Publications by authors named "Jung-Hyun Kim"

491 Publications

Abundance and diversity of antibiotic resistance genes and bacterial communities in the western Pacific and Southern Oceans.

Sci Total Environ 2022 Jan 24:153360. Epub 2022 Jan 24.

Korea Polar Research Institute, 26, Songdomirae-ro, Yeonsu-gu, Incheon 21990, South Korea.

This study investigated the abundance and diversity of antibiotic resistance genes (ARGs) and the composition of bacterial communities along a transect covering the western Pacific Ocean (36°N) to the Southern Ocean (74°S) using the Korean icebreaker R/V Araon (total cruise distance: 14,942 km). The relative abundances of ARGs and bacteria were assessed with quantitative PCR and next generation sequencing, respectively. The absolute abundance of ARGs was 3.0 × 10 ± 1.6 × 10 copies/mL in the western Pacific Ocean, with the highest value (7.8 × 10 copies/mL) recorded at a station in the Tasman Sea (37°S). The absolute abundance of ARGs in the Southern Ocean was 1.8-fold lower than that in the western Pacific Ocean, and slightly increased (0.7-fold) toward Terra Nova Bay in Antarctica, possibly resulting from natural terrestrial sources or human activity. β-Lactam and tetracycline resistance genes were dominant in all samples (88-99%), indicating that they are likely the key ARGs in the ocean. Correlation and network analysis showed that Bdellovibrionota, Bacteroidetes, Cyanobacteria, Margulisbacteria, and Proteobacteria were positively correlated with ARGs, suggesting that these bacteria are the most likely ARG carriers. This study highlights the latitudinal profile of ARG distribution in the open ocean system and provides insights that will help in monitoring emerging pollutants on a global scale.
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http://dx.doi.org/10.1016/j.scitotenv.2022.153360DOI Listing
January 2022

Growth Properties of Carbon Nanowalls on Nickel and Titanium Interlayers.

Molecules 2022 Jan 9;27(2). Epub 2022 Jan 9.

Department of Electrical Engineering, Hanbat National University, Daejeon 34158, Korea.

This research is conducted in order to investigate the structural and electrical characteristics of carbon nanowalls (CNWs) according to the sputtering time of interlayers. The thin films were deposited through RF magnetron sputtering with a 4-inch target (Ni and Ti) on the glass substrates, and the growth times of the deposition were 5, 10, and 30 min. Then, a microwave plasma-enhanced chemical vapor deposition (PECVD) system was used to grow CNWs on the interlayer-coated glass substrates by using a mixture of H and CH gases. The FE-SEM analysis of the cross-sectional and planar images confirmed that the thickness of interlayers linearly increased according to the deposition time. Furthermore, CNWs grown on the Ni interlayer were taller and denser than those grown on the Ti interlayer. Hall measurement applied to measure sheet resistance and conductivity confirmed that the electrical efficiency improved significantly as the Ni or Ti interlayers were used. Additionally, UV-Vis spectroscopy was also used to analyze the variations in light transmittance; CNWs synthesized on Ni-coated glass have lower average transmittance than those synthesized on Ti-coated glass. Based on this experiment, it was found that the direct growth of CNW was possible on the metal layer and the CNWs synthesized on Ni interlayers showed outstanding structural and electrical characterizations than the remaining interlayer type.
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http://dx.doi.org/10.3390/molecules27020406DOI Listing
January 2022

10-DEBC Hydrochloride as a Promising New Agent against Infection of .

Int J Mol Sci 2022 Jan 6;23(2). Epub 2022 Jan 6.

Center for Clinical Research, Masan National Tuberculosis Hospital, Changwon 51755, Korea.

() causes chronic pulmonary infections. Its resistance to current antimicrobial drugs makes it the most difficult non-tuberculous mycobacteria (NTM) to treat with a treatment success rate of 45.6%. Therefore, there is a need for new therapeutic agents against . We identified 10-DEBC hydrochloride (10-DEBC), a selective AKT inhibitor that exhibits inhibitory activity against . To evaluate the potential of 10-DEBC as a treatment for lung disease caused by , we measured its effectiveness in vitro. We established the intracellular activity of 10-DEBC against in human macrophages and human embryonic cell-derived macrophages (iMACs). 10-DEBC significantly inhibited the growth of wild-type and clinical isolates and clarithromycin (CLR)-resistant strains. 10-DEBC's drug efficacy did not have cytotoxicity in the infected macrophages. In addition, 10-DEBC operates under anaerobic conditions without replication as well as in the presence of biofilms. The alternative caseum binding assay is a unique tool for evaluating drug efficacy against slow and nonreplicating bacilli in their native caseum media. In the surrogate caseum, the mean undiluted fraction unbound () for 10-DEBC is 5.696. The results of an in vitro study on the activity of suggest that 10-DEBC is a potential new drug for treating infections.
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http://dx.doi.org/10.3390/ijms23020591DOI Listing
January 2022

Proliferation of Mouse Prostate Cancer Cells Inflamed by Trichomonas vaginalis.

Korean J Parasitol 2021 Dec 22;59(6):547-556. Epub 2021 Dec 22.

Department of Environmental Biology and Medical Parasitology, Hanyang University College of Medicine, Seoul 04763, Korea.

Our objective was to investigate whether inflammatory microenvironment induced by Trichomonas vaginalis infection can stimulate proliferation of prostate cancer (PCa) cells in vitro and in vivo mouse experiments. The production of CXCL1 and CCL2 increased when cells of the mouse PCa cells (TRAMP-C2 cell line) were infected with live T. vaginalis. T. vaginalis-conditioned medium (TCM) prepared from co-culture of PCa cells and T. vaginalis increased PCa cells migration, proliferation and invasion. The cytokine receptors (CXCR2, CCR2, gp130) were expressed higher on the PCa cells treated with TCM. Pretreatment of PCa cells with antibodies to these cytokine receptors significantly reduced the proliferation, mobility and invasiveness of PCa cells, indicating that TCM has its effect through cytokine-cytokine receptor signaling. In C57BL/6 mice, the prostates injected with T. vaginalis mixed PCa cells were larger than those injected with PCa cells alone after 4 weeks. Expression of epithelial-mesenchymal transition markers and cyclin D1 in the prostate tissue injected with T. vaginalis mixed PCa cells increased than those of PCa cells alone. Collectively, it was suggested that inflammatory reactions by T. vaginalis-stimulated PCa cells increase the proliferation and invasion of PCa cells through cytokine-cytokine receptor signaling pathways.
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http://dx.doi.org/10.3347/kjp.2021.59.6.547DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8721307PMC
December 2021

Multifunctional Composite Binder for Thick High-Voltage Cathodes in Lithium-Ion Batteries.

