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Haloarchaea represents a unique group of microorganisms that have adapted to thrive in high-salt environments. These microbes produce distinctive biomolecules, some of which exhibit extraordinary properties. One such biomolecule is bacterioruberin, a prominent red-pigmented C 50 carotenoid commonly found in halophilic archaea, renowned for its antioxidant properties and potential as a functional resource. This study aimed to enhance the culture conditions for optimal production of C 50 carotenoids, primarily bacterioruberin, using "Haloferax marinum" MBLA0078. The optimization process involved a combination of one-factor-at-a-time (OFAT) and statistical methodology. Under OFAT-optimized conditions, fed-batch fermentation, and response surface methodology (RSM) optimization, carotenoid production reached 0.954 mg/L, 2.80 mg/L, and 2.16 mg/L, respectively, in a 7-L laboratory-scale fermenter. Notably, RSM-optimized conditions led to a 12-fold increase in productivity (0.72 mg/L/day) compared to the basal DBCM2 medium (0.06 mg/L/day). These findings suggest that strain MBLA0078 holds significant promise for commercial-scale production of bacterioruberin., Competing Interests: Declarations. Ethical approval: Not applicable. Consent for publication: All authors read and approved the final manuscript and potential publication. Competing interests: The authors declare no competing interest in submitting the article., (© 2024. The Author(s).)

A novel extremely halophilic archaeon designated, MBLA0158 T , was isolated from a solar saltern in Sorae, Republic of Korea. The colonies are red-pigmented, Gram-stain-negative, pleomorphic, non-motile, and lysed in distilled water. The strain grows at 25-45 °C (optimum, 37 °C), in 15-30% (w/v) NaCl (optimum, 20%) and 0.1-1.0 M Mg 2+ (optimum, 0.2-0.3 M) at pH 6.0-10.0 (optimum, 7.0-8.0). Comparative analysis based on the 16S rRNA gene sequence revealed that this strain is most closely related to the Halobellus inordinatus YC20 T with a sequence identity of 96.0%. Strain MBLA0158 T contained phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester as major polar lipids. The genome size is 3.29 Mb and the DNA G + C content is 66.9 mol%. Phylogenomic analysis confirmed that strain MBLA0158 T is distinct from previously reported type strains of the genus Halobellus. Pan-genome analysis showed that strain MBLA0158 T contains 419 genes that are not present in other type strains of the genus Halobellus. Based on overall analyses, strain MBLA0158 T is considered to represent a new species of the genus Halobellus, for which the name Halobellus rubicundus sp. nov. is proposed. The type strain is MBLA0158 T (= KCTC 4318 T  = JCM 36642 T )., (© 2024. The Author(s), under exclusive licence to Springer Nature Japan KK.)

Amid rising environmental concerns, attempts have been made to produce glycolic acid (GA) using microbial processes with renewable carbon resources instead of using chemicals. The Dahms pathway for GA production uses xylose as a substrate and consists of relatively simple enzymatic steps. However, employing it leads to a decrease in cell growth and GA productivity. Systematically identifying and addressing metabolic bottlenecks in the Dahms pathway are essential for efficient glycolic acid (GA) production have not yet been performed. Through metabolic flux balance analysis, we found that insufficient aldehyde dehydrogenase (AldA) activity lowers GA production and negatively affects cell growth due to reduced energy production. Thus, we discovered a novel AldA isolated from Buttiauxella agrestis (BaAldA) demonstrated a 1.69-fold lower K M and a 1.49-fold higher turnover rate (k cat /K M ) than AldA from Escherichia coli (EcAldA). GA production in E. coli harboring BaAldA was 1.59 times higher than in the original strain. Fed-batch fermentation of E. coli harboring BaAldA produced 22.70 g/L GA with a yield of 0.497 g/g xylose (98.2% of the theoretical maximum yield in the Dahms pathway), showing a higher final yield for GA than previously reported in E. coli. Our novel BaAldA enzyme shows great potential for the production of GA using microorganisms or enzymes. Furthermore, our approach to identifying metabolic bottlenecks using flux balance analysis could be utilized to enhance the microbial production of various desirable products in future studies., Competing Interests: Declarations. Conflict of Interest: The authors declare that there are no competing financial interests. Ethical Approval: Not applicable., (© 2024. The Author(s), under exclusive licence to Microbiological Society of Korea.)

In this study, a novel species within the genus Paracoccus was isolated from the coastal soil of Dokdo (Seodo) Island and investigated. We elucidated the novel species, designated MBLB3053 T , through genomic analysis of novel functional microbial resources. Cells were gram-negative, non-motile, and coccoid, and the colony was light orange in color. Phylogenetic analysis based on the 16S rRNA gene showed that strain MBLB3053 T was related to the genus Paracoccus , with 98.5% similarity to Paracoccus aestuariivivens . Comparative genome analysis also revealed the strain to be a novel species of the genus Paracoccus by average nucleotide identity and in silico DNA-DNA hybridization values. Through secondary metabolite analysis, terpene biosynthetic gene clusters associated with carotenoid biosynthesis were found in strain MBLB3053 T . Using high-performance liquid chromatography, strain MBLB3053 T was confirmed to produce carotenoids, including all- trans -astaxanthin, by comparison to the standard compound. Notably, the isolate was also confirmed to produce carotenoids that other closely related species did not produce. Based on this comprehensive polyphasic taxonomy, strain MBLB3053 T represents a novel species within the genus Paracoccus , for which the name Paracoccus aurantius sp. nov is proposed. The type strain was MBL3053 T (=KCTC 8269 T =JCM 36634 T ). These findings support the research and resource value of this novel species, which was isolated from the Dokdo environmental microbiome.

The iminosugar class of carbohydrate-active enzyme inhibitors has therapeutic applications in metabolic syndrome conditions, viral infections and cancer. Compared to chemical synthesis, microbial iminosugar production has benefits of cost, sustainability and optimization. In this study, the 1-deoxynojirimycin (DNJ) biosynthetic gene cluster from Bacillus velezensis MBLB0692, and its individual genes, were cloned into Corynebacterium glutamicum (Cg). Characterizations of the encoded aminotransferase GabT1, phosphatase Yktc1, and dehydrogenase GutB1, were performed with purified enzymes and whole cell biocatalysts bearing individual and clustered (TYB) genes. GabT1 showed a variable pattern in its half-reaction with a slow turnover. GutB1 was an alkaline dehydrogenase with a broad substrate specificity and no divalent ion dependency while the zinc-dependent phosphatase Yktc1 had substrate specificity that was both pH- and ion-dependent. The CgYktc1 and CgGutB1 whole cells were viable biocatalysts with wider ranges of substrates than their enzyme counterparts. The CgTYB cells produced mannosidase-inhibiting iminosugars corresponding to mannojirimycin dehydrate (162 m/z) and deoxymannojirimycin (164 m/z). Mannosidase inhibitors have been found to be effective in treating orphan diseases, cancer and viral infections, and their biosynthesis by recombinant C. glutamicum can be optimized for industrial production and novel drug development., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

G protein-coupled receptor kinases (GRKs) are essential regulators of signaling pathways mediated by G protein-coupled receptors. Recent research suggests that GRK-mediated phosphorylation patterns dictate functional selectivity, leading to biased cellular responses. However, a comprehensive understanding of the structural mechanisms at the single-residue level remains elusive. This study aims to define the general conformational dynamics of GRKs with a particular focus on quantifying the transitions between the closed and open states. Specifically, we examined these transitions, classified based on the ionic lock between the regulatory G protein signaling homology domain and kinase domain. To facilitate a precise structural comparison, we assigned common labels to topologically identical positions across the 47 GRK structures retrieved from the Protein Data Bank. Our analysis identified both general and subfamily-specific dynamic movements within the networks and measured the conformational change scores between the two states. Elucidating these structural dynamics could provide significant insights into the regulatory mechanisms of GRK., Competing Interests: The authors declare no conflicts of interest.., (© 2024 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology.)

