Endoplasmic reticulum stress of endometrial mesenchymal stem cells in endometriosis.

OBJECTIVE: The human endometrium has significant regenerative abilities due to stem cells, which are vital in immunomodulation, immune tolerance, steroid hormone response, and inflammation. Endometriosis, an inflammatory gynecological disorder where endometrium-like tissue grows outside uterus, affects millions of women and often causes infertility. Recent research indicates that stem cells contribute to pathology of endometriosis. ER stress is implicated in various diseases, including endometriosis. This study aims to examine ER stress in eMSCs within endometriosis pathogenesis and uncover underlying disease mechanisms. METHODS: Samples were collected from healthy subjects and women with endometriosis in both proliferative and secretory phases. eMSCs were isolated and characterized via flow cytometry. ER stress protein levels were assessed using proteomic analysis, with validation through Western Blot and immunofluorescence staining. Gene expression was analyzed by RT-qPCR, and ultrastructural examination of eMSCs was conducted using TEM. ER stress markers in tissue samples were detected in SUSD2+ eMSCs through immunofluorescence staining and visualized using a confocal microscope. Statistical analysis was performed using SPSS program. RESULTS: The proteomics analysis uncovered ER stress-related proteins (DDRGK1, RTN3, ERp44, TMED2, TMEM33, TMX3) whose levels were significantly distinct from control group. Western Blot analysis and immunofluorescence staining results at protein level; RT-qPCR results at gene level supported these findings. TEM analysis also showed ultrastructural presence of ER stress in endometriosis groups. CONCLUSION: Presence of ER stress in eMSCs in pathogenesis of endometriosis has been demonstrated using various methods. Our research has potential to shed light on pathology of endometriosis and offer promising avenues for non-invasive diagnosis and potential treatment.

The effect of obesity on the outcomes of laparoscopic adrenal surgery in patients with Cushing syndrome.

Purpose: The aim of this study is to investigate the effect of obesity on the results of laparoscopic adrenal surgery in patients with Cushing syndrome. Methods: This retrospective study was performed in Department of General Surgery at Erciyes University School of Medicine between January 2010 and January 2023. Our analysis included Cushing syndrome patients who underwent unilateral laparoscopic adrenalectomy (LA) with the transabdominal lateral approach. All patients were evaluated in terms of age, sex, tumor diameter, body mass index (BMI), American Society of Anesthesiologists physical status classification, morbidities, surgery history, tumor side, operative time, conversion to open surgery, complications, and length of hospital stay. Results: A total of 90 patients (75 females and 15 males) underwent a transperitoneal LA. Patients were divided into 2 groups according to their BMI: obese group (≥30 kg/m2; n = 53, 58.8%) and nonobese group (<30 kg/m2; n = 37, 41.2%). All patients were classified into 3 subgroups: obese patient group, BMI ≥30-39 kg/m2 (n = 23); morbidly obese patient group, BMI ≥40 kg/m2 (n = 14); and nonobese patient group, BMI <30 kg/m2 (n = 53). There was no significant difference in intraoperative complications, conversion to open surgery, operative time, or length of hospital stay between the groups. Only conversion to open surgery was a risk factor for postoperative complications in univariate and multivariate analyses (odds ratio, 15.4; 95% confidence interval, 1.277-185.599; P = 0.031). Conclusion: Our results showed that LA is safe and effective in patients with Cushing syndrome with morbid obesity, allowing acceptable morbidity and length of hospital stay.

Enhanced anti-angiogenic effects of aprepitant-loaded nanoparticles in human umbilical vein endothelial cells.

Recent advancements in cancer therapy have led to the development of novel nanoparticle-based drug delivery systems aimed at enhancing the efficacy of chemotherapeutic agents. This study focuses on evaluating aprepitant-loaded PLGA and Eudragit RS 100 nanoparticles for their potential antiangiogenic effects. Characterization studies revealed that aprepitant-loaded nanoparticles exhibited particle sizes ranging from 208.50 to 238.67 nm, with monodisperse distributions (PDI < 0.7) and stable zeta potentials (between - 5.0 and - 15.0 mV). Encapsulation efficiencies exceeding 99% were achieved, highlighting the efficacy of PLGA and Eudragit RS 100 as carriers for aprepitant. Cellular uptake studies demonstrated enhanced internalization of aprepitant-loaded nanoparticles by HUVEC cells compared to free aprepitant, as confirmed by fluorescence microscopy. Furthermore, cytotoxicity assays revealed significant dose-dependent effects of aprepitant-loaded nanoparticles on HUVEC cell viability, with IC50 values at 24 h of 11.9 µg/mL for Eudragit RS 100 and 94.3 µg/mL for PLGA formulations. Importantly, these nanoparticles effectively inhibited HUVEC cell migration and invasion induced by M2c supernatant, as evidenced by real-time cell analysis and gene expression studies. Moreover, aprepitant-loaded nanoparticles downregulated VEGFA and VEGFB gene expressions and reduced VEGFR-2 protein levels in HUVEC cells, highlighting their potential as antiangiogenic agents. Overall, this research underscores the promise of nanoparticle-based aprepitant formulations in targeted cancer therapy, offering enhanced therapeutic outcomes through improved drug delivery and efficacy against angiogenesis.