ACS Appl Mater Interfaces 2022 Jan 29;14(1):861-872. Epub 2021 Dec 29.

Center for Automotive Research, Department of Mechanical and Aerospace Engineering, The Ohio State University, Columbus, Ohio 43210, United States.

High-voltage LiNiMnO (LNMO) spinel offers high specific energy and good rate capability with relatively low raw-material cost due to cobalt-free and manganese-rich chemical compositions. Also, increasing mass loading (mg/cm) by thickening cathodes has been one of the focused areas to greatly improve the energy density of lithium-ion batteries (LIBs) at the cell level. The LNMO cathode made with a polyvinylidene fluoride (PVdF) binder, however, suffers from an oxidative decomposition of liquid electrolytes and cathode delamination from a current collector. This problem is exacerbated with an increase in thickness. In this study, we developed a lithium polyacrylate (LiPAA)-sodium alginate (Na-Alg) composite binder series that offer positive multifunctions such as enhancing cathode adhesion and cohesion, improving cycle life, creating an effective passivating layer at the cathode-electrolyte interface (CEI), and lowering cell impedance. Comprehensive design of systematic experiments revealed a close chemo-mechano-electrochemical relationship in the thick high-voltage cathodes. Among the various binder compositions, the LiPAA (30 wt %)-Na-Alg (70 wt %) binder offered a strong adhesion property and positive multifunctions at the CEI layer, which consequently stabilized the solid-electrolyte interfacial (SEI) layer on the graphite anode and improved LIB performances. This novel composite binder will be applicable to various types of thick cathodes in future studies.
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http://dx.doi.org/10.1021/acsami.1c19554DOI Listing
January 2022

Nano-Sized Extracellular Matrix Particles Lead to Therapeutic Improvement for Cutaneous Wound and Hindlimb Ischemia.

Int J Mol Sci 2021 Dec 9;22(24). Epub 2021 Dec 9.

Center for Biomaterials, Korea Institute of Science and Technology (KIST), Seoul 02792, Korea.

Cell-derived matrix (CDM) has proven its therapeutic potential and been utilized as a promising resource in tissue regeneration. In this study, we prepared a human fibroblast-derived matrix (FDM) by decellularization of in vitro cultured cells and transformed the FDM into a nano-sized suspended formulation (sFDM) using ultrasonication. The sFDM was then homogeneously mixed with Pluronic F127 and hyaluronic acid (HA), to effectively administer sFDM into target sites. Both sFDM and sFDM containing hydrogel (PH/sFDM) were characterized via immunofluorescence, sol-gel transition, rheological analysis, and biochemical factors array. We found that PH/sFDM hydrogel has biocompatible, mechanically stable, injectable properties and can be easily administered into the external and internal target regions. sFDM itself holds diverse bioactive molecules. Interestingly, sFDM-containing serum-free media helped maintain the metabolic activity of endothelial cells significantly better than those in serum-free condition. PH/sFDM also promoted vascular endothelial growth factor (VEGF) secretion from monocytes in vitro. Moreover, when we evaluated therapeutic effects of PH/sFDM via the murine full-thickness skin wound model, regenerative potential of PH/sFDM was supported by epidermal thickness, significantly more neovessel formation, and enhanced mature collagen deposition. The hindlimb ischemia model also found some therapeutic improvements, as assessed by accelerated blood reperfusion and substantially diminished necrosis and fibrosis in the gastrocnemius and tibialis muscles. Together, based on sFDM holding a strong therapeutic potential, our engineered hydrogel (PH/sFDM) should be a promising candidate in tissue engineering and regenerative medicine.
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http://dx.doi.org/10.3390/ijms222413265DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8705579PMC
December 2021

Quality assessment of conventional and traditional oriental medicine clinical practice guidelines for knee osteoarthritis using AGREE II instrument.

Medicine (Baltimore) 2021 Dec;100(51):e28426

Joint Center, Kyung Hee University Hospital at Gangdong, Guangdong-gu, Seoul, Republic of Korea.

Introduction: Knee osteoarthritis is a degenerative disease and its prevalence tends to increase. Clinical practice guidelines (CPGs) are evidence-based recommendations for treatment that help policymakers, practitioners, and patients make more appropriate and efficient decisions during the course of management. This study aimed to evaluate the quality of knee osteoarthritis CPGs using the Appraisal of Guidelines for Research and Evaluation (AGREE II) instrument.

Method: The retrieval engines and websites were utilized from January 2010 to December 2020. The search words were "Clinical practice guideline" OR "Critical practice guideline" OR "guideline∗" AND "Osteoarthritis." The quality of the CPGs was independently examined by four appraisers using the AGREE II instrument. Consequently, the selected CPGs were graded as Classes A, B, and C according to the level of recommendation.

Result: In this study, 13 CPGs for knee osteoarthritis were selected and evaluated qualitatively using the AGREE II instrument. The overall quality percentage score was as follows: clarity of presentation, 72.6%, scope and purpose, 62.6%, rigor of development, 54.2%, stakeholder investment, 50.5%, editorial independence, 46.5%, applicability, 22.5%.

Conclusion: Auxiliary materials for the treatment process of knee OA should be supplemented in future revised versions for quality improvement of knee OA CPGs. Also, more evidence should be accumulated to support the recommendation of traditional oriental medical treatments in the clinical field. From the perspective of integrative medicine, along with conventional pharmacological treatment, exercise, weight loss, and acupuncture can be combined together in clinical situations.
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http://dx.doi.org/10.1097/MD.0000000000028426DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8702243PMC
December 2021

Efficacy of Phototherapy With 308-nm Excimer Light for Skin Microbiome Dysbiosis and Skin Barrier Dysfunction in Canine Atopic Dermatitis.

Front Vet Sci 2021 3;8:762961. Epub 2021 Dec 3.

Department of Veterinary Internal Medicine, College of Veterinary Medicine, Konkuk University, Seoul, South Korea.

