Lycopene prevents cell death in NRK-52E cells by inhibition of high glucose-activated DNA damage and apoptotic, autophagic, and necrotic pathways.

This study aims to investigate the effects of lycopene on apoptotic, autophagic, and necrotic pathways, oxidative status, and DNA damage in diabetic nephropathy at the molecular level. The sample of the study includes seven groups: lycopene (L), high glucose (G), high glucose + lycopene (GL), and control (C) groups tested at 12 and 24 h. The expression levels of genes in oxidative, apoptotic, autophagic, and necrotic cell death pathways are determined by reverse transcription-quantitative polymerase chain reaction analysis. The comet assay method is used for the analysis of DNA damage. It is observed that adding lycopene to high glucose for protective purposes reduces the expression of genes related to apoptosis, autophagy, and necrosis, as well as the DNA damage index, compared to cells given high glucose alone. Lycopene can be a safe and effective alternative agent.

Exploring the combined anti-cancer effects of sodium butyrate and celastrol in glioblastoma cell lines: a novel therapeutic approach.

Glioblastoma, a highly aggressive and lethal brain cancer, lacks effective treatment options and has a poor prognosis. In our study, we explored the potential anti-cancer effects of sodium butyrate (SB) and celastrol (CEL) in two glioblastoma cell lines. SB, a histone deacetylase inhibitor, and CEL, derived from the tripterygium wilfordii plant, act as mTOR and proteasome inhibitors. Both can cross the blood-brain barrier, and they exhibit chemo- and radiosensitive properties in various cancer models. GB cell lines LN-405 and T98G were treated with SB and CEL. Cell viability was assessed by MTT assay and IC50 values were obtained. Gene expression of DNA repair, apoptosis, and autophagy-related genes was analyzed by RT-PCR. Cell cycle distribution was determined using flow cytometry. Viability assays using MTT assay revealed IC50 values of 26 mM and 22.7 mM for SB and 6.77 µM, and 9.11 µM for CEL in LN-405 and T98G cells, respectively. Furthermore, we examined the expression levels of DNA repair genes (MGMT, MLH-1, MSH-2, MSH-6), apoptosis genes (caspase-3, caspase-8, caspase-9), and an autophagy gene (ATG-6) using real-time polymerase chain reaction. Additionally, flow cytometry analysis revealed alterations in cell cycle distribution following treatment with SB, CEL and their combination. These findings indicate that SB and CEL may act through multiple mechanisms, including DNA repair inhibition, apoptosis induction, and autophagy modulation, to exert their anti-cancer effects in glioblastoma cells. This is the first study providing novel insights into the potential therapeutic effects of SB and CEL in glioblastoma.

Lymphocele Outcomes After Renal Transplantations Performed by an Experienced Surgeon: Is Meticulously Performed Surgery and Experience Adequate to Prevent Lymphocele?

BACKGROUND The purpose of the present study was to analyze the rate of lymphoceles in kidney transplant operations meticulously performed by the same senior surgeon. MATERIAL AND METHODS The present study included 315 patients who were operated on in our organ transplantation center and followed up in the polyclinic after July 2013. The patients were retrospectively divided into 2 groups: patients with and without lymphocele. Symptomatic lymphocele (SL) has been defined as symptomatic fluid collection around the graft that necessitates an intervention for the graft or patient. RESULTS Lymphocele was observed in 82 (26%) patients. An intervention was needed in 16 (5.1%) of these cases. Demographic data such as age and sex of both groups were similar. Lymphocele cases were mostly asymptomatic, with a size <6 cm (75.6%). However, intervention was needed in 16 (75%) of the patients with a size ≥6 cm that were symptomatic. The length of time on dialysis in the pretansplant period was shorter in the group that developed lymphocele, and a lower rate of graft loss was observed in these patients. No statistically significant difference was found between the 2 groups in terms of rejection rates, serum albumin/globulin levels, and development of de novo DSA. CONCLUSIONS The risk factors reported in the literature related with lymphocele formation were not found to be statistically significant in our study. Complications, except lymphocele, were observed less frequently, but lymphocele formation was encountered in our patients despite meticulous surgery.