The accuracy of Gemini, GPT-4, and GPT-4o in ECG analysis: A comparison with cardiologists and emergency medicine specialists.

INTRODUCTION: GPT-4, GPT-4o and Gemini advanced, which are among the well-known large language models (LLMs), have the capability to recognize and interpret visual data. When the literature is examined, there are a very limited number of studies examining the ECG performance of GPT-4. However, there is no study in the literature examining the success of Gemini and GPT-4o in ECG evaluation. The aim of our study is to evaluate the performance of GPT-4, GPT-4o, and Gemini in ECG evaluation, assess their usability in the medical field, and compare their accuracy rates in ECG interpretation with those of cardiologists and emergency medicine specialists. METHODS: The study was conducted from May 14, 2024, to June 3, 2024. The book "150 ECG Cases" served as a reference, containing two sections: daily routine ECGs and more challenging ECGs. For this study, two emergency medicine specialists selected 20 ECG cases from each section, totaling 40 cases. In the next stage, the questions were evaluated by emergency medicine specialists and cardiologists. In the subsequent phase, a diagnostic question was entered daily into GPT-4, GPT-4o, and Gemini Advanced on separate chat interfaces. In the final phase, the responses provided by cardiologists, emergency medicine specialists, GPT-4, GPT-4o, and Gemini Advanced were statistically evaluated across three categories: routine daily ECGs, more challenging ECGs, and the total number of ECGs. RESULTS: Cardiologists outperformed GPT-4, GPT-4o, and Gemini Advanced in all three groups. Emergency medicine specialists performed better than GPT-4o in routine daily ECG questions and total ECG questions (p = 0.003 and p = 0.042, respectively). When comparing GPT-4o with Gemini Advanced and GPT-4, GPT-4o performed better in total ECG questions (p = 0.027 and p < 0.001, respectively). In routine daily ECG questions, GPT-4o also outperformed Gemini Advanced (p = 0.004). Weak agreement was observed in the responses given by GPT-4 (p < 0.001, Fleiss Kappa = 0.265) and Gemini Advanced (p < 0.001, Fleiss Kappa = 0.347), while moderate agreement was observed in the responses given by GPT-4o (p < 0.001, Fleiss Kappa = 0.514). CONCLUSION: While GPT-4o shows promise, especially in more challenging ECG questions, and may have potential as an assistant for ECG evaluation, its performance in routine and overall assessments still lags behind human specialists. The limited accuracy and consistency of GPT-4 and Gemini suggest that their current use in clinical ECG interpretation is risky.

Efficacy of cumulative cisplatin dose on survival in patients with locally advanced cervical cancer treated with definitive chemoradiotherapy: multicenter study by Turkish Oncology Group.

OBJECTIVE: To investigate the impact of cumulative cisplatin dose on clinical outcomes in locally advanced cervical cancer patients undergoing definitive chemoradiotherapy. METHODS: A retrospective analysis was conducted on 654 patients with stage IB3-IVA disease treated with definitive chemoradiotherapy. Radiotherapy was applied as external beam pelvic with or without para-aortic radiotherapy and brachytherapy. Concomitant chemotherapy was in the form of weekly or 3 weekly cisplatin. Data on demographics, treatment protocols, cumulative cisplatin dose, adverse effects, and survival outcomes were collected. Statistical analyses, including univariate and multivariate Cox regression models, were used to assess factors influencing progression free survival and overall survival. RESULTS: The median cumulative cisplatin dose was 210 mg (range 40-320), and ≥200 mg in 503 (76.9%) patients. Median follow-up was 35 months (range 1-150). The 5 year progression free survival and overall survival rates were 66.9% and 77.1%, respectively. Multivariate analysis identified poor performance status, non-squamous cell histology, presence of lymph node metastases, and hemoglobin <10 g/dL before chemoradiotherapy as poor prognostic factors for both progression free survival and overall survival in the whole group. When stage III cases were evaluated separately, the cumulative cisplatin dose <200 mg was found to be a significant poor prognostic factor in overall survival (hazard ratio 1.79, 95% confidence interval 1.1 to 3.0, p=0.031). CONCLUSION: Our study showed that a cumulative cisplatin dose >200 mg, particularly in patients with lymph node metastases, significantly improved overall survival. Factors such as anemia, toxicity related challenges, and comorbidities were identified as critical considerations in treatment planning. These findings emphasize the balance between maximizing therapeutic efficacy and managing toxicity, guiding personalized treatment approaches for locally advanced cervical cancer.