The management of canine atopic dermatitis, an allergic skin disorder, is challenging. To investigate the effect of phototherapy using a 308-nm excimer light as a topical treatment for canine atopic dermatitis, 10 dogs with canine atopic dermatitis and 10 with non-allergic skin were enrolled in this study. Phototherapy was applied every 7 days for a total of 2 months. The skin microbiome, skin barrier function, and clinical outcomes were evaluated after phototherapy. Phototherapy significantly changed the composition of the skin microbiome of dogs with atopic dermatitis and significantly increased the relative abundance of the phyla Actinobacteria and Cyanobacteria. It significantly alleviated the clinical signs of canine atopic dermatitis without serious adverse effects. Transepidermal water loss, as a measure of skin barrier function, significantly decreased after phototherapy. In addition, phototherapy increased microbial diversity and decreased the relative abundance of associated with the severity of canine atopic dermatitis. These results suggest that the excimer light therapy is a suitable and safe therapeutic option for canine atopic dermatitis, which is also a spontaneous animal model of atopic dermatitis.
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http://dx.doi.org/10.3389/fvets.2021.762961DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8677939PMC
December 2021

Clusters of Severe Eosinophilic Asthma in a Korean Asthma Cohort.

Respiration 2021 Dec 16:1-11. Epub 2021 Dec 16.

Department of Allergy and Clinical Immunology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea.

Background: Targeted therapies have broadened the available treatment options for patients with severe eosinophilic asthma (SEA). However, differences in the magnitude of treatment responses among patients indicate the presence of various underlying pathophysiological processes and patient subgroups.

Objectives: We aimed to describe the characteristics of SEA and identify its patient subgroups.

Methods: Clinical data from the Cohort for Reality and Evolution of Adult Asthma in Korea were analyzed. Cluster analysis was performed among those with SEA using 5 variables, namely, prebronchodilator forced expiratory volume in 1 s, body mass index, age at symptom onset, smoking amount, and blood eosinophil counts.

Results: Patients with SEA showed prevalent sensitization to aeroallergens, decreased lung function, and poor asthma control status. Cluster analysis revealed 3 distinctive subgroups among patients with SEA. Cluster 1 (n = 177) consisted of patients reporting the lowest blood eosinophils (median, 346.8 cells/μL) and modest severe asthma with preserved lung function during the 12-month treatment period. Cluster 2 (n = 42) predominantly included smoking males with severe persistent airway obstruction and moderate eosinophilia (median, 451.8 cells/μL). Lastly, cluster 3 (n = 95) included patients with the most severe asthma, the highest eosinophil levels (median, 817.5 cells/μL), and good treatment response in terms of improved lung function and control status.

Conclusions: Three subgroups were identified in SEA through the cluster analysis. The distinctive features of each cluster may help physicians predict patients who will respond to biologics with greater magnitude of clinical improvement. Further research regarding the underlying pathophysiology and clinical importance of each subgroup is warranted.
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http://dx.doi.org/10.1159/000520492DOI Listing
December 2021

Case Report: Non-traumatic Unilateral Forelimb Arterial Thrombosis Associated With Hyperadrenocorticism in a Dog.

Front Vet Sci 2021 25;8:795928. Epub 2021 Nov 25.

Department of Veterinary Internal Medicine, College of Veterinary Medicine, Konkuk University, Seoul, South Korea.

A 16-year-old spayed female Pomeranian dog was presented to the hospital with an acute onset of pain and non-weight-bearing lameness in the right forelimb. On physical examination, knuckling, coolness, pain, and cyanosis were observed in the affected forelimb. Peripheral blood glucose concentration and body surface temperature differed between the right and left forelimbs. Hypercoagulable thromboelastographic results and increased D-dimer levels were suggestive of thrombus. Accordingly, recombinant tissue plasminogen activator (rtPA) was administered intravenously. Prompt clinical improvements (including restored warmth of the affected limb) occurred, and rtPA was discontinued after two shots administered 2 h apart owing to concerns of bleeding side effects. The dog was discharged 6 days after admission, and outpatient treatment with clopidogrel was continued for the prevention of re-thrombosis. Following patient stabilization, further examinations for underlying diseases of hypercoagulability were conducted; hyperadrenocorticism (HAC) was diagnosed, and oral trilostane therapy was thus administered. Eight weeks later, the patient regained normal mobility. Finally, in the present canine patient with arterial thrombosis, thrombolysis with rtPA successfully improved clinical symptoms and the following administration of clopidogrel inhibited the formation of additional thrombus.
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http://dx.doi.org/10.3389/fvets.2021.795928DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8655981PMC
November 2021

Differential Cutaneous Thermal Sensitivity in Humans: Method of Limit vs. Method of Sensation Magnitude.

Int J Environ Res Public Health 2021 11 29;18(23). Epub 2021 Nov 29.

Department of Sports Medicine, Kyung Hee University, Yongin-si 17104, Korea.

Introduction: The method of limits (MLI) and method of level (MLE) are commonly employed for the quantitative assessment of cutaneous thermal sensitivity. Thermal sensation and thermal comfort are closely related and thermal sensations evoked from the peripheral thermoreceptors play an important role in thermoregulatory response to maintain normal body temperature. The purpose of this study was to compare the regional distribution of cutaneous warm and cold sensitivity between MLI and the method of sensation magnitude (MSM).

Method: Twenty healthy men completed MLI and MSM to compare the regional distribution of cutaneous warm and cold sensitivity in the thermal neutral condition. The subjects rested on a bed in a supine position for 20 min. Next, the cutaneous thermal sensitivity of ten body sites was assessed by the means of MLI and MSM for both warmth and cold stimuli.

Results: The absolute mean heat flux in MLI and thermal sensation magnitude in MSM showed significantly greater sensitivity to cold than to warm stimulation ( < 0.01), together with a similar pattern of regional differences across ten body sites. Both sensory modalities indicated acceptable reliability (SRD%: 6.29-8.66) and excellent reproducibility (ICC: 0.826-0.906; < 0.01). However, the Z-sore distribution in MSM was much narrower than in MLI, which may limit the test sensitivity for the detection of sensory disorders and/or comparison between individuals.

