The xenobiotic-metabolizing enzymes arylamine N-acetyltransferases in human lens epithelial cells: inactivation by cellular oxidants and UVB-induced oxidative stress

ABSTRACT The human arylamine N-acetyltransferases NAT1 and NAT2 are important xenobiotic-metabolizing enzymes involved in the detoxification and metabolic activation of numerous drugs and chemicals. NAT activity depends on genetic polymorphisms and on environmental factors. It has been shown that low NAT-acetylation activity could increase the risk of age-dependent cataract, suggesting that NAT detoxification function may be important for lens cells homeostasis. We report here that the NAT acetylation pathway may occur in human lens epithelial (HLE) cells. Functional NAT1 enzyme was readily detected in HLE cells by reverse transcription-polymerase chain reaction, Western blotting, and enzyme activity assays. NAT2 mRNA and enzymic activity were also detected. We investigated whether oxidants, known to be produced in HLE cells during oxidative stresses and involved in age-dependent cataract formation, decreased endogenous NAT1 and NAT2 activity. The exposure of HLE cells to peroxynitrite led to the dose-dependent irreversible inactivation of both NAT isoforms. Exposing HLE cells to continuously generated H 2 O 2 gave a dose-dependent inactivation of NAT1 and NAT2, reversible on addition of high concentrations of reducing agents. UVB irradiation also induced the reversible dose-dependent inactivation of endogenous NAT1 and NAT2, reversible on addition of reducing agents. Thus, our data suggest that functional NAT1 and NAT2 are present in HLE cells and may be impaired by oxidants produced during oxidative and photooxidative stresses. Oxidativedependent inhibition of NATs in these cells may increase exposure of lens to the harmful effects of toxic chemicals that could contribute to cataractogenesis over time

Molecular phenotyping of multiple mouse strains under metabolic challenge uncovers a role for <i>Elovl2</i> in glucose-induced insulin secretion.

In type 2 diabetes (T2D), pancreatic β cells become progressively dysfunctional, leading to a decline in insulin secretion over time. In this study, we aimed to identify key genes involved in pancreatic beta cell dysfunction by analyzing multiple mouse strains in parallel under metabolic stress. Male mice from six commonly used non-diabetic mouse strains were fed a high fat or regular chow diet for three months. Pancreatic islets were extracted and phenotypic measurements were recorded at 2 days, 10 days, 30 days, and 90 days to assess diabetes progression. RNA-Seq was performed on islet tissue at each time-point and integrated with the phenotypic data in a network-based analysis. A module of co-expressed genes was selected for further investigation as it showed the strongest correlation to insulin secretion and oral glucose tolerance phenotypes. One of the predicted network hub genes was &lt;i&gt;Elovl2&lt;/i&gt; , encoding Elongase of very long chain fatty acids 2. &lt;i&gt;Elovl2&lt;/i&gt; silencing decreased glucose-stimulated insulin secretion in mouse and human β cell lines. Our results suggest a role for &lt;i&gt;Elovl2&lt;/i&gt; in ensuring normal insulin secretory responses to glucose. Moreover, the large comprehensive dataset and integrative network-based approach provides a new resource to dissect the molecular etiology of β cell failure under metabolic stress

Advantages of sous-vide cooked red cabbage: structural, nutritional and sensory aspects

The comparison between equivalent cooking treatments should be applied in a systematic way. This study proposes a methodical way to provide cooked samples with similar firmness using two cooking treatments. In addition, the structural, nutritional and sensory properties of red cabbage cooked with sous-vide treatment in comparison with traditional cooking (boiling water) was evaluated. Changes in texture, color and anthocyanin content were measured in samples cooked with traditional cooking (for different times) and sous-vide (modifying time and temperature according to a Response Surface Methodology). Consumers described sensory properties and preferences between samples. Cryoscanning electron microscopy was used to study the samples microstructure. The firmness of samples, traditionally cooked for 11 min and preferred by consumers, was achieved in samples cooked with sous-vide treatment by optimizing of the cooking conditions (87 C/50 min or 91 C/30 min). Sous-vide treatment was preferred to traditional cooking by consumers. Sous-vide samples were more purple, more aromatic and tastier than traditionally cooked ones. The loss of anthocyanins in traditional cooking was twice that in sous-vide samples. Micrographs from different treatments showed different degrees of cell wall damage. Sous-vide treatment could be recommended as a treatment for the catering industry providing better quality products.Author Iborra-Bernad was supported by the Generalitat Valenciana under FPI (Researcher Formation Program) grant. Author Tarrega was financially supported by the Juan de la Cierva programme.Iborra Bernad, MDC.; Tárrega, A.; García Segovia, P.; Martínez Monzó, J. (2014). Advantages of sous-vide cooked red cabbage: structural, nutritional and sensory aspects. Food Science and Technology. 56(2):451-460. doi:10.1016/j.lwt.2013.12.027S45146056

Influence of genetic factors on toluene diisocyanate-related symptoms: evidence from a cross-sectional study

