Preclinical Characterization of Pharmacologic NAD+ Boosting as a Promising Therapeutic Approach in Rheumatoid Arthritis.

OBJECTIVE: We analyzed NAD+ metabolism in patients with rheumatoid arthritis (RA), its association with disease activity and clinical outcomes of RA, and the therapeutic potential of pharmacologic NAD+ boosting. METHODS: Our study included 253 participants. In the first cohort, comprising 153 RA patients and 56 healthy donors, we assessed NAD+ levels and NAD+ -related gene pathways. We analyzed 92 inflammatory molecules by proximity extension assay. In the second cohort, comprising 44 RA patients starting anti-tumor necrosis factor (anti-TNF) drugs, we evaluated changes in NAD+ levels and their association with clinical response after 3 months. Mechanistic studies were performed ex vivo on peripheral blood mononuclear cells (PBMCs) from patients with RA to test the beneficial effects of NAD+ boosters, such as nicotinamide and nicotinamide riboside. RESULTS: Reduced NAD+ levels were found in RA samples, in line with altered activity and expression of genes involved in NAD+ consumption (sirtuins, poly[ADP-ribose] polymerase, CD38), transport (connexin 43), and biosynthesis (NAMPT, NMNATs). Unsupervised clustering analysis identified a group of RA patients with the highest inflammatory profile, the lowest NAD+ levels, and the highest disease activity (as shown by the Disease Activity Score in 28 joints). NAD+ levels were modulated by anti-TNF therapy in parallel with the clinical response. In vitro studies using PBMCs from RA patients showed that nicotinamide riboside and nicotinamide increased NAD+ levels via NAMPT and NMNAT and reduced their prooxidative, proapoptotic, and proinflammatory status. CONCLUSION: RA patients display altered NAD+ metabolism, directly linked to their inflammatory and disease activity status, which was reverted by anti-TNF therapy. The preclinical beneficial effects of NAD+ boosters, as shown in leukocytes from RA patients, along with their proven clinical safety, might pave the way for the development of clinical trials using these compounds.

Electrically evoked stapedial reflex threshold and behavioral method usefulness in the cochlear implant programming in pediatric population.

OBJETIVE: To determine the utility of the electrically evoked stapedial reflex test (ESRT) and behavioral method in the CIs programming as an objective method to identify MCL levels in the CIs programming in pediatric patients. METHOD: Cross-sectional cohort study that included 20 pediatric patients with postlingual deafness and CI unilateral. They were performed clinical history, tympanometry, ESRT and by free field audiometry, before and after programming modifications according to MCL levels obtained by ESRT were performed. ESRT threshold was assessed with individual 300 ms stimuli on the 12 electrodes and recorded through manual decay. Likewise, the maximum comfort threshold (MCL) of each electrode was obtained through a behavioral analysis. RESULTS: No significant differences were found between the ESRT and behavioral method in MCLs levels in each of the electrodes evaluated. In addition, the correlation coefficients were significant and located in a range of 0.55-0.81, higher in electrodes 7, 8, and 9 (r = 0.77, 0.76, and 0.81, respectively). However, the median hearing threshold established by the ESRT was significantly lower compared to the behavioral threshold (36.0 vs. 47.0 dB, p < 0.0001), regardless of age (p = 0.249) or the etiology of hearing loss (p = 0.292). The difference between the tests was in the number of times to do it, the ESRT was done once and the behavioral on average 4 ± 1 times. CONCLUSION: Similar MCL thresholds were obtained in both ESRT and behavioral test, establishing that both methods are reliable for use in pediatric patients; however, ESRT allows shortening the time to achieve normal hearing and language acquisition thresholds in a more optimal time.

Mitochondrial adaptations to calorie restriction and bariatric surgery in human skeletal muscle: a systematic review with meta-analysis.

OBJECTIVE: We performed a meta-analysis to determine the changes induced by calorie restriction (CR) and bariatric surgery on human skeletal muscle mitochondria. METHODS: A systematic search of Medline and Web of Science was conducted. Controlled trials exploring CR (≥14 days) and mitochondrial function and/or content assessment were included. Moreover, studies analyzing weight loss following gastric surgery were included for comparison purposes. Human muscle data from 28 studies assessing CR (520 muscle samples) and from 10 studies assessing bariatric surgery (155 muscle samples) were analyzed in a random effect meta-analysis with three a priori chosen covariates. MAIN RESULTS: We report a decrease (p < 0.05) (mean (95 % CI)) in maximal mitochondrial state 3 respiration in response to CR (-0.44 (-0.85, -0.03)) but not in response to surgery (-0.33 (-1.18, 0.52)). No changes in mitochondrial content were reported after CR (-0.05 (-0.12, 0.13)) or in response to surgery (0.23 (-0.05, 0.52)). Moreover, data from CR subjects showed a reduction in complex IV (CIV) activity (-0.29 (-0.56, -0.03)) but not in CIV content (-0.21 (-0.63, 0.22)). Similar results were obtained when the length of the protocol, the initial body mass index, and the estimated energy deficit were included in the model as covariates. CONCLUSION: The observation of reduced maximal mitochondrial state 3, uncoupled respiration, and CIV activity without altering mitochondrial content suggests that, in human skeletal muscle, CR mainly modulates intrinsic mitochondrial function.

