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DNA Bacteriano , Filogenia , RNA Ribossômico 16S , Análise de Sequência de DNA , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , Mongólia , Técnicas de Tipagem Bacteriana , Minas de Carvão , Ácidos Graxos/química , Composição de Bases , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Microbiologia do SoloRESUMO
Cas12j, a hypercompact and efficient Cas protein, has potential for use in CRISPR diagnostics, but has not yet been used because the trans-cleavage activity of Cas12j is veiled. Here, the trans-cleavage behavior of Cas12j1, 2, and 3 variants and evaluate their suitability for nucleic acid detection is unveiled. The target preferences and mismatch specificities of the Cas12j variants are precisely investigated and the optimal Cas12j reaction conditions are determined. As a result, the EXP-J assay for miRNA detection by harnessing the robust trans-cleavage activity of Cas12j on short ssDNA is developed. The EXP-J method demonstrates exceptional detection capabilities for miRNAs, proving that Cas12j can be a pivotal component in molecular diagnostics. Furthermore, the translational potential of the EXP-J assay is validated by detecting oncogenic miRNAs in plasma samples from lung cancer patients. This investigation not only elucidates the trans-cleavage characteristics of Cas12j variants, but also advances the Cas12j-based diagnostic toolkit.
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Deep neural networks are increasingly used in medical imaging for tasks such as pathological classification, but they face challenges due to the scarcity of high-quality, expert-labeled training data. Recent efforts have utilized pre-trained contrastive image-text models like CLIP, adapting them for medical use by fine-tuning the model with chest X-ray images and corresponding reports for zero-shot pathology classification, thus eliminating the need for pathology-specific annotations. However, most studies continue to use the same contrastive learning objectives as in the general domain, overlooking the multi-labeled nature of medical image-report pairs. In this paper, we propose a new fine-tuning strategy that includes positive-pair loss relaxation and random sentence sampling. We aim to improve the performance of zero-shot pathology classification without relying on external knowledge. Our method can be applied to any pre-trained contrastive image-text encoder and easily transferred to out-of-domain datasets without further training, as it does not use external data. Our approach consistently improves overall zero-shot pathology classification across four chest X-ray datasets and three pre-trained models, with an average macro AUROC increase of 4.3%. Additionally, our method outperforms the state-of-the-art and marginally surpasses board-certified radiologists in zero-shot classification for the five competition pathologies in the CheXpert dataset.
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Redes Neurais de Computação , Humanos , Aprendizado Profundo , Processamento de Imagem Assistida por Computador/métodos , Radiografia Torácica/métodos , Raios X , AlgoritmosRESUMO
BACKGROUND: CSF1R-related leukoencephalopathy is a type of autosomal dominant leukodystrophy caused by mutations in the colony stimulating factor 1 receptor (CSF1R) gene. Subcortical ischemic vascular dementia (SIVaD), which is caused by cerebral small vessel disease, is similar to CSF1R-related leukoencephalopathy in that it mainly affects subcortical white matter. In this study, we compared the patterns of white matter hyperintensity (WMH) and cortical thickness in CSF1R-related leukoencephalopathy with those in SIVaD. METHODS: Fourteen patients with CSF1R-related leukoencephalopathy and 129 with SIVaD were retrospectively recruited from three tertiary medical centers. We extracted and visualized WMH data using voxel-based morphometry to compare the WMH distributions between the two groups. Cortical thickness was measured using a surface-based method. Statistical maps of differences in cortical thickness between the two groups were generated using a surface model, with age, sex, education, and intracranial volume as covariates. RESULTS: Predominant distribution of WMH in the CSF1R-related leukoencephalopathy group was in the bilateral frontal and parietal areas, whereas the SIVaD group showed diffuse WMH involvement in the bilateral frontal, parietal, and temporal areas. Compared with the SIVaD group, the CSF1R-related leukoencephalopathy group showed more severe corpus callosum atrophy (CCA) and widespread cortical thinning. CONCLUSIONS: To our knowledge, this is the first study using the automated MR measurement to capture WMH, cortical thinning, and CCA with signal changes in CSF1R-related leukoencephalopathy. It provides new evidence regarding differences in the patterns of WMH distribution and cortical thinning between CSF1R-related leukoencephalopathy and SIVaD.
