Rcut synthesized isoamyl butyrate (IAB) most efficiently. Docking model studies showed that butyric acid was the most suitable substrate in terms of binding energy and distance from the active site serine (Ser114) γ-oxygen. Up to 250 mM of IAB was synthesized by adjusting reaction conditions such as substrate concentration, reaction temperature, and reaction time. When the enzyme reaction was performed by reusing the immobilized enzyme, the enzyme activity was maintained at least six times. These results demonstrate that the immobilized Rcut enzyme can be used in the food industry to synthesize a variety of fruity flavor compounds, including IAB.

Association between the utilization of senior centers and participation in health check-ups.

The global older adult population is increasing. Early detection and intervention through health check-ups are crucial for successful aging, as they play a significant role in identifying and addressing diseases. This study explored the relationship between the utilization of senior centers and the promotion of health check-ups. It utilized data from 10,097 individuals aged 65 years and above, sourced from the 2020 Elderly Survey in South Korea. The primary variable of interest was classified into two groups: those who utilized senior centers and those who did not. Subgroups were further categorized based on the frequency of usage and the presence of family members among senior centers users. Logistic regression analyses were conducted to assess the association between the utilization of senior centers and participation in health check-ups. Both men and women utilizing senior centers demonstrated a higher likelihood of participating in health check-ups compared with those who did not use senior centers. Participants visiting senior centers in a week exhibited a progressively higher likelihood of engaging in health check-ups compared with those who visited such senior centers zero times a week. Senior centers can serve as effective intervention methods to enhance health check-ups among older adults. Furthermore, this can contribute to fostering successful aging among older adults.

Assessment of Patient-Derived Xenograft Growth and Antitumor Activity: The NCI PDXNet Consensus Recommendations.

Although patient-derived xenografts (PDX) are commonly used for preclinical modeling in cancer research, a standard approach to in vivo tumor growth analysis and assessment of antitumor activity is lacking, complicating the comparison of different studies and determination of whether a PDX experiment has produced evidence needed to consider a new therapy promising. We present consensus recommendations for assessment of PDX growth and antitumor activity, providing public access to a suite of tools for in vivo growth analyses. We expect that harmonizing PDX study design and analysis and assessing a suite of analytical tools will enhance information exchange and facilitate identification of promising novel therapies and biomarkers for guiding cancer therapy.

Association between Preoperative C-Reactive Protein-to-Albumin Ratio and Mortality after Plastic and Reconstructive Surgery.

Background: Prognostic markers have not been extensively studied in plastic and reconstructive surgery. Objective: We aimed to evaluate the prognostic value of preoperative C-reactive protein (CRP)-to-albumin ratio (CAR) in plastic and reconstructive surgery and to compare it with the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and modified Glasgow prognostic score (mGPS). Methods: From January 2011 to July 2019, we identified 2519 consecutive adult patients who were undergoing plastic and reconstructive surgery with available preoperative CRP and albumin levels. The receiver operating characteristic (ROC) curve was generated to evaluate predictability and estimate the threshold. The patients were divided according to this threshold, and the risk was compared. The primary outcome was one-year mortality, and the overall mortality was also analyzed. Results: The one-year mortality was 4.9%. The CAR showed an area under the ROC curve of 0.803, which was higher than those of NLR, PLR, and mGPS. According to the estimated threshold of 1.05, the patients were divided into two groups; 1585 (62.9%) were placed in the low group, and 934 (37.1%) were placed in the high group. After inverse probability weighting, the mortality rate during the first year after plastic and reconstructive surgery was significantly increased in the high group (1.3% vs. 10.9%; hazard ratio, 2.88; 95% confidence interval, 2.17-3.83; p < 0.001). Conclusions: In this study, high CAR was significantly associated with one-year mortality of patients after plastic and reconstructive surgery. Further studies are needed on prognostic markers in plastic and reconstructive surgery.

Pulmonary function characteristics in children with suspected asthma: implications for asthma diagnosis.