Metabarcoding data of prokaryotes and eukaryotes inhabiting the phosphogypsum stockpiles on the salt marshes of Huelva (SW Spain).

Around 100 Mt of phosphogypsum (PG) of extreme acidity and with high concentrations of heavy metals and radionuclides have been deposited on the salt marshes of the Tinto River estuary in Huelva (SW Spain) for more than forty years. The microbial community able to thrive in these adverse conditions remains totally unknown, despite the fact that it can highly influence the biogeochemical cycle of the phosphogypsum components and include new species with biotechnological interest. High throughput sequencing of 16S/18S rRNA encoding genes is a potent tool to uncover the microbial diversity of extreme environments. This data article describes for the first time the prokaryotic and eukaryotic diversity of two water samples collected in the Huelva phosphogypsum stacks. The raw amplicons of the 16S/18S rRNA maker genes for the two phosphogypsum samples and two reference samples (seawater and the Tinto River water) obtained after sequencing on MiSeq platform are provided. The operational taxonomic units (OTUs) obtained after the treatment and clustering of the obtained reads with the QIIME2 pipeline and their taxonomic assignation performed by comparison with the SILVA database are also presented to complete the information of the article "Exploring the microbial community inhabiting the phosphogypsum stacks of Huelva (SW, Spain) by a high throughput 16S/18S rDNA Sequencing approach".

Exploring the microbial community inhabiting the phosphogypsum stacks of Huelva (SW SPAIN) by a high throughput 16S/18S rDNA sequencing approach.

Around 100 Mt of phosphogypsum (PG) have been deposited in large stacks on the salt marshes of the Tinto River estuary in Huelva (SW Spain), covering about 1000 ha. These stacks contain extremely acidic water (pH < 2) with high concentrations of pollutants which can cause emissions into their surroundings, generating important environmental concerns. Despite many chemical, geological or hydrological studies have been conducted to characterize the PG stacks of Huelva, the microbial community inhabiting this extreme environment remains unexplored. Using a 16S/18S-rRNA-high throughput sequencing approach, we have uncovered the main taxonomic groups able to live in the acidic metal-contaminated water, which is in direct contact with the PG, demonstrating for the first time the existence of a huge diversity of microbial species in these extreme conditions. In addition, the physicochemical characteristics of the water sampled have been analyzed. These studies have revealed that the most abundant bacteria found in two different leachate samples of the PG stacks belong to the genera Acidiphilium, Pseudomonas, Leptosprillum, Acidithrix, or Acidithiobacillus, typically found in acid mine drainage (AMD) environments, which in total represent around 50% of the total bacterial community. Biodiversity of eukaryotes in PG water is lower than that of prokaryotes, especially in the water collected from the perimeter channel that surrounds the PG stacks, where the pH reaches a value of 1.5 and the activity concentrations exceed 300 Bq L-1 for 238U or 20 Bq L-1 for 210Po, values which are from four to five orders of magnitude higher than those usually found in unperturbed surface waters. Even so, an unexpected diversity of algae, fungi, and ciliates have been found in the PG stacks of Huelva, where chlorophyte microalgae and basidiomycetes fungi are the most abundant eukaryotes. Additional bioinformatics tools have been used to perform a functional analysis and predict the most common metabolic pathways in the PG microbiota. The obtained data indicate that the extreme conditions of these PG stacks hide an unexpected microbial diversity, which can play an important role in the dynamics of the contaminating compounds of the PG and provide new strains with unique biotechnological applications.