Conclusion: The present results showed that both MLI and MSM are effective means for evaluating regional cutaneous thermal sensitivity to innocuous warm and cold stimulations to a strong degree of reliability and reproducibility.
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http://dx.doi.org/10.3390/ijerph182312576DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8657088PMC
November 2021

Clinical Impact of the Bronchiectasis with Chronic Bronchitis Symptoms in COPD: Analysis of a Longitudinal Cohort.

Int J Chron Obstruct Pulmon Dis 2021;16:2997-3008. Epub 2021 Nov 1.

Department of Pulmonology, Allergy and Critical Care Medicine, CHA Bundang Medical Center, CHA University, Seongnam, Republic of Korea.

Purpose: Bronchiectasis (BE) is a poor prognostic factor in COPD. However, it is not clear whether the poor prognosis is a result of BE alone or accompanying chronic bronchitis symptoms. Therefore, we investigated the effect of chronic bronchitis symptoms on clinical outcomes in COPD patients with BE.

Patients And Methods: We analyzed data of COPD patients from the Korean Obstructive Lung Disease (KOLD) cohort. The presence of BE was verified by chest computed tomography. Chronic bronchitis symptoms were determined using items in the symptomatic domain of the SGRQ, which is also used as an alternative definition of chronic bronchitis (CB). Patients were divided into four groups according to the presence of BE and CB symptoms: BE/CB, BE-only, CB-only, and no BE/CB. Demographic features and clinical outcomes were compared among these groups.

Results: In total, 389 COPD patients were included in the analysis. BE was present in 148 (38%) patients and CB symptoms were found in 123 patients (33.2%). The patients were divided according to BE and CB symptoms, and the numbers and percentages of each group were as follows: BE/CB, 52 (13.4%); BE-only, 96 (24.7%); CB-only, 77 (19.8%); no BE/CB, 164 (42.2%). No significant differences were observed in baseline characteristics of lung function, radiological findings, and inflammatory markers among the four groups. The proportion of annual exacerbators was higher in the BE/CB and CB-only groups than the other two groups. After adjusting other parameters, the BE/CB group was significantly associated with acute exacerbation of COPD (AE-COPD) (OR = 2.110, p = 0.045).

Conclusion: BE accompanying CB symptoms is associated with AE-COPD, while BE alone was not significantly associated. This finding suggests that it is more important to examine chronic bronchitis symptoms of BE to predict acute exacerbation than simply to identify BE in COPD patients.
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http://dx.doi.org/10.2147/COPD.S332299DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8570290PMC
November 2021

Generation of a TLR2 knockout human induced pluripotent stem cell line using CRISPR/Cas9.

Stem Cell Res 2021 Oct 19;57:102578. Epub 2021 Oct 19.

Division of Intractable Diseases, Department of Chronic Diseases Convergence Research, Korea National Institute of Health, Cheongju 28160, Republic of Korea; National Stem Cell Bank of Korea, Korea National Institute of Health, Cheongju 28160, Republic of Korea. Electronic address:

The TLR family plays a fundamental function in the recognition of pathogens and activation of innate immunity. TLR2 is a membrane protein that is expressed on the surface of certain cells and recognizes foreign substances; it initiates a cascade of innate immune responses. TLR2 recognizes many bacterial, fungal, and viral components, as well as certain endogenous substances. We generated biallelic mutants of the TLR2 gene in human induced pluripotent stem cells (hiPSCs) using a CRISPR-Cas9 genome editing method. The TLR2 heterozygous-knockout hiPSCs retained normal morphology, gene expression, and in vivo differentiation potential.
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http://dx.doi.org/10.1016/j.scr.2021.102578DOI Listing
October 2021

FAK Activation Promotes SMC Dedifferentiation via Increased DNA Methylation in Contractile Genes.

Circ Res 2021 12 27;129(12):e215-e233. Epub 2021 Oct 27.

Departments of Biochemistry and Molecular Biology (K.J., J.M.M., J.-H.K., H.P., Y.A.R.R., J.-S.K., R.E.H., S.-T.S.L.), University of South Alabama, College of Medicine, Mobile.

Rationale: Vascular smooth muscle cells (SMCs) exhibit remarkable plasticity and can undergo dedifferentiation upon pathological stimuli associated with disease and interventions.

Objective: Although epigenetic changes are critical in SMC phenotype switching, a fundamental regulator that governs the epigenetic machineries regulating the fate of SMC phenotype has not been elucidated.

Methods And Results: Using SMCs, mouse models, and human atherosclerosis specimens, we found that FAK (focal adhesion kinase) activation elicits SMC dedifferentiation by stabilizing DNMT3A (DNA methyltransferase 3A). FAK in SMCs is activated in the cytoplasm upon serum stimulation in vitro or vessel injury and active FAK prevents DNMT3A from nuclear FAK-mediated degradation. However, pharmacological or genetic FAK catalytic inhibition forced FAK nuclear localization, which reduced DNMT3A protein via enhanced ubiquitination and proteasomal degradation. Reduced DNMT3A protein led to DNA hypomethylation in contractile gene promoters, which increased SMC contractile protein expression. RNA-sequencing identified SMC contractile genes as a foremost upregulated group by FAK inhibition from injured femoral artery samples compared with vehicle group. DNMT3A knockdown in injured arteries reduced DNA methylation and enhanced contractile gene expression supports the notion that nuclear FAK-mediated DNMT3A degradation via E3 ligase TRAF6 (TNF [tumor necrosis factor] receptor-associated factor 6) drives differentiation of SMCs. Furthermore, we observed that SMCs of human atherosclerotic lesions exhibited decreased nuclear FAK, which was associated with increased DNMT3A levels and decreased contractile gene expression.

Conclusions: This study reveals that nuclear FAK induced by FAK catalytic inhibition specifically suppresses DNMT3A expression in injured vessels resulting in maintaining SMC differentiation by promoting the contractile gene expression. Thus, FAK inhibitors may provide a new treatment option to block SMC phenotypic switching during vascular remodeling and atherosclerosis.
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http://dx.doi.org/10.1161/CIRCRESAHA.121.319066DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8639767PMC
December 2021

Clomiphene Citrate Shows Effective and Sustained Antimicrobial Activity against .

Int J Mol Sci 2021 Oct 13;22(20). Epub 2021 Oct 13.

Center for Clinical Research, Masan National Tuberculosis Hospital, Korea Centers for Disease Control and Prevention, Changwon 51755, Korea.