BACKGROUND: In children suspected of asthma, diagnosis is confirmed via variable expiratory airflow limitation. However, there is no single gold standard test for diagnosing asthma. OBJECTIVE: This study aimed to evaluate the pulmonary function characteristics in children suspected of asthma without bronchodilator response (BDR) and bronchial hyperresponsiveness (BHR). METHODS: We utilized two separate real-world retrospective observational cohorts of children who underwent both spirometry and bronchial provocation testing for asthma. Spirometry parameters were collected and compared between definite asthma, probable asthma, and non-asthma groups. The original cohort comprised 1199 children who visited the Severance Hospital (Seoul, Korea) between January 2017 and December 2019. The external cohort included 105 children who visited the Gangnam Severance Hospital between January 2019 and December 2019. RESULTS: Probable asthma accounted for 16.8% and 32.4% of the original and external cohorts, respectively. This group showed a significantly higher FeNO level and prevalence of allergic sensitization. Baseline forced expiratory volume in 1 second (FEV1), FEV1/forced vital capacity (FVC), forced expiratory flow at 25-75% of FVC (FEF25-75), and FEF75 showed stepwise decrements from non-asthma, probable asthma, to definite asthma patients (P < 0.001). The probable asthma group showed significantly higher odds of abnormal FEV1/FVC (OR, 2.24 [95%CI, 1.43-3.52])and FEF25-75 (2.05 [1.13-3.73]) than the non-asthma group and lower odds of abnormal FEV1(0.05 [0.01-0.19]),FEV1 /FVC (0.27 [0.18-0.41]), FEF25-75 (0.17 [0.11-0.28]), and FEF75 (0.14 [0.08-0.24]) compared to the definite asthma group. The external cohort was consistent with the original cohort. CONCLUSION: We show evidence of airway dysfunction in children for whom a high clinical suspicion of asthma exists without evidence of BDR and BHR. Repeated pulmonary function tests that closely monitor for subtle lung function impairments and active utilization of additional tests, such as allergic screening and FeNO, should be considered to close the gap in diagnosing asthma.

Effect of dynamic neuromuscular stabilization training using the inertial load of water on functional movement and postural sway in middle-aged women: a randomized controlled trial.

BACKGROUND: Chronic stress and diseases occur more frequently in middle-aged compared to younger women and this is often the result of physical, psychological and socio-economic changes. These health consequences reduce lower body muscle mass and flexibility, leading to generalized impairments in functional movement and balance. Dynamic Neuromuscular Stabilization (DNS) training using the inertial load of water is known for its positive impact on functional strength improvement and muscle stabilization. OBJECTIVE: This study aimed to determine the effect of DNS training using inertial water loads on functional movement and postural sway in middle-aged women. METHOD: A sample of 24 middle-aged women participated in the study and were randomly divided into two groups: the experimental group, n = 12 (age: 58.33 ± 1.48 yrs, height: 162.16 ± 1.27 cm, weight: 61.77 ± 2.21 kg) and control group, n = 12 (age: 59.58 ± 1.13 yrs, height: 160.1 ± 1.13 cm, weight: 57.51 ± 1.12 kg). Center of Pressure (COP), moving distance, Root Mean Square (RMS), movement area and Functional Movement Screen (FMS) were conducted and analyzed pre- and post-examination. Participants engaged in the DNS training regimen, which utilized the inertial load of water, for 60 min each session, conducted twice weekly for 12 weeks. RESULTS: There were significant differences in the COP distance (p < 0.001), RMS (p < 0.05), COP area and FMS test (p < 0.001) in the pre-post comparison of each group. And significant differences were found in COP distance (p < 0.05), RMS (p < 0.05), COP area (p < 0.05) and FMS test (p < 0.05) between groups. The DNS training improved the dynamic stability of single-leg standing, torso stability and functional movement in middle-aged women. CONCLUSION: DNS training programs using the inertial load of water have been shown to be effective in movement improvement and posture retention ability, which is beneficial for functional movement, equilibrium strategy, and dynamic stability of middle-aged women. Furthermore, the DNS training method designed in this study can be useful for trainees who require posture correction in a safe and effective way regardless of their age and gender.