Carotenoids are natural pigments utilized as colourants and antioxidants across food, pharmaceutical and cosmetic industries. They exist in carbon chain lengths of C 30 , C 40 , C 45 and C 50 , with C 40 variants being the most common. Bacterioruberin (BR) and its derivatives are part of the less common C 50 carotenoid group, synthesized primarily by halophilic archaea. This study analysed the compositional characteristics of BR extract (BRE) isolated from 'Haloferax marinum' MBLA0078, a halophilic archaeon isolated from seawater near Yeoungheungdo Island in the Republic of Korea, and investigated its antioxidant activity and protective effect on lipopolysaccharide (LPS)-induced C2C12 myotube atrophy. The main components of BRE included all-trans-BR, monoanhydrobacterioruberin, 2-isopentenyl-3,4-dehydrorhodopin and all-trans-bisanhydrobacterioruberin. BRE exhibited higher antioxidant activity and DNA nicking protection activity than other well-known C 40 carotenoids, such as β-carotene, lycopene and astaxanthin. In C2C12 myotubes, LPS treatment led to a reduction in myotube diameter and number, as well as the hypertranscription of the muscle-specific ubiquitin ligase MAFbx and MuRF1. BRE mitigated these changes by activating the Akt/mTOR pathway. Furthermore, BRE abolished the elevated cellular reactive oxygen species levels and the inflammation response induced by LPS. This study demonstrated that 'Hfx. marinum' is an excellent source of natural microbial C 50 carotenoids with strong antioxidant capacity and may offer potential protective effects against muscle atrophy., (© 2024 The Author(s). Microbial Biotechnology published by John Wiley & Sons Ltd.)

Purpose: In the emergency department (ED), triage significantly impacts patient safety. Therefore, triage nurses must make decisions accurately and timeously. This study aims to investigate how South Korean pediatric emergency nurses perceive urgency and classify severity using the Q methodology, which examines individuals' subjectivity., Methods: We collected 84 statements from a Q population based on a literature review and interviews and selected 33 Q samples. The P samples included 30 pediatric emergency nurses at a Seoul tertiary care hospital. The principal component factor analysis method was used to analyze data using the PC-QUANL program., Results: Four urgency perception types were identified among pediatric ED nurses-Type 1: "Experiential coping"; Type 2: "Careful reasoning"; Type 3: "Patient-centered thinking"; and Type 4: "Intuitive prediction." These types appear to be an integrated process of knowledge and clinical experience that considers children's characteristics and developmental stages., Conclusion: This study may serve as a basis for future education to improve pediatric ED nurses' urgency judgment and severity classification skills., (Copyright © 2024. Published by Elsevier B.V.)

Insulin resistance is a common pathophysiology in patients with type 2 diabetes mellitus, cardiovascular disease, and non-alcoholic fatty liver disease. Thus, screening for the risk of insulin resistance is important to prevent disease progression. We evaluated the alanine aminotransferase/aspartate aminotransferase (ALT/AST) ratio to predict insulin resistance in the general population, regardless of comorbidities. Datasets from the 2015, 2019, and 2020 Korea National Health and Nutrition Examination Surveys were used, and the following four indices were implemented to indicate insulin resistance: fasting serum glucose, insulin, homeostatic model assessment for insulin resistance (HOMA-IR), and β-cell function. We analyzed the degree of association between the liver enzyme profile and insulin resistance indices using Pearson's correlation coefficient and determined the associations using linear or logistic regression analysis. Accordingly, ALT levels in both sexes were positively and consistently correlated with the four aforementioned insulin resistance indices in stratification analyses based on diabetes, dyslipidemia, alcohol consumption, and obesity status. In multivariate linear regression, when comparing with ALT levels, the ALT/AST ratio exhibited superior predictive performance for fasting serum glucose and HOMA-β in Korean men and improved outcomes for all insulin resistance indices in Korean women. In this analysis that included a large community-based population, the ALT/AST ratio was a more useful predictive marker than the HOMA-IR. Regarding the predicted presence or absence of insulin resistance, the ALT/AST ratio could better predict HOMA-IR than the ALT level alone in Koreans. A simple, precise marker that represents the ALT/AST ratio could be a practical method to screen for insulin resistance in the general population, regardless of diabetes mellitus, alcohol intake, and sex., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Han et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Cardiopulmonary resuscitation (CPR) is a life-saving technique used in emergencies when the heart stops beating, typically involving chest compressions and ventilation. Current adult CPR guidelines do not differentiate based on age beyond infancy and childhood. This oversight increases the risk of fatigue fractures in the elderly due to decreased bone density and changes in thoracic structure. Therefore, this study aimed to investigate the correlation and impact of factors influencing rib fatigue fractures for safer out-of-hospital manual cardiopulmonary resuscitation (OHMCPR) application. Using the finite element analysis (FEA) method, we performed fatigue analysis on rib cage models incorporating chest compression conditions and age-specific trabecular bone properties. Fatigue life analyses were conducted on three age-specific rib cage models, each differentiated by trabecular bone properties, to determine the influence of four explanatory variables (the properties of the trabecular bone (a surrogate for the age of the subject), the site of application of the compression force on the breastbone, the magnitude of applied compression force, and the rate of application of the compression force) on the fatigue life of the model. Additionally, considering the complex interaction of chest compression conditions during actual CPR, we aimed to predict rib fatigue fractures under conditions simulating real-life scenarios by analyzing the sensitivity and interrelation of chest compression conditions on the model's fatigue life. Time constraints led to the selection of optimal analysis conditions through the use of design of experiments (DOE), specifically orthogonal array testing, followed by the construction of a deep learning-based metamodel. The predicted fatigue life values of the rib cage model, obtained from the metamodel, showed the influence of the four explanatory variables on fatigue life. These results may be used to devise safer CPR guidelines, particularly for the elderly at a high risk of acute cardiac arrest, safeguarding against potential complications like fatigue fractures.

Background: Distinguishing between large-vessel diseases such as acute aortic syndrome (AAS) and pulmonary embolism (PE), and non-large-vessel diseases, such as acute coronary syndrome (ACS), heart failure (HF), and neurogenic diseases, in patients presenting with chest symptoms remains a challenge, which can result in a significant number of misdiagnoses. Simultaneously distinguishing both AAS and PE is essential because large-vessel diseases require angio-computed tomography (CT) during initial presentation whereas, non-large-vessel diseases do not. This study aimed to determine the optimal method for differentiating between large-vessel and non-large-vessel diseases using D-dimer, troponin I, and pretest probability scores., Methods: From the 11683 patients who presented with chest symptoms including chest pain, discomfort, or dyspnea, this retrospective observational study included 1817 patients who had complete data for essential biomarkers; 105 with AAS, 139 with PE, 1093 with ACS, 451 with HF, and 83 with neurogenic diseases., Results: D-dimer, D-dimer/troponin I ratio (DT ratio), and troponin I results distinguished the 2 groups: D-dimer (>2.38 μg/mL), AUC 0.935; DT ratio, AUC 0.827; and troponin I, AUC 0.653. For predicting AAS, the performances of D-dimer level and aortic dissection detection risk score (ADD-RS) were AUCs of 0.915 (p < 0.0001) and 0.67 (p = 0.0004), respectively; for predicting PE, the AUCs of D-dimer level and modified Wells score were 0.95 (p = 0.0001) and 0.857 (p < 0.0001), respectively., Conclusions: The D-dimer levels proved to be a crucial discriminator for identifying AAS and PE, even when compared with the ADD-RS and modified Wells scores. Moderately elevated D-dimer levels suggest the need to consider AAS and PE diagnoses via angio-CT for patients with chest symptoms., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

The bioconversion of 4-hydroxy-2-keto acid derivatives via aldol condensation of formaldehyde and pyruvate has received substantial attention as potential source of chemicals for production of amino acids, hydroxy carboxylic acids, and chiral aldehydes. We developed an environmentally friendly biocatalyst consisting of a novel thermostable class II pyruvate aldolase from Deinococcus radiodurans with maltose-binding protein (MBP-DrADL), which has specific activity of 46.3 µmol min -1  mg -1 . Surprisingly, MBP-DrADL maintained over 60% of enzyme activity for 4 days at 50 to 65 °C, we used MBP-DrADL as the best candidate enzyme to produce 2-keto-4-hydroxybutyrate (2-KHB) from formaldehyde and pyruvate via aldol condensation. The optimum reaction conditions for 2-KHB production were 50 °C, pH 8.0, 5 mM Mg 2+ , 100 mM formaldehyde, and 200 mM pyruvate. Under these optimized conditions, MBP-DrADL produced 76.5 mM (8.94 g L -1 ) 2-KHB over 60 min with a volumetric productivity of 8.94 g L -1  h -1 and a specific productivity of 357.6 mg mg-enzyme -1  h -1 . Furthermore, 2-KHB production was improved by continuous addition of substrates, which produced approximately 124.8 mM (14.6 g L -1 ) of 2-KHB over 60 min with a volumetric productivity and specific productivity of 14.6 g L -1  h -1 and 583.4 mg mg-enzyme -1  h -1 , respectively. MBP-DrADL showed the highest specific productivity for 2-KHB production yet reported. Our study provides a highly efficient biocatalyst for the synthesis of 2-KHB and lays the foundation for large-scale production and application of high-value compounds from formaldehyde., (© 2024. The Author(s).)