Factors Affecting Recurrence and Survival in Stage IIA Colon Cancer Patients.

INTRODUCTION: Our study delves into the intricate interplay of risk factors and the strategic selection of adjuvant therapy, scrutinizing their influence on recurrence and survival outcomes in stage IIA (T3N0M0) colon cancer patients. MATERIALS AND METHODS: The study examined the medical records of patients who underwent surgery for stage IIA colon cancer. Identification of stage IIA (pT3N0M0) colon cancer involved a comprehensive review of postoperative clinical records and histological reports. Parameters such as demographic data, tumor characteristics, microsatellite instability status, tumor locations, recurrence risk factors, preoperative carcinoembryonic antigen levels, and adjuvant treatments were systematically evaluated. RESULTS: In our study involving 220 patients, 138 were male (62.7%), with a median age of 62 years and a median body mass index of 25.1 kg/m2. In the patient group without risk factors, no statistically significant difference was detected in disease-free survival (DFS) rates between those who received treatment and those who did not (p = 0.546). DFS rates of patients with >1 risk factor were statistically significantly lower than those with a single risk factor (p = 0.017). In patients with >1 risk factor, the DFS of those who did not receive adjuvant treatment was significantly lower than those who received adjuvant treatment (p < 0.001). In the patient group with recurrence, when adjuvant treatments were considered, recurrence was significantly higher in the group receiving capecitabine (p = 0.01). CONCLUSION: The decision for adjuvant chemotherapy in stage IIA colon cancer patients involves careful consideration of various parameters and risk factors. The evolving landscape of research may refine recommendations, ensuring optimal treatment outcomes while minimizing unnecessary toxicity.

Comparison of aquaporin profile of advanced passage mesenchymal stem cells with early passage mesenchymal stem cells and determination of its effect on adipogenic differentiation efficiency.

OBJECTIVE: Our study aimed to compare aquaporin profiles in advanced and early passage bone marrow-derived mesenchymal stem cells (BM-MSCs) and assess the impact of aquaporin changes after adipogenic differentiation. Aquaporins are crucial for stem cell survival and differentiation during their life cycle. We focused on the role of aquaporins in the cell structures of advanced and early passage stem cells. METHODS: In our study, BM-MSCs were used for our objectives. Characterization of the cells was evaluated via flow cytometry using stem cell surface markers. The characterized BM-MSCs were divided into control and differentiation groups at passages 3 (P3) and 8 (P8). AQP1, AQP3, AQP7, AQP9, and AQP10 expression levels on days 0, 1, 3, 7, 14, and 21 were evaluated using Real Time-PCR, ELISA, and immunofluorescence studies. RESULTS: The cells were characterized by flow cytometry and confirmed to exhibit BM-MSC characteristics. At P3 and P8, differentiation was initiated, and AQP protein expression was observed to initially increase and then decrease on subsequent days. The increase in AQP protein expression at P3 occurred earlier than that at P8. Gene expression analysis demonstrated a statistically significant increase in AQP gene expression on days when AQP protein expression decreased. Moreover, statistical differences were observed between late and early passage AQP profiles. CONCLUSION: Our study examined the composition of AQPs in BM-MSCs in association with cell passage, and found that AQPs play a role in the differentiation process. The connection between the AQP profile and aging might be related to differentiation capacity, which could have implications for slowing down cellular aging and developing new therapeutic approaches.

Comparative analysis of ChatGPT, Gemini and emergency medicine specialist in ESI triage assessment.