() causes chronic pulmonary infections and is the most difficult non-tuberculous mycobacteria (NTM) to treat due to its resistance to current antimicrobial drugs, with a treatment success rate of 45.6%. Thus, novel treatment drugs are needed, of which we identified the drug clomiphene citrate (CC), known to treat infertility in women, to exhibit inhibitory activity against . To assess the potential of CC as a treatment for pulmonary diseases, we measured its efficacy in vitro and established the intracellular activity of CC against in human macrophages. CC significantly inhibited the growth of not only wild-type strains but also clinical isolate strains and clarithromycin (CLR)-resistant strains of . CC's drug efficacy did not have cytotoxicity in the infected macrophages. Furthermore, CC worked in anaerobic non-replicating conditions as well as in the presence of biofilm. The results of this in vitro study on activity suggest the possibility of using CC to develop new drug hypotheses for the treatment of infections.
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http://dx.doi.org/10.3390/ijms222011029DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8537717PMC
October 2021

Establishment of a human-induced pluripotent stem cell line, KSCBi014-A, from a long QT syndrome type 2 patient harboring a KCNH2 mutation.

Stem Cell Res 2021 Oct 13;57:102570. Epub 2021 Oct 13.

Division of Intractable Diseases Research, Department of Chronic Diseases Convergence Research, Korea National Institute of Health, Osong Health Technology, Administration Complex 202, South Korea. Electronic address:

Long QT syndrome type 2 (LQT2) is a heart disorder caused by a loss-of-function mutation in the KCNH2 gene that is an essential factor in cardiac repolarization and affects the heart rate. This study has generated a human-induced stem cell line (KSCBi014-A) carrying the KCNH2 (c.453delC) mutation from an LQT2 patient. The non-integrative Sendai virus-mediated induced pluripotent stem cell (iPSC) reprogramming method was used for iPSC line generation. The KSCBi014-A line maintained stem cell-like morphology, normal karyotype, and pluripotency, and could differentiate into three germ layers in vitro.
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http://dx.doi.org/10.1016/j.scr.2021.102570DOI Listing
October 2021

When river water meets seawater: Insights into primary marine aerosol production.

Sci Total Environ 2022 Feb 7;807(Pt 2):150866. Epub 2021 Oct 7.

Korea Polar Research Institute, 26 Songdomirae-ro, Yeonsu-gu, Incheon 21990, South Korea.

The impact of inorganic salts and organic matter (OM) on the production of primary marine aerosols is still under debate. To constrain their impact, we investigated primary aerosols generated by a sea-spray generator chamber using surface water samples from rivers, estuaries, and seas that were collected along salinity gradients in two temperate Korean coastal systems and one Arctic coastal system. Salinity values showed an increasing trend along the river-estuary-coastal water transition, indicating the lowest amount of inorganic salts in the river but the highest amount in the sea. In river samples, the lowest number concentration of primary aerosol particles (1.01 × 10 cm) was observed at the highest OM content, suggesting that low salinity controls aerosol production. Moreover, the number concentration of primary aerosols increased drastically in estuarine (1.13 × 10 cm) and seawater (1.35 × 10 cm) samples as the OM content decreased. Our results indicate that inorganic salts associated with increasing salinity play a much larger role than OM in aerosol production in river-dominated coastal systems. Laboratory studies using NaCl solution supported the conclusion that inorganic salt is a critical factor in modulating the particles produced from river water and seawater. Accordingly, this study highlights that inorganic salts are a critical factor in modulating the production of primary marine aerosols.
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http://dx.doi.org/10.1016/j.scitotenv.2021.150866DOI Listing
February 2022

Thirty-Year Trends in Graft Survival After Heart Transplant: Modeled Analyses of a Transplant Registry.

Ann Thorac Surg 2021 Sep 21. Epub 2021 Sep 21.

Department of Medicine, Rutgers Robert Wood Johnson Medical School, New Brunswick, New Jersey; Robert Wood Johnson University Hospital, New Brunswick, New Jersey; Rutgers Institute for Health, Health Policy, and Aging Research, New Brunswick, New Jersey. Electronic address:

Background: Heart failure is an epidemic in the United States, and transplantation remains the most definitive therapy. We describe multidecade trends in posttransplant graft survival, adjusted for concurrent changes in the population, over the 30 years antecedent to the most recent heart allocation policy change.

Methods: Scientific Registry of Transplant Recipients data were used to identify all primary adult heart recipients from 1989 to 2017. We described temporal changes in population characteristics (recipient and donor demographics and comorbidities, pretransplant interventions, clinical transplant measures, and providers). The primary outcome was graft survival, defined as freedom from all-cause death and graft failure, within 6 months posttransplant. Modified Poisson logistic regression estimated relative changes in risk of outcomes compared with 1989, with and without adjustment for changing population characteristics. We identified risk factors, quantified by associated risk ratios.

Results: Among 56,488 primary adult heart recipients, we observed 5529 (9.8%) all-cause deaths and 1933 (3.4%) graft failure events within 6 months posttransplant. Prevalence of known recipient risk factors increased over time. Unadjusted modeling demonstrated a significant 30-year improvement in graft survival, averaging 2.6% per year (95% confidence interval, 2.4-2.9; P for trend < .001). After adjusting for population changes the 30-year trend remained significant and graft survival improved on average 3.0% per year (95% confidence interval, 2.6-3.3). Regression modeling identified multiple predictors of graft survival. Modeling 2 additional outcomes of 6-month mortality and 6-month graft failure produced similar results.

Conclusions: Short-term graft survival after heart transplantation has improved significantly leading up to the 2018 heart allocation policy change, despite concurrent increase in prevalence of higher risk population characteristics.
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http://dx.doi.org/10.1016/j.athoracsur.2021.08.023DOI Listing
September 2021

SON drives oncogenic RNA splicing in glioblastoma by regulating PTBP1/PTBP2 switching and RBFOX2 activity.

Nat Commun 2021 09 21;12(1):5551. Epub 2021 Sep 21.

Department of Pathology, Division of Molecular and Cellular Pathology, University of Alabama at Birmingham, Birmingham, AL, USA.