Valosin-containing protein (VCP)/p97, an AAA+ ATPase critical for maintaining proteostasis, emerges as a promising target for cancer therapy. This study reveals that targeting VCP selectively eliminates breast cancer cells while sparing non-transformed cells by inducing paraptosis, a non-apoptotic cell death mechanism characterized by endoplasmic reticulum and mitochondria dilation. Intriguingly, oncogenic HRas sensitizes non-transformed cells to VCP inhibition-mediated paraptosis. The susceptibility of cancer cells to VCP inhibition is attributed to the non-attenuation and recovery of protein synthesis under proteotoxic stress. Mechanistically, mTORC2/Akt activation and eIF3d-dependent translation contribute to translational rebound and amplification of proteotoxic stress. Furthermore, the ATF4/DDIT4 axis augments VCP inhibition-mediated paraptosis by activating Akt. Given that hyperactive Akt counteracts chemotherapeutic-induced apoptosis, VCP inhibition presents a promising therapeutic avenue to exploit Akt-associated vulnerabilities in cancer cells by triggering paraptosis while safeguarding normal cells., (© 2024. The Author(s).)

Haloarchaea produce bacterioruberin, a major C 50 carotenoid with antioxidant properties that allow for its potential application in the food, cosmetic, and pharmaceutical industries. This study aimed to optimize culture conditions for total carotenoid, predominantly comprising bacterioruberin, production using Halorubrum ruber MBLA0099. A one-factor-at-a-time and statistically-based experimental design were applied to optimize the culture conditions. Culture in the optimized medium caused an increase in total carotenoid production from 0.496 to 1.966 mg L - 1 Maximal carotenoid productivity was achieved in a 7-L laboratory-scale fermentation and represented a 6.05-fold increase (0.492 mg L -1 d -1 ). The carotenoid extracts from strain MBLA0099 exhibited a 1.8-10.3-fold higher antioxidant activity in vitro, and allowed for a higher survival rate of Caenorhabditis elegans under oxidative stress conditions. These results demonstrated that Hrr. ruber MBLA0099 has significant potential as a haloarchaon for the commercial production of bacterioruberin., (© 2023. The Author(s).)

A lipoxygenase from Pleurotus sajor-caju (PsLOX) was cloned, expressed in Escherichia coli, and purified as a soluble protein with a specific activity of 629 μmol/min/mg for arachidonic acid (AA). The native PsLOX exhibited a molecular mass of 146 kDa, including a 73-kDa homodimer, as estimated by gel-filtration chromatography. The major products converted from polyunsaturated fatty acids (PUFAs), including AA, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA), were identified as trioxilins (TrXs), namely 13,14,15-TrXB 3 , 13,14,15-TrXB 4 , and 15,16,17-TrXB 5 , respectively, through high-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses. The enzyme displayed its maximum activity at pH 8.0 and 20 °C. Under these conditions, the specific activity and catalytic efficiency of PsLOX for PUFAs exhibited the following order: AA>EPA>DHA. Based on HPLC analysis and substrate specificity, PsLOX was identified as an arachidonate 15-LOX. PsLOX efficiently converted 10 mM of AA, EPA, and DHA to 8.7 mM of 13,14,15-TrXB 3 (conversion rate: 87 %), 7.9 mM of 13,14,15-TrXB 4 (79 %), and 7.2 mM of 15,16,17-TrXB 5 (72 %) in 15, 20, and 20 min, respectively, marking the highest conversion rates reported to date. Collectively, our results demonstrate that PsLOX is an efficient TrXs-producing enzyme., (© 2023 Wiley-VCH GmbH.)

Type A acute aortic syndrome (urgent AAS, UAAS) has a low incidence and high mortality rate; however, it is often missed or diagnosed late. Our aim was to create a new tool for distinguishing UAAS by using multiple modalities to select patients for CT aortography. This study included 75 patients with UAAS, 77 with acute coronary syndrome (ACS), and 81 with heart failure (HF) who received urgent treatment after propensity matching. Specific symptoms, past medical history, mediastinal width, region of interest (ROI) ratio in the lung base/apex, D-dimers, and troponin I were investigated to differentiate UAAS from ACS and HF. The most significant variables were selected to create a new scoring system. The UAAS score exhibited a performance AUC of 0.982. A simple UAAS score >1, excluding ROI ratios in lung base/apex, showed an AUC of 0.977, a sensitivity of 96%, and specificity of 92.41%. The results were validated using an external data set of 292 patients (simple UAAS score > 1: AUC of 0.966, sensitivity 93.33%, and specificity 95.36%). The simple UAAS score may be a valuable tool for suspecting UAAS and may reduce the likelihood of misdiagnosis or performing unnecessary CT aortography.

The novel bacterial strain MBLB1776 T was isolated from marine mud in Uljin, the Republic of Korea. Cells were Gram-positive, spore-forming, non-motile, and non-flagellated rods. Growth was observed at a temperature range of 10-45 °C, pH range of 6.0-8.0, and NaCl concentrations of 0-4% ( w / v ). Phylogenetic analysis of the 16S rRNA gene sequence revealed that MBLB1776 T belonged to the genus Paenibacillus and was closely related to Paenibacillus cavernae C4-5 T (94.83% similarity). Anteiso-C 15:0 , iso-C 16:0 , C 16:0 , and iso-C 15:0 were the predominant fatty acids. Menaquinone 7 was identified as the major isoprenoid quinone. The major polar lipids included diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. Its whole genome was 6.3 Mb in size, with a G+C content of 55.8 mol%. Average nucleotide identity and in silico DNA-DNA hybridization values were below the species delineation threshold. Gene function analysis revealed the presence of a complete C 30 carotenoid biosynthetic pathway. Intriguingly, MBLB1776 T harbored carotenoid pigments, imparting an orange color to whole cells. Based on this comprehensive polyphasic taxonomy, the MBLB1776 T strain represents a novel species within the genus Paenibacillus , for which the name Paenibacillus aurantius sp. nov is proposed. The type strain was MBLB1776 T (=KCTC 43279 T = JCM 34220 T ). This is the first report of a carotenoid-producing Paenibacillus sp.

It is essential to detect pressure from a robot's fingertip in every direction to ensure efficient and secure grasping of objects with diverse shapes. Nevertheless, creating a simple-designed sensor that offers cost-effective and omnidirectional pressure sensing poses substantial difficulties. This is because it often requires more intricate mechanical solutions than when designing non-omnidirectional pressure sensors of robot fingertips. This paper introduces an innovative pressure sensor for fingertips. It utilizes a uniquely designed dynamic focusing cone to visually detect pressure with omnidirectional sensitivity. This approach enables cost-effective measurement of pressure from all sides of the fingertip. The experimental findings demonstrate the great potential of the newly introduced sensor. Its implementation is both straightforward and uncomplicated, offering high sensitivity (0.07 mm/N) in all directions and a broad pressure sensing range (up to 40 N) for robot fingertips.