INTRODUCTION: The term Artificial Intelligence (AI) was first coined in the 1960s and has made significant progress up to the present day. During this period, numerous AI applications have been developed. GPT-4 and Gemini are two of the best-known of these AI models. As a triage system The Emergency Severity Index (ESI) is currently one of the most commonly used for effective patient triage in the emergency department. The aim of this study is to evaluate the performance of GPT-4, Gemini, and emergency medicine specialists in ESI triage against each other; furthermore, it aims to contribute to the literature on the usability of these AI programs in emergency department triage. METHODS: Our study was conducted between February 1, 2024, and February 29, 2024, among emergency medicine specialists in Turkey, as well as with GPT-4 and Gemini. Ten emergency medicine specialists were included in our study but as a limitation the emergency medicine specialists participating in the study do not frequently use the ESI triage model in daily practice. In the first phase of our study, 100 case examples related to adult or trauma patients were extracted from the sample and training cases found in the ESI Implementation Handbook. In the second phase of our study, the provided responses were categorized into three groups: correct triage, over-triage, and under-triage. In the third phase of our study, the questions were categorized according to the correct triage responses. RESULTS: In the results of our study, a statistically significant difference was found between the three groups in terms of correct triage, over-triage, and under-triage (p < 0.001). GPT-4 was found to have the highest correct triage rate with an average of 70.60 (±3.74), while Gemini had the highest over-triage rate with an average of 35.2 (±2.93) (p < 0.001). The highest under-triage rate was observed in emergency medicine specialists (32.90 (±11.83)). In the ESI 1-2 class, Gemini had a correct triage rate of 87.77%, GPT-4 had 85.11%, and emergency medicine specialists had 49.33%. CONCLUSION: In conclusion, our study shows that both GPT-4 and Gemini can accurately triage critical and urgent patients in ESI 1&2 groups at a high rate. Furthermore, GPT-4 has been more successful in ESI triage for all patients. These results suggest that GPT-4 and Gemini could assist in accurate ESI triage of patients in emergency departments.

Effect of tumor-infiltrating immune cells (mast cells, neutrophils and lymphocytes) on neoadjuvant chemotherapy response in breast carcinomas.

INTRODUCTION: Despite screening, the incidence of breast cancer is increasing worldwide. Neoadjuvant chemotherapy (NAC) response is one of the most important parameters taken into consideration in surgery, optimal adjuvant chemotherapy planning and prognosis prediction. Research on predictive markers for the response to NAC is still ongoing. In our study, we investigated the relationship between tumor-infiltrating neutrophils/mast cells/lymphocytes and NAC response in breast carcinomas. MATERIAL AND METHOD: Study included 117 patients who were diagnosed with invasive breast carcinoma using core needle biopsy. In these biopsies tumor-infiltrating neutrophils/mast cells/lymphocytes were evaluated and Miller Payne Score was used for NAC response. RESULT: 53 patients exhibited high TILs, 36 had high TINs, and 46 showed high TIMs. While pathological complete response was 27 % in all patients, it was 38 % in high TINs patients, 35 % in high TILs patients, and 28 % in high TIMs patients. High TIMs were observed to be statistically associated with survival. TILs, TINs, nuclear grade, ER, PR and HER2 expression, Ki-67 proliferation index were found to be associated with the Miller - Payne score. In multivariate analysis, TINs, nuclear grade, pathological stage, and molecular subtype were found to be independent risk factors for treatment response. CONCLUSION: TINs have better prognostic value to predict neoadjuvant treatment than TILs. High TIMs are associated with increased overall survival. The inclusion of TINs in NAC response and TIMs in overall survival in pathology reports and treatment planning is promising in breast carcinomas as they are simple to use and reproducible markers.

Comparison of emergency medicine specialist, cardiologist, and chat-GPT in electrocardiography assessment.