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Demência Vascular , Leucoencefalopatias , Imageamento por Ressonância Magnética , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos , Substância Branca , Humanos , Masculino , Feminino , Substância Branca/patologia , Substância Branca/diagnóstico por imagem , Leucoencefalopatias/diagnóstico por imagem , Leucoencefalopatias/patologia , Demência Vascular/patologia , Demência Vascular/diagnóstico por imagem , Pessoa de Meia-Idade , Idoso , Estudos Retrospectivos , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/genética , Córtex Cerebral/patologia , Córtex Cerebral/diagnóstico por imagem , Receptor de Fator Estimulador de Colônias de MacrófagosRESUMO
Alternative pre-mRNA splicing (AS) is a biological process that results in proteomic diversity. However, implications of AS alterations in cancer remain poorly understood. Herein, we performed a comprehensive AS analysis in cancer driver gene transcripts across fifteen cancer types and found global alterations in inclusion rates of the PBAF SWI/SNF chromatin remodeling complex subunit Polybromo 1 (PBRM1) exon 27 (E27) in most types of cancer tissues compared with those in normal tissues. Further analysis confirmed that PBRM1 E27 is excluded by the direct binding of RBFOX2 to intronic UGCAUG elements. In addition, the E27-included PBRM1 isoform upregulated PD-L1 expression via enhanced PBAF complex recruitment to the PD-L1 promoter. PBRM1 wild-type patients with clear cell renal cell carcinoma were resistant to PD-1 blockade therapy when they expressed low RBFOX2 mRNA levels. Overall, our study suggests targeting of RBFOX2-mediated AS of PBRM1 as a potential therapeutic strategy for immune checkpoint blockade.
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BACKGROUND: Little is known about the quality of life (QOL) of patients with myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD). We compared QOL and associated factors in patients with MOGAD and aquaporin4 IgG (AQP4-IgG) positive neuromyelitis optica spectrum disorder (NMOSD). METHODS: This multicenter questionnaire study compared the QOL of 41 patients with MOGAD and 78 with AQP4-IgG positive NMOSD. Patients who were positive for AQP4-IgG or MOG antibodies were included. WHO Quality of Life Scale Brief Version was used to assess QOL in physical, psychological, social, and environmental domains. QOL, sleep quality, pain, fatigue, and depression were compared between the two groups. The factors associated with QOL in each group and the entire cohort were analyzed. RESULTS: The proportion of patients with poor QOL was not significantly different between MOGAD (51.22 %) and AQP4-IgG positive NMOSD (58.97 %, p = 0.054). In the MOGAD group, the pain score (ß=-1.032, p = 0.001) and depression score (ß=-0.694, p = 0.007) were negatively associated with physical and psychological QOL, respectively. Sleep quality was negatively associated with physical (ß=-1.506, p = 0.034) and psychological (ß =-2.064, p = 0.033) QOL. When the entire cohort was analyzed, a positive MOG antibody was independently associated with worse psychological QOL (ß=-8.998, p = 0.013) compared to positive AQP4-Ab after adjustment for sleep quality, depression, fatigue, and pain. CONCLUSIONS: The overall QOL of the patients of MOGAD was comparable to that of AQP4-IgG positive NMOSD. Patients with MOGAD were experiencing sleep disorder, fatigue, and depression at similar degrees to those of patients with AQP4-IgG positive NMOSD. Further consideration of sleep quality and psychological QOL is required to improve QOL in patients with MOGAD.
