Rare-earth orthovanadate nanoparticles trigger Ca2+-dependent eryptosis.

Introduction. Rare-earth orthovanadate nanoparticles (ReVO4:Eu3+, Re = Gd, Y or La) are promising agents for photodynamic therapy of cancer due to their modifiable redox properties. However, their toxicity limits their application.Objective. The aim of this research was to elucidate pro-eryptotic effects of GdVO4:Eu3+and LaVO4:Eu3+nanoparticles with identification of underlying mechanisms of eryptosis induction and to determine their pharmacological potential in eryptosis-related diseases.Methods. Blood samples (n= 9) were incubated for 24 h with 0-10-20-40-80 mg l-1GdVO4:Eu3+or LaVO4:Eu3+nanoparticles, washed and used to prepare erythrocyte suspensions to analyze the cell membrane scrambling (annexin-V-FITC staining), cell shrinkage (forward scatter signaling), reactive oxygen species (ROS) generation through 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) staining and intracellular Ca2+levels via FLUO4 AM staining by flow cytometry. Internalization of europium-enabled luminescent GdVO4:Eu3+and LaVO4:Eu3+nanoparticles was assessed by confocal laser scanning microscopy.Results.Both nanoparticles triggered eryptosis at concentrations of 80 mg l-1. ROS-mediated mechanisms were not involved in rare-earth orthovanadate nanoparticles-induced eryptosis. Elevated cytosolic Ca2+concentrations were revealed even at subtoxic concentrations of nanoparticles. LaVO4:Eu3+nanoparticles increased intracellular calcium levels in a more pronounced way compared with GdVO4:Eu3+nanoparticles. Our data disclose that the small-sized (15 nm) GdVO4:Eu3+nanoparticles were internalized after a 24 h incubation, while the large-sized (∼30 nm) LaVO4:Eu3+nanoparticles were localized preferentially around erythrocytes.Conclusions.Both internalized GdVO4:Eu3+and non-internalized LaVO4:Eu3+nanoparticles (80 mg l-1) promote eryptosis of erythrocytes after a 24 h exposurein vitrovia Ca2+signaling without involvement of oxidative stress. Eryptosis is a promising model for assessing nanotoxicity.

The Saliva Cortisol and Amylase Levels Related with Stress Response Compared by Different Analytical Methods.

Sampling of salivary cortisol and amylase is a non-invasive method and important for the evaluation of the hypothalamic-pituitary-adrenal axis function and stress levels. This study aimed to compare the values of the salivary cortisol and amylase levels which were measured by three different analytical methods to discuss the alterations of stress levels of samples. The saliva samples of young adults (n = 23) were collected between 08.00 and 09.00 a.m., noon at 12.00 (before exam) and between 14.00 and 15.00 p.m. (after unaware exam). The samples were measured within the first 48 h, and no freezing/thawing was done. Salivary cortisol and amylase levels of subjects were measured by three different analytical methods as ELISA, chemiluminescence and biosensor methods. Comparison of ELISA and biosensor methods in order to determine the salivary cortisol levels showed a good correlation y = 2.971 + 0.748x (R2 = 0.839). Salivary amylase concentrations were only detected by ELISA method. Biosensor can be offered as an alternative analytic method to the conventional determination method ELISA. It can be preferred because of the detection/information effectiveness, low cost, fast results and specificity characteristics.

UV Light-Activated GdYVO4:Eu3+ Nanoparticles Induce Reactive Oxygen Species Generation in Leukocytes Without Affecting Erythrocytes In Vitro.

Nanoparticles (NPs) have been reported to be promising enhancement agents for radiation therapy. The aim of the study was to assess the cytotoxicity of UV non-treated and UV pretreated GdYVO4:Eu3+ nanoparticles against erythrocytes and leukocytes by detecting eryptosis and reactive oxygen species (ROS) generation. Levels of intracellular ROS in erythrocytes and leukocytes using a ROS-sensitive dye 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA), as well as eryptosis rate utilizing annexin V staining, following direct exposure to UV-activated and nonactivated NPs were detected by flow cytometry. Blood cells were collected from 9 intact WAG rats. Neither the UV light-untreated GdYVO4:Eu3+ NPs nor the treated ones promoted eryptosis and ROS generation in erythrocytes. Low concentrations of UV light-untreated NPs did not induce oxidative stress in leukocytes, evidenced by unaffected intracellular ROS levels. UV light treatment grants prooxidant properties to NPs, confirmed by NP-induced ROS overproduction in leukocytes. High concentrations of both UV light-treated and untreated NPs altered the redox state of leukocytes. UV light treatment imparts prooxidant properties to GdYVO4:Eu3+ NPs, making them promising radiosensitizing agents in cancer radiation therapy.

