Identification and functional analysis of missense mutations in the lecithin cholesterol acyltransferase gene in a Chilean patient with hypoalphalipoproteinemia.

BACKGROUND: Lecithin-cholesterol acyltransferase (LCAT) is a plasma enzyme that esterifies cholesterol in high- and low-density lipoproteins (HDL and LDL). Mutations in LCAT gene causes familial LCAT deficiency, which is characterized by very low plasma HDL-cholesterol levels (Hypoalphalipoproteinemia), corneal opacity and anemia, among other lipid-related traits. Our aim is to evaluate clinical/biochemical features of a Chilean family with a proband showing clinical signs of familial LCAT deficiency, as well as to identify and assess the functional effects of LCAT mutations. METHODS: An adult female proband with hypoalphalipoproteinemia, corneal opacity and mild anemia, as well as her first-degree relatives, were recruited for clinical, biochemical, genetic, in-silico and in-vitro LCAT analysis. Sequencing of exons and intron-exon boundaries was performed to identify mutations. Site-directed mutagenesis was carried out to generate plasmids containing cDNA with wild type or mutant sequences. Such expression vectors were transfected to HEK-239 T cells to asses the effect of LCAT variants in expression, synthesis, secretion and enzyme activity. In-silico prediction analysis and molecular modeling was also used to evaluate the effect of LCAT variants. RESULTS: LCAT sequencing identified rare p.V333 M and p.M404 V missense mutations in compound heterozygous state in the proband, as well the common synonymous p.L363 L variant. LCAT protein was detected in proband's plasma, but with undetectable enzyme activity compared to control relatives. HEK-293 T transfected cells with vector expression plasmids containing either p.M404 V or p.V333 M cDNA showed detectable LCAT protein expression both in supernatants and lysates from cultured cells, but with much lower enzyme activity compared to cells transfected with the wild-type sequence. Bioinformatic analyses also supported a causal role of such rare variations in LCAT lack of function. Additionally, the proband carried the minor allele of the synonymous p.L363 L variant. However, this variant is unlikely to affect the clinical phenotype of the proband given its relatively high frequency in the Chilean population (4%) and its small putative effect on plasma HDL-cholesterol levels. CONCLUSION: Genetic, biochemical, in vitro and in silico analyses indicate that the rare mutations p.M404 V and p.V333 M in LCAT gene lead to suppression of LCAT enzyme activity and cause clinical features of familial LCAT deficiency.

Removal of arsenic from semiarid area groundwater using a biosorbent from watermelon peel waste.

Groundwater is one of the most important reservoirs in semi-arid and arid zones of the world, particularly in Mexico. The aims of this work were to produce a biosorbent from watermelon peel waste and a biosorbent with citric acid treatment and to evaluate both biosorbents with different concentrations of arsenic in groundwater. The biosorbents were produced with watermelon peel residues, which were observed by SEM microscopy to evaluate their physical morphology. Its removal potential was tested at concentrations of 0, 1, 13, 22, and 65 µg/L of arsenic, and both adsorption capacity and removal percentage were analyzed by final measurement obtained by atomic absorption spectrometry. The pH was measured throughout the experimentation maintaining ranges between 5.5 and 7.5. The biosorbent without treatment presented clearer and more compact flakes. At the microscopic level, the biosorbent without treatment presented pores with a more circular shape, and the biosorbent with treatment was more polygonal, similar to a honeycomb. The highest removal percentage was 99.99%, for both treatments at 4 h. The biosorbent without treatment at 4 h with arsenic concentrations of 65 µg/L presented the highest adsorption capacity (2.42 µg/g). It is concluded that watermelon peel biosorbent is a material that has the potential to remove arsenic from groundwater. This type of biosorbent is effective to remove arsenic and could be used in the field, however, it still needs to be optimized to convert it into a material completely suitable for large-scale use.

Characterization of Scale Deposits in a Drinking Water Network in a Semi-Arid Region.