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Background: The treatment of chronic scapholunate dissociation (SLD) can be challenging due to several factors such as poor quality of ligament, malalignment of the carpus, limited surgical options, and risk of recurrent instability. Various surgical techniques have been developed, but there is ongoing debate regarding the optimal surgical technique. This study aimed to report the clinical and radiological outcomes after dorsal scapholunate (SL) ligament complex reconstruction using suture tape-augmented autologous tendon graft. Methods: The study included patients with Garcia-Elias stage 3-4 chronic SLD, SL advanced collapse (SLAC) stage 1, and a follow-up period exceeding 1 year. Pre- and postoperative SL gap, SL angle (SLA), radiolunate angle (RLA), and dorsal scaphoid translation (DST) were measured, and wrist active range of motion, Modified Mayo Wrist Score (MMWS), and visual analog scale (VAS) were evaluated. Results: Nine patients were included in this study with a mean follow-up period of 17 months (range, 15-31 months). All patients were male, with a mean age of 49 years (range, 30-62 years). Eight patients were classified as Garcia-Elias stage 4, while one was classified as SLAC 1. The median (range) of preoperative, immediate postoperative, and final follow-up measurements for SL gap, SLA, RLA, and DST were 5.4 mm (4.5-5.9), 2.1 mm (1.8-2.5), and 2.5 mm (2.0-2.8) (p = 0.008); 76° (69°-88°), 50° (32°-56°), and 54° (50°-64°) (p = 0.008); 22° (11.5°-33°), 2.8° (0.5°-3.8°), and 3.8° (2.2°-5.6°) (p = 0.008); and 2.8 mm (2.0-3.4), 0.8 mm (0.1-1.2), and 1.0 mm (0.1-2.0) (p = 0.008), respectively. Immediately after surgery, all radiological measurements showed significant improvement, which persisted up to 15 months postoperatively. The preoperative and final follow-up measurements of active flexion, extension, radial deviation, and ulnar deviation of the wrist showed significant improvement. The median preoperative and final follow-up values of MMWS were 51.1 (range, 40-60) and 88.3 (range, 85-95) (p = 0.007), respectively, and those of VAS were 7 (range, 6-8) and 2 (range, 1-3) (p = 0.007), respectively. Conclusions: Dorsal SL ligament complex reconstruction using suture tape-augmented autologous free tendon graft could be regarded as a feasible and straightforward technique for addressing irreparable chronic SLD.
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Ligamentos Articulares , Osso Semilunar , Osso Escafoide , Tendões , Humanos , Masculino , Adulto , Pessoa de Meia-Idade , Ligamentos Articulares/cirurgia , Osso Escafoide/cirurgia , Tendões/transplante , Tendões/cirurgia , Osso Semilunar/cirurgia , Instabilidade Articular/cirurgia , Amplitude de Movimento Articular , Transplante Autólogo , Articulação do Punho/cirurgia , Fita Cirúrgica , Estudos Retrospectivos , Procedimentos de Cirurgia Plástica/métodosRESUMO
Background: Irreparable anterosuperior rotator cuff tears (IASRCTs) present significant challenges, especially in young, active patients with limited joint-preserving options. Recently, latissimus dorsi (LD) transfer and combined latissimus dorsi and teres major (LDTM) transfer have gained attention as a potential surgical option. We aimed to compare the clinical and radiological outcomes of LD versus combined LDTM transfer in IASRCTs. Methods: In this retrospective comparative study, 53 patients with IASRCTs were analyzed after undergoing either LD transfer attached to lesser tuberosity (LT) (LD group, n = 23) or combined LDTM transfer attached to greater tuberosity (GT) (LDTM group, n = 30). Clinical evaluations included the visual analog scale score for pain, active shoulder range of motion (ROM), University of California Los Angeles Shoulder Score, American Shoulder and Elbow Surgeons score, activities of daily living that require active internal rotation (ADLIR) scores, and subscapularis (SSC)-specific examinations. Radiographic analyses involved assessing acromiohumeral distance (AHD), Hamada grade, the rate of anterior glenohumeral subluxation reduction, and integrity of the transferred tendon. Results: Postoperatively, both groups demonstrated significant improvements in pain and clinical scores (p < 0.001). At the 2-year follow-up, the LDTM group showed superior internal rotation strength (p < 0.001), ADLIR score (p = 0.017), and SSC-specific physical examination results (belly-press, p = 0.027; bear-hug, p = 0.031; lift-off, p = 0.032). No significant changes in AHD or Hamada grade were observed in either group. At final follow-up, no significant differences were found between the 2 groups in terms of AHD (p = 0.539) and Hamada grade (p = 0.974). Although preoperative anterior glenohumeral subluxation was improved in both groups, the LDTM group showed a statistically significantly higher rate of restoration compared to the LD group (p = 0.015). Conclusions: While both LD and combined LDTM transfers for IASRCTs improved postoperative pain relief, clinical scores, and active ROM, the combined LDTM transfer attached to GT was superior to LD transfer attached to LT in terms of internal rotational strength, ADLIR score, and SSC-specific examinations. Neither group showed significant progress in cuff tear arthropathy or decreased AHD at 2-year follow-up; however, the combined LDTM transfer notably improved preoperative anterior glenohumeral subluxation.