Trastornos vestibulares y del equilibrio en niños y adolescentes mexicanos: revisión de registros clínicos de 8 años / Vestibular and balance disorders in Mexican children and adolescents: Review of 8-year clinical records

Introducción: El vértigo es un síntoma poco estudiado en la población pediátrica, donde la prevalencia es inferior al 1%. Los trastornos vestibulares en el niño no son equiparables a los del adulto y a menudo son subdiagnosticados. Identificar las principales causas de estos trastornos proporcionaría datos fidedignos y precisos de las enfermedades en ciertos grupos de edad, mejorando el proceso de diagnóstico.Material y métodosEstudio observacional, transversal y retrospectivo. Se revisaron los expedientes consecutivos de pacientes, de entre 3 y 17años, atendidos por síntomas vestibulares, del equilibrio y asociados (vértigo, mareo, hipoacusia, alteraciones del equilibrio y cefalalgia) en el servicio de Otoneurología de un hospital de tercer nivel de atención entre septiembre de 2010 y septiembre de 2018. Se utilizó estadística descriptiva e inferencial para el análisis. Todos los valores de p informados de estos análisis fueron de dos colas con un nivel de significación <0,05.ResultadosSe registraron 212 casos entre 6.444 consultas, por lo que la frecuencia fue del 3,3%. La proporción entre hombres y mujeres fue similar. La edad media del grupo fue de 14,5 (±3,9) años, la mediana de la edad de inicio de los síntomas fue de 11 (RIC 8-14) años y la de la primera consulta fue de 13 (RIC 10-15) años. Como síntoma cardinal, el 51,9% acudieron por vértigo, el 25,5% por mareo, el 9,9% por inestabilidad de la marcha, el 7,5% por hipoacusia y el 5,2% por cefalalgia. El 61,8% de los episodios de vértigo tuvieron un origen periférico, el 32,5% central y el 5,7% fuera del sistema vestibular. El vértigo paroxístico benigno de la infancia, la disfunción y migraña vestibular representaron más del 65% de los casos. No se encontraron diferencias entre los diagnósticos con respecto al sexo y la edad de los participantes (p>0,05). (AU)

Introduction: Vertigo is a seldom studied symptom in the paediatric population, where the prevalence is less than 1%. Vestibular disorders in children are not comparable to those of adults and are often underdiagnosed. Identifying the main causes of these disorders would provide reliable and accurate data of the diseases in certain age groups, improving the diagnostic process.Material and methodsObservational, cross-sectional, and retrospective study. Consecutive records of patients, aged 3 to 17years, attended due to vestibular, balance and associated symptoms (vertigo, dizziness, hearing loss, balance disorders and headache), attended by the otoneurology service of a tertiary hospital, between September 2010 and September 2018, were included. Descriptive and inferential statistics were used for the analysis. All p-values reported from these analyses were two-tailed with a significance level <.05.Results212 cases were registered from 6,444 consultations (3.3%). The proportion between males and females was similar. The mean age of the group was 14.5 (±3.9) years, the median age of onset of symptoms was 11 (IQR 8-14) years and that of the first consultation was 13 (IQR 10-15) years. As a cardinal symptom, 51.9% attended due to vertigo, 25.5% due to dizziness, 9.9% due to gait instability, 7.5% due to hearing loss and 5.2% due to headache. Of the episodes of vertigo, 61.8% were of peripheral origin, 69 32.5% central and 12 5.7% outside the vestibular system. Benign paroxysmal vertigo of childhood, dysfunction and vestibular migraine accounted for more than 65% of cases. No differences were found between the diagnoses regarding the sex and age of the participants (P>.05).ConclusionVestibular and balance disorders are rare in the study population and most of the causes are benign and potentially self-limited disorders. (AU)

Vestibular and balance disorders in Mexican children and adolescents: Review of 8-year clinical records. / Trastornos vestibulares y del equilibrio en niños y adolescentes mexicanos: revisión de registros clínicos de 8 años.

INTRODUCTION: Vertigo is a seldom studied symptom in the paediatric population, where the prevalence is less than 1%. Vestibular disorders in children are not comparable to those of adults and are often underdiagnosed. Identifying the main causes of these disorders would provide reliable and accurate data of the diseases in certain age groups, improving the diagnostic process. MATERIAL AND METHODS: Observational, cross-sectional, and retrospective study. Consecutive records of patients, aged 3 to 17years, attended due to vestibular, balance and associated symptoms (vertigo, dizziness, hearing loss, balance disorders and headache), attended by the otoneurology service of a tertiary hospital, between September 2010 and September 2018, were included. Descriptive and inferential statistics were used for the analysis. All p-values reported from these analyses were two-tailed with a significance level <.05. RESULTS: 212 cases were registered from 6,444 consultations (3.3%). The proportion between males and females was similar. The mean age of the group was 14.5 (±3.9) years, the median age of onset of symptoms was 11 (IQR 8-14) years and that of the first consultation was 13 (IQR 10-15) years. As a cardinal symptom, 51.9% attended due to vertigo, 25.5% due to dizziness, 9.9% due to gait instability, 7.5% due to hearing loss and 5.2% due to headache. Of the episodes of vertigo, 61.8% were of peripheral origin, 69 32.5% central and 12 5.7% outside the vestibular system. Benign paroxysmal vertigo of childhood, dysfunction and vestibular migraine accounted for more than 65% of cases. No differences were found between the diagnoses regarding the sex and age of the participants (P>.05). CONCLUSION: Vestibular and balance disorders are rare in the study population and most of the causes are benign and potentially self-limited disorders.