INTRODUCTION: ChatGPT, developed by OpenAI, represents the cutting-edge in its field with its latest model, GPT-4. Extensive research is currently being conducted in various domains, including cardiovascular diseases, using ChatGPT. Nevertheless, there is a lack of studies addressing the proficiency of GPT-4 in diagnosing conditions based on Electrocardiography (ECG) data. The goal of this study is to evaluate the diagnostic accuracy of GPT-4 when provided with ECG data, and to compare its performance with that of emergency medicine specialists and cardiologists. METHODS: This study has received approval from the Clinical Research Ethics Committee of Hitit University Medical Faculty on August 21, 2023 (decision no: 2023-91). Drawing on cases from the "150 ECG Cases" book, a total of 40 ECG cases were crafted into multiple-choice questions (comprising 20 everyday and 20 more challenging ECG questions). The participant pool included 12 emergency medicine specialists and 12 cardiology specialists. GPT-4 was administered the questions in a total of 12 separate sessions. The responses from the cardiology physicians, emergency medicine physicians, and GPT-4 were evaluated separately for each of the three groups. RESULTS: In the everyday ECG questions, GPT-4 demonstrated superior performance compared to both the emergency medicine specialists and the cardiology specialists (p < 0.001, p = 0.001). In the more challenging ECG questions, while Chat-GPT outperformed the emergency medicine specialists (p < 0.001), no significant statistical difference was found between Chat-GPT and the cardiology specialists (p = 0.190). Upon examining the accuracy of the total ECG questions, Chat-GPT was found to be more successful compared to both the Emergency Medicine Specialists and the cardiologists (p < 0.001, p = 0.001). CONCLUSION: Our study has shown that GPT-4 is more successful than emergency medicine specialists in evaluating both everyday and more challenging ECG questions. It performed better compared to cardiologists on everyday questions, but its performance aligned closely with that of the cardiologists as the difficulty of the questions increased.

A Comparison of 18 F-FDG PET/CT and 68 Ga-PSMA PET/CT in Detecting Osteonecrosis of the Jaw in a Patient With Prostate Cancer.

ABSTRACT: Medication-related osteonecrosis of the jaw (ONJ) is rare adverse effect of zoledronic acid. We present ONJ detected in a 65-year-old man with prostate cancer who underwent 18 F-FDG PET/CT for metabolic characterization and to exclude a second primary malignancy of the liver lesion observed in 68 Ga-PSMA PET/CT. ONJ has high metabolic activity on 18 F-FDG PET/CT and no PSMA receptor activation on the 68 Ga-PSMA PET/CT. In this case, we wanted to underline the importance of 18 F-FDG PET/CT imaging of medication-related ONJ in patients receiving zoledronic acid therapy and to emphasize that rapid and appropriate treatment can be provided.

Production of alginate macrocapsule device for long-term normoglycaemia in the treatment of type 1 diabetes mellitus with pancreatic cell sheet engineering.

Type 1 diabetes-mellitus (T1DM) is characterized by damage of beta cells in pancreatic islets. Cell-sheet engineering, one of the newest therapeutic approaches, has also been used to create functional islet systems by creating islet/beta cell-sheets and transferring these systems to areas that require minimally invasive intervention, such as extrahepatic areas. Since islets, beta cells, and pancreas transplants are allogeneic, immune problems such as tissue rejection occur after treatment, and patients become insulin dependent again. In this study, we aimed to design the most suitable cell-sheet treatment method and macrocapsule-device that could provide long-term normoglycemia in rats. Firstly, mesenchymal stem cells (MSCs) and beta cells were co-cultured in a temperature-responsive culture dish to obtain a cell-sheet and then the cell-sheets macroencapsulated using different concentrations of alginate. The mechanical properties and pore sizes of the macrocapsule-device were characterized. The viability and activity of cell-sheets in the macrocapsule were evaluatedin vitroandin vivo. Fasting blood glucose levels, body weight, and serum insulin & C-peptide levels were evaluated after transplantation in diabetic-rats. After the transplantation, the blood glucose level at 225 mg dl-1on the 10th day dropped to 168 mg dl-1on the 15th day, and remained at the normoglycemic level for 210 days. In this study, an alginate macrocapsule-device was successfully developed to protect cell-sheets from immune attacks after transplantation. The results of our study provide the basis for future animal and human studies in which this method can be used to provide long-term cellular therapy in T1DM patients.

Prognostic Value of Inflammation And Nutrition-Based Scores in Non-Small Cell Lung Cancer.

OBJECTIVE: In studies conducted on non-small cell lung cancer (NSCLC) patients, many factors such as age, stage, weight loss, lymph node, and pleural involvement have been shown to affect survival. On the other hand, systemic inflammation plays a critical role in proliferation, migration, invasion, and metastasis. Inflammation and nutrition-based prognostic scores are reported to be associated with survival in patients with NSCLC. The aim of our study is to show the effects of these scores on survival and disease progression in NSCLC patients. SUBJECTS AND METHODS: Neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), modified Glasgow prognostic score (mGPS), and prognostic nutritional index (PNI) values in 102 patients with stage 1,2 and 3A NSCLC were analyzed retrospectively. RESULTS: NLR (p < 0.001), PLR (p = 0.001), PNI (p < 0.001), and mGPS (p = 0.001) variables showed a statistically significant difference according to mortality groups. NLR and PLR values were higher in exitus patients. However, PNI values were higher in surviving patients. NLR (p < 0.001), PLR (p = 0.004), PNI (p = 0.001), and mGPS (p = 0.015) variables showed a statistically significant difference in terms of locoregional recurrence. PNI (p = 0.001) and mGPS (p = 0.001) in terms of distant metastasis development during follow-up and treatment, showed a statistically significant difference. CONCLUSION: NLR, PLR, PNI, and mGPS are easily accessible and non-invasive parameters and provide predictive information about survival and disease course. We showed the effect of these parameters on the prognosis.