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Amplitude de Movimento Articular , Lesões do Manguito Rotador , Músculos Superficiais do Dorso , Transferência Tendinosa , Humanos , Lesões do Manguito Rotador/cirurgia , Estudos Retrospectivos , Transferência Tendinosa/métodos , Masculino , Feminino , Pessoa de Meia-Idade , Músculos Superficiais do Dorso/transplante , Idoso , AdultoRESUMO
The fusion of autophagosomes and lysosomes is essential for the prevention of nonalcoholic fatty liver disease (NAFLD). Here, we generate a hepatocyte-specific CHIP knockout (H-KO) mouse model that develops NAFLD more rapidly in response to a high-fat diet (HFD) or high-fat, high-fructose diet (HFHFD). The accumulation of P62 and LC3 in the livers of H-KO mice and CHIP-depleted cells indicates the inhibition of autophagosome-lysosome fusion. AAV8-mediated overexpression of CHIP in the murine liver slows the progression of NAFLD induced by HFD or HFHFD feeding. Mechanistically, CHIP induced K63- and K27-linked polyubiquitination at the lysine 198 residue of STX17, resulting in increased STX17-SNAP29-VAMP8 complex formation. The STX17 K198R mutant was not ubiquitinated by CHIP; it interfered with its interaction with VAMP8, rendering STX17 incapable of inhibiting steatosis development in mice. These results indicate that a signaling regulatory mechanism involving CHIP-mediated non-degradative ubiquitination of STX17 is necessary for autophagosome-lysosome fusion.
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Autofagossomos , Lisossomos , Camundongos Knockout , Hepatopatia Gordurosa não Alcoólica , Ubiquitina-Proteína Ligases , Ubiquitinação , Animais , Lisossomos/metabolismo , Hepatopatia Gordurosa não Alcoólica/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , Hepatopatia Gordurosa não Alcoólica/genética , Autofagossomos/metabolismo , Camundongos , Humanos , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina-Proteína Ligases/genética , Dieta Hiperlipídica/efeitos adversos , Masculino , Proteínas Qa-SNARE/metabolismo , Proteínas Qa-SNARE/genética , Hepatócitos/metabolismo , Modelos Animais de Doenças , Fígado/metabolismo , Fígado/patologia , Camundongos Endogâmicos C57BL , Proteínas R-SNARE/metabolismo , Proteínas R-SNARE/genética , Fusão de Membrana , Autofagia , Fator de Transcrição TFIIHRESUMO
We investigated the feasibility of magnetic resonance elastography (MRE) using a pelvic acoustic driver for the detection and classification of prostate cancer (PCa). A total of 75 consecutive patients (mean age, 70; range, 56-86) suspected of having PCa and who underwent multi-parametric MRI including MRE and subsequent surgical resection were included. The analyzed regions consisted of cancer (n = 69), benign prostatic hyperplasia (BPH) (n = 70), and normal parenchyma (n = 70). A histopathologic topographic map served as the reference standard for each region. One radiologist and one pathologist performed radiologic-pathologic correlation, and the radiologist measured stiffness values in each region of interest on elastograms automatically generated by dedicated software. Paired t-tests were used to compare stiffness values between two regions. ROC curve analysis was also used to extract a cutoff value between two regions. The stiffness value of PCa (unit, kilopascal (kPa); 4.9 ± 1.1) was significantly different to that of normal parenchyma (3.6 ± 0.3, p < 0.0001) and BPH (4.5 ± 1.4, p = 0.0454). Under a cutoff value of 4.2 kPa, a maximum accuracy of 87% was estimated, with a sensitivity of 73%, a specificity of 99%, and an AUC of 0.839 for discriminating PCa from normal parenchyma. Between PCa and BPH, a maximum accuracy of 62%, a sensitivity of 70%, a specificity of 56%, and an AUC of 0.598 were estimated at a 4.5 kPa cutoff. The stiffness values tended to increase as the ISUP grade increased. In conclusion, it is feasible to detect and classify PCa using pelvic MRE. Our observations suggest that MRE could be a supplement to multi-parametric MRI for PCa detection.