Vitamin D may assist the UPR against sodium fluoride-induced damage by reducing RIPK1, ATG5, BECN1, oxidative stress and increasing caspase-3 in the osteoblast MC3T3-E1 cell line.

BACKGROUND: Out of all measure systemic exposure to fluorides can cause defect of skeletal and dental fluorosis. Endoplasmic reticulum (ER) stress is caused by fluorine-induced oxidative stress and importance of vitamin D in its prevention is not known enough in bone cells. This study was carried out to investigate fluorine-induced oxidative stress, ER stress, and death pathways and the effect of vitamin D on them. METHODS: MC3T3-E1 mouse osteoblast cell line was used as the material of the study. The NaF and vitamin D concentrations were determined by the MTT assay. NaF treatments and vitamin D supplementation (pre-add, co-add, and post-add) was administered in the cell line at 24th and 48th hours. The expression of the genes in oxidative stress, ER stress, and death pathways was determined using RT-qPCR and Western blotting techniques. RESULTS: Vitamin D significantly reduced mRNA expression levels of SOD2, CYGB, ATF6, PERK, IRE1, ATG5 and BECN1 whereas caused an increase in levels GPX1, SOD1, NOS2 and Caspase-3 in MC3T3-E1 mouse osteoblast cell line of NaF-induced. In addition, GPX1, SOD1, ATF6, PERK, IRE1, BECN1, Caspase-3 and RIPK1 protein levels were examined by Western blot analysis, and it was determined that vitamin D decreased IRE1 and PERK protein levels, but increased GPX1, SOD1, ATF6 and Caspase-3 protein levels. CONCLUSION: The findings of the study suggest that vitamin D has protective potential against NaF-induced cytotoxicity reasonably through the attenuation of oxidative stress, ER stress, ATG5, IRE1 and by increasesing caspase-3 in vitro conditions.

Can a novel drug dose be used for T3 suppression test?

PURPOSE: This study aimed to investigate whether 25 µg/day dose of triiodothyronine (T3) can also suppress thyroid stimulating hormone (TSH) level, as well as the routine dose of 50-100 µg/day in T3 suppression test, which is used to the distinguish between resistance to thyroid hormone (RTH) and TSH secreting pituitary adenoma. METHODS: In this prospective study, 26 patients with genetically proven RTH were randomly divided into two groups: Group 1 comprised 13 patients who were administered 50-100 µg/day T3 for 3-9 days, while Group 2 also comprised 13 patients who were administered 25 µg/day T3 for 7 days for T3 suppression test. The two groups' responses to T3 suppression tests were compared. RESULTS: The comparison of the mean percentage changes in TSH values by the T3 suppression tests showed no significant differences between the groups, and a ≥80% decrease was detected in all patients. Nine patients in Group 1 and one patient in Group 2 reported that they had to use propranolol due to tachycardia developed during the test. CONCLUSION: As higher doses of T3 can increase the risk of severe tachycardia during T3 suppression test, a low dose with 25 mcg/day for a week appears to be safer and more useful.

The Effects of Vitamin D Application on NaF-Induced Cytotoxicity in Osteoblast Cells (hFOB 1.19).

This study was planned to evaluate the effect of vitamin D administration on cytotoxicity due to fluoride exposure in vitro. NaF (IC50) and vitamin D (proliferative) were applied to human osteoblast (hFOB 1.19) cells. The major genes of apoptotic, autophagic, and necrotic pathways were determined by RT-PCR. 2-∆∆Ct formulation was used for expression analysis. In the NaF group, caspase 3, Bax, Bad, Bak, Bclx, Atg3, Atg5, Atg6, pG2, LC3-I, LC3-II, RIP1, and RIP3 genes were increased (2.6-15 times). It was observed that the expressions of these genes approached the control when vitamin D was given together with NaF. The Bcl2 gene increased significantly (sixfold) with the effect of NaF, and was down-regulated to some extent with additional vitamin D administration, but still more than in the control. As a result, it was determined that apoptotic, necrotic, and autophagic pathways were activated as the molecular basis of the damage in the bone tissue, which was most affected by fluorine, and these genes were down-regulated and approached the control group with the addition of vitamin D. It was concluded that this is an important data to explain the molecular basis of the protective and therapeutic effect of vitamin D against fluorine toxicity.

