Identification of the central role of RNA polymerase mitochondrial for angiogenesis.

Mitochondria are central to endothelial cell activation and angiogenesis, with the RNA polymerase mitochondrial (POLRMT) serving as a key protein in regulating mitochondrial transcription and oxidative phosphorylation. In our study, we examined the impact of POLRMT on angiogenesis and found that its silencing or knockout (KO) in human umbilical vein endothelial cells (HUVECs) and other endothelial cells resulted in robust anti-angiogenic effects, impeding cell proliferation, migration, and capillary tube formation. Depletion of POLRMT led to impaired mitochondrial function, characterized by mitochondrial depolarization, oxidative stress, lipid oxidation, DNA damage, and reduced ATP production, along with significant apoptosis activation. Conversely, overexpressing POLRMT promoted angiogenic activity in the endothelial cells. In vivo experiments demonstrated that endothelial knockdown of POLRMT, by intravitreous injection of endothelial specific POLRMT shRNA adeno-associated virus, inhibited retinal angiogenesis. In addition, inhibiting POLRMT with a first-in-class inhibitor IMT1 exerted significant anti-angiogenic impact in vitro and in vivo. Significantly elevated expression of POLRMT was observed in the retinal tissues of streptozotocin-induced diabetic retinopathy (DR) mice. POLRMT endothelial knockdown inhibited pathological retinal angiogenesis and mitigated retinal ganglion cell (RGC) degeneration in DR mice. At last, POLRMT expression exhibited a substantial increase in the retinal proliferative membrane tissues of human DR patients. These findings collectively establish the indispensable role of POLRMT in angiogenesis, both in vitro and in vivo.

Estimation of rice yield using multivariate analysis techniques based on meteorological parameters.

This study aims to develop predictive models for rice yield by applying multivariate techniques. It utilizes stepwise multiple regression, discriminant function analysis and logistic regression techniques to forecast crop yield in specific districts of Haryana. The time series data on rice crop have been divided into two and three classes based on crop yield. The yearly time series data of rice yield from 1980-81 to 2020-21 have been taken from various issues of Statistical Abstracts of Haryana. The study also utilized fortnightly meteorological data sourced from the Agrometeorology Department of CCS HAU, India. For comparing various predictive models' performance, evaluation of measures like Root Mean Square Error, Predicted Error Sum of Squares, Mean Absolute Deviation and Mean Absolute Percentage Error have been used. Results of the study indicated that discriminant function analysis emerged as the most effective to predict the rice yield accurately as compared to logistic regression. Importantly, the research highlighted that the optimum time for forecasting the rice yield is 1 month prior to the crops harvesting, offering valuable insight for agricultural planning and decision-making. This approach demonstrates the fusion of weather data and advanced statistical techniques, showcasing the potential for more precise and informed agricultural practices.

Cross-sectional study on dysphagia evaluation in community-dwelling older adults using the Eating Assessment Tool (EAT) -10, EAT-2, and Water Swallow Test.

BACKGROUND: Dysphagia is a health concern that causes severe complications and affects the life quality of the older population. This study aimed to determine the diagnostic performance of the Eating Assessment Tool (EAT)-2 compared with the EAT-10 and the Water Swallow Test (WST) in screening for dysphagia. METHODS: A cross-sectional study was conducted among 5,090 community-dwelling older adults. Dysphagia was evaluated using both a subjective measure, the 10-item EAT (EAT-10) and an objective measure, the WST. The kappa index in pairs were analyzed. The validity and reliability of EAT-2 were also assessed. RESULTS: The sensitivity and specificity of the EAT-2 were 96.3 % and 94.8 %, respectively. The kappa index between the EAT-2 and EAT-10 was 0.64, whereas it was 0.11 between the EAT-10 and WST. CONCLUSIONS: The EAT-2 was a simpler screening tool for dysphagia. Combining the subjective questionnaire (EAT-10 or EAT-2) and the objective test (WST) is recommended.

