Investigation of the effects of the toll-like receptor 4 pathway on immune checkpoint vista in pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most common malignant tumors of the pancreas. Preclinical studies show that it evades the immune system with immune checkpoints and promotes tumor development. V-domain Ig suppressor of T cell activation (VISTA) is a new immune-check point from the B7 family and is highly expressed in cancer cells. Overexpression of toll like receptor 4 (TLR4) in pancreatic adenocarcinoma is associated with induced tumorigenesis, tumor growth, resistancy to chemotherapy. Naloxone is an opioid and inhibits TLR4-ligand association. In this study, we investigated the relation of TLR4 and downstream pathways with immune-check point VISTA in pancreatic cancer proliferation. We initially collected pancreatic cancer-related datasets using the GEPIA2 and UALCAN databases. Based on this data obtained the effect of various concentrations and incubation times of naloxone were used on PANC-1 cells proliferation. A combination of naloxone and VISTA-siRNA were applied, and the effect of both naloxone and combined treatment on TLR4, Interleukin 1 receptor associated kinase 4 (IRAK4) and VISTA gene expression were analyzed in pancreatic cancer cells. As a result of analysis with Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR), gene expression levels of TLR4, IRAK4 and VISTA were significantly suppressed and cell proliferation was significantly reduced. We found that administration of naloxone and VISTA-siRNA in combination with PDAC cells suppressed signaling. Therefore, we considered that the relationship between VISTA and TLR4 signaling pathways and the other possible associated signal molecules may be an important marker in determining the response of immune checkpoint inhibitors in cancer treatment.

Artemisinin May Disrupt Hyphae Formation by Suppressing Biofilm-Related Genes of Candida albicans: In Vitro and In Silico Approaches.

This study aimed to assess the antifungal and antibiofilm efficacy of artemisinin against Candida (C.) species, analyze its impact on gene expression levels within C. albicans biofilms, and investigate the molecular interactions through molecular docking. The antifungal efficacy of artemisinin on a variety of Candida species, including fluconazole-resistant and -susceptible species, was evaluated by the microdilution method. The effect of artemisinin on C. albicans biofilm formation was investigated by MTT and FESEM. The mRNA expression of the genes related to biofilm was analyzed by qRT-PCR. In addition, molecular docking analysis was used to understand the interaction between artemisinin and C. albicans at the molecular level with RAS1-cAMP-EFG1 and EFG1-regulated genes. Artemisinin showed higher sensitivity against non-albicans Candida strains. Furthermore, artemisinin was strongly inhibitory against C. albicans biofilms at 640 µg/mL. Artemisinin downregulated adhesion-related genes ALS3, HWP1, and ECE1, hyphal development genes UME6 and HGC1, and hyphal CAMP-dependent protein kinase regulators CYR1, RAS1, and EFG1. Furthermore, molecular docking analysis revealed that artemisinin and EFG1 had the highest affinity, followed by UME6. FESEM analysis showed that the fluconazole- and artemisinin-treated groups exhibited a reduced hyphal network, unusual surface bulges, and the formation of pores on the cell surfaces. Our study suggests that artemisinin may have antifungal potential and showed a remarkable antibiofilm activity by significantly suppressing adhesion and hyphal development through interaction with key proteins involved in biofilm formation, such as EFG1.

Gene expression of MTATP6 and cytochrome P450 in MCF-7 and MDA-MB -231 breast cancer cell lines with juglone and curcumin supplemented.

It is aimed to determine the effects of naphthoquinones as juglone and curcumin application on cell viability and expression analyzes of CYP3A4 and MTATP6 genes in MCF-7 and MDA-MB-231 human breast cancer cell lines. MCF-7 and MDA-MB-231 cells were incubated, were replaced with containing various concentrations of 5, 10, 15 µM curcumin and 5, 10, 15 µM juglone for MCF-7 and 1, 5, 10 µM curcumin and 1, 2, 3 µM juglone for MDA-MB-231 for 24 h. CYP3A4 and MTATP6 gene expression levels in both cell lines were determined by quantitative real-time polymerase chain reaction (qPCR) method and western blot method. IC50 values for 24 h were found as 22.41 µM for curcumin, and 16.27 µM for juglone in MCF-7, and 10.43 µM for curcumin, and 3.42 µM for juglone in MDA-MB-231 cells. Curcumin showed anti-proliferative, and antioxidant effects. CYP3A4 and MTATP6 gene expressions were decreased in MCF-7 breast cancer cell line when the cells treated with juglone or curcumin. CYP3A4 and MTATP6 gene expressions were decreased at all application doses of juglone in MDA-MB-231 cells whereas CYP3A4 and MTATP6 protein levels were only decreased at 10 µM curcumin compared with the control group.

In vitro antifungal and antibiofilm activities of novel sulfonyl hydrazone derivatives against Candida spp.