In Silico Study of the Resistance to Organophosphorus Pesticides Associated with Point Mutations in Acetylcholinesterase of Lepidoptera: B. mandarina, B. mori, C. auricilius, C. suppressalis, C. pomonella, H. armígera, P. xylostella, S. frugiperda, and S. litura.

An in silico analysis of the interaction between the complex-ligands of nine acetylcholinesterase (AChE) structures of Lepidopteran organisms and 43 organophosphorus (OPs) pesticides with previous resistance reports was carried out. To predict the potential resistance by structural modifications in Lepidoptera insects, due to proposed point mutations in AChE, a broad analysis was performed using computational tools, such as homology modeling and molecular docking. Two relevant findings were revealed: (1) Docking results give a configuration of the most probable spatial orientation of two interacting molecules (AChE enzyme and OP pesticide) and (2) a predicted ΔGb. The mutations evaluated in the form 1 acetylcholinesterase (AChE-1) and form 2 acetylcholinesterase (AChE-2) structures of enzymes do not affect in any way (there is no regularity of change or significant deviations) the values of the binding energy (ΔGb) recorded in the AChE-OPs complexes. However, the mutations analyzed in AChE are associated with a structural modification that causes an inadequate interaction to complete the phosphorylation of the enzyme.

The sequences of MinE responsible for its subcellular localization analyzed by competitive binding method in Escherichia coli.

The subcellular localization of a protein is important for its proper function. Escherichia coli MinE is a small protein with clear subcellular localization, which provides a good model to study protein localization mechanism. In the present study, a series of recombinant minEs truncated in one end or in the middle regions, fused with egfp, was constructed, and these recombinant proteins could compete to function with the chromosomal MinE. Our results showed that the sequences related to the subcellular localization of MinE span several functional domains, demonstrating that MinE positioning in cells depends on multiple factors. The eGFP fusions with some truncated MinE from N-terminal resulted in different cell phenotypes and localization features, implying that these fusions can interfere chromosomal MinE's function, similar to MinE36-88 phenotype in the previous report. The amino acid in the region (32-48) is sensitive to change MinE conformation and influence its dimerization. Some truncated protein structure could be unstable. Thus, the MinE localization is prerequisite for its proper anti-MinCD function and some new features of MinE were demonstrated. This approach can be extended for subcellular localization research for other essential proteins.

Time series analysis of onchocerciasis data from Mexico: a trend towards elimination.

BACKGROUND: In Latin America, there are 13 geographically isolated endemic foci distributed among Mexico, Guatemala, Colombia, Venezuela, Brazil and Ecuador. The communities of the three endemic foci found within Mexico have been receiving ivermectin treatment since 1989. In this study, we predicted the trend of occurrence of cases in Mexico by applying time series analysis to monthly onchocerciasis data reported by the Mexican Secretariat of Health between 1988 and 2011 using the software R. RESULTS: A total of 15,584 cases were reported in Mexico from 1988 to 2011. The data of onchocerciasis cases are mainly from the main endemic foci of Chiapas and Oaxaca. The last case in Oaxaca was reported in 1998, but new cases were reported in the Chiapas foci up to 2011. Time series analysis performed for the foci in Mexico showed a decreasing trend of the disease over time. The best-fitted models with the smallest Akaike Information Criterion (AIC) were Auto-Regressive Integrated Moving Average (ARIMA) models, which were used to predict the tendency of onchocerciasis cases for two years ahead. According to the ARIMA models predictions, the cases in very low number (below 1) are expected for the disease between 2012 and 2013 in Chiapas, the last endemic region in Mexico. CONCLUSION: The endemic regions of Mexico evolved from high onchocerciasis-endemic states to the interruption of transmission due to the strategies followed by the MSH, based on treatment with ivermectin. The extremely low level of expected cases as predicted by ARIMA models for the next two years suggest that the onchocerciasis is being eliminated in Mexico. To our knowledge, it is the first study utilizing time series for predicting case dynamics of onchocerciasis, which could be used as a benchmark during monitoring and post-treatment surveillance.