Panax ginseng total protein promotes proliferation and secretion of collagen in NIH/3T3 cells by activating extracellular signal-related kinase pathway.

BACKGROUND: Recently, protein from ginseng was studied and used for the treatment of several kinds of diseases. However, the effect of ginseng total protein (GTP) on proliferation and wound healing in fibroblast cells remains unclear. METHODS: In this study, cell viability was analyzed using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Cell cycle distribution was analyzed by flow cytometer. The levels of transforming growth factor ß1, vascular endothelial growth factor, and collagens were analyzed by enzyme-linked immunosorbent assay and immunofluorescence staining. The expressions of cyclin A, phosphorylation of extracellular signal-related kinase (p-ERK1/2), and ERK1/2 were analyzed by Western blotting. RESULTS: Our results showed that GTP promoted cell proliferation and increased the percentage of cells in S phase through the upregulation of cyclin A in NIH/3T3 cells. We also found that GTP induced the secretion of type I collagen, and promoted the expression of other factors that regulate the synthesis of collagen such as transforming growth factor ß1 and vascular endothelial growth factor. In addition, the phosphorylation of ERK1/2 at Thr202/Tyr204 was also increased by GTP. CONCLUSION: Our studies suggest that GTP promoted proliferation and secretion of collagen in NIH/3T3 cells by activating the ERK signal pathway, which shed light on a potential function of GTP in promoting wound healing.

Baclofen Protects Primary Rat Retinal Ganglion Cells from Chemical Hypoxia-Induced Apoptosis Through the Akt and PERK Pathways.

Retinal ganglion cells (RGCs) consume large quantities of energy to convert light information into a neuronal signal, which makes them highly susceptible to hypoxic injury. This study aimed to investigate the potential protection by baclofen, a GABAB receptor agonist of RGCs against hypoxia-induced apoptosis. Cobalt chloride (CoCl2) was applied to mimic hypoxia. Primary rat RGCs were subjected to CoCl2 with or without baclofen treatment, and RNA interference techniques were used to knock down the GABAB2 gene in the primary RGCs. The viability and apoptosis of RGCs were assessed using cell viability and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assays, Hoechst staining, and flow cytometry. The expression of cleaved caspase-3, bcl-2, bax, Akt, phospho-Akt, protein kinase RNA (PKR)-like ER kinase (PERK), phospho-PERK, eIF2α, phospho-eIF2α, ATF-4 and CCAAT/enhancer-binding protein homologous protein (CHOP) were measured using western blotting. GABAB2 mRNA expression was determined using quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Our study revealed that CoCl2 significantly induced RGC apoptosis and that baclofen reversed these effects. CoCl2-induced reduction of Akt activity was also reversed by baclofen. Baclofen prevented the activation of the PERK pathway and the increase in CHOP expression induced by CoCl2. Knockdown of GABAB2 and the inactivation of the Akt pathway by inhibitors reduced the protective effect of baclofen on CoCl2-treated RGCs. Taken together, these results demonstrate that baclofen protects RGCs from CoCl2-induced apoptosis by increasing Akt activity and by suppressing the PERK pathway and CHOP activation.

Motivating protégés' personal learning in teams: a multilevel investigation of autonomy support and autonomy orientation.

This study examined the roles of 3 multilevel motivational predictors in protégés' personal learning in teams: an autonomy-supportive team climate, mentors' autonomy support, and protégés' autonomy orientation. The authors followed 305 protégés in 58 teams for 12 weeks and found that all 3 predictors were positively related to the protégés' personal learning in teams and that an autonomy-supportive team climate augmented the effects of mentors' autonomy support and protégés' autonomy orientation on protégés' personal learning in teams. Protégés' personal learning in teams mediated the interactive effects of an autonomy-supportive team climate with mentors' autonomy support or protégés' autonomy orientation on protégés' behavioral and attitudinal outcomes, including their organizational citizenship behaviors and job involvement. The findings of this study provide business researchers and practitioners with valuable insights into the management of autonomy.

