Mass spectrometry-based identification and characterization of protein and peptide adducts of lipoxidation-derived aldehydes

Oxidative stress is recognized as an important underlying factor in the pathogenesis of many degenerative diseases as well as normal senescence. The free radicals, reactive oxygen species (ROS) and electrophiles produced during oxidative stress are capable of modifying nucleic acids, lipids and proteins. There are a variety of oxidative modifications that occur to proteins including: cleavage of the protein backbone, direct oxidation of amino acid side chains by ROS, and adduction by electrophilic species such as lipid peroxidation products. Many of these oxidative modifications result in the introduction of carbonyl groups into the proteins. Protein carbonylation levels are commonly used as a biomarker to assess the degree of oxidative damage to a system. However the most commonly employed methods for measuring oxidative modifications to proteins, typically fail to provide any information about the identity of the modified protein, site of modification, or the chemical nature of the modification. In the present study we develop an analytical technique based on affinity labeling with N'-aminooxymethylcarbonylhydrazino-D-biotin (aldehyde reactive probe, ARP), along with mass spectrometric analysis which allows for the full characterization of protein carbonylation modifications. The ability of the ARP method was first demonstrated for the case of oxylipid peptide and protein conjugates formed by Michael addition-type conjugation reactions with α,β- unsaturated aldehydic lipid peroxidation products with nucleophilic amino acid residue side chains. ARP was used to label a 4-hydroxy-2-nonenal (HNE) modified cysteine containing model peptide, and HNE modified E. coli thioredoxin, which were characterized using ESI-MS/MS and MALDI-MS/MS. ARP was also used to label the oxidative modifications alpha-aminoadipic semialdehyde (AAS) and gamma-glutamic semialdehyde (GGS), formed during the metal catalyzed oxidation of GAPDH. After demonstrating the utility of the technique on model systems, it was then applied to complex biological systems. In one case, subsarcolemmal mitochondria (SSM) isolated from rat cardiac tissue. Mitochondria are well known to be a major source of ROS within the cell. They are therefore important mediators of oxidative stress, as well as regulators of cell death. We were able to identify 39 unique sites on 27 mitochondrial proteins which were modified by six different α,β-unsaturated aldehydes, including acrolein, β-hydroxyacrolein, crotonaldehyde, 4-hydroxy-2-hexenal, 4-hydroxy-2-nonenal and 4-oxo-2- nonenal. Additionally we identified nine Lys residues on four mitochondrial proteins that were oxidized to AAS and subsequently labeled with ARP. The proteins identified with oxidative modifications include members of the mitochondrial electron transport chain, TCA cycle, membrane transport, lipid metabolism, and other important mitochondrial enzymes. The ARP technique was also applied to identify protein targets of 4-hyroxy-2- nonenal in human monocytic THP-1 cells that were exogenously exposed to HNE. It was shown previously that exposure of THP-1 cells to HNE resulted in apoptosis, necrosis and protein carbonylation. We applied a multi-pronged proteomic approach involving electrophoretic, immunoblotting and mass spectrometric analysis to unequivocally identify eighteen sites of HNE modification on sixteen proteins. It was also demonstrated in this study that pretreatment of THP-1 cells with ascorbic acid resulted in decreased levels of HNE-protein conjugate formation

Visualization of positive and negative sense viral RNA for probing the mechanism of direct-acting antivirals against hepatitis C virus

RNA viruses are highly successful pathogens and are the causative agents for many important diseases. To fully understand the replication of these viruses it is necessary to address the roles of both positive-strand RNA ((+)RNA) and negative-strand RNA ((-)RNA), and their interplay with viral and host proteins. Here we used branched DNA (bDNA) fluorescence in situ hybridization (FISH) to stain both the abundant (+)RNA and the far less abundant (-)RNA in both hepatitis C virus (HCV)- and Zika virus-infected cells, and combined these analyses with visualization of viral proteins through confocal imaging. We were able to phenotypically examine HCV-infected cells in the presence of uninfected cells and revealed the effect of direct-acting antivirals on HCV (+)RNA, (-)RNA, and protein, within hours of commencing treatment. Herein, we demonstrate that bDNA FISH is a powerful tool for the study of RNA viruses that can provide insights into drug efficacy and mechanism of action