Examining the effect of activated cytotoxic (CD8+) T-cell exosomes to the lung cancer.

Lung cancer continues to be a major health problem worldwide owing to its incidence, and causes physical, psychological, social, and economic problems. Activated cytotoxic T cells (ACTC) are positively correlated with the tumor microenvironment (TME), improving the prognosis of cancer patients. Recently, ACTC-derived exosomes (ACTC-dExo) were implicated in this effect by inhibiting mesenchymal stem cells, which may promote metastasis in the TME. Exosomes are thought to be advantageous for the specific delivery of drugs to cancer cells because they have the characteristics of natural liposomes, are nanosized, and remain largely stable in the blood due to the protein and lipid content they carry on their membranes. In this study, we aimed to determine the cytotoxic and metastatic inhibitory effects of ACTC-dExo in A549 cells in vitro. Cytotoxic CD8+ T cells were isolated from whole blood obtained from healthy individuals and cultured for 5-7 days after stimulation. The ACTC-dExo serum-free culture medium was collected by ultracentrifugation. Characterization and quantification of the isolated exosomes were performed using flow cytometry, electron microscopy, zeta-sizer measurements, and bicinchoninic acid (BCA) assays. We co-cultured ACTC and ACTC-dExo with A549 cells for 48 h. The viability of A549 cells was evaluated using a WST-1 assay. The metastasis-related genes MMP2, MMP9, TWIST, SNAI1, and CDH1 were detected by qRT-PCR, and MMP2 and MMP9 proteins were evaluated by confocal microscopy. In addition, changes in cell migration were investigated using a scratch assay. ACTC-dExo were found to have anti-proliferative and anti-metastatic effects and reduced cancer cell proliferation and metastatic properties.

Investigation of impacts of decellularized heart extracellular matrix and VEGF on cardiomyogenic differentiation of mesenchymal stem cell through Notch/Hedgehog signaling pathways.

OBJECTIVE: Decellularization is the process to obtain natural scaffolds with tissue integrity and extracellular matrix components, and recellularization is used to produce tissue-like constructs with specific cell types. In this study, rat bone marrow-derived mesenchymal stem cells (rBM-MSCs) were cultured on decellularized heart extracellular matrix. These cells were then induced to differentiate into cardiomyogenic cells under the stimulatory effect of vascular endothelial growth factor (VEGF) and other chemicals. This study aimed to investigate the effect of the cardiac extracellular matrix and VEGF on cardiomyogenic differentiation in the context of the Notch and Hedgehog signaling pathways. METHODS: Heart samples extracted from rats were decellularized by serial application of detergent to remove cells from the tissue, and then recellularized with rBM-MSCs. The recellularized tissue matrices were then analyzed for cardiomyogenesis. Cardiomyogenic differentiation was performed on decellularized heart extracellular matrix (ECM; three-dimensional scaffolds) and culture plates (two-dimensional cell culture system) for 28 days to understand the effects of the heart extracellular matrix. In addition, differentiation was induced with and without the stimulatory effect of VEGF to understand the effect of VEGF on cardiomyogenic differentiation of rBM-MSCs. RESULTS: Immunofluorescence staining showed that decellularization of the heart was performed effectively and successfully. After decellularization process, the heart extracellular matrix was completely free of cells. It was observed that rBM-MSCs transplanted onto the heart extracellular matrix remained viable and proliferated for 21 days after recellularization. The rBM-MSCs promoted cardiomyogenic differentiation in the conventional differentiation medium but were inversely affected by both VEGF and heart extracellular matrix proteins. Lower expression of connexin43 and cardiac troponin I genes was observed in cells induced by either matrix proteins or VEGF, compared to cells differentiated by chemical agents alone. CONCLUSION: In this study, we investigated the effect of decellularized heart extracellular matrix and VEGF on cardiomyogenic differentiation of rBM-MSCs. On the decellularized cardiac extracellular matrix, rBM-MSCs maintained their viability by adhering to the matrix and proliferating further. The adhesion of the cells to the matrix also produced a physical stimulus that led to the formation of histological structures resembling myocardial layers. Chemical stimulation of the decellularized heart extracellular matrix and cardiomyogenic differentiation supplements resulted in increased expression of cardiomyogenic biomarkers through modulation of the Notch and Hedgehog signaling pathways.