The quantity and quality of the supply of fresh water to households, commercial areas, small industries, green spaces irrigation and public and private institutions in large cities face challenges from the supply sources availability and suitable distribution network performance to the full satisfaction of the established drinking water guidelines. In Mexico, the main source of water comes from groundwater. Most of the Mexican aquifers are located in arid and semi-arid weather conditions. The groundwater's physical-chemical properties are closely related to geology. This study was carried out at the north-central part of the country in which igneous and sedimentary rocks predominate, with high calcium carbonate (CaCO3) concentrations. The accumulation of CaCO3 in the pipelines is also known as scale deposit that decreases the fluid flow, causing a deficiency in the water supply. The main objectives of this study were determining the physical-chemical groundwater parameters and saturation indexes injected into the drinking water networks and characterizing the scale deposits by scanning electron microscopy (SEM) and X-ray diffraction (XRD). The results indicate that the scale deposits are mainly calcium carbonate and silica oxide crystals, caused by the water aggressiveness according to the saturation indexes and the lack of control over the saturation pH.

Evaluation of Groundwater Quality for Human Consumption and Irrigation in Relation to Arsenic Concentration in Flow Systems in a Semi-Arid Mexican Region.

The supply of drinking water to the population is an important challenge facing humanity, since both surface and underground sources present a great variability of water storage with respect to space and time. This problem is further aggravated in arid and semi-arid areas where rainfall is low and torrential, which makes groundwater the main source of supply; therefore, it is necessary to carry out studies that allow evaluating the evolution of the quantity and quality of water. This study addresses the behavior of groundwater in a semi-arid region, considering the theory of flow systems to identify movement as well as water quality, es determined by a water quality index (WQI), calculated considering arsenic and fluorine. In addition, a quality irrigation classification is used, employing the norms in accordance with international standards and the Mexican Norm, which allows for a comparison. Local, regional, intermediate and mixed flow systems are identified, and the evolution of cations and anions in addition to temperature is examined. It is observed that the drinking water quality index classifies them as excellent in most of the monitored wells (<50), but with a negative evolution. Regarding irrigation, most of the water samples are classified without restriction for the establishment of any type II crop (C2S1) and with restrictions for horticultural crops. It is observed that arsenic had values between 0.49 and 61.40 (µg/L) in 2005, while in 2015 they were between 0.10 and 241.30 (µg/L). In addition, fluoride presented values between 0.00 and 2.6 (mg/L) in 2005, while in 2015 they were between 0.28 and 5.40 (mg/L). The correlations between arsenic and fluorine are noted as well as WQI and SAR. A finding in this research was to include arsenic and fluorine in the calculation of the WQI allowing a better interpretation of the quality of water for both human consumption and for agricultural use to based on this make the best decision to control any harmful effects for the population, in addition to identifying the appropriate purification treatment required to control pollutants. It is concluded that arsenic is an element of utmost importance when considering water quality, so it is necessary to examine its evolution and continue to monitor its levels constantly.

Inositol Monophosphatase: A Bifunctional Enzyme in Mycobacterium smegmatis.

Inositol monophosphatase (IMPase) is a crucial enzyme for the biosynthesis of phosphatidylinositol, an essential component in mycobacterial cell walls. IMPase A (ImpA) from Mycobacterium smegmatis is a bifunctional enzyme that also functions as a fructose-1,6-bisphosphatase (FBPase). To better understand the bifunctional nature of this enzyme, point mutagenesis was conducted on several key residues and their enzyme activity was tested. Our results along with active site models support the fact that ImpA is a bifunctional enzyme with residues Gly94, Thr95 hypothesized to be contributing to the FBPase activity and residues Trp220, Asp221 hypothesized to be contributing to the IMPase activity. Double mutants, W220A + D221A reduced both FBPase and IMPase activity drastically while the double mutant G94A + T95A surprisingly partially restored the IMPase activity compared to the single mutants. This study establishes the foundation toward obtaining a better understanding of the bifunctional nature of this enzyme.

Spatio-Temporal Analysis of Natural and Anthropogenic Arsenic Sources in Groundwater Flow Systems.