While dysregulation of RNA splicing has been recognized as an emerging target for cancer therapy, the functional significance of RNA splicing and individual splicing factors in brain tumors is poorly understood. Here, we identify SON as a master regulator that activates PTBP1-mediated oncogenic splicing while suppressing RBFOX2-mediated non-oncogenic neuronal splicing in glioblastoma multiforme (GBM). SON is overexpressed in GBM patients and SON knockdown causes failure in intron removal from the PTBP1 transcript, resulting in PTBP1 downregulation and inhibition of its downstream oncogenic splicing. Furthermore, SON forms a complex with hnRNP A2B1 and antagonizes RBFOX2, which leads to skipping of RBFOX2-targeted cassette exons, including the PTBP2 neuronal exon. SON knockdown inhibits proliferation and clonogenicity of GBM cells in vitro and significantly suppresses tumor growth in orthotopic xenografts in vivo. Collectively, our study reveals that SON-mediated RNA splicing is a GBM vulnerability, implicating SON as a potential therapeutic target in brain tumors.
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http://dx.doi.org/10.1038/s41467-021-25892-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8455679PMC
September 2021

Elevated Neutrophil-to-Lymphocyte Ratio in Perioperative Periods is Suggestive of Poor Prognosis in Patients with Colorectal Cancer.

J Inflamm Res 2021 7;14:4457-4466. Epub 2021 Sep 7.

Department of Surgery, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea.

Background: Recent data suggest that alterations in the neutrophil-to-lymphocyte ratio (NLR) in the perioperative periods can serve as prognostic factors. However, research on the clinical impact has been limited and even discordant in patients with colorectal cancer (CRC).

Patients And Methods: The optimal cut-off value of preoperative NLR (NLR-pre), postoperative NLR (NLR-post), and its change (NLR-delta) were determined to maximize differences in overall survival (OS) between groups. Patients were categorized into four groups (NLR-trend) as follows: G1, low NLR-pre and NLR-post; G2, low NLR-pre and high NLR-post; G3, high NLR-pre and low NLR-post; and G4, high NLR-pre and NLR-post. Discriminatory performance was compared using integrated AUC (iAUC) between all indicators.

Results: A total of 576 patients diagnosed with stage I-IV CRC were included. The cut-off points were determined as 2.33 for NLR-pre, 2.06 for NLR-post, and -1.08 for NLR-delta. Subgroup dichotomization using NLR-pre, NLR-post, NLR-delta and NLR-trend were all identified as significant prognostic factors by univariate analysis. However, NLR-trend was only remained as an independent prognostic factor in the multivariate analysis. The iAUC of the NLR-trend was superior to that of NLR-pre (bootstrap iAUC mean difference=0.036; 95% CI 0.013-0.073), NLR-post (bootstrap iAUC mean difference=0.045; 95% CI 0.019-0.081) and NLR-delta (bootstrap iAUC mean difference=0.061; 95% CI 0.025-0.104).

Conclusion: Risk stratification and combining of preoperative and postoperative NLR (NLR-trend) can improve prognostic discrimination compared with single measurements or simple changes in NLR in patients with CRC.
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http://dx.doi.org/10.2147/JIR.S327443DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8434909PMC
September 2021

Establishing the phenotypic spectrum of ZTTK syndrome by analysis of 52 individuals with variants in SON.

Eur J Hum Genet 2021 Sep 15. Epub 2021 Sep 15.

Department of Human Genetics, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, P.O. Box 9101, 6500 HB, Nijmegen, The Netherlands.

Zhu-Tokita-Takenouchi-Kim (ZTTK) syndrome, an intellectual disability syndrome first described in 2016, is caused by heterozygous loss-of-function variants in SON. Its encoded protein promotes pre-mRNA splicing of many genes essential for development. Whereas individual phenotypic traits have previously been linked to erroneous splicing of SON target genes, the phenotypic spectrum and the pathogenicity of missense variants have not been further evaluated. We present the phenotypic abnormalities in 52 individuals, including 17 individuals who have not been reported before. In total, loss-of-function variants were detected in 49 individuals (de novo in 47, inheritance unknown in 2), and in 3, a missense variant was observed (2 de novo, 1 inheritance unknown). Phenotypic abnormalities, systematically collected and analyzed in Human Phenotype Ontology, were found in all organ systems. Significant inter-individual phenotypic variability was observed, even in individuals with the same recurrent variant (n = 13). SON haploinsufficiency was previously shown to lead to downregulation of downstream genes, contributing to specific phenotypic features. Similar functional analysis for one missense variant, however, suggests a different mechanism than for heterozygous loss-of-function. Although small in numbers and while pathogenicity of these variants is not certain, these data allow for speculation whether de novo missense variants cause ZTTK syndrome via another mechanism, or a separate overlapping syndrome. In conclusion, heterozygous loss-of-function variants in SON define a recognizable syndrome, ZTTK, associated with a broad, severe phenotypic spectrum, characterized by a large inter-individual variability. These observations provide essential information for affected individuals, parents, and healthcare professionals to ensure appropriate clinical management.
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http://dx.doi.org/10.1038/s41431-021-00960-4DOI Listing
September 2021

Establishment of a human induced pluripotent stem cell line, KSCBi015-A, from a long QT syndrome type 1 patient harboring a KCNQ1 mutation.

Stem Cell Res 2021 10 2;56:102521. Epub 2021 Sep 2.

Division of Intractable Diseases Research, Department of Chronic Diseases Convergence Research, Korea National Institute of Health, Osong Health Technology, Administration Complex 202, South Korea; Korea National Stem Cell Bank, South Korea. Electronic address:

Long QT syndrome type 1 (LQT1) is a genetic cardiac disorder caused by a loss-of-function mutation in the KCNQ1 gene. In this study, we generated a human induced stem cell line (KSCBi015-A) from an LQT1 patient with a heterozygous mutation located in the KCNQ1 gene, c.569G > A. The KSCBi015-A cell line showed the maintenance of stem cell-like morphology, normal karyotype, and pluripotency, and could differentiate into three germ layers in vitro.
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http://dx.doi.org/10.1016/j.scr.2021.102521DOI Listing
October 2021

Establishment of Canine Transitional Cell Carcinoma Cell Lines Harboring BRAF V595E Mutation as a Therapeutic Target.

Int J Mol Sci 2021 Aug 25;22(17). Epub 2021 Aug 25.

