Juglone-ascorbate treatment enhances reactive oxygen species mediated mitochondrial apoptosis in pancreatic cancer.

BACKGROUND: Treatment of Pancreatic Cancer (PC) is challenging due to its aggressiveness and acquired resistance to conventional chemotherapy and radiotherapy. Therefore, the discovery of new therapeutic agents and strategies is essential. Juglone, a naphthoquinone, is a secondary metabolite produced naturally in walnut-type trees having allelopathic features in its native environment. Juglone was shown to prevent cell proliferation and induce ROS-mediated mitochondrial apoptosis. Ascorbate with both antioxidant and oxidant features, shows selective cytotoxicity in cancer cells. METHODS AND RESULTS: In this study, we evaluated the anticancer effects of Juglone in combination with ascorbate in PANC-1 and BxPC-3 PC cells. The MTT assay was used to determine the IC50 dose of Juglone with 1 mM NaAscorbate (Jug-NaAsc). Subsequently, the cells were treated with 5, 10, 15 and 20 µM Jug-NaAsc for 24 h. Apoptotic effects were evaluated by analyzing the following genes using qPCR; proapoptotic Bax, antiapoptotic Bcl-2 related to the mitochondrial apoptotic pathway and apoptosis inhibitor Birc5 (Survivin). Immunofluorescence analysis was performed using Annexin V-FITC in PC cells. As an antioxidant enzyme, Trx2 protein levels were determined by a commercial ELISA test kit. Jug-NaAsc treatment decreased the expressions of antiapoptotic genes Bcl-2 and Birc5 while the apoptotic gene Bax expression increased at all doses. Additionally, a dose-dependently increase of apoptosis according to immunofluorescence analysis and the decreases of Trx2 enzyme levels at all treatments in both cell lines supported gene expression results. CONCLUSION: Our results suggest that Juglone is a potential anticancer agent especially when combined with ascorbate.

The Effects of Adipose-Derived Mesenchymal Stem Cells and Adipose-Derived Mesenchymal Stem Cell-Originating Exosomes on Nerve Allograft Regeneration: An Experimental Study in Rats.

INTRODUCTION: Nerve regeneration has been the subject of many studies because of its complex mechanism and functional outcome. Mesenchymal stem cells and exosomes are promising factors in regeneration in many areas. Reconstruction of nerve defects is a controversial issue, and nerve allografts are promising alternatives with many advantages. In this study, it is aimed to evaluate the nerve regeneration in cellularized and decellularized nerve allografts and whether it is possible to accelerate this process with adipose-derived mesenchymal stem cells (ad MSC) or ad MSC-originating exosomes. METHOD: This study was performed with 36 Lewis and 18 Brown Norway isogenic male rats aged 10 to 12 weeks and weighing 300 to 350 g. The Lewis rats were divided into 6 groups. Nerve allografts at a length of 12 mm that were obtained from the Brown Norway rats' proximal portion of both sciatic nerve branching points were coapted as cellularized in group A and decellularized in group B to the sciatic nerve defects of the Lewis rats. Group A received oral tacrolimus (0.2 mg/kg) for 30 days. Perineural saline (A1-B1), ad MSC (A2-B2), or ad MSC-originating exosomes (A3-B3) were applied to these groups. Walking track analysis, pinch-prick test and electromyelography were applied at the 8th and 16th weeks following surgery. Nerves were examined histopathologically at the 16th week. RESULTS: Between cellularized groups, better results were shown in A3 about axon-myelin regeneration/organization (P = 0.001), endoneural connective tissue (P = 0.005), and inflammation (P = 0.004). Better results were shown in the B2 and B3 groups electromyelographicaly about latency period (P = 0.033) and action potential (P = 0.008) at late period, and histomorphologicaly at vascularization (P = 0.012). DISCUSSION: It is argued that regeneration is accelerated with decellularization of nerve allografts by removing the chondroidin sulfate proteoglycans. The positive effects of stem cells are derived by exosomes without the cell-related disadvantages. In this study, better results were obtained by decellularization and perineural application of ad MSC and/or ad MSC exosome.