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Two-dimensional (2D) halide perovskites have become a promising class of memristive materials due to their low power consumption, compositional versatility, and microstructural anisotropy in electronics. However, implementing high-performance resistive random-access memory requires a higher reliability and moisture resistance. To address these issues, component studies and attempts to improve the phase stability have been reported but have not been able to achieve sufficient reliability. Here, highly textured thin films grown perpendicular to the substrate in Ruddlesden-Popper 2D perovskites exhibited highly stable and reliable binary memory performance. We further built a flexible crossbar array to verify data storage capability, achieving a high device yield, robust endurance, long retention, reliability to operate under bending conditions, and moisture stability over a year. These device performances are attributed to preformed vertically oriented nanocrystals that allow the conductive filaments to operate reliably. Our finding provides the material design strategy that can be extended to the development of semiconductor materials for next-generation memory devices.
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As electronic circuit integration intensifies, there is a rising demand for dielectric insulators that provide both superior insulation and high dielectric constants. This study focuses on developing high-k dielectric insulators by controlling the phase of the Hf0.5Zr0.5O2 (HZO) film with additional doping, utilizing yttrium (Y), tantalum (Ta), gallium (Ga), silicon (Si), and aluminum (Al) as dopants. Doping changes the ratio of tetragonal to monoclinic phases in doped HZO films, and Y-doped HZO (Y:HZO) films specifically exhibit a high tetragonal phase ratio and a dielectric constant of 40.9, indicating superior insulating properties compared to undoped HZO films. To clarify the fundamental mechanism driving the enhancement in dielectric properties, we have carried out various analyses combined with density functional theory (DFT) calculations. Through the optimization of the post-deposition annealing (PDA) process and the heterojunction structure with Al2O3, an Al2O3/Y:HZO heterojunction with a high dielectric constant and even lower leakage current compared to a single layer was developed. The thin-film transistor (TFT) with the Au/Ti/amorphous InGaZnO4 (a-IGZO)/Al2O3/YHZO/TiN heterojunction structure exhibits low subthreshold swing (SS) values within a narrow gate-source voltage (Vsg) range. This study advances knowledge on how the controlled-phase doped HZO films affect the dielectric constant and leakage current and will contribute to semiconductor technology advancements by overcoming the limitations of conventional high-k dielectric insulators.
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Men taking antioxidant vitamin E supplements have increased prostate cancer (PC) risk. However, whether pro-oxidants protect from PC remained unclear. In this work, we show that a pro-oxidant vitamin K precursor [menadione sodium bisulfite (MSB)] suppresses PC progression in mice, killing cells through an oxidative cell death: MSB antagonizes the essential class III phosphatidylinositol (PI) 3-kinase VPS34-the regulator of endosome identity and sorting-through oxidation of key cysteines, pointing to a redox checkpoint in sorting. Testing MSB in a myotubular myopathy model that is driven by loss of MTM1-the phosphatase antagonist of VPS34-we show that dietary MSB improved muscle histology and function and extended life span. These findings enhance our understanding of pro-oxidant selectivity and show how definition of the pathways they impinge on can give rise to unexpected therapeutic opportunities.
