Erhan Sahin

Background/Objectives: Duchenne muscular dystrophy (DMD) is a genetic disorder caused by mutations in the dystrophin gene. DMD is characterized by exon deletions in about 76% of cases, with common deletions in exons 47, 48, 51, and 52. We evaluated the effectiveness of an mRNA-based therapy targeting these exon deletions, which are frequently seen in DMD patients. Methods: The current study involved two protocols: 1. applying the therapy to cells from patients diagnosed with DMD, and 2. applying the therapy to genetically modified transgenic mdx/d2 mice. After treatment, dystro-phin was detected in all experimental groups. Results: Our study showed that, both in vitro and in vivo analyses demonstrated that the mRNA-based therapy successfully restored dystrophin expression in dystrophic muscle cells and tissues. Gene expression analysis, together with protein-level assessments, including Western blot, immunoflu-orescence (IF), ELISA, and immunohistochemistry (IHC), confirmed a significant in-crease in dystrophin levels in the treated groups compared with the control group. In addition to dystrophin restoration, other key sarcolemmal proteins involved in main-taining muscle membrane stability, such as γ-sarcoglycan, β-dystroglycan, and β-actin, were also highly expressed. These findings suggest an overall improvement in muscle cell membrane integrity. Consistent with the molecular results, behavioral analyses performed in the animal model revealed significant functional improvements, includ-ing enhanced mobility, motor coordination, longer walking and resting durations, and a reduced risk of falls. Overall, the results indicate that mRNA-mediated dystrophin replacement improves both muscle structural integrity and functional performance. Conclusions: Our study proved that the mRNA complex successfully produced func-tional dystrophin in transgenic mdx/d2 mice without causing allergic reactions or damage to the kidney, intestines, muscles, or brain.

Introduction: Sodium Selenite (NaSe) is a molecule with various biological activities. Bone fractures and osteoporotic diseases are increasingly common health issues, prompting the search for alternative treatments. Therefore, the purpose of this study was to examine the antioxidant and osteogenic properties of NaSe. Methods: The experiments were conducted using the hFOB1.19 osteoblast cell line. The MTT assay was used to assess the effects of NaSe on cell viability, while cytotoxicity was evaluated with Lactate Dehydrogenase (LDH) assays. Osteogenic differentiation was assessed by alizarin red staining, and Alkaline Phosphatase (ALP) activity and intracellular Reactive Oxygen Species (ROS) levels were also analyzed. Results: The results showed that NaSe significantly enhanced cell viability in a dose-dependent manner at low doses (0.01-1μM), with the most effective dose being 1μM (p<0.05). LDH activity remained similar to the control within the 0.01-1μM range but increased significantly at higher concentrations (5-50 μM) in both 24- and 48-hour experiments (p<0.05). NaSe reduced intracellular ROS levels significantly between 0.01-1 μM, with 1 μM being the most effective concentration (p<0.05). The highest ALP activity was observed at 0.1 μM NaSe (p < 0.05), and calcium deposition increased in a concentration- dependent manner (p<0.05). The most effective dose for enhancing mineralization was 0.1 μM (p<0.05). Conclusion: This study demonstrates that NaSe has antioxidant and osteogenic effects at low doses in hFOB cells. These positive effects suggest that NaSe could be a promising candidate for in-vitro, in-vivo, and clinical trials, providing hope for new treatments for bone diseases.

Background/Objectives: Cholesterolosis is a relatively common, yet often incidental, histopathological finding in cholecystectomy specimens, frequently associated with chronic cholecystitis. Although its clinical significance remains unclear, possible associations with glycemic regulation have been proposed. This study aimed to examine the relationship between cholesterolosis, glycemic parameters, and inflammatory markers in patients with chronic cholecystitis. Methods: We retrospectively analyzed gallbladder specimens from patients who underwent cholecystectomy for chronic cholecystitis between 2014 and 2023. Histopathology assessed cholesterolosis presence, distribution, and inflammatory features. Patients were grouped according to diabetes status (diabetic vs. non-diabetic) as well as the presence or absence of cholesterolosis. Demographic data, gallstone status, one-year mean fasting glucose, HbA1c levels, and hematological/inflammatory indices were compared between groups. Results: Among diabetic patients, those with cholesterolosis had significantly lower fasting glucose than those without cholesterolosis (129.04 ± 28.02 vs. 158.41 ± 54.23 mg/dL, p < 0.05). The presence of cholesterolosis was not significantly associated with diabetes status (p > 0.05). Inflammatory indices, including neutrophil count, did not differ significantly between groups (p > 0.05), although fasting glucose correlated positively with neutrophil count (r = 0.167, p < 0.05). Gallstones were less frequent in cholesterolosis cases compared with non-cholesterolosis cases (59.4% vs. 75%, p < 0.05), suggesting a distinct pathophysiology. Conclusions: This is the first study to assess one-year glycemic profiles in diabetic and non-diabetic patients with and without cholesterolosis. The findings suggest that cholesterolosis may occur independently of poor glycemic control and systemic inflammation, supporting the concept of a distinct underlying mechanism.