The effects of Juglone-Selenium combination on invasion and metastasis in pancreatic cancer cell lines.

Background: Pancreatic cancer does not show any symptoms in the early period and metastatic process is already passed when the diagnosis is done. Therefore, in the battle with pancreatic cancer, novel treatment strategies, particularly antiinvasive and antimetastatic strategies, are needed. The cytotoxic and anticancer effects of juglone and sodium selenite (NaSe) have been showed in various cancer cells. Objectives: In this study, it is aimed to investigate the synergistic effects of juglone and selenium on PANC-1 and BxPC-3 pancreatic cancer cells. Methods: Antimetastatic effects of juglone-NaSe were carried out by adhesion and invasion assays and the genes and protein expressions. Expression analysis of the CDH1, ITGB3 and COL4A3 genes and their proteins E-cadherin, ß3 integrin and tumstatin which play role in metastasis and angiogenesis processes, were done by qPCR and immunohistochemical analysis, respectively. Results: Study findings have provided evidences that the juglone-selenium has a cytotoxic and dose dependent suppressive effect on invasion and metastasis in PANC-1 and BxPC-3 cells. Conclusion: The juglone-NaSe has the potential to be a promising agent especially to inhibit invasion and metastasis in pancreatic cancer treatment. However, more in depth studies are needed to more clearly demonstrate the effects of juglone-selenium.

Evaluation of sialic acid, malondialdehyde and glutathione levels in infertile male.

INTRODUCTION AND OBJECTIVES: Around 15% of productive couples in the world are infertile. Recent years, biochemical mechanisms leads to male infertility are started to research. Redox regulation and oxidative stress (OS) show importance in the pathogenesis of infertility in male. Malondialdehyde (MDA) and Glutathione (GSH) are biochemical indicatives of sperm damage by reactive oxygen species (ROS). In addition, sperm are coated with a thick glycocalyx rich in sialic acids. It is aimed to determine and evaluate the differences between normozoospermic and oligozoospermic individuals according to sialic acid, MDA and GSH concentrations and correlations between spermyogram and these parameters. MATERIAL AND METHODS: This study was carried out on seminal plasma of individuals who admitted to Selcuk University Faculty of Medicine IVF Unit Andrology Laboratory. The groups were divided into two as normozoospermics (n=30, sperm concentration≥15million/mL), and oligozoospermics (n=30, sperm concentration<15million/mL). Spermyogram were evaluated regarding WHO (2010) Kruger criteria. GSH, MDA and sialic acid concentrations were analyzed in seminal plasma. Diagnostic performance of sialic acid has been determined with ROC curve analysis. RESULTS: Sialic acid levels were significantly lower in Normozoospermic than Oligozoospermic individuals (p<0.0001), MDA and GSH levels were not differ significantly in both groups (p>0.05). Sialic acid correlated significantly with most of the spermyogram findings. When diagnostic performance of sialic acid was evaluated, the cut off value of sialic acid found as 4.175nmol/mL by ROC curve. CONCLUSION: High seminal plasma sialic acid levels may be used as a biomarker and sialic acid is important determinant in oligozoospermia.

Is there a relation between serum methylarginine levels and infertility?

OBJECTIVES: Infertility is defined as the absence of pregnancy within the reproductive period despite regular sexual intercourse. Methylarginines are formed as a result of methylation of arginine residues in proteins and formed in three forms as asymmetric dimethyl arginine (ADMA), symmetrical dimethyl arginine (SDMA) and monomethylarginine (L-NMMA). So, here, we aimed to evaluate arginine and their derivatives levels in fertile and infertile individuals. METHODS: Present study were consist of 30 oligozoospermia patients (proven by spermiogram analysis) and 30 healthy individuals with normozoospermia group who were applied to the urology department. With blood samples taken from individuals, serum methylarginine and its derivatives levels were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Clinic data and demographic characteristics of individuals were also recorded at the same time. RESULTS: The serum ADMA level (0.38 ± 0.07) of the oligozoospermia group was found to be significantly higher than the normozoospermia group (0.35 ± 0.05) (p=0.046). A positive correlation were observed between ADMA and SDMA (r=0.686, p=0.000), HArg and SDMA (r=0.611, p=0.001), citrulline and L-NMMA (r=0.595, p=0.001) in patients with oligosospermia. The increase in SDMA, arginine and HArg levels and a decrease in L-NMMA and citrulline levels were not significant as statistically. Also, the ADMA level was found to be high in individuals with low sperm concentration. CONCLUSIONS: Consequently, serum ADMA levels of individuals with oligozoospermia were statistically significantly higher than those with normozoospermia. As proposal, determination of ADMA levels may be a potential biomarker parameter in terms of early diagnosis of fertility and infertility.