Targeting DNA2 overcomes metabolic reprogramming in multiple myeloma.

DNA damage resistance is a major barrier to effective DNA-damaging therapy in multiple myeloma (MM). To discover mechanisms through which MM cells overcome DNA damage, we investigate how MM cells become resistant to antisense oligonucleotide (ASO) therapy targeting Interleukin enhancer binding factor 2 (ILF2), a DNA damage regulator that is overexpressed in 70% of MM patients whose disease has progressed after standard therapies have failed. Here, we show that MM cells undergo adaptive metabolic rewiring to restore energy balance and promote survival in response to DNA damage activation. Using a CRISPR/Cas9 screening strategy, we identify the mitochondrial DNA repair protein DNA2, whose loss of function suppresses MM cells' ability to overcome ILF2 ASO-induced DNA damage, as being essential to counteracting oxidative DNA damage. Our study reveals a mechanism of vulnerability of MM cells that have an increased demand for mitochondrial metabolism upon DNA damage activation.

Effect of the peripartum depressive symptoms on the Internet use disorder of their offspring in late childhood: retrospective longitudinal study.

Internet use disorder (IUD) is an emerging social and mental health concern. This study aimed to analyze the relative risk of IUD in late childhood among children whose mothers experienced peripartum depressive symptoms. This study included 762 participants (397 boys and 365 girls) and was conducted in 2017 (aged 9) and 2019 (aged 11). We analyzed the adjusted relative risk of being at high risk for IUD based on whether the mother experienced depressive symptoms during pregnancy or one month after delivery. We also considered the persistence of depressed mood for 4 months after delivery and the severity of peripartum depressive symptoms. From 2017, 20.7% of boys and 14.0% of girls were at high risk of developing IUD. Compared to the non-peripartum depressive group, girls whose mothers experienced peripartum depressive symptoms and those that persisted for 4 months were 1.084 and 1.124 times more likely to be at high risk of IUD (95% confidence interval = 1.005-1.170 and 1.013-1.248), respectively. There were no statistically significant differences among boys. Peripartum depressed mood could be one of risk factors of IUD. IUD needs to be monitored in children whose mothers experienced peripartum depressive symptoms.

Unveiling Neurocognitive Disparities in Encoding and Retrieval between Paper and Digital Tablet-Based Learning.

The widespread use of mobile devices and laptops has replaced traditional paper-based learning and the question of how the brain efficiency of digital tablet-based learning differs from that of paper-based learning remains unclear. The purpose of this study was to investigate the difference in brain efficiency for learning between paper-based and digital tablet-based learning by measuring activity in the prefrontal cortex (PFC) using functional near-infrared spectroscopy. Thirty-two subjects were randomly assigned to the paper-based learning or the digital tablet-based learning group. Subjects in each group performed a memory task that required memorizing a three-minute novel (encoding phase) on a paper or digital tablet, followed by a test in which they answered four multiple-choice questions based on the novel's content. To compare both groups, behavioral performance on the test (retrieval phase) and activity in the PFC were measured. As a result, no significant difference in behavioral performance between both groups was observed (p > 0.05). However, the paper-based learning group showed significantly lower activity in the PFC in the encoding phase than the digital tablet-based learning group (p < 0.05) but not in the retrieval phase. The current study demonstrated that brain efficiency in encoding is higher in subjects with paper-based learning than those with digital tablet-based learning. This finding has important implications for education, particularly in terms of the pros and cons of electronic document-based learning.

Mild exercise improves executive function with increasing neural efficiency in the prefrontal cortex of older adults.