The effect of quinoa (Chenopodium quinoa) on apoptotic, autophagic, antioxidant and inflammation markers in glucocorticoid-induced insulin resistance in rats.

BACKGROUND: Insulin resistance plays an important role in predicting type 2 diabetes that may develops. This study was planned in order to investigate the beneficial effects of quinoa (Chenopodium quinoa) use in glucocorticoid induced-insulin resistance. METHODS AND RESULTS: Forty-two rats were used as the material (experimental) groups: the control group (C), the quinoa-administered group (Q), the insulin resistance-created group (IR), the IR + metformin group (IM), the IR + quinoa for treatment group (IQ) and the quinoa + IR for prophylaxis group (QI). Blood glucose, insulin levels and HOMA-IR were found to be highest (p < 0.05) in the IR group (p < 0.05). Glucose levels decreased significantly with the administration of quinoa and approached the levels of the control, but the insulin levels and the HOMA-IR did not significantly change. It was also observed that other biochemical parameters (ALT, AST, ALP, total cholesterol, total protein, urea and creatinine) changed significantly in the IR group and approached the levels of the control group with the administration of quinoa. Apoptotic (BCL2 5, BAX 9, CAS 3), autophagic (SQSTM1 7, ATG5) and inflammation (IL-1ß, TNF-α) genes were upregulated by 5-11-fold in the IR group. In the groups in which quinoa was administered for treatment and protection, all these genes were found to be upregulated to a lower extent than the IR group. Antioxidant genes (GPX1, SOD1) increased by nine to tenfold in the quinoa groups. CONCLUSION: As a result, after administration of quinoa, it was determined that the glucose level increased due to experimental insulin resistance and the liver and kidney damage indicators decreased. It was determined that quinoa (Chenopodium quinoa) had significant beneficial effects on biochemical parameters and apoptotic, autophagic, antioxidant and inflammatory markers in experimental glucocorticoid-induced insulin resistance.

MYELOLIPOMA COEXISTENCE WITH GLUCOCORTICOID AND ANDROGEN SECRETING ADRENOCORTICAL CARCINOMA: SLOW AND BENIGN CLINICAL COURSE.

We present a case of androgen and glucocorticoid secreting adrenocortical carcinoma with concomitant myelolipoma. A giant adrenal tumor which was initially nonfunctional was reassessed four years later due to the patient's refusal to treat. The patient was a 48-year-old woman with hypertension and acne lesions on the face. Laboratory findings were consistent with glucocorticoid and androgen hypersecretion. Computed tomography revealed a heterogeneously contrasting mass of 145x118x100 mm with lobular contour and soft tissue areas. The patient underwent left laparoscopic transperitoneal adrenalectomy with three port technique. There were no complications in the perioperative period. The resected specimen weighed 850 grams. Pathological findings showed a combination of myelolipoma-adrenal cortical cancer. In the postoperative period, hypertension improved and the hormone panel was normalized. Postoperative computed tomography and PET-CT showed no residual mass and metastasis. Although imaging is compatible with benign masses such as myelolipoma, coexistence of ACC-myelolipoma should be kept in mind and functional evaluation should be performed.

Apoptotic and Oxidative Mechanisms in Liver and Kidney Tissues of Sheep with Fluorosis.

This study was planned to determine the molecular basis and causes of damage to the kidney and the liver, which are the most affected tissues in sheep exposed to chronic fluoride. For this purpose, liver and kidney tissues were obtained from sheep with signs of fluorosis in the age range of 4-6 years. The control group consisted of clinically healthy sheep without fluorosis. The apoptotic and oxidative genes expression of target genes was determined using the real qRT-PCR method. According to the control gene (Gapdh) that was detected that in the liver, the apoptotic genes caspase-8, caspase-9, and Bim increased and caspase-3, Bcl-2, and Bak decreased, while in the kidney, caspase-3 and Bax and caspase-8, Bcl-2, Bcl2l-1, Bim, and Bak decreased. According to the 2-ΔCt values of the oxidative stress genes, it was determined that Cygb, Gstp1, and Ncf1 genes increased significantly in the fluorosis group and Gpx1, sod1, and sod2 genes decreased significantly. In the kidney tissue, Cygb and Gpx1 increased in the fluorosis group, while sod1, sod2, Gstp1, Ncf1 and Ccs, and Nos2 were found to decrease significantly. As a result, it was shown that apoptosis and oxidative mechanisms are activated in the liver and the kidney tissues of sheep with fluorosis and these parameters have an important role in understanding the molecular basis of tissue damage in fluorosis.