Individuals with end-stage kidney disease (ESKD) on dialysis are at a high risk of developing foot ulcerations and undergoing subsequent lower extremity amputation (LEA), which can exert significant impacts on their quality of life and contribute to rising healthcare costs. We aimed to identify risk factors associated with LEA in patients with ESKD to predict LEA progression and eventually prevent it. We used 18 years (2002-2019) of data from the Korean National Health Insurance Service (KNHIS). Data were collected from patients with ESKD who underwent renal replacement therapy (RRT) and had no history of amputation caused by trauma or toxins. The risk factors were compared between patients with or without LEA. We collected data from 220,838 patients newly diagnosed with ESKD, including 6348 in the LEA group and 214,490 in the non-LEA group. The total incidence of LEA was 2.9%. Older age, male gender, lower income, non-metropolitan residence, diabetes mellitus, dialysis treatment (compared to kidney transplantation), microvascular disease, peripheral vascular disease, endovascular procedure, and endovascular operation were associated with an increased risk of LEA. Thus, individuals with ESKD who are at a higher risk for LEA should be closely monitored, and kidney transplantation should be considered as a preventative measure.

A total of 187 lactic acid bacteria were isolated from four types of grains collected in South Korea. The bacterial strains were assigned as members of Levilactobacillus brevis, Latilactobacillus curvatus, Lactiplantibacillus plantarum, Lactococcus taiwanensis, Pediococcus pentosaceus, and Weissella paramesenteroides based on the closest similarity using 16S rRNA gene sequence analysis. The strains belonging to the same species were analyzed using RAPD-PCR, and one or two among strains showing the same band pattern were selected. Finally, 25 representative strains were selected for further functional study. Inhibitory effects of lipid accumulation were observed in the strains tested. Pediococcus pentosaceus K28, Levilactobacillus brevis RP21 and Lactiplantibacillus plantarum RP12 significantly reduced lipid accumulation and did not show cytotoxicity in C3H10T1/2 cells at treatment of 1-200 μg/mL. The three LAB strains decreased significantly expression of six adipogenic marker genes, PPARγ, C/EBPα, CD36, LPL, FAS and ACC, in C3H10T1/2 adipocytes. The three strains survived under strong acidity and bile salt conditions. The three strains showed adhesion to Caco-2 cells similar to a reference strain LGG. The resistance of the three strains to several antibiotics was also assessed. Strains RP12 and K28 were confirmed not to produce harmful enzymes based on API ZYM kit results. Based on these results, strains K28, RP21 and RP12 isolated from grains had the ability to inhibit adipogenesis in adipocytes and potentially be useful as probiotics., (© 2023. The Author(s).)

Various risk factors and the multifactorial pathophysiology of cardiovascular disease (CVD) have been studied. We focused on the interaction between insulin resistance and inflammatory processes. Thus, we analyzed the association of triglyceride/high-density lipoprotein cholesterol (TG/HDL) ratio with subclinical inflammation and CVD risk in male smokers. We retrospectively reviewed medical records from the Health Promotion Center of Severance Hospital in Korea between 2015 and 2017. Subjects were currently smoking men aged 30 to 59, with leukocyte counts within the normal range, for a total of 1,566. We grouped participants into four groups using the median TG/HDL ratio and the median leukocyte count. The odds ratio (OR) of high Framingham 10-year CVD risk (≥10%) was calculated using multiple logistic regression. The median Framingham 10-year CVD risk increased significantly from Group 1 (low leukocyte count, low TG/HDL ratio) to Group 4 (high leukocyte count, high TG/HDL ratio). The OR for Group 4 was 2.46 compared with Group 1 after adjusting for various CVD risk factors. In subgroup analysis, the OR of the highest leukocyte group was 1.57 compared to the lowest leukocyte group after adjusting for other variables. In conclusion, TG/HDL ratio and subclinical inflammation were positively related to CVD risk in Korean male smokers.

Objective: This study investigated the effects of corn silage as a source of microbial inoculant containing antifungal and carboxylesterase-producing bacteria on fermentation, aerobic stability, and nutrient digestibility of fermented total mixed ration (FTMR) with different energy levels., Methods: Corn silage was used as a bacterial source by ensiling for 72 d with an inoculant mixture of Lactobacillus brevis 5M2 and L. buchneri 6M1 at a 1:1 ratio. The corn silage without or with inoculant (CON vs MIX) was mixed with the other ingredients to formulate for low and high energy diets (LOW vs HIGH) for Hanwoo steers. All diets were ensiled into 20 L mini silo (5 kg) for 40 d in quadruplicate., Results: The MIX diets had lower (p<0.05) acid detergent fiber with higher (p<0.05) in vitro digestibilities of dry matter and neutral detergent fiber compared to the CON diets. In terms of fermentation characteristics, the MIX diets had higher (p<0.05) acetate than the CON diets. The MIX diets had extended (p<0.05) lactic acid bacteria growth at 4 to 7 d of aerobic exposure and showed lower (p<0.05) yeast growth at 7 d of aerobic exposure than the CON diets. In terms of rumen fermentation, the MIX diets had higher (p<0.05) total fermentable fraction and total volatile fatty acid, with lower (p<0.05) pH than those of CON diets. The interaction (p = 0.036) between inoculant and diet level was only found in the immediately fermentable fraction, which inoculant was only effective on LOW diets., Conclusion: Application of corn silage with inoculant on FTMR presented an antifungal effect by inhibiting yeast at aerobic exposure and a carboxylesterase effect by improving nutrient digestibility. It also indicated that fermented feedstuffs could be used as microbial source for FTMR. Generally, the interaction between inoculant and diet level had less effect on this FTMR study.

One-carbon chemicals (C 1s) are potential building blocks as they are cheap, sustainable, and abiotic components. Methanol-derived formaldehyde can be another versatile building block for the production of 2-keto-4-hydroxyacid derivatives that can be used for amino acids, hydroxy carboxylic acids, and chiral aldehydes. To produce 2-keto-4-hydroxybutyrate from C 1s in an environment-friendly way, we characterized an aldolase from Pseudomonas aeruginosa PAO1 ( Pa ADL), which showed much higher catalytic activity in condensing formaldehyde and pyruvate than the reported aldolases. By applying a structure-based rational approach, we found a variant ( Pa ADL V121A/L241A ) that exhibited better catalytic activities than the wild-type enzyme. Next, we constructed a one-pot cascade biocatalyst system by combining Pa ADL and a methanol dehydrogenase (MDH) and, for the first time, effectively produced 2-keto-4-hydroxybutyrate as the main product from pyruvate and methanol via an enzymatic reaction. This simple process applied here will help design a green process for the production of 2-keto-4-hydroxyacid derivatives.

Microbial biocatalysts are evolving technological tools for glycosylation research in food, feed and pharmaceuticals. Advances in bioengineered Leloir and non-Leloir carbohydrate-active enzymes allow for whole-cell biocatalysts to curtail production costs of purified enzymes while enhancing glucan synthesis through continued enzyme expression. Unlike sugar nucleotide-dependent Leloir glycosyltransferases, non-Leloir enzymes require inexpensive sugar donors and can be designed to match the high value, yield and selectivity of the former. This review addresses the current state of bacterial cell-based production of glucans and glycoconjugates via transglycosylation, and describes how alterations made to microbial hosts to surpass purified enzymes as the preferred mode of catalysis are steadily being acquired through genetic engineering, rational design and process optimization. A comprehensive exploration of relevant literature has been summarized to describe whole-cell biocatalysis in non-Leloir glycosylation reactions with various donors and acceptors, and the characterization, application and latest developments in the optimization of their use., Competing Interests: Competing interestAuthors declare no conflicts of interest., (© The Korean Society of Food Science and Technology 2023, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

In the era of digital transformation, a memristor and memristive circuit can provide an advanced computer architecture that efficiently processes a vast quantity of data. With the unique characteristic of memristor, a memristive crossbar array has been utilized for realization of nonvolatile memory, logic-in-memory circuit, and neuromorphic system. However, the crossbar array architecture suffers from leakage of current, known as the sneak current, which causes a cross-talk interference problem between adjacent memristor devices, leading to an unavoidable operational error and high power consumption. Here, we present an amorphous In-Sn-Zn-O (a-ITZO) oxide semiconductor-based selector device to address the sneak current issue. The a-ITZO-selector device is realized with the back-to-back Schottky diode with nonlinear current-voltage (I-V) characteristics. Its nonlinearity is dependent on the oxygen plasma treatment process which can suppress the surface electron accumulation layer arising on the a-ITZO surface. The a-ITZO-selector device shows reliable characteristics against electrical stress and high temperature. In addition, the selector device allows for a stable read margin over 1 Mbit of memristor crossbar array. The findings may offer a feasible solution for the development of a high-density memristor crossbar array.