Investigation of the effect of pancreatic decellularized matrix on encapsulated Islets of Langerhans with mesenchymal stem cells.

OBJECTIVE: In this study, it was aimed to provide a therapeutic approach for T1DM by encapsulating the pancreatic islets with mesenchymal stem cells and decellularized pancreatic extracellular matrix to support the survival of islets while maintaining their cellular activity. METHOD: Pancreatic extracellular matrix was decellularized using different concentrations of detergent series. After the preparation of the protein-based tissue extracellular matrix was shown to be free of cells or any genetic material by molecular, immunofluorescence and histochemical techniques. Following the homogenization of the decellularized pancreatic extracellular matrix and the analysis of its protein composition by LC-MS, the matrix proteins were incorporated with pancreatic islets and rat adipose tissue-derived MSCs (rAT-MSCs) in alginate microcapsules. Glucose-stimulated insulin secretion property of the islet cells in the microbeads was evaluated by insulin ELISA. The gene expression profile of the encapsulated cells was analyzed by Real-Time PCR. RESULTS: Unlike the protein composition of whole pancreatic tissue, the decellularized pancreas matrix was free of histone proteins or proteins originated from mitochondria. The protein matrix derived from pancreatic tissue was shown to support the growth and maintenance of the islet cells. When compared to the non-encapsulated pancreatic islet, the encapsulated cells demonstrate to be more efficient in terms of insulin expression. CONCLUSION: The extracellular pancreatic matrix obtained in this study was directly used as supplementary in the alginate-based microcapsule enhancing the cell survival. The tissue matrix protein and alginate had a synergistic effect on total insulin secretion, which might have the potential to overcome the insulin deficiency. Despite the improvement in the cell viability and the number, the efficiency of the insulin secretion in response to glucose stimulation from the alginate microcapsules did not meet the expectation when compared with the non-encapsulated pancreatic islets.

The anti-scar effect of tyrosine-kinase inhibitor nintedanib in experimental glaucoma filtration surgery in rabbits.

PURPOSE: To investigate the efficacy of nintedanib on preventing postoperative scar in formation following glaucoma filtering surgery (GFC) in rabbits in comparison with Mitomycin-C (MMC). DESIGN: Experimental Animal Study. METHODS: 24 New Zealand rabbits were divided randomly into 3 groups as Sham, Nindetanib and MMC(n = 8). Limbal-based trabeculectomy was performed on the right eyes of the rabbits. Left eyes that did'nt undergo surgery were included in the control group (n = 8). Following surgery, Intraocular pressures (IOP), postoperative complications and morphological changes in the bleb were evaluated. On the 28th day, eight eyes from each group were enucleated and histologically and immunohistochemically analyzed. Matrix metalloproteinase-2 (MMP-2), Transforming Growth Factor-1 (TGF-B1) and alpha-smooth muscle actin (a-SMA) were evaluated. RESULTS: It was observed that nintedanib has no side effects and reduces subconjunctival fibrosis. Postoperative IOP values in the Nindetanib group were lower than the other groups (p < 0.05). The longest bleb survival was observed in the Nintedanib group and the shortest in the Sham group (p < 0.001). Conjunctival vascularity and inflammation was reduced in the Nintedanib group compared to the Sham group (p < 0.05). The highest subconjunctival fibrosis was observed in the Sham group and the least in the Nintedanib group (p < 0.05). Although the fibrosis score was found lower in the Nintedanib group compared to the MMC(p > 0.05). α-SMA TGF-ß1, MMP-2 expressions were similar in Nintedanib and MMC groups (p > 0.05), however, it was observed that significantly decreased in both groups compared to Sham group (p < 0.05). CONCLUSION: It has been observed that Nindetanib suppress fibroblast proliferation Thus, It may be a drug that can prevent subconjunctival fibrosis in GFC.

Differentiation of Benign and Malignant Thyroid Nodules with ANFIS by Using Genetic Algorithm and Proposing a Novel CAD-Based Risk Stratification System of Thyroid Nodules.