Efficacy comparison of oral rosuvastatin versus oral progesterone and bevacizumab on regression of surgically endometriotic implants in rats.

This study hypothesizes that oral rosuvastatin, oral dienogest and intraperitoneal bevacizumab might improve endometriosis in randomly selected female Wistar albino rats with surgically endometriotic implants. Thirty female Wistar albino rats with surgically endometriotic implants were randomized into three treatment groups: oral rosuvastatin (20 mg kg/day; oral rosuvastatin group 1; n = 10), oral progesterone (dienogest group 2; n = 10) and intraperitoneal bevacizumab (2.5 mg/kg of single intraperitoneal injection of bevacizumab; bevacizumab group 3; n = 10), for 10 days. Post-treatment variables were compared. The oral rosuvastatin group showed higher reduction for the glandular epithelium and uterine vessels of histopathological scores values than the oral progesterone group (both, p < 0.017, respectively). The median glandular epithelium and uterine vessels and histopathological scores values did not show a statistically significant difference between group 1 and group 3 (p > 0.017). Endometrial thickness values and uterine volume values were more significantly reduced in the oral rosuvastatin group than the oral progesterone group (both, p < 0.017, respectively). Moreover, endometrial thickness and uterine volume values were not different in groups wecompared with group 3 (p > 0.017). In conclusion, oral rosuvastatin and intraperitoneal injection of bevacizumab may cause more significant regression of surgically endometriotic implants in rats than oral progesterone medications.

Antiangiogenic effect of itraconazole on corneal neovascularization: a pilot experimental investigation.

PURPOSE: To investigate the antiangiogenic effect of itraconazole for the prevention of experimentally induced corneal neovascularization and whether the efficacy depends on the route of administration. MATERIALS AND METHODS: Thirty-six rats were randomly divided into 6 groups with 6 rats in each group. Chemical cauterization of the cornea was performed using silver nitrate/potassium nitrate sticks, and the rats were subsequently treated daily with topical (10 mg/ml), subconjunctival (10 mg/ml) or intraperitoneal (19 mg/kg) itraconazole for 7 days. Control rats received topical, subconjunctival or intraperitoneal 0.9% saline. On the 8th day of the experiment, the rat corneas were photographed to determine the percentage area of the cornea covered by neovascularization. The maximum density of corneal neovascularization was determined by microscopy. RESULTS: The median percentage of corneal neovascularization for group 1 was 31.5% (95% confidence interval, 27.5-35.5%); in group 3, it was 32% (23.5-39.8%); in group 5, it was 47% (36.3-60.0%). The percentages of corneal neovascularization in groups 2, 4 and 6 (the control groups) were 70% (95% confidence interval, 60.7-77.3%), 69% (63.0-77.7%) and 68% (56.5-78.5%), respectively. The area of neovascularization was smaller after itraconazole treatment as compared to saline treatment. Further, the area of neovascularization was smaller after topical and subconjunctival administration than after intraperitoneal administration. Histological evaluation of the corneas showed the most extensive corneal neovascularization in the control group. No local or systemic adverse effects were seen from either treatment group. CONCLUSION: Itraconazole reduces corneal neovascularization shortly after chemical burn. However, a larger experimental study is necessary to confirm the data of this investigation.