Auranofin (AF), a gold (I)-containing phosphine compound, is being investigated for oncological application as a repurposed drug. We show here that 4~5 µM AF induces paraptosis, a non-apoptotic cell death mode characterized by dilation of the endoplasmic reticulum (ER) and mitochondria, in breast cancer cells. Although the covalent inhibition of thioredoxin reductase (TrxR), an enzyme that critically controls intracellular redox homeostasis, is considered the primary mechanism of AF's anticancer activity, knockdown of TrxR1 did not induce paraptosis. Instead, both TrxR1 knockdown plus the proteasome inhibitor (PI), bortezomib (Bz), and 2 μM AF plus Bz induced paraptosis, thereby mimicking the effect of 5 μM AF. These results suggest that the paraptosis induced by 5 μM AF requires the inhibition of both TrxR1 and proteasome. We found that TrxR1 knockdown/Bz or subtoxic doses of AF and Bz induced paraptosis selectively in breast cancer cells, sparing non-transformed MCF10A cells, whereas 4~5 μM AF killed both cancer and MCF10A cells. GSH depletion was found to be more critical than ROS generation for the paraptosis induced by dual TrxR1/proteasome inhibition. In this process, the ATF4/CHAC1 (glutathione-specific gamma-glutamylcyclotransferase 1) axis leads to GSH degradation, contributing to proteotoxic stress possibly due to the accumulation of misfolded thiol-containing proteins. These results suggest that the paraptosis-inducing strategy of AF plus a PI may provide an effective therapeutic strategy against pro-apoptotic therapy-resistant cancers and reduce the potential side effects associated with high-dose AF., (© 2023. The Author(s).)

Background: Febrile seizure (FS) is one of the most common neurological manifestations of coronavirus disease-2019 (COVID-19) in children. We compared the clinical characteristics of FS in patients with and without COVID-19 during the pandemic period., Methods: This retrospective single-center study included patients aged 0-18 years who visited the pediatric emergency department (ED) with FS from January 1, 2022, to April 30, 2022., Results: A total of 186 patients visited the pediatric ED with FS during the study period: 123 (66.1%) were positive for COVID-19 and 63 (33.9%) were negative. Patients with COVID-19 were predominantly male (70.7% vs. 50.8%, p = 0.007) and older (2.4 vs. 1.8 years, p = 0.005) than those without COVID-19. A higher proportion of patients with COVID-19 were of atypical age (age > 5 years or <6 months) than those without COVID-19 (26.8% vs. 9.5%, p = 0.006). This was especially true for those aged >5 years (22% vs. 4.8%, p = 0.003). Patients with COVID-19 had a higher probability of multiple episodes of convulsion within 24 h than those without COVID-19 (10.6% vs. 1.6%, p = 0.037). Among patients with COVID-19, males had a shorter fever-to-seizure duration than females (3 h vs. 6.5 h, p = 0.045)., Conclusions: Patients with FS with COVID-19 tend to be predominantly male and have older age of onset than those without COVID-19. Because of the atypical age of onset and probability of multiple convulsion episodes, vigilance for FS is needed in patients with COVID-19, especially males., (© 2023 Japan Pediatric Society.)

Carotenoids, a group of isoprenoid pigments, are naturally synthesized by various microorganisms and plants, and are industrially used as ingredients in food, cosmetic, and pharmaceutical product formulations. Although several types of carotenoids and diverse microbial carotenoid producers have been reported, studies on lactic acid bacteria (LAB)-derived carotenoids are relatively insufficient. There is a notable lack of research focusing on C 30 carotenoids, the functional characterizations of their biosynthetic genes and their mass production by genetically engineered microorganisms. In this study, the biosynthesis of 4,4'-diaponeurosporene in Escherichia coli harboring the core biosynthetic genes, dehydrosqualene synthase (crtM) and dehydrosqualene desaturase (crtN), from Lactiplantibacillus plantarum subsp. plantarum KCCP11226 was constructed to evaluate and enhance 4,4'-diaponeurosporene production and antioxidant activity. The production of 4,4'-diapophytoene, a substrate of 4,4'-diaponeurosporene, was confirmed in E. coli expressing only the crtM gene. In addition, recombinant E. coli carrying both C 30 carotenoid biosynthesis genes (crtM and crtN) was confirmed to biosynthesize 4,4'-diaponeurosporene and exhibited a 6.1-fold increase in carotenoid production compared to the wild type and had a significantly higher antioxidant activity compared to synthetic antioxidant, butylated hydroxytoluene. This study presents the discovery of an important novel E. coli platform in consideration of the industrial applicability of carotenoids., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

This study focuses on the development of functional probiotics using caroteonid-producing lactic acid bacteria (LAB) with antioxidant properties. Thirty LAB strains were evaluated for their probiotic properties. Carotenoid biosynthesis gene cluster ( crtMN operon) was detected using polymer chain reaction (PCR). The carotenoid identified as 4,4'-diaponeurosporene was analyzed via UV visible absorption spectra and HPLC. Five carotenoid-producing strains showed antioxidant activities. Lactiplantibacillus plantarum MGB0112, which showed the highest carotenoid production measuring at 470 nm of absorbance per ml of culture broth (0.014 A 470nm /ml), showed low pH (56.5%) and bile salt (97.8%) tolerance with high adhesion properties (55.1% for toluene). Furthermore, this strain and 4,4'-diaponeurosporene extract exhibited antioxidant activity (99.5 and 40.1%, respectively) against DPPH free radicals in vitro. Their antioxidant properties were confirmed in vivo (45.6 and 55.2% survival rates in Caenorhabditis elegans ). Therefore, C 30 carotenoid-producing strain MGB0112 demonstrates outstanding antioxidant effects and can be a potential functional probiotics., Competing Interests: Conflict of interestThe authors declare that there is no conflict of interest., (© The Korean Society of Food Science and Technology 2022, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Context: Zeaxanthin is a yellow‑coloured dietary carotenoid widely recognized as an essential component of the macula. It exerts blue light filtering and antioxidant activities, offering eye health and vision benefits., Objective: This study explores the oral absorption and systemic disposition of zeaxanthin from biopharmaceutical and pharmacokinetic perspectives., Materials and Methods: In vivo intravenous (5 and 10 mg/kg) and intraportal (5 mg/kg) pharmacokinetic studies were performed to determine intrinsic tissue‑blood partition coefficient, elimination pathway, and hepatic clearance, of zeaxanthin in rats. Moreover, in vitro physicochemical property test, in situ closed loop study, in vivo oral pharmacokinetic study (20 and 100 mg/kg), and in vivo lymphatic absorption study (100 mg/kg) were conducted to investigate the gut absorption properties of zeaxanthin and assess the effects of several lipids on the lymphatic absorption of zeaxanthin in rats., Results: Zeaxanthin exhibited poor solubility (≤144 ng/mL) and stability (6.0-76.9% of the initial amount remained at 24 h) in simulated gut luminal fluids. Gut absorption of zeaxanthin occurred primarily in the duodenum, but the major fraction (≥84.7%) of the dose remained unabsorbed across the entire gut tract. Considerable fractions of intravenous zeaxanthin accumulated in the liver, lung, and spleen (21.3, 11.7, and 2.0%, respectively). It was found that the liver is the major eliminating organ of zeaxanthin, accounting for 53.5-90.1% of the total clearance process (hepatic extraction ratio of 0.623)., Discussion and Conclusions: To our knowledge, this is the first systematic study to report factors that determine the oral bioavailability and systemic clearance of zeaxanthin.