SATCON2: Community Engagement Working Group Report

The SATCON2 Community Engagement Working Group aimed to engage a broad and diverse swath of stakeholders in dark skies and near-Earth space who are impacted by large mega-constellations of tens of thousands of low-Earth orbit (LEO) satellites, beyond professional astronomy alone. The working group consisted of 22 members across 23 time zones including professional and amateur astronomers, members of sovereign Indigenous/First Nations communities, dark-sky advocates, planetarium professionals, and environmental/ecological non-governmental organizations. We set out to work together towards a new and effective conceptual, ethical, legal, and regulatory framework for the protection and sustainability of space and the night sky as a global cultural, natural and scientific commons. Community Engagement Working Group members invested thousands of volunteer hours in working group meetings, listening sessions with impacted constituencies, numerous conversations, developing, conducting and analyzing surveys, and finalizing our results and recommendations

Glaucomatous Optic Neuropathy Associated with Nocturnal Dip in Blood Pressure: Findings from the Maracaibo Aging Study

Purpose—To determine which nocturnal blood pressure (BP) parameters (low levels or extreme dipper status) are associated with an increased risk of glaucomatous damage in Hispanics. Design—Observational cross-sectional study. Participants—A subset (n=93) of the participants from the Maracaibo Aging Study (MAS) who met the study eligibility criteria were included. These participants — who were at least 40 years of age — had measurements for optical tomography coherence, visual field tests, 24-hour BP, office BP, and intraocular pressureHg. Methods—Univariate and multivariate logistic regression analyses under the generalized estimating equations (GEE) framework were used to examine the relationships between glaucomatous damage and BP parameters, with particular attention to drops in nocturnal BP. Main Outcome Measures—Glaucomatous optic neuropathy (GON) based on the presence of optic nerve damage and visual field defects. Results—The mean age was 61.9 years, and 87.1% were women. Of 185 eyes evaluated, 50 (27.0%) had signs of GON. Individuals with GON had significantly lower 24-hour and nighttime diastolic BP levels than those without. However, results of the multivariate GEE models indicated that the glaucomatous damage was not related to the average systolic or diastolic BP levels measured over 24 hours, daytime, or nighttime. In contrast, extreme drops in nighttime systolic and diastolic BP (\u3e20% compared with daytime BP) were significant risk factors for glaucomatous damage (odds ratio=19.78 and 5.55, respectively). Conclusions—In this population, the link between nocturnal BP and GON is determined by extreme dipping effects rather than low nocturnal BP levels alone. Further studies considering extreme drops in nocturnal BP in individuals at high risk of glaucoma are warranted

Preventive Conservation and Restoration Monitoring of Heritage Buildings Based on Fuzzy Logic

This article discusses the usability of the Art-Risk 3.0 software for research on the conservation of heritage buildings. It is a new and free software based on fuzzy logic, which enables the assessment of preventive conservation and surveillance of the restoration of heritage buildings over a period of time. This artificial intelligence-based tool considers the vulnerability of buildings, their environ ments, and their management to evaluate the necessity of their restoration or preventive con servation. To validate the Art-Risk 3.0, 500 theoretical case studies were analyzed, and a 14th century Mudejar-Gothic-style Church in Seville, Spain was studied both before and after its restora tion to identify post-restoration changes. This proof of concept demonstrates the capability of the Art-Risk 3.0 software to analyze environmental impacts on the vulnerability, risk, and functional service life of buildings, and assess the effectiveness of restoration activities. Additionally, this software identifies the most problematic factors and the necessity of restoration.Ministerio de Economía y Competitividad BIA2015- 64878-R (RETOS)Ministerio de Ciencia e Innovación PID2019-107257RB-I00 (FENIX)Ministerio de Ciencia, Innovación y Universidades EQC2019-005780-P (Ambulab-LAB)Junta de Andalucía PYC20 RE 034 UPO RESILIENT-TOURISMMinisterio de Ciencia e Innovación PTA2019-01688