Department of Veterinary Biochemistry, College of Veterinary Medicine, Konkuk University, Seoul 05029, Korea.

Transitional cell carcinoma (TCC) is the most common malignant tumor of the canine urinary tract and tends to have a poor prognosis due to its invasive potential. Recent studies have reported that up to 80% of canine urothelial carcinoma has the BRAF V595E mutation, which is homologous to the human V600E mutation. Activating the BRAF mutation is an actionable target for developing effective therapeutic agents inhibiting the BRAF/mitogen-activated protein kinase (MAPK) pathway in canine cancer as well as human cancer. We established novel canine TCC cell lines from two tumor tissues and one metastatic lymph node of canine TCC patients harboring the BRAF V595E mutation. Tumor tissues highly expressed the BRAF mutant and phosphorylated extracellular signal-related kinases (ERK)1/2 proteins. The derived cell lines demonstrated activated MAPK pathways. We also evaluated the cell lines for sensitivity to BRAF inhibitors. Sorafenib, a multiple kinase inhibitor targeting RAF/vascular endothelial growth factor receptor (VEGFR), successfully inhibited the BRAF/MAPK pathway and induced apoptosis. The established canine TCC cell lines responded with greater sensitivity to sorafenib than to vemurafenib, which is known as a specific BRAF inhibitor in human cancer. Our results demonstrated that canine TCC cells showed different responses compared to human cancer with the BRAF V600E mutation. These cell lines would be valuable research materials to develop therapeutic strategies for canine TCC patients.
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http://dx.doi.org/10.3390/ijms22179151DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8430554PMC
August 2021

Serum Folliculin Is Related to Lower Pulmonary Function in Patients With Asthma.

Allergy Asthma Immunol Res 2021 Sep;13(5):822-826

Department of Allergy and Clinical Immunology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea.

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http://dx.doi.org/10.4168/aair.2021.13.5.822DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8419642PMC
September 2021

Morphological Analysis of PSMA/PEI Core-Shell Nanoparticles Synthesized by Soap-Free Emulsion Polymerization.

Nanomaterials (Basel) 2021 Jul 29;11(8). Epub 2021 Jul 29.

Research Institute of Sustainable Manufacturing System, Intelligent Sustainable Materials R&D Group, Korea Institute of Industrial Technology, 89 Yangdaegiro-gil, Ipjang-myeon, Seobuk-gu, Cheonan-si 31056, Korea.

Emulsion polymerization presents the disadvantage that the physical properties of polymer particles are altered by surfactant adsorption. Therefore, in the soap-free emulsion polymerization method, a hydrophilic initiator is utilized while inducing repulsion among particles on the polymer particle surface, resulting in stable polymer particle production. In this study, we developed a methodology wherein spherical and uniform poly(styrene--maleic anhydride) (PSMA)/polyethyleneimine (PEI) core-shell nanoparticles were prepared. Further, their morphology was analyzed. During PSMA polymerization, the addition of up to 30% maleic anhydride (MA) resulted in stable polymerization. In PSMA/PEI nanoparticle fabrication, the number of reactants increased with increased initial monomer feed amounts; consequently, the particle size increased, and as the complete monomer consumption time increased, the particle distribution widened. The styrene (St) copolymer acted as a stabilizer, reducing particle size and narrowing particle distribution. Furthermore, the monomers were more rapidly consumed at high initiator concentrations, irrespective of the initiator used, resulting in increased particle stability and narrowed particle distribution. The shell thickness and particle size were PEI feed ratio dependent, with 0.08 being the optimal PEI-to-MA ratio. The fabricated nanoparticles possess immense potential for application in environmental science and in chemical and health care industries.
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http://dx.doi.org/10.3390/nano11081958DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8402240PMC
July 2021

Efficacy of Tigecycline and Linezolid Against Pan-Drug-Resistant Bacteria Isolated From Companion Dogs in South Korea.

Front Vet Sci 2021 6;8:693506. Epub 2021 Aug 6.

Department of Veterinary Internal Medicine, College of Veterinary Medicine, Konkuk University, Seoul, South Korea.

The emergence of multidrug-resistant bacteria in companion animals is an increasing concern in view of the concept of One Health. The antimicrobials linezolid (LZD) and tigecycline (TGC) are effective against multidrug-resistant bacteria isolated from humans; however, thus far, no previous study has evaluated the efficacy of these drugs against bacteria isolated from companion animals. This study aimed to evaluate the efficacy of LZD and TGC against bacteria that were isolated from companion dogs and showed resistance to all classes of antimicrobial agents. Clinical samples (auditory channel, eye, skin, and urine) were collected from dogs that visited the Veterinary Medical Teaching Hospital of Konkuk University (Seoul, South Korea) from October 2017 to September 2020. In total, 392 bacterial isolates were obtained, of which 85 were resistant to all classes of antimicrobial agents tested and were, therefore, considered potentially pan-drug resistant (PDR). The susceptibility of isolates to LZD and TGC was determined by the disk diffusion method and interpreted using the Clinical Laboratory Standards Institute guidelines. In total, 95.6% (43/45) and 97.8% (44/45) of gram-positive isolates were susceptible to LZD and TGC, respectively, whereas 82.5% (33/40) of gram-negative isolates were sensitive to TGC. In conclusion, both agents showed favorable efficacy, with the susceptibility rates for all potential PDR bacteria, except spp., ranging from 72.7 to 100%. Thus, these drugs may serve as excellent antimicrobial options for veterinary medicine in the future.
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http://dx.doi.org/10.3389/fvets.2021.693506DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8377367PMC
August 2021

Establishing a critical pathway for Korean medical management of lumbar disc herniation: A modified Delphi consensus process.

Medicine (Baltimore) 2021 Aug;100(33):e26991

Department of Acupuncture & Moxibustion, College of Korean Medicine, Kyung Hee University, 26, Kyungheedae-ro, Dongdaemun-gu, Seoul, Republic of Korea.