Adiponectin activation of AMPK disrupts leptin-mediated hepatic fibrosis via suppressors of cytokine signaling (SOCS-3).

Adiponectin is an adipocytokine that was recently shown to be anti-fibrogenic in hepatic fibrosis. Leptin, on the other hand, promotes hepatic fibrosis. The purpose of the present study was to elucidate a mechanism (or mechanisms) whereby adiponectin dampens leptin signaling in activated hepatic stellate cells (HSCs), and prevents excess extracellular matrix production. Activated HSCs, between passages 2 and 5, were cultured and exposed to recombinant human adiponectin and recombinant leptin. Immunoblot analysis for SOCS-3, TIMP-1, and the phosphorylated species of Stat3 and adenosine monophosphate-activated protein kinase (AMPK) were conducted. We also examined MMP-1 activity by immunosorbant fluorimetric analysis. In HSCs, adiponectin-induced phosphorylation of AMPK, and subsequently suppressed leptin-mediated Stat3 phosphorylation and SOCS-3 induction. Adiponectin also blocked leptin-stimulated secretion of TIMP-1, and significantly increased MMP-1 activity, in vitro. To extend this study, we treated adiponectin knockout mice (Ad-/-) daily with 5 mg/kg recombinant leptin and/or carbon tetrachloride (2 ml/kg) for 6 weeks. Post-necropsy analysis was performed to examine for inflammation, and histological changes in the Ad-/- and wild-type mice. There was no significant difference in inflammation, or aminotransferases, between mice receiving carbon tetrachloride and leptin versus carbon tetrachloride alone. As anticipated, the combination of leptin and CCl(4) enhanced hepatic fibrosis in both wild-type and Ad-/- mice, as estimated by amount of collagen in injured livers, but wild-type mice had significantly higher levels of SOCS-3 and significantly lower levels of TIMP-1 mRNA and protein than did adiponectin KO mice exposed to both CCl(4) and leptin. We therefore conclude that the protective effects of adiponectin against liver fibrosis require AMPK activation, and may occur through inhibition of the Jak-Stat signal transduction pathway.

Enforced expression of MCAM/MUC18 increases in vitro motility and invasiveness and in vivo metastasis of two mouse melanoma K1735 sublines in a syngeneic mouse model.

Human MCAM/MUC18 has been shown to increase metastasis of human melanoma cells in xenograft mouse systems. To be more relevant to understanding the progression of clinical melanoma and for designing better preclinical therapeutic trials, it is highly desirable to establish a syngeneic mouse model for studying the mechanisms of MCAM/MUC18-mediated tumorigenesis and metastasis of melanoma cells. To reach this goal, we transfected the mouse MCAM/MUC18 (moMCAM/MUC18) cDNA into two MCAM/MUC18-minus, low-metastatic mouse melanoma K1735 sublines, K1735-10 (tumor(-)/met(low)) and K1735-3 (tumor(+)/met(low)), and selected for G418-resistant clones, which expressed different levels of moMCAM/MUC18, and used for testing the effect of MCAM/MUC18 overexpression on their in vitro growth rate, motility, and invasiveness and in vivo subcutaneous tumor growth and pulmonary metastasis in syngeneic mice. Enforced expression of moMCAM/MUC18 did not significantly affect in vitro growth rate, but it increased the in vitro motility and invasiveness of clones derived from both sublines. Ectopic expression of moMCAM/MUC18 did not alter the nontumorigenicity of the K1735-10 clones per cells nor significantly affect the subcutaneous tumor growth of the K1735-3 clones per cells. The moMCAM/MUC18-expressing K1735-10 clones were able to establish only microscopic lung modules in 86% of the mice. In contrast, the moMCAM/MUC18-expressing K1735-3 clones could induce numerous large lung nodules (3-4 mm in diameter) in all the mice. We concluded that increased moMCAM/MUC18 expression in the two K1735 sublines minimally affected their tumorigenicity, but it augmented their in vitro motility and invasiveness and increased their pulmonary metastasis in the syngeneic C3H mice.