A novel haloarchaeal species designated as MBLA0099 T was isolated from seawater near Yeongheung Island. Cells were Gram-negative, non-motile, red-pigmented, and rod-shaped. They grew at 10-45°C, within pH 5.5-9.0, and between 7.5% and 30% NaCl concentrations. Cells were able to grow without Mg 2+ and were lysed in distilled water. The size of the whole-genome and G + C content of DNA was 3.02 Mb and 68.9 mol%, respectively. Phylogenetic analysis shows that the strain MBLA0099 T belongs to the genus Halorubrum. The average nucleotide and amino acid identity, and in silico DNA-DNA hybridization values were below the species delineation threshold. Pan-genomic analysis revealed that 3.2% of all genes present in strain MBLA0099 T were unique to the strain. The red carotenoid produced by strain MBLA0099 T was subjected to spectrometric and chromatographic analyses and confirmed to be bacterioruberin as C50 carotenoid. Mevalonic acid, terpenoid backbone, and carotenoid biosynthesis pathway were annotated for strain MBLA0099 T . The C50 carotenoid production by strain MBLA0099 T was also enhanced under various stress conditions including relatively netural pH, high oxidative and salinity conditions. Additionally, the strain MBLA0099 T -derived bacterioruberin showed the antioxidant activity with EC 50 value of 12.29 µg/ml, based on the evaluation of DPPH free radical scavenging activity. The present study would be the first report on the identification of C 50 carotenoid from the strain MBLA0099 T representing a novel species of the genus Halorubrum, for which the name Halorubrum ruber sp. nov. is proposed. The typestrain used was MBLA0099 T (= KCTC 4296 T = JCM 34701 T )., (© 2022. Author(s).)

Carotenoids are lipophilic tetraterpenoid pigments produced by plants, algae, arthropods, and certain bacteria and fungi. These biologically active compounds are used in the food, feed, and nutraceutical industries for their coloring and the physiological benefits imparted by their antioxidant properties. The current global carotenoid market is dominated by synthetic carotenoids; however, the rising consumer demand for natural products has led to increasing research and development in the mass production of carotenoids from alternative natural sources, including microbial synthesis and plant extraction, which holds a significant market share. To date, microbial research has focused on C 40 carotenoids, but studies have shown that C 30 carotenoids contain similar-and in some microbial strains, greater-antioxidant activity in both the physical and chemical quenching of reactive oxygen species. The discovery of carotenoid biosynthetic pathways in different microorganisms and advances in metabolic engineering are driving the discovery of novel C 30 carotenoid compounds. This review highlights the C 30 carotenoids from microbial sources, showcasing their antioxidant properties and the technologies emerging for their enhanced production. Industrial applications and tactics, as well as biotechnological strategies for their optimized synthesis, are also discussed.

A Gram-negative, aerobic, non-motile, and rod-shaped bacterial strain, designated BSSL-BM10 T , was isolated from sand of a dune that was collected from the Yellow Sea, Republic of Korea. It was subjected to a polyphasic taxonomic study. 16S rRNA gene sequence analysis showed that strain BSSL-BM10 T fell phylogenetically within the radiation comprising type strains of Devosia species. The 16S rRNA gene sequence of strain BSSL-BM10 T shared sequence similarities of 98.2% with the type strain of D. naphthalenivorans and 93.5-97.7% with type strains of other Devosia species. ANI and dDDH values between strain BSSL-BM10 T and type strains of 18 Devosia species were 71.0-78.4% and 18.8-21.5%, respectively. The DNA G + C content of strain BSSL-BM10 T was 60.9% based on its genomic sequence data. Strain BSSL-BM10 T contained Q-10 as the predominant ubiquinone and 11-methyl C 18:1 ω7c, C 18:1 ω7c, summed feature 3 (C 16:1 ω7c and/or C 16:1 ω6c), and C 16:0 as its major fatty acids. Major polar lipids of strain BSSL-BM10 T were phosphatidylglycerol and two unidentified glycolipids. Strain BSSL-BM10 T showed distinguishable phenotypic properties with its phylogenetic and genetic distinctiveness separated from recognized Devosia species. Based on data presented in this study, strain BSSL-BM10 T should be placed in the genus Devosia. The name Devosia litorisediminis sp. nov. is proposed for strain BSSL-BM10 T (= KACC 21633 T  = NBRC 115152 T )., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: This study using multi-center health examination data from Korean adults was conducted to confirm changes in weight, and their related cardiometabolic parameters, before and after strengthening of social distancing regulations., Methods: A retrospective cohort study was conducted using health check-up data from 13 university hospitals. The study period was from January 2018 to July 2020. To examine the effect of systematic social distancing measures, participants who underwent a health check-up (Visit 3) between July 2020 and July 2021 (during full scale social distancing), and had undergone two previous health check-ups (Visits 1 and 2) between January 2018 and June 2020 (before social distancing), were selected. In total, data from 7,875 participants were analyzed. Linear mixed-effect models were used to calculate estimates of anthropometric indices and metabolic markers measured on Visits 2 and 3, compared with measurements from Visit 1., Results: There were no significant differences in body weight, body mass index, waist circumference, and body composition on Visit 3 than on Visits 1 and 2. However, the odds of metabolic syndrome and its components, including hypertension, high glucose, diabetes, hypercholesterolemia, hypertriglyceridemia, hyper-non-high-density lipoprotein cholesterolemia, and dyslipidemia were significantly higher on Visit 3 than on Visits 1 and 2. The increase in metabolic complications was marked, particularly in relatively young adults who visited health check-up centers located in the capital area., Conclusion: Metabolic syndrome and its components were significantly worse after high level social distancing, although there were no significant increases in anthropometric indices and body fat levels. Healthcare providers need to prevent and manage worsening of metabolic parameters in subpopulations prone to be more sedentary and eat unhealthy food during the COVID-19 pandemic and associated social distancing measures., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ko, Cho, Kim, Kang, Kim, Haam, Hur, Choi, Lee, Park, Cho, Kim, Lee, Seo, Yoon, Seo, Nam and Kim.)

Recombinant Escherichia coli cells expressing 8,11-linoleate diol synthase (LDS) from Penicillium chrysogenum convert oleic and palmitoleic acids to 8-hydroperoxy-9(Z)-octadecenoic acid (HPOME) and 8-hydroperoxy-9(Z)-hexadecenoic acid (HPHME) only, respectively. However, recombinant E. coli cells expressing Q889A variant 6,8-LDS from Penicillium oxalicum as an 8,11-LDS converted oleic and palmitoleic acids to 8,11-dihydroxy-9(Z)-octadecenoic acid (DiHOME) and 8,11-dihydroxy-9(Z)-hexadecenoic acid (DiHHME), respectively, which were identified using liquid chromatography-tandem mass spectrometry analysis. To select suitable variants for producing these compounds, position 889 of 6,8-LDS from P. oxalicum was substituted with other amino acids, and recombinant E. coli cells expressing Q889L and Q889A variants were chosen as the best biocatalysts for producing 8,11-DiHOME and 8,11-DiHHME, respectively. The optimal conditions for producing 8,11-DiHOME or 8,11-DiHHME using cells expressing Q889L or Q889A variant 6,8-LDS were pH 6.5 and 30 °C with 5% (v/v) dimethyl sulfoxide, 60 g L -1 cells, and 10 g L -1 oleic acid or 7.5 g L -1 palmitoleic acid, respectively. Under these conditions, 10.7 g L -1 8,11-DiHOME and 8.1 g L -1 8,11-DiHHME were produced for 1.5 h with molar yields of 96.4% and 96.2% and productivities of 7.1 and 5.4 g L -1  h -1 , respectively. The molar yields and concentrations of 8,11-DiHOME and 8,11-DiHHME were highest among those of other reported DiHOMEs and DiHHMEs. To the best of our knowledge, this is the first quantitative production of 8,11-DiHOME and 8,11-DiHHME., (© 2022 American Institute of Chemical Engineers.)