BACKGROUND: The aim of this study was to investigate the antifungal and antibiofilm activity of the new sulfonyl hydrazones compound derived from sulphonamides. METHODS: In this study, new sulfonyl hydrazone series were synthesized via a green chemistry method. The structures of the synthesized compounds were characterized by elemental analyses and spectroscopic methods. The antifungal activities of the Anaf compounds against Candida strains under planktonic conditions were tested. The biofilm-forming ability of Candida strains was determined and the inhibitory effects of Anaf compounds on Candida biofilms compared with fluconazole were measured by MTT assay. Expression analysis of biofilm-related genes was investigated with qRT-PCR. The statistical analysis was performed using a one-way ANOVA test. CANDIDA: strains was determined and the inhibitory effects of Anaf compounds on Candida biofilms compared with fluconazole were measured by MTT assay. Expression analysis of biofilm-related genes was investigated with qRT-PCR. The statistical analysis was performed using a one-way ANOVA test. RESULTS: A total of 16 (45.7%) out of 35 Candida isolates were determined as strong biofilm producers in this study. C. albicans was the most biofilm producer, followed by C. krusei and C. lusitaniae. The Anaf compounds had a broad spectrum of activity with MIC values ranging from 4 µg/ml to 64 µg/ml. Our data indicated that the Anaf compound had a significant effect on inhibiting biofilm formation in both fluconazole-susceptible and -resistant strains. The expression levels of hypha-specific genes als3, hwp1, ece1 and sap5 were downregulated by Anaf compounds. CONCLUSIONS: Our study revealed that the Anaf compounds had antifungal activity and inhibited fungal biofilms, which may be related to the suppression of C. albicans adherence and hyphal formation. These results suggest that Anaf compounds may have therapeutic potential for the treatment and prevention of biofilm-associated Candida infections.

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.

Regenerative effects of hyperbaric oxygen therapy and platelet-rich plasma on the osteochondral defects of rats.

OBJECTIVES: This study aims to investigate the effects of hyperbaric oxygen (HBO) therapy and platelet-rich plasma (PRP) on the regeneration of osteochondral defects of the rats, and the synergistic effect of this combined treatment. MATERIALS AND METHODS: This randomized, controlled, and interventional animal study was conducted between May 2014 and August 2014 Osteochondral regeneration was evaluated in four treatment groups (control, PRP, HBO, and HBO+PRP groups) at the 30th day after iatrogenic injury. Thirty-two female Wistar albino rats (weighing 248-305 g) underwent arthrotomy and osteochondral surgery on left knees. The regenerations of defects were then examined histologically by the modified version of O'Driscoll score. RESULTS: Groups that were treated with either HBO or PRP alone regenerated significantly better than the control group (p=0.01), while no significant difference was found between the HBO- and PRP-treated groups (p>0.05). The defects in group 4 (treated with both HBO and PRP) regenerated significantly better than the control group, the HBO-treated group alone, and the PRP-treated group alone (p=0.01). CONCLUSION: The results of this study showed a synergistic effect of HBO and PRP on knee cartilage regeneration. However, the possible underlying mechanisms should be the subject of future researches. The aggregation and activation of growth factors released from platelets whose activation is increased in the hyperbaric environment may explain this effect. This may result in a better regeneration than the effect of PRP or HBO alone.

Homocysteine levels in patients with masked hypertension.

OBJECTIVE: Masked hypertension is a clinical condition, the importance of which is agreed in recent years and which is characterized by increased cardiovascular mortality and morbidity and is thought to be important endothelial dysfunction in the pathophysiology. Plasma total homocysteine levels are accepted as a major independent biomarker for endothelial dysfunction and/or a contributor to hypertension and coronary artery disease. In this study, we aimed to measure the level of serum homocysteine and to evaluate the relationship between the parameters of ambulatory blood pressure monitoring in patients with masked hypertension. METHODS: This cross-sectional observational study included 37 subjects with normal blood pressure, 30 with masked-hypertension and 27 patients with obvious hypertension. Masked hypertension (MHT) was defined as office blood pressure <140/90 mm Hg and mean daytime ambulatory systolic blood pressure in 24 hours monitoring ≥135/85 mm Hg. Homocysteine levels of the subjects were measured by using HPLC system with fluorescent detector. Lipid parameters were measured by routine methods. Mann-Whitney U test was used for statistical analysis. RESULTS: In the analysis of homocysteine, it was observed that there was no difference between the control group and patients with MHT. Patients with high blood pressure showed higher homocysteine levels when compared to MHT (p=0.02). Homocysteine levels showed a weak positive correlation with average systolic blood pressure (r=0.335, p=0.043). Homocysteine levels were higher in smokers than non-smokers. compared with non-smokers group in all participants (p=0.036). CONCLUSION: We have reached the opinion that in the individuals with no obvious health problems but with MHT, homocysteine levels may not have any significant effect upon high blood pressure levels.

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. (AU)