The thyroid nodule risk stratification guidelines used in the literature are based on certain well-known sonographic features of nodules and are still subjective since the application of these characteristics strictly depends on the reading physician. These guidelines classify nodules according to the sub-features of limited sonographic signs. This study aims to overcome these limitations by examining the relationships of a wide range of ultrasound (US) signs in the differential diagnosis of nodules by using artificial intelligence methods. An innovative method based on training Adaptive-Network Based Fuzzy Inference Systems (ANFIS) by using Genetic Algorithm (GA) is used to differentiate malignant from benign thyroid nodules. The comparison of the results from the proposed method to the results from the commonly used derivative-based algorithms and Deep Neural Network (DNN) methods yielded that the proposed method is more successful in differentiating malignant from benign thyroid nodules. Furthermore, a novel computer aided diagnosis (CAD) based risk stratification system for the thyroid nodule's US classification that is not present in the literature is proposed.

The Effects of Transcranial Focused Ultrasound Stimulation of Nucleus Accumbens on Neuronal Gene Expression and Brain Tissue in High Alcohol-Preferring Rats.

We investigated the effect of low-intensity focused ultrasound (LIFU) on gene expression related to alcohol dependence and histological effects on brain tissue. We also aimed at determining the miRNA-mRNA relationship and their pathways in alcohol dependence-induced expression changes after focused ultrasound therapy. We designed a case-control study for 100 days of observation to investigate differences in gene expression in the short-term stimulation group (STS) and long-term stimulation group (LTS) compared with the control sham group (SG). The study was performed in our Experimental Research Laboratory. 24 male high alcohol-preferring rats 63 to 79 days old, weighing 270 to 300 g, were included in the experiment. LTS received 50-day LIFU and STS received 10-day LIFU and 40-day sham stimulation, while the SG received 50-day sham stimulation. In miRNA expression analysis, it was found that LIFU caused gene expression differences in NAc. Significant differences were found between the groups for gene expression. Compared to the SG, the expression of 454 genes in the NAc region was changed in the STS while the expression of 382 genes was changed in the LTS. In the LTS, the expression of 32 genes was changed in total compared to STS. Our data suggest that LIFU targeted on NAc may assist in the treatment of alcohol dependence, especially in the long term possibly through altering gene expression. Our immunohistochemical studies verified that LIFU does not cause any tissue damage. These findings may lead to new studies in investigating the efficacy of LIFU for the treatment of alcohol dependence and also for other psychiatric disorders.

Triponderal mass index is as strong as body mass index in the determination of obesity and adiposity.

OBJECTIVE: In determining obesity and body adiposity, triponderal mass index (TMI) is as strong an anthropometric measurement as body mass index (BMI). The aim of this study was to develop TMI reference values for Turkish children and adolescents and compare TMI with BMI according to body adiposity and obesity indices. METHODS: Data from the DAMTCA-II (Determination of Anthropometric Measurements of Turkish Children and Adolescents II) study were used in this cross-sectional study. Data from 4330 children (1931 boys, 2399 girls) ages 6 to 17 y were evaluated, and the TMI percentile values were produced. The predictive power of TMI and BMI for obesity and overweight were done for waist circumference, waist/height ratio, body fat percentage, and upper arm fat area, which are different parameters used to determine body adiposity. RESULTS: The 3rd, 5th, 10th, 25th, 50th, 75th, 85th, 90th, 95th, and 97th TMI percentiles and mean values were calculated for all children's age and sex. TMI cutoff values were calculated by receiver operating characteristic analysis regarding waist/height ratio 0.5, waist circumference ≥90 percentile, arm fat area ≥85 percentile, and body fat percentage ≥85. TMI and BMI area under the curve values were similar for each of these four measurements. TMI was as robust an index as BMI in demonstrating obesity and adiposity for all age groups in boys and girls. It was concluded that the values >90th percentile (median 15.8 kg/m3) in girls aged ≤10 y, 95th percentile (median 16.2 kg/m3) in girls aged >10 y, >85th percentile (median 14.9 kg/m3) in boys aged ≤12 y and 75th percentile (median value 14.5 kg/m3) in boys aged >12 y are critical values for TMI when evaluating adiposity and obesity. CONCLUSIONS: We considered that TMI is as effective as BMI in terms of waist/height ratio, waist circumference, arm fat area, and body fat percentage in determining overweight and obesity in children. The ages at which TMI showed distinct variation were determined for both sexes.