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Classe III de Fosfatidilinositol 3-Quinases , Doenças Musculares , Oxidantes , Neoplasias da Próstata , Vitamina K 3 , Animais , Humanos , Masculino , Camundongos , Classe III de Fosfatidilinositol 3-Quinases/antagonistas & inibidores , Cisteína/metabolismo , Suplementos Nutricionais , Longevidade/efeitos dos fármacos , Oxidantes/administração & dosagem , Oxidantes/farmacologia , Oxirredução , Neoplasias da Próstata/tratamento farmacológico , Proteínas Tirosina Fosfatases não Receptoras/metabolismo , Vitamina K 3/administração & dosagem , Vitamina K 3/farmacologia , Doenças Musculares/tratamento farmacológicoRESUMO
Background: Glucagon-like peptide-1 receptor agonists (GLP-1 RAs) are increasingly used for the treatment of type 2 diabetes mellitus (T2DM) given their extra-pancreatic effects. However, there are concerns about carcinogenesis in the pancreas and thyroid gland. We aimed to evaluate the site-specific incidence of cancer in patients with T2DM-treated GLP-1 RAs using a nationwide cohort. Methods: This study included data obtained from the Korean National Health Insurance Service (between 2004 and 2021). The primary outcome was newly diagnosed cancer, and the median follow-up duration for all participants was 8 years. Results: After propensity score matching, 7,827 participants were analyzed; 2,609 individuals each were included in the GLP-1 RA, diabetes mellitus (DM) control, and non-DM control groups. The incidence rate ratio (IRR) of subsequent cancer in patients with T2DM was 1.73, which was higher than that of individuals without DM, and it increased in both men and women. Analysis of patients with T2DM showed no increased cancer risk associated with the use of GLP-1 RA, and similar results were observed in both men and women. The IRRs of pancreatic cancer (0.74), thyroid cancer (1.32), and medullary thyroid cancer (0.34) did not significantly increase in the GLP-1 RA group compared with those in the DM control group. Conclusion: There was a 73% higher risk of cancer in patients with T2DM compared with the general population. However, among patients with T2DM, there was no association between the use of GLP-1 RAs and new-onset cancers, including pancreatic and medullary thyroid cancers.
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High-resolution electrogastrography (HR-EGG) presents a new paradigm in diagnosing and treating functional gastroduodenal disorders. Unlike traditional electrogastrography, HR-EGG allows for a more precise analysis of the gastric electrical activity, offering improved diagnostic accuracy. Recent studies have revealed the clinical potential of HR-EGG, particularly in detecting abnormal electrical patterns in patients with functional dyspepsia and gastroparesis, supporting the development of novel therapeutic strategies. The non-invasive HR-EGG method has shown promise in identifying new biomarkers. Moreover, further integration of artificial intelligence, is expected to enhance diagnostic efficiency and develop more refined treatment models for functional gastrointestinal disorders.
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Gastroparesia , Humanos , Gastroparesia/diagnóstico , Eletrodiagnóstico , Dispepsia/diagnóstico , EletromiografiaRESUMO
Chronic kidney disease (CKD) is a global health problem characterized by progressive renal fibrosis and excessive extracellular matrix deposition. Oxidative stress and epigenetic regulation, particularly through microRNAs (miRNAs), play crucial roles in the pathogenesis of CKD. In this study, we investigated the role of urinary miR-144-3p, which is upregulated in rats with CKD induced by diabetes and hypertension, in renal fibrosis progression, particularly its regulation of the nuclear factor erythroid-2-related factor 2 (NRF2) pathway. Our findings revealed elevated miR-144-3p levels and reduced NRF2 and target gene levels in kidney tissues of streptozotocin-treated spontaneously hypertensive rats. In vitro experiments demonstrated that miR-144-3p directly binds to the 3'-untranslated region of nrf2, suppressing the NRF2 pathway in renal tubular epithelial cells. Additionally, the profibrogenic factor transforming growth factor (TGF)-ß1 increased miR-144-3p expression. TGF-ß1-induced NRF2 suppression and reactive oxygen species elevation were found to be mediated through miR-144-3p upregulation. In vivo, cilostazol, an antiplatelet drug with an NRF2-activating effect, ameliorated renal injury in diabetic hypertensive rats by decreasing TGF-ß1 and miR-144-3p levels while increasing NRF2 and its target gene levels in the kidneys. These findings highlight the potential therapeutic value of targeting the miR-144-3p/NRF2 pathway to attenuate CKD progression in hypertensive diabetic conditions.
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Oyster aquaculture is one of the fastest-growing food production industries worldwide; however, it faces a significant challenge from the protist Perkinsus marinus, particularly in the USA. Although several quantitative molecular diagnostic methodologies are available for identifying diseases caused by P. marinus, the primer pairs used therein led to non-specific identification of other Perkinsus spp. Hence, a quantitative real-time PCR (Pm-qPCR) assay specific for P. marinus was developed using a TaqMan-based probe with the internal quencher in this study. A primer pair and probe specific to P. marinus were designed from a hypothetical protein of P. marinus collected from the whole-genome shotgun sequence database of the National Center for Biotechnology Information (NCBI). In silico analysis using homologous sequences of P. olseni and P. chesapeaki confirmed the high specificity of primers designed in this study. The Pm-qPCR assay was performed using seven different strains of P. marinus, P. olseni, and P. chesapeaki, revealing high specificity and sensitivity for detecting only P. marinus strains. In conclusion, it was demonstrated that Pm-qPCR can effectively and accurately diagnose P. marinus with high specificity and sensitivity. This assay is promising for monitoring oyster health and disease management in ecosystems and aquaculture.