Increased expression of MUC18 correlates with the metastatic progression of mouse prostate adenocarcinoma in the TRAMP model.

PURPOSE: The transgenic adenocarcinoma mouse prostate (TRAMP) model is a paradigm that closely mimics the progression of clinical prostate cancer. We have previously reported that MUC18, a cell adhesion molecule in the Ig gene superfamily, is a marker as well as an important mediator for the metastatic potential of human prostate cancer cells. In this study we investigated the possible correlation of increased MUC18 expression with the malignant progression of prostate cancer in the TRAMP model. MATERIALS AND METHODS: We used immunohistochemistry, Western blot and reverse transcriptase-polymerase chain reaction analyses to determine MUC18 expression in the prostate gland of 178 to 282-day-old TRAMP positive males with a prostate tumor size of 0.4 to 12.7 gm. Eight normal prostates, 10 prostates with high grade prostatic intraepithelial neoplasia (PIN), 24 prostates with primary prostate cancer, 10 metastatic lesions from 50 pure C57BL/6 TRAMP mice (Wu colony) and 2 normal prostates, 2 prostates with high grade PIN, 6 prostates with primary prostate cancer and 4 metastatic lesions from 10 [C57BL/6 TRAMP x FVB] F1 mice (NMG colony) were used. RESULTS: We found that mouse MUC18 was expressed in all (100%) high grade PIN, adenocarcinomas and metastatic lesions. All mice bearing primary prostate tumors had prostate cancer metastatic to the peri-aortic lymph nodes and some had it to other organs (liver, lung, kidney, testes, seminal vesicles and abdominal cavity). In contrast, prostates from 10 nontransgenic littermates did not have detectable MUC18 expression. CONCLUSIONS: MUC18 expression is up-regulated in the TRAMP model and it correlates with the malignant progression of mouse prostate adenocarcinoma in this transgenic model. This further strengthens the hypothesis that MUC18 has an important role in increasing the metastatic potential of prostate cancer cells.

Ectopical expression of human MUC18 increases metastasis of human prostate cancer cells.

MUC18, a cell adhesion molecule (CAM), has been reported to be a diagnostic marker for the early detection of the metastatic potential of prostate cancers as well as implicated to be an important determinant for mediating the tumorigenesis and metastasis of prostate cancer. To test the hypothesis, we further investigated the possible role of MUC18 in the malignant progression of human prostate cancer. The human MUC18-minus, non-metastatic human prostate cancer LNCaP cells were transfected with the human cytomegalovirus immediate-early gene (HCMV-IE) promoter-driven human MUC18 (huMUC18) cDNA. The G418-resistant (G418R)-LNCaP clones that expressed a high level of huMUC18 were selected and used for testing the effect of huMUC18 expression on the in vitro growth, motility, and invasiveness as well as on the in vivo metastasis (via orthotopical injection) in a xenograft nude mouse model. HuMUC18 expression increased by four- to fivefold of in vitro motility and invasiveness of LNCaP cells. Anti-huMUC18 antibody significantly inhibited the in vitro motility and invasiveness of huMUC18-expressing LNCaP clones, but not the control clones. We suggest that huMUC18 expression is responsible for increasing these behaviors of LNCaP cells. HuMUC18 expression also directly increased the in vivo metastatic abilities of the LNCaP cells from the prostate gland to multiple distant organs. Western blot and immunohistochemistry analyses showed that the prostatic tumors as well as metastatic lesions expressed high levels of MUC18, indicating that they originated from the injected huMUC18-expressing LNCaP cells. We therefore conclude that HuMUC18 is an important determinant in increasing metastasis of human prostate cancer LNCaP cells to distant organs in a nude mouse model.