The protective effect of Ferula elaeochytris on age-related erectile dysfunction.

ETHNOPHARMACOLOGICAL RELEVANCE: Ferula elaeochytris Korovin (FE) is a perennial medicinal plant of Apiaceae family. Ferula elaeochytris Korovin, known as 'Çaksir' in Anatolia, is widely used as an aphrodisiac as well as antioxidant, anti-inflammatory, and anti-diabetic. AIM OF THE STUDY: Erectile dysfunction (ED) is a serious public health problem that has a high prevalence and negatively affects the quality of life in elderly men. In the treatment and prophylaxis of many diseases, because of widely increasing use of plant extracts as therapeutic agents, preclinical studies related to plant extracts are becoming more important by the day. In this study, we aimed to investigate the protective effect of Ferula elaeochytris Korovin (FE) root extract on age-related ED. MATERIALS AND METHODS: Seventy-two male Wistar albino rats were equally divided into four groups: 4-month aged rats (Y), 24-month aged rats (AG), and FE-administered (20 and 40 mg/kg/day; oral gavage; over 8 weeks) 24-month aged rats (AG + FE). The measurements included: changes in smooth muscle cells and collagen fibrils, tumor necrosis factor alpha (TNF-α), penile neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) expression, serum testosterone concentrations (ST), neurogenic- and endothelial-dependent relaxations of the corpus cavernosum (CC), intracavernosal pressure/mean arterial pressure (ICP/MAP), area under the curve (total ICP), total antioxidant status (TAS), and total oxidant status (TOS) on corpus cavernosal tissue. RESULTS: These results have an important role in the development of ED. ICP/MAP, total ICP, eNOS/nNOS expressions and ST levels increased in AG+40 mg FE group compared to the AG group, whereas TNF-α levels decreased and oxidative and antioxidant parameters balanced. CONCLUSIONS: Our findings show that FE may have a useful effect on decelerating the development of age-related ED.

A new biosensor for noninvasive determination of fetal RHD status in maternal blood of RhD negative pregnant women.

Prenatal detection of the fetal RHD status can be useful in the management of RhD incompatibility to identify fetuses at risk of hemolytic disease. Hemolytic disease causes morbidity and mortality of the fetus in the neonatal period. The routine use of antenatal and postnatal anti-D prophylaxis has reduced the incidence of hemolytic disease of the fetus and newborn. This study describe the detection of fetal RhD antigens in blood of RhD negative pregnant women using a nanopolymer coated electrochemical biosensor for medical diagnosis. Cell free fetal DNA in maternal plasma was also used to genotyping fetal RHD status using multiplex real-time PCR. Twenty-six RhD negative pregnant women in different gestational ages were included in the study. RhD positive fetal antibodies detected with a developed biosensor in maternal blood of RhD negative mothers. The electrochemical measurements were performed on a PalmSens potentiostat, and corundum ceramic based screen printed gold electrode combined with the reference Ag/AgCl electrode, and the auxiliary Au/Pd (98/2%) electrode. Fetal RHD genotyping performed using fluorescence-based multiplex real-time PCR exons 5 and 7 of the RHD gene. The fetal RHD status of 26 RhD negative cases were detected 21 as RhD positive and 5 as RhD negative with electrochemical biosensor. Fetal RHD status confirmed with extracted fetal DNA in maternal plasma using multiplex real-time PCR RHD genotyping and by serological test after delivery. The new method for fetal RhD detection in early pregnancy is useful and can be carry out rapidly in clinical diagnosis. Using automated biosensors are reproducible, quick and results can be generated within a few minutes compared to noninvasive fetal RHD genotyping from maternal plasma with real-time PCR-based techniques. We suggest the biosensor techniques could become an alternative part of fetal RHD genotyping from maternal plasma as a prenatal screening in the management of RhD incompatibility.