Since the cartilage tissue is fed by the vessels in the surrounding connective tissue, it is a very difficult tissue to repair. Therefore, cardiovascular diseases primarily affect the cartilage tissue. In this study, the possible effects of high blood pressure on discus intervertebralis were examined histopathologically and histomorphometrically. Discus intervertebralis of 13 normotensive and 13 hypertensive patients were compared histopathologically and histomorphometrically. Demographic information of the patients was obtained from the patient registration system of the hospital. Hematoxylin-eosin-stained slices were used for histopathological evaluation. In histological sections, chondrocytes and lacunae diameters were measured at 400x magnification in 10 different areas with a computer morphometric measurement program. Discus intervertebralis of normotensive and hypertensive patients did not differ in terms of chondrocyte count and lacunae diameter. The mean age of hypertensive patients (61.69±13.55) was higher than the mean age of normotensive patients (42±8.84) (p0.05). In this study, it was shown that the discus intervertebralis of normotensive and hypertensive patients who underwent lumbar discectomy were not different in terms of histopathologically and histomorphometrically (in terms of chondrocyte number and lacuna diameter).

Introduction: Pancreatic cancer is a challenging disease to diagnose and treat due to its asymptomatic progression and high mortality rate. It is often identified at an advanced stage, where it typically metastasizes to other organs. To address these challenges, scientific studies explore various treatments, including natural and anticancer compounds like resveratrol. This study investigates the effects of resveratrol on pancreatic cancer cells. Methods: The study used PANC-1 pancreatic cancer cells cultured in vitro. Cell viability in resveratrol-containing media was measured using the MTT assay. Safe, toxic, and IC50 doses for a 24-hour period were determined. Morphological changes were analyzed using hematoxylin-eosin staining. In invasion experiments, a wound-healing assay was performed by scraping cells in a 6-well plate and photographing their movement in resveratrol media. Results: Resveratrol increased cell viability at concentrations up to 10 µM, showed similar viability to the control at 50 µM, and reduced viability below 50% at 200 µM. The IC50 dose was calculated as 189.5 µM. At concentrations above 100 µM, significant disruptions in nuclear and cytoplasmic structures were observed. Additionally, cell movement ceased at 100, 150, and 189 µM doses, showing dose-dependent inhibition of cell invasion (p<0.05). Conclusions: Resveratrol, a naturally occurring compound found in many fruits and plants, demonstrated cytotoxic and anti-metastatic effects on PANC-1 cells. This study highlights its potential role in pancreatic cancer treatment, emphasizing the importance of continued research to combat this aggressive disease effectively.

Objective: Mast cells play pivotal roles in tumorigenesis dynamics, yet the specific factors driving hyperplastic gastric polyp formation remain unclear. This study aims to elucidate this gap by examining mast cell quantities in incidental gastric polyps, with a specific focus on hyperplastic lesions, and exploring their correlation with polypogenesis and metaplasia. Methods: 45 endoscopy specimens exhibiting anterior non-atrophic chronic inactive gastritis (2014-2023) were examined. Following histological evaluations, the cases were classified into three groups: hyperplastic gastric polyps with or without intestinal metaplasia, and chronic inactive gastritis as the control. Mast cell density per unit area was assessed from toluidine blue-stained sections. Polyp diameter and demographic information were also recorded for statistical analysis. Results: The average ages of patients with hyperplastic polyps, metaplasia, and gastritis were 51.2±12.84, 55.93±13.58, and 56.2±14.68, respectively (p>0.05). In each group, the proportion of women was higher. Mast cell density, which varied within each group, did not show a statistically significant difference (p>0.05). The average polyp diameters for the metaplasia and hyperplastic polyp groups were 0.26 and 0.30 mm, respectively (p>0.05). There was no correlation between mast cell density and polyp diameter (p>0.05). Conclusions: Intestinal metaplasia did not significantly influence mast cell density in non-dysplastic, low-malignant gastric polyps. Mast cell density and diameter in hyperplastic gastric polyps of 1 cm or smaller showed no correlation. Overall, non-cancerous gastric polypogenesis and mast cell count lack a unidirectional relationship.