The presence of arsenic in groundwater constitutes a hazard for the environment and human health, and the determination of its source has become a global challenge, which can be approached by defining the natural background levels (NBL) in conjunction with the indicator kriging method, with the aim of delineating anthropogenically contaminated areas. However, having a unique value of NBL for large areas can generate interpretation errors. This research integrates the determination of the flow systems present in the Calera Aquifer, and the definition of the natural background levels in each flow system by making estimation maps in ArcGIS using two databases, 10 years apart, to evaluate the spatio-temporal variation of arsenic in groundwater. The results indicate a notable increase in the probability of exceeding the arsenic NBL, mainly in the intermediate flow, which may be due to movement resulting from mining activities as well as a mixture of regional and intermediate flows caused by the extraction of water for agriculture and drinking water supplies. The presented values exceed the maximum limits allowed for human consumption, as stated by the World Health Organization.

Trastornos de la conducta alimentaria en el embarazo / Eating disorders in pregnancy

Los trastornos de la conducta alimentaria afectan a mujeres en edad fértil, sin embargo, la intersección de éstos y el embarazo es aún pobremente comprendida, existiendo aún datos limitados en la literatura. Esta revisión narrativa resume la evidencia actual acerca de los trastornos alimentarios durante el embarazo: su prevalencia, curso, el impacto que éstos tienen en la salud materna, fetal y en su descendencia, así como posibles estrategias de intervención.

Eating disorders affect women of childbearing age, however, their intersection with pregnancy is still poorly understood, while there is still limited data in the literature. This narrative review summarizes the current evidence about eating disorders during pregnancy: their prevalence, course, the impact they have on maternal, fetal and offspring health, as well as possible intervention strategies.

Antioxidative stress effect of epicatechin and catechin induced by Aß25-35 in rats and use of the electrostatic potential and the Fukui function as a tool to elucidate specific sites of interaction.

Alzheimer's disease (AD) is a neurodegenerative disorder caused by the aggregation of the amyloid-beta peptide (Aß) in senile plaques and cerebral vasculature. The Aß25-35 fraction has shown the most toxicity; its neurotoxic mechanisms are associated with the generation of oxidative stress and reactive astrogliosis that induce neuronal death and memory impairment. Studies indicate that pharmacological treatment with flavonoids reduces the rate of AD, in particular, it has been shown that antioxidants are compounds that could interact with this peptide due to their antioxidant proprieties. In this study, experimental and computational tools were used to calculate the molecular electrostatic potential and the Fukui function with the Gaussian 09 computational program, to predict the most reactive parts of these molecules and make the complex between Aß25-35 and two flavonoids (catechin and epicatechin) in the absolute gas-phase, where a possible interaction between them was observed. This is important for understanding the Aß25-35-Flavonoid (A-F) interaction as a therapeutic strategy to inhibit the neurotoxic effects that this peptide causes in AD, which currently is still considered an ambiguous process.

Evaluation of disk diffusion method compared to broth microdilution for antifungal susceptibility testing of 3 echinocandins against Aspergillus spp.

We tested the susceptibility of caspofungin, micafungin, and anidulafungin against Aspergillus spp. isolates by the new Clinical and Laboratory Standards Institute M51-A disk diffusion (DD) and the broth microdilution methods. A total of 65 clinical isolates of Aspergillus spp. were evaluated. The DD assay was performed on nonsupplemented Müeller-Hinton agar using caspofungin 2-µg, micafungin 1-µg, and anidulafungin 2-µg disks. Echinocandin minimal effective concentrations (MECs) and inhibition zones (IZs) were read after 24 to 48 (A. terreus) h at 35 °C. Caspofungin MECs for all Aspergillus spp. strains tested were ≤ 0.25 µg/mL; IZs were ≥ 15 mm for most species except for A. terreus (11-22 mm). Both micafungin and anidulafungin MECs were ≤ 0.015 µg/mL, but micafungin IZs were ≥ 14 mm while anidulafungin IZs were ≥ 22 mm. As for caspofungin, the DD method could be a useful method for susceptibility testing of micafungin and anidulafungin against Aspergillus spp.