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The ubiquitin kinase and ligase PINK1 and PRKN together label damaged mitochondria for their elimination in lysosomes by selective autophagy (mitophagy). This cytoprotective quality control pathway is genetically linked to familial Parkinson disease but is also altered during aging and in other neurodegenerative disorders. However, the molecular mechanisms of these mitophagy changes remain uncertain. In healthy mitochondria, PINK1 protein is continuously imported, cleaved, and degraded, but swiftly accumulates on damaged mitochondria, where it triggers the activation of the mitophagy pathway by phosphorylating its substrates ubiquitin and PRKN. Levels of PINK1 protein can therefore be used as a proxy for mitochondrial damage and mitophagy initiation. However, validated methodologies to sensitively detect and quantify PINK1 protein are currently not available. Here, we describe the development and thorough validation of a novel immunoassay to measure human PINK1 on the Meso Scale Discovery platform. The final assay showed excellent linearity, parallelism, and sensitivity. Even in the absence of mitochondrial stress (i.e. at basal conditions), when PINK1 protein is usually not detectable by immunoblotting, significant differences were obtained when comparing samples from patient fibroblasts or differentiated neurons with and without PINK1 expression. Of note, PINK1 protein levels were found increased in human postmortem brain with normal aging, but not in brains with Alzheimer disease, suggesting that indeed different molecular mechanisms are at play. In summary, we have developed a novel sensitive PINK1 immunoassay that will complement other efforts to decipher the roles and biomarker potential of the PINK1-PRKN mitophagy pathway in the physiological and pathological context. Abbreviations : AD: Alzheimer disease; CCCP: carbonyl cyanide 3-chlorophenylhydrazone; ECL: electrochemiluminescence; ELISA: enzyme-linked immunosorbent assay; iPSC: induced pluripotent stem cell; KO: knockout; LLOQ: lower limit of quantification; MSD: Meso Scale Discovery; PD: Parkinson disease; p-S65-Ub: serine-65 phosphorylated ubiquitin; Ub: ubiquitin; ULOQ: upper limit of quantification; WT: wild-type.

The ubiquitin kinase-ligase pair PINK1-PRKN recognizes and transiently labels damaged mitochondria with ubiquitin phosphorylated at Ser65 (p-S65-Ub) to mediate their selective degradation (mitophagy). Complete loss of PINK1 or PRKN function unequivocally leads to early-onset Parkinson disease, but it is debated whether impairments in mitophagy contribute to disease later in life. While the pathway has been extensively studied in cell culture upon acute and massive mitochondrial stress, basal levels of activation under endogenous conditions and especially in vivo in the brain remain undetermined. Using rodent samples, patient-derived cells, and isogenic neurons, we here identified age-dependent, brain region-, and cell type-specific effects and determined expression levels and extent of basal and maximal activation of PINK1 and PRKN. Our work highlights the importance of defining critical risk and therapeutically relevant levels of PINK1-PRKN signaling which will further improve diagnosis and prognosis and will lead to better stratification of patients for future clinical trials.

The ubiquitin kinase-ligase pair PINK1-PRKN identifies and selectively marks damaged mitochondria for elimination via the autophagy-lysosome system (mitophagy). While this cytoprotective pathway has been extensively studied in vitro upon acute and complete depolarization of mitochondria, the significance of PINK1-PRKN mitophagy in vivo is less well established. Here we used a novel approach to study PINK1-PRKN signaling in different energetically demanding tissues of mice during normal aging. We demonstrate a generally increased expression of both genes and enhanced enzymatic activity with aging across tissue types. Collectively our data suggest a distinct regulation of PINK1-PRKN signaling under basal conditions with the most pronounced activation and flux of the pathway in mouse heart compared to brain or skeletal muscle. Our biochemical analyses complement existing mitophagy reporter readouts and provide an important baseline assessment in vivo, setting the stage for further investigations of the PINK1-PRKN pathway during stress and in relevant disease conditions.