Colorectal cancer poses a significant global health challenge, with a considerable proportion arising from colon adenomas. Understanding the molecules involved in the carcinogenesis process is crucial for improving colon cancer diagnosis and prognosis. While research on the role of epiplakin in cancer remains limited compared to other plakin group proteins, comprehending its expression patterns and correlations can offer valuable insights into colon carcinogenesis. In this study, we analyzed 60 tissue samples, including colon adenocarcinomas, tubular adenomas (low malignancy risk group), tubulovillous adenomas (high malignancy risk group), and adjacent normal colon tissues. Classification and grading were reevaluated by histological examination. Immunohistochemistry was performed to assess epiplakin and Ki67 expression. Epiplakin optical density and the Ki67 proliferation index were calculated using ImageJ. Statistical analyses were conducted to evaluate correlations and significance. Epiplakin expression was significantly decreased in colon adenocarcinomas [optical density median 4.04 (95% CI, 3.98 to 4.24)] and tubulovillous adenomas [4.32 (95% CI, 4.08 to 4.32)] compared to normal colon tissues [4.61 (95% CI, 4.50 to 4.67)] and tubular adenomas [4.87 (95% CI, 4.67 to 4.88)] (P < 0.05). Moreover, adenoma groups exhibited higher proliferation indices (P < 0.05), and a positive correlation was found between epiplakin expression and the Ki67 proliferation index (r = 0.317, P < 0.05). Our study highlights the potential significance of epiplakin in colorectal cancer. Decreased epiplakin expression is associated with colon malignancy progression, suggesting its role as a potential marker.

Osteoporosis and resulting fractures affect a significant group of people in the world. It has been shown in many studies that selenium has positive effects on bone metabolism. Based on this information, the aim of this study is to investigate whether bone differentiation will start in a shorter time by applying selenomethionine (SeMet) to hFOB cells. First, hFOB 1.19 cells were cultured. Safe doses of SeMet were determined by MTT and LDH tests. Ossification levels were determined by alizarin red staining and measurement of alkaline phosphatase enzyme levels. The results were analyzed with statistical tests. It was observed that SeMet increased cell viability at concentrations of 10, 25, 50, 100, and 200 µM in 24 h. At these concentrations, cell viability increased above the control, the viabilities were as follows: 109.4%, 104.9%, 104.3%, 103.15%, and 100.27%. High doses of SeMet significantly reduce cell viability. According to Alizarin red staining, SeMet increases the amount of calcium deposits in hFOB cells in a dose-dependent manner. In the experimental groups, the highest ALP enzyme was determined in the 7-day SeMet application. The most effective dose was measured as 15 µM. It was determined that SeMet, which is found as a trace element in living things in nature, increases the viability of hFOB cells, which are osteoblast cell precursors, and increases osteoblastic differentiation and osteoblastic activity in these cells. Our results are at a level that sheds light on an important problem in public health.

5-Fluorouracil (5-FU), an antineoplastic agent, causes intestinal mucositis (IM) that limits the drug efficacy. The aim of this study was to reveal the protective and/or therapeutic effects of berberine (BER), an isoquinoline quaternary alkaloid, on IM induced by 5-FU in rats. Forty Wistar albino male rats were randomly divided into five groups: Control, 5-FU, 5-FU+BER75 (75 mg/kg/day), 5-FU+BER150 (150 mg/kg/day) and 5-FU+BER300 (300 mg/kg/day). 5-FU was intraperitoneally (i.p) injected at a dose of 60 mg/kg on alternate days for 4 days, while other groups were orally received BER daily for 7 days. By the end of study, intracardiac blood samples were collected and intestinal tissues were removed for histological examination. TNF-α, IL-6 and NF-κB levels were measured in serum using commercial ELISA kits. Results showed that BER at increasing doses ameliorated body weight loss and histopathological damage. It was concluded that BER may be effective in the treatment and/or prophylaxis of the damage in the gastrointestinal system caused by 5-FU in a dose-dependent manner.

Hyperthyroidism influences the development of cardiac hypertrophy. Transient receptor potential canonical channels (TRPCs) and endoplasmic reticulum (ER) stress are regarded as critical pathways in cardiac hypertrophy. Hence, we aimed to identify the TRPCs associated with ER stress in hyperthyroidism-induced cardiac hypertrophy. Twenty adult Wistar albino male rats were used in the study. The control group was fed with standard food and tap water. The group with hyperthyroidism was also fed with standard rat food, along with tap water that contained 12 mg/L of thyroxine (T4) for 4 weeks. At the end of the fourth week, the serum-free triiodothyronine (T3), T4, and thyroid-stimulating hormone (TSH) levels of the groups were measured. The left ventricle of each rat was used for histochemistry, immunohistochemistry, Western blot, total antioxidant capacity (TAC), and total oxidant status (TOS) analysis. As per our results, activating transcription factor 6 (ATF-6), inositol-requiring kinase 1 (IRE-1), and TRPC1, which play a significant role in cardiac hypertrophy caused by hyperthyroidism, showed increased activation. Moreover, TOS and serum-free T3 levels increased, while TAC and TSH levels decreased. With the help of the literature review in our study, we could, for the first time, indicate that the increased activation of ATF-6, IRE-1, and TRPC1-induced deterioration of the Ca²⁺ ion balance leads to hypertrophy in hyperthyroidism due to heart failure.