The Effect of Lycopene on DNA Damage and Repair in Fluoride-Treated NRK-52E Cell Line.

Exposure of fluorine at toxic concentrations causes serious damage by accumulating in especially bones, kidneys, and other soft tissues. Fluorine at cytotoxic concentrations may cause DNA damage. This study aims to determine the level of DNA damage due to sodium fluoride (NaF) at different hours (3rd, 12th, and 24th hours) and in IC50 concentrations designated for each hour and reveal the protective effect of lycopene on possible damage. The best enhancer concentrations (1 µM) of microtitration (MTT) viability test and proliferation of lycopene and IC50 values of NaF at the 3rd, 12th, and 24th hour were 9600, 5500, and 3200 µM, respectively. DNA damage significantly increased in all NaF-treated groups in comparison with the control group (p < 0.05). DNA damage due to NaF+LYC application significantly decreased in comparison with the control group (p < 0.05). Lycopene application significantly increased the expression levels of the Ku70 and Ku80 genes which have a part in DNA repair (p < 0.05). The statistical data showed that application of lycopene which is an important antioxidant molecule may be beneficial for decreasing NaF-induced DNA damage. In conclusion, applying lycopene for cytotoxicity due to fluorine in NRK-52E cell line had different effects based on the dosage and time; thus, it can be a potential option for preventing fluorosis-induced toxicity and developing new treatment approaches.

The effect of vitamin E and selenium combination in repairing fluoride-induced DNA damage to NRK-52E cells.

Prolonged and excessive fluoride exposure can lead to fluorosis. The kidney is one of the organs that are injured mostly due to fluoride-induced damage. Fluoride can induce DNA damage at cytotoxic concentrations. This study aims to determine the extent of NaF-induced DNA damage and to investigate the effect of vitamin E and selenium combination (ES) in preventing and repairing this damage. For this purpose, we administered different combinations of NaF and ES to NRK-52E cells and determined the effective concentrations of ES and the NaF IC50 values associated with different incubation times (3, 12, and 24 h) by using the MTT assay. The determined quantities of NaF IC50 in association with time and the NaF IC50 + ES combination were administered to the cells. The extent of DNA damage was determined with the comet assay and the expression levels of the Ku70/80 and PARP-1 genes were determined with the RT-qPCR method. DNA damage significantly increased in all experimental groups compared to the control group (p < 0.05). It was found out that the NaF and ES combination statistically reduced the DNA damage compared to the damage observed in the NaF-treated groups (p < 0.05). Treatment of the ES combination significantly increased the expressions of Ku70 and Ku80 genes involved in DNA repair (p < 0.05). We concluded that vitamin E and selenium can potentially be effective in the repair of fluoride-induced DNA damage based on the results of this in vitro study. Our results may shed light on the prevention of DNA damage associated with fluorosis.

The role and importance of auxiliary tests in differential diagnosis in patients with mildly high basal 17-OH-progesterone levels in the evaluation of hirsutism