In response to the COVID-19 pandemic, the Korean government implemented policies including the systematic social distancing (SSD) system which started on 28 June 2020. The present study investigated the development and aggravation of fatty liver measured using ultrasonography during the transition period (from pre-SSD to SSD) compared to the fatty liver changes during the pre-SSD period. Changes in fatty liver and liver enzymes were assessed in different groups stratified by alcohol consumption. Our retrospective cohort analysis included 5668 participants who underwent health checkups at 13 university hospitals during the SSD period and two or more checkups before the SSD period. Fatty liver developed and aggravated more in the transition period (13.6% development and 12.0% aggravation) than in the pre-SSD period (10.8% development and 10.1% aggravation) in the alcohol consumption group. This finding was more prominent in women than in men. Abnormal alanine transaminase levels were more often developed in the transition period than in the pre-SSD period, especially in men (11.1% vs. 8.6% in each period). In conclusion, the SSD system may contribute to fatty liver changes in individuals who regularly consume alcohol. Further research of the post-pandemic period is needed to assess long-term changes in fatty liver disease.

Fluorescent protein-DNA-binding peptides or proteins (FP-DBP) are a powerful means to stain and visualize large DNA molecules on a fluorescence microscope. Here, we constructed 21 kinds of FP-DBPs using various colors of fluorescent proteins and two DNA-binding motifs. From the database of fluorescent proteins (FPbase.org), we chose bright FPs, such as RRvT, tdTomato, mNeonGreen, mClover3, YPet, and mScarlet, which are four to eight times brighter than original wild-type GFP. Additionally, we chose other FPs, such as mOrange2, Emerald, mTurquoise2, mStrawberry, and mCherry, for variations in emitting wavelengths. For DNA-binding motifs, we used HMG (high mobility group) as an 11-mer peptide or a 36 kDa tTALE (truncated transcription activator-like effector). Using 21 FP-DBPs, we attempted to stain DNA molecules and then analyzed fluorescence intensities. Most FP-DBPs successfully visualized DNA molecules. Even with the same DNA-binding motif, the order of FP and DBP affected DNA staining in terms of brightness and DNA stretching. The DNA staining pattern by FP-DBPs was also affected by the FP types. The data from 21 FP-DBPs provided a guideline to develop novel DNA-binding fluorescent proteins.

An isolate, designated MBLB2552 T , was isolated from the gut of the honey bees (Apis mellifera L.) and identified as a member of the genus Paenibacillus based on the sequences of the 16S rRNA gene sequences. The most closely related species to strain MBLB2552 T were Paenibacillus timonensis 2301032  T , Paenibacillus barengoltzii NBRC 101215  T , and Paenibacillus macerans IAM 12467  T , with similarity values of 98.1, 97.21 and 97.0%, respectively, based on 16S rRNA gene sequences. The genome size and G + C content of MBLB2552 T were 5.2 Mb and 52.4%, respectively. The Ortho average nucleotide identity (OrthoANI) and in silico DNA-DNA hybridization (isDDH) values between strain MBLB2552 T and the type strains of the closest species were below the species delineation threshold. Comparative genomic analysis showed that most core POGs of strain MBLB2552 T and other related taxa were related to translation, ribosomal structure and biogenesis (J) and carbohydrate metabolism in the COG category and KEGG pathways, respectively. Strain MBLB2552 T was Gram stain-positive, spore-forming, rod-shaped, facultative anaerobic, motile, and grew at 20‒45 °C in 0‒2% (w/v) NaCl at pH 6.0‒9.0. The major polar lipids identified were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylserine, unidentified polar lipids, and an unidentified glycolipid. We propose that strain MBLB2552 T represents the type strain of the genus Paenibacillus and its name Paenibacillus mellifer sp. nov. is proposed. The type of strain was MBLB2552 T (= JCM 35371  T  = KCTC 43386  T )., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The rising demand for carotenoids can be met by microbial biosynthesis as a promising alternative to chemical synthesis and plant extraction. Several species of lactic acid bacteria (LAB) specifically produce C 30 carotenoids and offer the added probiotic benefit of improved gut health and protection against chronic conditions. In this study, the recently characterized Lactiplantibacillus plantarum subsp. plantarum KCCP11226 T produced the rare C 30 carotenoid, 4,4'-diaponeurosporene, and its yield was optimized for industrial production. The one-factor-at-a-time (OFAT) method was used to screen carbon and nitrogen sources, while the abiotic stresses of temperature, pH, and salinity, were evaluated for their effects on 4,4'-diaponeurosporene production. Lactose and beef extract were ideal for optimal carotenoid production at 25°C incubation in pH 7.0 medium with no salt. The main factors influencing 4,4'-diaponeurosporene yields, namely lactose level, beef extract concentration and initial pH, were enhanced using the Box-Behnken design under response surface methodology (RSM). Compared to commercial MRS medium, there was a 3.3-fold increase in carotenoid production in the optimized conditions of 15% lactose, 8.3% beef extract and initial pH of 6.9, producing a 4,4'-diaponeurosporene concentration of 0.033 A 470 /ml. To substantiate upscaling for industrial application, the optimal aeration rate in a 5 L fermentor was 0.3 vvm. This resulted in a further 3.8-fold increase in 4,4'-diaponeurosporene production, with a concentration of 0.042 A 470 /ml, compared to the flask-scale cultivation in commercial MRS medium. The present work confirms the optimization and scale-up feasibility of enhanced 4,4'-diaponeurosporene production by L. plantarum subsp. plantarum KCCP11226 T .

An isolate, designated strain KIGAM418 T was isolated from marine mud below 192 m depth in the Hupo Basin, Republic of Korea. Strain KIGAM418 T was Gram-stain positive, spore-forming, rod-shaped, facultatively anaerobic, and grew at 10‒45 °C, in 0‒2% (w/v) NaCl at pH 4.0‒12.0. The strain tested positive for catalase, oxidase, and motility. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain KIGAM418 T was related to the genus Fictibacillus. The strain showed the highest similarity to Fictibacillus rigui WPCB074 T (98.0-98.1%) and Fictibacillus solisalsi YC1 T (97.2-97.8%). The diagnostic diamino acid of the cell wall was meso-diaminopimelic acid. The major fatty acids were characterized as anteiso-C 15:0 and iso-C 15:0 . Strain KIGAM418 T possessed diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine as the major polar lipids and menaquinone-7 as the predominant menaquinone. The genome size and G + C content were 4.56 Mb and 43.2 mol%, respectively. According to predicted functional genes of the genome, the category of amino acid transport and metabolism was mainly distributed. Based on the polyphasic taxonomic data, strain KIGAM418 T represents a novel species of the genus Fictibacillus, for which the name Fictibacillus marinisediminis sp. nov. is proposed. The type strain is KIGAM418 T (= KCTC 43291  T  = JCM 34437  T )., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

β-Carotene 15,15'-oxygenase (BCO1) and β-carotene 9',10'-oxygenase (BCO2) are potential producers of vitamin A derivatives, since they can catalyze the oxidative cleavage of dietary provitamin A carotenoids to retinoids and derivative such as apocarotenal. Retinoids are a class of chemical compounds that are vitamers of vitamin A or are chemically related to it, and are essential nutrients for humans and highly valuable in the food and cosmetics industries. β-carotene oxygenases (BCOs) from various organisms have been overexpressed in heterogeneous bacteria, such as Escherichia coli , and their biochemical properties have been studied. For the industrial production of retinal, there is a need for increased production of a retinal producer and biosynthesis of retinal using biocatalyst systems improved by enzyme engineering. The current review aims to discuss BCOs from animal, plants, and bacteria, and to elaborate on the recent progress in our understanding of their functions, biochemical properties, substrate specificity, and enzyme activities with respect to the production of retinoids in whole-cell conditions. Moreover, we specifically propose ways to integrate BCOs into retinal biosynthetic bacterial systems to improve the performance of retinal production.

The newly isolated strain KIGAM252 T was found to be facultatively anaerobic, Gram-stain-positive, spore-forming, and rod-shaped. They grew at 10-45 °C, pH 6.0-10.0, and were able to tolerate up to 6% NaCl in the growth medium. Phylogenetic analysis indicated that the KIGAM252 T strain was related to the genus Metabacillus . The cell membrane fatty acid composition of strain KIGAM252 T included C 15:0 anteiso and C 15:0 iso (25.6%) as the major fatty acids, and menaquinone 7 was the predominant isoprenoid quinone. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The size of the whole genome was 4.30 Mbp, and the G + C content of the DNA was 43.8%. Average nucleotide and amino acid identity and in silico DNA-DNA hybridization values were below the species delineation threshold. Pan-genomic analysis revealed that 15.8% of all genes present in strain KIGAM252 T was unique to the strain. The analysis of the secondary biosynthetic pathway predicted the carotenoid synthetic gene cluster in the strain KIGAM252 T . Based on these current polyphasic taxonomic data, strain KIGAM252 T represents a novel species of the genus Metabacillus that produces carotenoids, for which we propose the name Metabacillus flavus sp. nov. The type of strain was KIGAM252 T (=KCTC 43261 T = JCM 34406 T ).