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Alveolados , Ostreidae , Reação em Cadeia da Polimerase em Tempo Real , Animais , Reação em Cadeia da Polimerase em Tempo Real/métodos , Alveolados/genética , Alveolados/isolamento & purificação , Ostreidae/parasitologia , Sensibilidade e Especificidade , Aquicultura , Primers do DNA/genéticaRESUMO
Muscle atrophy due to prolonged immobilization leads to severe dysfunction and progression of disease and injury. This highlights the necessity for early rehabilitation, even during the non-ambulatory stages. As manifestations vary among individuals, target-specific rehabilitation is crucial for achieving optimal outcomes. Conventional bicycle ergometers, despite their wide usage in rehabilitation and sports training, fail to adequately recruit the muscles that predominantly operate in the coronal or axial plane. This is thought to be due to the restriction of pedaling trajectory in the sagittal plane. This study introduces a new bicycle ergometer capable of tilting the pedaling plane and adjusting the pedal orientation, in order to alter muscle recruitment patterns and limb alignments. A biomechanical analysis of eight healthy volunteers suggests that our device can elicit dynamic valgus/varus alignment and alter the axial rotations of the lower extremities. An electromyography analysis revealed a significant increase in the activation of specific muscle groups, particularly the abductors and adductors, suggesting promising results for targeted muscle recruitment. We expect this ergometer to provide a new method for target-specific rehabilitation and the treatment of bedridden patients or those with weak muscles.
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Ciclismo , Eletromiografia , Estudos de Viabilidade , Extremidade Inferior , Humanos , Masculino , Adulto , Extremidade Inferior/fisiopatologia , Feminino , Fenômenos Biomecânicos , Ergometria/instrumentação , Ergometria/métodos , Músculo Esquelético/fisiopatologia , Músculo Esquelético/fisiologia , Voluntários Saudáveis , Atrofia Muscular/reabilitação , Atrofia Muscular/fisiopatologia , Adulto Jovem , Teste de Esforço/métodosRESUMO
BACKGROUND: The genus Robinsonia DC. (tribe Senecioneae, Asteraceae) endemic to the Juan Fernández Islands in Chile is one of the most conspicuous insular plant groups in the world. Unlike typical herbaceous Asteraceae plants, these plants demonstrate spectacular and unusual rosette tree growth forms as shown by the alpine giant senecios (genus Dendrosenecio, tribe Senecioneae) endemic to the East African mountains. However, monophyly of the genus and phylogenetic relationships among species of Robinsonia as well as their plastome evolution remain elusive. This study aims to explore their phylogeny, species diversification, and molecular evolution based on the complete plastome sequences in the context of adaptive radiation on oceanic islands. RESULTS: The insular Robinsonia plastomes are highly conserved in their structures and organization of contents. Five divergence hotspots as potential chloroplast markers and five positively selected coding genes (accD, ndhF, rpoA, ycf1, and ycf2) are identified. Robinsonia plastomes has an overall nucleotide diversity higher than that of the sky island Dendrosenecio, but much lower than herbaceous Senecio. Phylogenetic analysis demonstrates the monophyly of Robinsonia and identifies two major infrageneric lineages. Both Robinsonia and Dendrosenecio are deeply nested within large genus Senecio. CONCLUSIONS: While plastid genomes of Robinsonia are highly conserved, their sequences strongly demonstrated the monophyly of the genus and inferred robust interspecific relationships, including herbaceous Senecio and woody Dendrosenecio. Different sets of positively selected chloroplast genes, five for Robinsonia and two for Dendrosenecio, may play an important role in the adaptation strategies of these fascinating woody species in insular and continental sky island habitats. Overall phylogenetic positions and sister lineages of Robinsonia and Dendrosenecio require additional study based on broader sampling of Senecio.