Neuropsychiatric Symptoms Among Hispanics: Results of the Maracaibo Aging Study

Background: Neuropsychiatric symptoms play an important role in diagnosing and clinical follow-up of cognitive impairment and dementia. Objective: We investigated the relationship between neuropsychiatric symptoms, cognitive impairment, and dementia in Hispanics. Methods: We included 529 participants (age ≥40 years) from the Maracaibo Aging Study with standardized neuropsychiatric assessments, including the Neuropsychiatric Inventory (NPI). Based on the Clinical Dementia Rating and the Mini-Mental State Examination scores, participants’ cognitive status was categorized into normal cognition, mild/moderate, and severe cognitive impairment. Diagnosis of dementia was established in a consensus conference. Statistical analyses included multivariable logistic regression models and area under the curve (AUC). Results: The mean age of participants was 59.3 years, and 71.8% were women. The proportion of dementia was 6.8%. Disturbed sleep, anxiety, and depression were the most common neuropsychiatric symptoms in the study sample. In crude analyses, the proportions of hallucinations, aberrant motor behavior, agitation/aggression, apathy, delusions, irritability, eating disturbance, depression, and euphoria were differently distributed among cognitive status groups (p \u3c 0.05). After accounting for confounders, aberrant motor behavior and agitation/aggression remained significantly associated with cognitive impairment and dementia (p \u3c 0.05). The inclusion of the NPI domains significantly improved the AUC to discriminate severe cognitive impairment and dementia above of a basic model that included sex, age, education, alcohol, obesity, serum glucose, total cholesterol, hypertension, and stroke. Conclusion: Neuropsychiatric symptoms are associated with severe cognitive impairment and dementia. The addition of NPI items to the global cognitive assessment might help early detection of dementia in primary care settings

Protocolo para la Incorporación de Cartas Temáticas de la Dirección de Geología Ambiental y Aplicada de la República de Argentina

Fil: Marquínez García, Jorge. SADIM: Sociedad Asturiana de Diversificación Minera; España.Fil: García Manteca, Pilar. SADIM: Sociedad Asturiana de Diversificación Minera; España.Fil: Sánchez, D. SADIM: Sociedad Asturiana de Diversificación Minera; España.Fil: Colina, A. SADIM: Sociedad Asturiana de Diversificación Minera; España.Fil: Fernández Iglesias, Juan Carlos. SADIM: Sociedad Asturiana de Diversificación Minera; España.Fil: Tobio, María Inés. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Chavez, Roxana. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Candaosa, Norberto Gabriel. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Ferpozzi, Federico Javier. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Chavez, Silvia Beatríz. SEGEMAR: Servicio Geológico Minero Argentino; Argentina

Producción de biodiesel por esterificación y transesterificación simultánea de aceite de pulpa de coco (Acrocomia aculeata) utilizando metanol y etanol en condiciones de fluido supercrítico.

El objetivo general del proyecto es contribuir al conocimiento del sistema de colecta y almacenamiento de los frutos de Acrocomia aculeata y aprovechamiento del aceite de la pulpa con miras a su utilización como materia prima para la obtención de biodiesel.CONACYT - Consejo Nacional de Ciencias y TecnologíaPROCIENCI

Manual de Usuario de las Herramientas del Módulo de Salidas Gráficas del SIGAM

Fil: Marquínez García, Jorge. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: García Manteca, Pilar. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Sánchez, D. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Fernández Iglesias, Juan Carlos. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Candaosa, Norberto Gabriel. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Chavez, Silvia Beatríz. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Ferpozzi, Federico Javier. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Colina, A. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España