Background: Toluene diisocyanate (TDI) is a highly reactive compound used in the production of, e. g., polyurethane foams and paints. TDI is known to cause respiratory symptoms and diseases. Because TDI causes symptoms in only a fraction of exposed workers, genetic factors may play a key role in disease susceptibility. Methods: Workers (N = 132) exposed to TDI and a non-exposed group ( N = 114) were analyzed for genotype (metabolising genes: CYP1A1*2A, CYP1A1*2B, GSTM1*O, GSTM3*B, GSTP1 1105V, GSTP1 A114V, GSTT1*O, MPO -463, NAT1*3, *4, *10, *11, *14, *15, NAT2*5, *6, *7, SULT1A1 R213H; immune-related genes: CCL5 -403, HLA-DQB1* 05, TNF-308, TNF-863) and symptoms of the eyes, upper and lower airways ( based on structured interviews). Results: For three polymorphisms: CYP1A1*2A, CYP1A1*2B, and TNF -308 there was a pattern consistent with interaction between genotype and TDI exposure status for the majority of symptoms investigated, although it did reach statistical significance only for some symptoms: among TDI-exposed workers, the CYP1A1 variant carriers had increased risk (CYP1A1*2A and eye symptoms: variant carriers OR 2.0 95% CI 0.68-6.1, p-value for interaction 0.048; CYP1A1*2B and wheeze: IV carriers OR = 12, 1.4-110, p-value for interaction 0.057). TDI-exposed individuals with TNF-308 A were protected against the majority of symptoms, but it did not reach statistical significance. In the non-exposed group, however, TNF -308 A carriers showed higher risk of the majority of symptoms ( eye symptoms: variant carriers OR = 2.8, 1.1-7.1, p-value for interaction 0.12; dry cough OR = 2.2, 0.69-7.2, p-value for interaction 0.036). Individuals with SULT1A1 213H had reduced risk both in the exposed and non-exposed groups. Other polymorphisms, showed associations to certain symptoms: among TDI-exposed, NAT1*10 carriers had a higher risk of eye symptoms and CCL5 -403 AG+AA as well as HLA-DQB1 *05 carriers displayed increased risk of symptoms of the lower airways. GSTM1, GSTM3 and GSTP1 only displayed effects on symptoms of the lower airways in the non-exposed group. Conclusion: Specific gene-TDI interactions for symptoms of the eyes and lower airways appear to exist. The results suggest different mechanisms for TDI- and non- TDI-related symptoms of the eyes and lower airways

From arylamine N-acetyltransferase to folate-dependent acetyl CoA hydrolase : impact of folic acid on the activity of (HUMAN)NAT1 and its homologue (MOUSE)NAT2

Acetyl Coenzyme A-dependent N-, O- and N,O-acetylation of aromatic amines and hydrazines by arylamine N-acetyltransferases is well characterised. Here, we describe experiments demonstrating that human arylamine N-acetyltransferase Type 1 and its murine homologue (Type 2) can also catalyse the direct hydrolysis of acetyl Coenzyme A in the presence of folate. This folate-dependent activity is exclusive to these two isoforms; no acetyl Coenzyme A hydrolysis was found when murine arylamine N-acetyltransferase Type 1 or recombinant bacterial arylamine N-acetyltransferases were incubated with folate. Proton nuclear magnetic resonance spectroscopy allowed chemical modifications occurring during the catalytic reaction to be analysed in real time, revealing that the disappearance of acetyl CH3 from acetyl Coenzyme A occurred concomitantly with the appearance of a CH3 peak corresponding to that of free acetate and suggesting that folate is not acetylated during the reaction. We propose that folate is a cofactor for this reaction and suggest it as an endogenous function of this widespread enzyme. Furthermore, in silico docking of folate within the active site of human arylamine N-acetyltransferase Type 1 suggests that folate may bind at the enzyme's active site, and facilitate acetyl Coenzyme A hydrolysis. The evidence presented in this paper adds to our growing understanding of the endogenous roles of human arylamine N-acetyltransferase Type 1 and its mouse homologue and expands the catalytic repertoire of these enzymes, demonstrating that they are by no means just xenobiotic metabolising enzymes but probably also play an important role in cellular metabolism. These data, together with the characterisation of a naphthoquinone inhibitor of folate-dependent acetyl Coenzyme A hydrolysis by human arylamine N-acetyltransferase Type 1/murine arylamine N-acetyltransferase Type 2, open up a range of future avenues of exploration, both for elucidating the developmental role of these enzymes and for improving chemotherapeutic approaches to pathological conditions including estrogen receptor-positive breast cancer

Increased intestinal permeation and modulation of presystemic metabolism of resveratrol formulated into self-emulsifying drug delivery systems

International audienceDespite various beneficial biological properties, resveratrol lacks therapeutic applications because of poor bioavailability due to variable absorption and extensive metabolism. The present study aims at evaluating the capability of self-emulsifying drug delivery systems (SEDDS) to enhance resveratrol permeation across rat intestine and to modulate its presystemic metabolism. For that purpose, semi-solid (SS) and liquid (L) SEDDS were prepared and dispersed in an aqueous buffer to produce nanoemulsions (NE). The jejunal absorptive transepithelial fluxes (J(ms)) of resveratrol elicited by these formulations (SSNE and L-NE) and presystemic metabolization were determined on Ussing chambers. The absorptive fluxes through the intestinal epithelium from the nanoemulsions (J(ms) =20.5 +/- 3.1 mu gh (1) cm (2) SS-NE; 28.9 +/- 2.9 mu gh (1) cm (2) L-NE) were significantly increased compared to an ethanolic control solution (J(ms) = 3.4 +/- 0.3 mu gh (1) cm (2), p < 0.05). No significant variations of conductance were observed after two hours of contact between the formulations and the mucosa. Simultaneously, the presystemic metabolization pattern was modified in the case of the nanoemulsions compared to the control solution. In conclusion, our data suggests that oil-in-water nanoemulsions prepared from SEDDS dispersions of medium-chain lipids could be promising formulations for enhancing oral delivery of resveratrol. (C) 2017 Elsevier B.V. All rights reserved

Structure of Mesorhizobium loti arylamine N-acetyltransferase 1

The crystal structure of a M. loti arylamine N-acetyltransferase 1 has been determined at 2.0 Å resolution

Alterations in the Serotonin and Dopamine Pathways by Cystathionine Beta Synthase Overexpression in Murine Brain

International audienc