This study examined whether a 3-month mild-exercise intervention could improve executive function in healthy middle-aged and older adults in a randomized control trial. Ultimately, a total of 81 middle-aged and older adults were randomly assigned to either an exercise group or a control group. The exercise group received 3 months of mild cycle exercise intervention (3 sessions/week, 30-50 min/session). The control group was asked to behave as usual for the intervention period. Before and after the intervention, participants did color-word matching Stroop tasks (CWST), and Stroop interference (SI)-related reaction time (RT) was assessed as an indicator of executive function. During the CWST, prefrontal activation was monitored using functional near-infrared spectroscopy (fNIRS). SI-related oxy-Hb changes and SI-related neural efficiency (NE) scores were assessed to examine the underlying neural mechanism of the exercise intervention. Although the mild-exercise intervention significantly decreased SI-related RT, there were no significant effects of exercise intervention on SI-related oxy-Hb changes or SI-related NE scores in prefrontal subregions. Lastly, changes in the effects of mild exercise on NE with advancing age were examined. The 81 participants were divided into two subgroups (younger-aged subgroup [YA], older-aged subgroup [OA], based on median age [68 years.]). Interestingly, SI-related RT significantly decreased, and SI-related NE scores in all ROIs of the prefrontal cortex significantly increased only in the OA subgroup. These results reveal that a long-term intervention of very light-intensity exercise has a positive effect on executive function especially in older adults, possibly by increasing neural efficiency in the prefrontal cortex.

Production of octyl butyrate using psychrophilic mutant lipase from Croceibacter atlanticus LipCA lipase developed by a molecular evolution technique.

Lipases are used to synthesize a variety of industrially useful compounds. Among them, psychrophilic lipase can be used to synthesize thermo-labile compounds at low temperatures. In this study, random mutagenesis was introduced into Antarctic Croceibacter atlanticus lipase gene using error-prone PCR, resulting in changes in its protein sequence. Through two rounds of mutagenesis and screening, we found that a mutant R1 showed an enhanced activity at low temperatures. Mutant R1 had five mutations (F43L, S48G, S49G, D141K, and K297R) and higher kcat/KM value than the wild type (WT) at 10 °C. We immobilized this enzyme on methacrylate divinylbenzene resin and used it to synthesize octyl butyrate, a flavor compound. The esterification reaction proceeded even at 10 °C. Mutant R1 synthesized the ester compound faster than the WT. To determine which amino acids were responsible for the increase of activity, site-directed mutagenesis was performed to introduce five back mutations into mutant R1. Three back mutants (L43F, G48S, G49S) showed significant decreases of activity at low temperatures, indicating that these amino acids were closely related to the increase in activity. This psychrophilic mutant R1 is expected to be used in low-temperature enzyme conversion reactions in the food industry.

Diversity and pathogenic characteristics of the Fusarium species isolated from minor legumes in Korea.

Legumes are primarily grown agriculturally for human consumption, livestock forage, silage, and as green manure. However, production has declined primarily due to fungal pathogens. Among them, this study focused on Fusarium spp. that cause Fusarium wilt in minor legumes in Korea. Diseased legume plants were collected from 2020 to 2021, and diverse fungal genera were isolated from the internal tissues of the plant roots and stems. Fusarium spp. were the most dominant, accounting for 71% of the isolates. They were identified via morphological characteristics and molecular identification. In the pathogenicity test, Fusarium oxysporum and Fusarium fujikuroi generally exhibited high virulence. The host range investigation revealed that the NC20-738, NC20-739, and NC21-950 isolates infected all nine crops, demonstrating the widest host range. In previous studies, the focus was solely on Fusarium wilt disease in soybeans. Therefore, in this study, we aimed to investigate Fusarium wilt occurred in minor legumes, which are consumed as extensively as soybeans, due to the scarcity of data on the diversity and characteristics of Fusarium spp. existing in Korea. The diverse information obtained in this study will serve as a foundation for implementing effective management strategies against Fusarium-induced plant diseases.