Abstract: A modified Delphi method was used to establish a consensus. Stakeholders and experts were invited to participate in the expert panel. Best practice statements and decision-making questionnaires were distributed to the panel. Panel members were asked to mark "Strongly disagree" to "Strongly agree" after a series of statements over several rounds until either a consensus was reached or the decision-making method was deemed unsuitable for reaching a consensus.The most common cause of lumbar pain is intervertebral degeneration, which leads to degenerative disc disease and lumbar disc herniation. There is a lack of unanimity regarding appropriate patient protocols and rehabilitation expectations for Korean medical care. The long-term viability of Korean medical treatment, further adoption in the institutional setting, and specific patient outcomes are contingent on the existence of appropriate Korean medical programs.A Korean medical expert panel of 17 practitioners employed a modified Delphi method to achieve consensus on Korean medical care for lumbar disc herniation. The panel first reviewed the literature and guidelines relevant to Korean medical treatment for lumbar disc herniation. The panel members considered questionnaires intended to determine "standardized" Korean medical care recommendations for patients with a wide range of symptoms of lumbar disc herniation. Each panel member participated in a round of voting, which was followed by an opinion-collecting session online. Consensus was defined as a ≥75% agreement among the respondents.In the first round, 144 questionnaires across 5 domains were administered to the expert panels. After reviewing the responses and open-ended comments collected in the first round, the authors modified the questionnaires to 53 items and proceeded. In round 2, consensus was achieved in all 53 survey questions. The final treatment pathway comprised a standardized and comprehensive care approach for lumbar disc herniations in 4 types of medical institutions.This study identified a core set of evidence- and consensus-based principles that are essential to a comprehensive model of care, incorporating identification, referral, and management of patients with lumbar disc herniation.
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http://dx.doi.org/10.1097/MD.0000000000026991DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8376369PMC
August 2021

A bibliometric analysis of research on the treatment of facial nerve palsy.

Medicine (Baltimore) 2021 Aug;100(33):e26984

Department of Acupuncture & Moxibustion, College of Korean Medicine, Kyung Hee University, Dongdaemun-gu, Seoul, Republic of Korea.

Background: There are various treatments for facial nerve palsy, and research into this topic is ongoing. In the present study, we carried out bibliometric and visualized analyses to identify the trends of research into facial nerve palsy treatment.

Methods: To identify articles, the SCOPUS database was searched for articles published from its inception to December 27, 2020. The search was conducted twice, with Search 1 investigating general treatment trends and Search 2 narrowing the scope to complementary and alternative treatment. The extracted keywords were analyzed using the Visualization Of Similarities (VOS) viewer. Through analysis of keywords, research hotspots in the treatment of facial nerve palsy were identified.

Results: A total of 1609 and 223 articles were identified in Searches 1 and 2, respectively. The number of articles published each year showed a tendency to increase, and most of the studies were only conducted in a few countries. In terms of subject area, "medicine" was overwhelmingly the most common(77.6%). Based on the analysis of 316 keywords in Search1, "medication treatment," and "complementary and alternative treatment" were the hotspots of research.

Conclusion: This study provides the overall trends of facial nerve palsy treatment. To date, research on medication treatment has been main focus, and antiviral use among medication treatment and complementary and alternative treatment has emerged in recent years.
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http://dx.doi.org/10.1097/MD.0000000000026984DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8376370PMC
August 2021

Complete plastid genome sequence of (Laurales: Lauraceae), an evergreen broad-leaved tree endemic to East Asia.

Mitochondrial DNA B Resour 2021 2;6(9):2553-2555. Epub 2021 Aug 2.

Department of Biology Education, Chonnam National University, Gwangju, Republic of Korea.

In this study, we sequenced the complete plastid genome (plastome) of , an evergreen broad-leaved tree endemic to East Asia, a woody component of East Asian warm-temperate and subtropical forests across China, Korea, and Japan. The plastome of is assembled as a single contig (152,722 bp). A large and a small single copy (93,785 and 18,795 bp, respectively) of the genome are separated by a pair of inverted repeats (20,071 bp). The genome consists of 126 genes, including 80 protein-coding, eight ribosomal RNA, and 36 transfer RNA genes. Two genes in the IR region (1 and 2) are pseudogenized. Our phylogenetic analysis revealed the phylogenetic position of in a highly supported clade of the genus along with other two congeners, and
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http://dx.doi.org/10.1080/23802359.2021.1959457DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8330707PMC
August 2021

Highly Selective, CRISPR/Cas9-Mediated Isolation of Genes and Genomic Loci from Complex Genomes by TAR Cloning in Yeast.

Curr Protoc 2021 Aug;1(8):e207

Developmental Therapeutics Branch, National Cancer Institute, NIH, Bethesda, Maryland.

Here we describe an updated TAR cloning protocol for the selective and efficient isolation of any genomic fragment or gene of interest up to 280 kb in size from genomic DNA. The method exploits the special recombination machinery of the yeast Saccharomyces cerevisiae. TAR cloning is based on the high level of in vivo recombination that occurs between a specific genomic DNA fragment of interest and targeting sequences (hooks) in a TAR vector that are homologous to the 5' and 3' ends of the targeted region. Upon co-transformation into yeast, this results in the isolation of the chromosomal region of interest as a circular YAC molecule, which then propagates and segregates in yeast cells and can be selected for. In the updated TAR cloning protocol described here, the fraction of region-positive clones typically obtained is increased from 1% up to 35% by pre-treatment of the genomic DNA with specifically designed CRISPR/Cas9 endonucleases that create double-strand breaks (DSBs) bracketing the target genomic DNA sequence, thereby making the ends of the chromosomal region of interest highly recombinogenic. In addition, a new TAR vector was constructed that contains YAC and BAC cassettes, permitting direct transfer of a TAR-cloned DNA from yeast to bacterial cells. Once the TAR vector with the hooks is constructed and genomic DNA is prepared, the entire procedure takes 3 weeks to complete. The updated TAR protocol does not require significant yeast experience or extensively time-consuming yeast work because screening only about a dozen yeast transformants is typically enough to find a clone with the region of interest. TAR cloning of chromosomal fragments, individual genes, or gene families can be used for functional, structural, and population studies, for comparative genomics, and for long-range haplotyping, and has potential for gene therapy. Published 2021. This article is a U.S. Government work and is in the public domain in the USA. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Preparation of CRISPR/Cas9-treated genomic DNA for TAR cloning Basic Protocol 2: Isolation of a gene or genomic locus by TAR cloning Basic Protocol 3: Transfer of TAR/YAC/BAC isolates from yeast to E. coli.
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http://dx.doi.org/10.1002/cpz1.207DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8363120PMC
August 2021
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