The current study was performed to review the conducted studies regarding the concentration of radon 222 in the tap drinking water; furthermore, by estimation of ingestion and inhalation effective dose, the health risk assessment in the adults and children using MCS technique was assessed. All related studies published among January 1990 to October 2016; were screened in the available databases such as Web of Science, PubMed, Science Direct, Scopus, SID, and Irandoc. The total effective dose was estimated by calculating E ing (Effective dose of ingestion) and E inh (Effective dose of inhalation) by Monte Carlo simulation (MCS) method. The range of ND ─ 40.9 Bq/L for radon 222 in water resources was proposed after evaluation of data collected from 13 studies with 1079 samples. The overall concentration of radon 222 in drinking water in Iran was 3.98: 95%CI (3.79 ─ 4.17 Bq/L). Also, the effective ingestion dose of radon 222 in adults age groups was 1.35 times higher than children. The rank order of drinking water resources based on the concentration of radon 222 was Spring > Spring and Well > Well > Spring and Qanat > Tap water. The overall concentration of radon 222 in drinking water in Iran was lower than WHO and EPA standard limits. Also, the rank order regarding area studied based on the concentration of radon 222 was Gillan > Mashhad > Mazandaran > Kerman > Yazd > Tehran > Kermanshah > Golestan > Hormozgan. The effective ingestion dose of radon 222 to consumers in the Gillan, Mashhad, Mazandaran, and Kerman were higher than WHO guidance (0.1 mSv/y). Also except consumers in the Hormozgan, inhalation effective dose radon 222, in the other investigated areas were higher than WHO guidance (0.1 mSv/y). Therefore, it is recommended to conduct the required programs regarding control and elimination of radon 222 concentration in Iranian drinking water supply., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The ingestion of heavy metals through contaminated seafood can causes significant outcomes on human health. In recent years, consume fishes and shrimps has increased in Iran, and several study about heavy metals content in fishes and shrimps from Persian Gulf were carried out to check their food safety. The aims of these systematic reviews and meta-analysis was to summarize the evidence on the relation of the intakes of Arsenic (As) and lead (Pb) levels, based on the origin and sub-groups of shrimp species consumed, Hence that we can estimate the risk of oral cancer induced by Pb and As in these groups of shrimp from the persian gulf. We carried out a search of all suitable studies published between 1995 and 2017 in Scopus, Science Direct, PubMed and Web of Science databases. Since the heterogeneity among studied was significant, we used the random effect model (REM) to perform meta-analysis of data. Data were obtained from 9 articles (14 studies), with 511 samples, and it was reported that pooled levels of As and Pb in the muscle shrimps were 1.37 (95% CI: 0.66-2.08 mg/kg d.w.) and 0.58 (95% CI: 0.33-0.82 mg/kg d.w.), respectively. This pooled levels in muscle shrimps were higher than safe dose reported on Food and Agriculture Organization/World Health Organization guidelines (FAO/WHO). The rank order of shrimps species based on As was Panulirus homarus > Penaeus semisulcatus and for the Pb levels was Litopenaeus vannamei > Panulirus homarus > Fenneropenaeus indicus > Metapenaeus affinis. The lowest and highest risk levels of oral cancer, divided by consumers age groups, were respectively 45-54 (6.94E-04) and 15-24 (8.42E-04) for the Pb, and 45-54 (2.87E-01) and 15-24 (3.51E-01) for arsenic. Incremental Lifetime Cancer Risk (ILCR) of Pb and As was higher than 10-4 and 10-3, respectively. All groups (age) of consumers are subject to the cancer risk of due to the consumption of shrimps contaminated by Pb and As, therefore, should be started a control plan for the reduction of the heavy metal bioaccumulation levels in shrimps of the Persian Gulf coupled to a capillary food safety communication., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Exposure to heavy metals such as arsenic (As), lead (Pb), and cadmium (Cd) in either the short or the long term can cause cancers in humans. Dietary intake and consumption of rice (Oryza sativa L.) is increasing in Iran, and several studies on the concentration of heavy metals in rice have been carried out in this country in recent years. In this perspective, the main objective of the present study was to investigate, even via a meta-analysis of the existing literature, the presence of As and Pb in rice from many geographical areas in Iran, as well as to estimate the carcinogenic risk of these heavy metals in rice consumers. The results of the present ten years-spanning systematic review indicate that 21 reports, collecting a total of 2088 samples, were performed between 2008 and October 2017. The minimum and maximum concentration of As was observed in the Golestan area (0.01 ± 0.01 mg/kg d.w) and the Gillan region (3 mg/kg d.w); and Pb in the Shahrekord (0.07 ± 0.02 mg/kg d.w) and Mazandaran (35 mg/kg d.w). The meta-analysis of data showed that pooled concentration of As in the rice was 0.04 (95%CI: 0.02-0.06 mg/kg d.w), which resulted lower than the National Standard (NS) limits. However, the pooled concentration of Pb in the rice was 0.38 (95%CI: 0.25-0.5 mg/kg d.w), i.e., higher than NS limits. The heterogeneity was significant between As (I 2  = 63%, P value = .003) and Pb (I 2  = 96%, P value < .001) studies. The carcinogenic risk assessment showed that minimum and maximum incremental lifetime cancer risk (ILCR) of As was in the 45-54 (4.53 × 10 -2 ) and 15-24 (5.50 × 10 -2 ) year age groups consumers; and Pb, 45-54 (2.442 × 10 -3 ) and 15-24 (2.96 × 10 -3 ), respectively. The overall carcinogenesis risk of As (4.864 × 10 -2 ) was 18.5 times higher than Pb (2.623 × 10 -3 ). All age groups consumers of rice content of As and Pb are at considerable carcinogenesis risk (ILCR > 10 -3 ). Therefore a decreased level of heavy metals in rice cultivation should be encouraged and performed in next planning., (Copyright © 2018 Elsevier Ltd. All rights reserved.)