Mammalian N1-adenosine PARylation is a reversible DNA modification.

Poly-ADP-ribosylation (PARylation) is regarded as a protein-specific modification. However, some PARPs were recently shown to modify DNA termini in vitro. Here, we use ultrasensitive mass spectrometry (LC-MS/MS), anti-PAR antibodies, and anti-PAR reagents to show that mammalian DNA is physiologically PARylated and to different levels in primary tissues. Inhibition of PAR glycohydrolase (PARG) increases DNA PARylation, supporting that the modification is reversible. DNA PARylation requires PARP1 and in vitro PARP1 PARylates single-stranded DNA, while PARG reverts the modification. DNA PARylation occurs at the N1-position of adenosine residues to form N1-Poly(ADP-ribosyl)-deoxyadenosine. Through partial hydrolysis of mammalian gDNA we identify PAR-DNA via the diagnostic deamination product N1-ribosyl-deoxyinosine to occur in vivo. The discovery of N1-adenosine PARylation as a DNA modification establishes the conceptual and methodological framework to elucidate its biological relevance and extends the role of PARP enzymes.

Early Toxic Effects in a Central American Native Fish (Parachromis dovii) Exposed to Chlorpyrifos and Difenoconazole.

In Costa Rica, agriculture is one of the most important economic activities. Chlorpyrifos and difenoconazole have been identified as agrochemicals widely used in banana and pineapple crops in the Caribbean area of the country and are constantly recorded in aquatic ecosystems. The toxicity of these pesticides in Parachromis dovii was studied. Median lethal concentrations (LC50s) for each substance were obtained from 96-h acute tests. Then, fish were exposed to sublethal concentrations of both substances (10% of LC50), individually and in mixture, to evaluate biomarker responses. Ethoxyresorufin-O-deethylase (EROD), catalase, and glutathione S-transferase activities as well as lipid peroxidation were measured in liver and gill tissues as markers of biotransformation and oxidative stress processes. Cholinesterase activity in brain and muscle tissue was also quantified as a biomarker of toxicity. The LC50s were 55.34 µg/L (95% confidence interval [CI] 51.06-59.98) for chlorpyrifos and 3250 µg/L (95% CI 2770-3810) for difenoconazole. Regarding the biomarkers, a significant inhibition of brain and muscle cholinesterase activity was recorded in fish exposed to 5.50 µg/L of chlorpyrifos. This activity was not affected when fish were exposed to the mixture of chlorpyrifos with difenoconazole. Significant changes in lactate dehydrogenase activity were observed in fish exposed to 325 µg/L of difenoconazole, whereas fish exposed to the mixture showed a significant increase in EROD activity in the liver. These results suggest harmful effects of chlorpyrifos insecticide at environmentally relevant concentrations. There is also evidence for an interaction of the 2 substances that affects the biotransformation metabolism at sublethal levels of exposure. Environ Toxicol Chem 2021;40:1940-1949. © 2021 SETAC.

Biogardens as constructed wetlands in tropical climate: A case study in the Central Pacific Coast of Costa Rica.

In Costa Rica, <10% of wastewater is treated before its discharge. This generates a significant impact on the environment, public health, and tourism industry, which is one of the country's main economic activities. Biogardens, subsurface flow artificial wetlands, are alternative systems for the treatment of wastewater. The present study evaluated the removal of organic matter and nutrients in a biogarden located at a hotel in the Central Pacific Coast of Costa Rica between 2012 and 2017. Pretreatment involved septic tanks and grease traps for sewage and gray water, respectively. The biogarden, which is composed of seven wetlands with an average area of 12 m2 and a depth of 0.7 m, contains river cobble as support material, gravel as bed, and Cyperus papyrus and Heliconia sp. plants. Removal of the biochemical oxygen demand (BOD), the chemical oxygen demand (COD), and the total suspended solids (TSS) on average were 80%, 66%, and 72%, respectively, thus producing an effluent in compliance with current national legislation. Furthermore, the biogarden did not emit noxious odors or display an excessive presence of mosquitoes. The results showed consistent and efficient removal of organic matter and nutrients from the wastewater throughout different seasons and pollutant loads, verifying that such systems can be used in decentralized locations (e.g., tourist areas) in tropical climates.

Impaired DNA demethylation of C/EBP sites causes premature aging.

Changes in DNA methylation are among the best-documented epigenetic alterations accompanying organismal aging. However, whether and how altered DNA methylation is causally involved in aging have remained elusive. GADD45α (growth arrest and DNA damage protein 45A) and ING1 (inhibitor of growth family member 1) are adapter proteins for site-specific demethylation by TET (ten-eleven translocation) methylcytosine dioxygenases. Here we show that Gadd45a/Ing1 double-knockout mice display segmental progeria and phenocopy impaired energy homeostasis and lipodystrophy characteristic of Cebp (CCAAT/enhancer-binding protein) mutants. Correspondingly, GADD45α occupies C/EBPß/δ-dependent superenhancers and, cooperatively with ING1, promotes local DNA demethylation via long-range chromatin loops to permit C/EBPß recruitment. The results indicate that enhancer methylation can affect aging and imply that C/EBP proteins play an unexpected role in this process. Our study suggests a causal nexus between DNA demethylation, metabolism, and organismal aging.

Lipoprotein (a) and other prothrombotic risk factors in Caucasian women with unexplained recurrent miscarriage. Results of a multicentre case-control study.

From 1998 to 2003, 133 Caucasian women aged 17-40 years (median 29 years) suffering from unexplained recurrent miscarriage (uRM) were consecutively enrolled. In patients and 133 age-matched healthy controls prothrombotic risk factors (factor V (FV) G1691A, factor II (FII) G20210A, MTHFR T677T, 4G/5G plasminogen activator inhibitor (PAI)-1, lipoprotein (Lp) (a), protein C (PC), protein S (PS), antithrombin (AT), antiphospholipid/anticardiolipin (APA/ACA) antibodies) as well as associated environmental conditions (smoking and obesity) were investigated. 70 (52.6%) of the patients had at least one prothrombotic risk factor compared with 26 control women (19.5%; p<0.0001). Body mass index (BMI; p=0.78) and smoking habits (p=0.44) did not differ significantly between the groups investigated. Upon univariate analysis the heterozygous FV mutation, Lp(a) > 30 mg/dL, increased APA/ACA and BMI > 25 kg/m(2) in combination with a prothrombotic risk factor were found to be significantly associated with uRM. In multivariate analysis, increased Lp(a) (odds ratio (OR): 4.7/95% confidence interval (CI): 2.0-10.7), the FV mutation (OR:3.8/CI:1.4-10.7), and increased APA/ACA (OR: 4.5/CI: 1.1-17.7) had independent associations with uRM.