Background/aim: In the differential diagnosis of hirsutism, early follicular basal 17-OH-progesterone levels sometimes overlap with the diagnosis of late onset congenital adrenal hyperplasia (LOCAH) and other causes of hyperandrogenism. This study aims to investigate the role of some common tests and clinical findings in differential diagnosis in such cases. Materials and methods: One hundred seventy-five female patients with hirsutism and mildly high initial 17-OH-progesterone levels (2-10 ng/mL) were included in the study. The cases were divided into three groups according to their diagnosis: LOCAH (n = 16, mean age = 26.1 ± 6.9), polycystic ovary syndrome (PCOS) (n = 122, mean age = 23.9 ± 5.1), and intracranial hypertension (IH) (n = 37, mean age = 25.2 ± 7.3). Clinical signs and symptoms, such as menstrual irregularity and hirsutism score, and hormone levels including total testosterone and dehydroepiandrosterone sulfate (DHEAS), were compared between the groups. Results: There was no difference between the groups with PCOS, LOCAH, and IH for total testosterone level results (P = 0.461). The DHEAS level was higher in the PCOS group than in the LOCAH group (449.6 ± 151.14 vs. 360.31 ± 152.40, P = 0.044). While there was no difference between the PCOS and LOCAH groups in terms of menstrual irregularity (P = 0.316), the hirsutism score for IH was significantly lower than those of PCOS and LOCAH (9.2 vs. 12.2 and 11.1, respectively; P < 0.001). Basal 17-OH-progesterone levels were higher in the LOCAH group than in the other groups (P = 0.016). Conclusion: While DHEAS level was lower in LOCAH than in PCOS, it was not different from that in IH. While the severity of hirsutism was higher in LOCAH than in IH, it was not different from that in PCOS. Menstrual irregularity was similar between PCOS and LOCAH. According to these results, although the auxiliary tests and clinical findings for the diagnosis of LOCAH contribute to the clinical interpretation, they are not superior to the 17-OH-progesterone level for diagnosis.

Hemogram parameters in the patients with subacute thyroiditis.

BACKGROUND &OBJECTIVE: Subacute Granulomatous Thyroiditis (De Quervain's Thyroiditis) is an acute painful inflammatory disease of the thyroid. We aimed to investigate easily accessible and cheap hemogram based parameters of Neutrophil to Lymphocyte Ratio (NLR), Platelet to Lymphocyte Ratio (PLR), and Mean Platelet Volume (MPV) in the follow up of inflammatory thyroid disease. METHODS: Patients admitted to Sakarya University Education and Research Hospital Endocrinology and Metabolism Outpatient Clinic and diagnosed as Subacute Granulomatous Thyroiditis between May 2017 and November 2018 were included in the study. Hemogram, thyroid function tests and acute phase values of these patients were recorded and compared with the values after treatment and disease recovery. On the sixth month, thyroid function tests were repeated and the rate of permanent hypothyroidism was screened. The relationships between initial hemogram parameters and acute phase reactants were evaluated. RESULTS: Total 71 patients were included in our study. 60 (84.5%) were female and 11 (15.5%) were male. The F/M ratio was found to be 6/1. Mean age was 43 ± 9.95 years. Receiver Operating Characteristics (ROC) Curve Analysis was performed and values for Area Under the Curve (AUC) for NLR and PLR, respectively, were 0.739 (95% CI 0.657-0.820 p<0.0001) and 0.772 (95% CI 0.694-0.850 p<0.0001), which are significant and associated with disease activity. However, the AUC for MPV parameter was: 0.578 (95% CI 0.484-0.672 P: 0.10) and was not significant. The cut off values defined as 2.4 (80% sensitivity and 51% specificity) for NLR and 146.84 (83% sensitivity and 54% specificity) for PLR for the acute phase of the disease. In the Correlation Analysis, NLR and PLR values were significantly correlated with ESR and CRP parameters, which are the most commonly used acute phase reactants. CONCLUSION: According to the present study, we believe that the NLR and PLR parameters will be of benefit in the follow-up the disease, accurately demonstrate the inflammatory load in the acute phase of the disease, and correlate with the common acute phase reactants.

In Vitro evaluation of thymoquinone on apoptosis and oxidative DNA damage in high glucose condition.

The study was planned to investigate the effects of thymoquinone (TQ), which is a compound in N. sativa, on caspase dependent apoptosis and oxidative DNA damage in high glucose treated PC12 cells. PC12 cells were treated with high glucose (G1-150 mM, G2-250 mM, G3-350 mM), TQ (20 µM), and their combinations. Oxidative DNA damage (8-OHdG (8-Oxo-2'-deoxyguanosine)), and apoptosis (caspase 3, caspase 8, caspase 9 enzymes and M30 protein) parameters were analyzed with ELISA. The 8-OHdG levels decreased in all combination groups compared to the control (p≤0.001). There was no statistically significant difference between caspase 3 and 9. Caspase 8 in TQ, G3, TQG1, TQG2 groups were higher than the control (p≤0.002). Low M30 levels were observed in TQG1 group (p≤0.002). In conclusion, it was observed that in PC12 cell line treated with the high glucose concentrations, TQ administration had a statistically significant effect on oxidative DNA damage and some apoptotic parameters (caspase 8 and M30 protein).