Background: Smad4 and p53 mutations are the most common mutations in human colorectal cancers (CRCs). We evaluated whether and how they are synergistic in intestinal carcinogenesis using novel autochthonous mouse models., Method: To recapitulate human CRCs, we generated Villin-Cre;Smad4 F / F ;Trp53 F / F mice. We then compared the intestinal phenotype of Villin-Cre;Smad4 F / F ;Trp53 F / F mice (n = 40) with Villin-Cre;Smad4 F / F (n = 30) and Villin-Cre;Trp53 F / F mice (n = 45)., Results: Twenty-week-old Villin-Cre;Smad4 F / F ;Trp53 F / F mice displayed spontaneous highly proliferative intestinal tumors, and 85% of mice developed adenocarcinomas. p21 was downregulated in the intestinal mucosa in Villin-Cre;Smad4 F / F ;Trp53 F / F mice than in Villin-Cre;Smad4 F / F and Villin-Cre;Trp53 F / F mice. Villin-Cre;Smad4 F / F ;Trp53 F / F mice displayed multistep intestinal tumorigenesis and Wnt activation. Long-term CWP232291 (small-molecule Wnt inhibitor) treatment of Villin-Cre;Smad4 F / F ;Trp53 F / F mice suppressed intestinal tumorigenesis and progression. CWP232291 treatment downregulated cancer stem cell (CSC) tumor markers including CD133, Lgr-5, and Sca-1. CWP232291 treatment reduced the CSC frequency. Small-molecule Wnt inhibitors reduced intestinal CSC populations and inhibited their growth, along with Bcl-X L downregulation. Furthermore, BH3I-1, a Bcl-X L antagonist, increasingly inhibited intestinal CSCs than bulk tumor cells., Conclusion: Smad4 loss and p53 loss are synergistic in autochthonous intestinal carcinogenesis, by downregulating p21 and activating Wnt/β-catenin pathway., (© 2022 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)

Polymerase chain reaction (PCR) is a powerful tool to diagnose infectious diseases. Uracil DNA glycosylase (UDG) is broadly used to remove carryover contamination in PCR. However, UDG can contribute to false negative results when not inactivated completely, leading to DNA degradation during the amplification step. In this study, we designed novel thermolabile UDG derivatives by supercomputing molecular dynamic simulations and residual network analysis. Based on enzyme activity analysis, thermolability, thermal stability, and biochemical experiments of Escherichia coli-derived UDG and 22 derivatives, we uncovered that the UDG D43A mutant eliminated the false negative problem, demonstrated high efficiency, and offered great benefit for use in PCR diagnosis. We further obtained structural and thermodynamic insights into the role of the D43A mutation, including perturbed protein structure near D43; weakened pairwise interactions of D43 with K42, N46, and R80; and decreased melting temperature and native fraction of the UDG D43A mutant compared with wild-type UDG., (Copyright © 2022 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

One- or two-carbon (C1 or C2) compounds have been considered attractive substrates because they are inexpensive and abundant. Methanol and ethanol are representative C1 and C2 compounds, which can be used as bio-renewable platform feedstocks for the biotechnological production of value-added natural chemicals. Methanol-derived formaldehyde and ethanol-derived acetaldehyde can be converted to 3-hydroxypropanal (3-HPA) via aldol condensation. 3-HPA is used in food preservation and as a precursor for 3-hydroxypropionic acid and 1,3-propanediol that are starting materials for manufacturing biocompatible plastic and polytrimethylene terephthalate. In this study, 3-HPA was biosynthesized from formaldehyde and acetaldehyde using deoxyribose-5-phosphate aldolase from Thermotoga maritima (DERA Tma ) and cloned and expressed in Escherichia coli for 3-HPA production. Under optimum conditions, DERA Tma produced 7 mM 3-HPA from 25 mM substrate (formaldehyde and acetaldehyde) for 60 min with 520 mg/L/h productivity. To demonstrate the one-pot 3-HPA production from methanol and ethanol, we used methanol dehydrogenase from Lysinibacillus xylanilyticus (MDH Lx ) and DERA Tma . One-pot 3-HPA production via aldol condensation of formaldehyde and acetaldehyde from methanol and ethanol, respectively, was investigated under optimized reaction conditions. This is the first report on 3-HPA production from inexpensive alcohol substrates (methanol and ethanol) by cascade reaction using DERA Tma and MDH Lx .

Despite the success of proteasome inhibitors (PIs) in treating hematopoietic malignancies, including multiple myeloma (MM), their clinical efficacy is limited in solid tumors. In this study, we investigated the involvement of the integrated stress response (ISR), a central cellular adaptive program that responds to proteostatic defects by tuning protein synthesis rates, in determining the fates of cells treated with PI, bortezomib (Bz). We found that Bz induces ISR, and this can be reversed by ISRIB, a small molecule that restores eIF2B-mediated translation during ISR, in both Bz-sensitive MM cells and Bz-insensitive breast cancer cells. Interestingly, while ISRIB protected MM cells from Bz-induced apoptosis, it enhanced Bz sensitivity in breast cancer cells by inducing paraptosis, the cell death mode that is accompanied by dilation of the endoplasmic reticulum (ER) and mitochondria. Combined treatment with ISRIB and Bz may shift the fate of Bz-insensitive cancer cells toward paraptosis by inducing translational rescue, leading to irresolvable proteotoxic stress., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Human gut-originated lactic acid bacteria were cultivated, and high γ-aminobutyric acid (GABA)-producing Lactococcus garvieae MJF010 was identified. To date, despite the importance of GABA, no studies have investigated GABA-producing Lactococcus species, except for Lc. lactis. A recombinant glutamate decarboxylase of the strain MJF010 (rLgGad) was successfully expressed in Escherichia coli BL21(DE3) with a size of 53.9 kDa. rLgGad could produce GABA, which was verified using the silylation-derivative fragment ions of GABA. The purified rLgGad showed the highest GABA-producing activity at 35 °C and pH 5. rLgGad showed a melting temperature of 43.84 °C. At 30 °C, more than 80% of the activity was maintained even after 7 h; however, it rapidly decreased at 50 °C. The kinetic parameters, K m , V max , and k cat , of rLgGad were 2.94 mM, 0.023 mM/min, and 12.3 min - 1 , respectively. The metal reagents of CaCl 2 , MgCl 2 , and ZnCl 2 significantly had positive effects on rLgGad activity. However, most coenzymes including pyridoxal 5'-phosphate showed no significant effects on enzyme activity. In conclusion, this is the first report of Gad from Lc. garvieae species and provides important enzymatic information related to GABA biosynthesis in the Lactococcus genus., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

PSMD14, a subunit of the 19S regulatory particles of the 26S proteasome, was recently identified as a potential prognostic marker and therapeutic target in diverse human cancers. Here, we show that the silencing and pharmacological blockade of PSMD14 in MDA-MB 435S breast cancer cells induce paraptosis, a non-apoptotic cell death mode characterized by extensive vacuolation derived from the endoplasmic reticulum (ER) and mitochondria. The PSMD14 inhibitor, capzimin (CZM), inhibits proteasome activity but differs from the 20S proteasome subunit-inhibiting bortezomib (Bz) in that it does not induce aggresome formation or Nrf1 upregulation, which underlie Bz resistance in cancer cells. In addition to proteasome inhibition, the release of Ca 2+ from the ER into the cytosol critically contributes to CZM-induced paraptosis. Induction of paraptosis by targeting PSMD14 may provide an attractive therapeutic strategy against cancer cells resistant to proteasome inhibitors or pro-apoptotic drugs.