Levels of Trace Elements in Muscle and Kidney Tissues of Sheep with Fluorosis.

In this study, we report the concentrations of four trace elements in muscle and kidney tissues of sheep grown in an area with endemic fluorosis. Fifteen 3- to 4-year-old fluorotic sheep were selected for the study. Ten age-matched sheep with no sign of fluorosis were used as the control group. The animals were killed in a slaughterhouse in the village of Dogubeyazit, located in the Agri province of Eastern Turkey, where kidney and muscle tissue samples were surgically obtained to be analyzed for copper, zinc, nickel, and iron. In muscle tissue of the fluorotic sheep, the copper levels were higher than those of the control group (p < 0.05). In the case of zinc, its levels were significantly higher in the controls than in the sheep with fluorosis (p < 0.01). No statistically significant differences were found in the muscle contents of nickel and zinc (p > 0.05). Compared to controls, the concentrations of zinc (0.01), iron (p < 0.05), and nickel (p < 0.05) were significantly higher in kidney tissues of fluorotic sheep, but there were no significant differences of the copper levels (p > 0.05). These results suggest that fluorosis significantly alters the mineral metabolism in muscle and kidney resulting in higher levels of mineral accumulation and excretion caused by fluoride intoxication. Further research shall focus on the enzymatic and metabolic activities of these and other trace elements.

Clinical presentation and management of Fasciola hepatica infection: single-center experience.

AIM: To identify the characteristic clinical, laboratory and radiological findings and response to treatment in patients with fascioliasis. METHODS: Patients who were diagnosed with Fasciola hepatica infection were included in this prospective study. Initial clinical, laboratory and radiological findings were recorded. All patients were followed until a complete response was achieved or for 6 mo after treatment discontinuation. RESULTS: Fasciola hepatica infection was diagnosed in 30 patients (24 females; mean age: 42.6 years) between January 2008 and February 2011. Twenty-two (73%) patients had hepatic phase fascioliasis, 5 patients had biliary phase, and 3 patients had biliary phase associated with acute pancreatitis. Of the 8 patients with biliary phase fascioliasis, 2 patients displayed features that overlapped with both hepatic and biliary phase. Abdominal pain and right upper abdominal tenderness were the most prominent signs and symptoms in all patients. Eosinophilia was the most prominent laboratory abnormality in both patients with hepatic and biliary phase (100% and 50%, respectively). Multiple nodular lesions like micro-abscesses on abdominal computerized tomography were the main radiological findings in patients with hepatic phase. Small linear filling defects in the distal choledochus were the main endoscopic retrograde cholangiopancreatography (ERCP) findings in patients with biliary phase. Patients with hepatic phase were treated with triclabendazole alone, and patients with biliary phase were treated with triclabendazole and had live Fasciola hepatica extracted from the bile ducts during ERCP. CONCLUSION: Fasciola hepatica infection should be considered in the differential diagnosis of patients with hepatic or biliary disease and/or acute pancreatitis associated with eosinophilia.

Experimental and theoretical studies on the identification of p-biphenyloxycarbonylphenyl acrylate.

This study presents the identification of a title compound, p-biphenyloxycarbonylphenyl acrylate by means of experimental and theoretical evidences. The spectroscopic properties of the compound were experimentally investigated by Fourier transformation-infrared spectra (in the region 400-4000 cm(-1)) and nuclear magnetic resonance (NMR) chemical shifts (with a frequency of 400 MHz). Moreover, the optimized molecular structures, vibrational frequencies including infrared intensities and Raman activities, corresponding vibrational spectra interpreted with the aid of normal coordinate analysis based on scaled density functional force field, thermodynamic properties, atomic charges and ultraviolet-visible (UV-vis) spectra were analyzed utilizing ab initio Hartree-Fock (HF) and Density Functional Theory (B3LYP) methods at 6-31G(d,p) calculation level. It was found that the vibrational frequencies and chemical shifts obtained were shown to have a good agreement with available experimental results. We not only simulated frontier molecular orbitals (FMO) and molecular electrostatic potential (MEP) but also evaluated the transition state and energy band gap clearly.