Current status and future trends of microbial and nematode-based biopesticides for biocontrol of crop pathogens.

The increasing public demand to avoid the use of synthetic pesticides and fertilizers in agricultural production systems, causing serious environmental damages, has challenged industry to develop new and effective solutions to manage and control phytopathogens. Biopesticides, particularly microbial-based biopesticides, are a promising new alternative with high biodegradability, specificity, suitability for incorporation into integrated pest management practices, low likelihood of resistance development, and practically no known human health risks. However: expensive production methods, narrow action spectra, susceptibility to environmental conditions, short shelf life, poor storage stability, legislation registry constraints, and general lack of knowledge are slowing down their adoption. In addition to regulatory framework revisions and improved training initiatives, improved preservation methods, thoughtfully designed formulations, and field test validations are needed to offer new microbial- and nematode-based biopesticides with improved efficacy and increased shelf-life. During the last several years, substantial advancements in biopesticide production have been developed. The novelty part of this review written in 2023 is to summarize (i) mechanisms of action of beneficial microorganisms used to increase crop performance and (ii) successful formulation including commercial products for the biological control of phytopathogens based on microorganisms, nematode and/or metabolites.

IBD sharing patterns as intra-breed admixture indicators in small ruminants.

In this study, we investigated how IBD patterns shared between individuals of the same breed could be informative of its admixture level, with the underlying assumption that the most admixed breeds, i.e. the least genetically isolated, should have a much more fragmented genome. We considered 111 goat breeds (i.e. 2501 individuals) and 156 sheep breeds (i.e. 3304 individuals) from Europe, Africa and Asia, for which beadchip SNP genotypes had been performed. We inferred the breed's level of admixture from: (i) the proportion of the genome shared by breed's members (i.e. "genetic integrity level" assessed from ADMIXTURE software analyses), and (ii) the "AV index" (calculated from Reynolds' genetic distances), used as a proxy for the "genetic distinctiveness". In both goat and sheep datasets, the statistical analyses (comparison of means, Spearman correlations, LM and GAM models) revealed that the most genetically isolated breeds, also showed IBD profiles made up of more shared IBD segments, which were also longer. These results pave the way for further research that could lead to the development of admixture indicators, based on the characterization of intra-breed shared IBD segments, particularly effective as they would be independent of the knowledge of the whole genetic landscape in which the breeds evolve. Finally, by highlighting the fragmentation experienced by the genomes subjected to crossbreeding carried out over the last few generations, the study reminds us of the need to preserve local breeds and the integrity of their adaptive architectures that have been shaped over the centuries.

Biochemical and microbial characterization of a forest litter-based bio-fertilizer produced in batch culture by fermentation under different initial oxygen concentrations.

This work focused on the physico-chemical, biochemical and microbiological characterization of a new organic fertilizer based on fermented forest litter (FFL) mixed with agro-industrial by-products, on the culture realized in airtight glass bottle. Under strict anaerobiosis (0% initial oxygen concentration (IOC)), after a 16-day batch culture, the bottle-headspace analysis showed that the specific CO2 production rate was low (0.014 mL/h.g dry matter) compared to those reached under aerobic conditions (e.g. 0.464 mL/h.g dm at 21% IOC). Moreover, the culture displayed a slight fermented fruity odour, mainly due to ethanol and ethyl acetate detected in the headspace (335 µL and 58.6 µL accumulated, respectively). The FFL organic matter degradation followed by infrared spectroscopy and catabolic potential and diversity characterized by BIOLOG® EcoPlates were poor and pH dropped to 4.54. The microbiome's metabolism was oriented toward lactic fermentation with medium acidification, enrichment in lactic acid bacteria (LAB), depletion in fungi and absence of pathogens. By increasing IOC from 0 to 21%, the respirometric activity, and the catabolic potential and diversity increased. However, some enterobacteria were detected above 5% IOC. Ethanol and ethyl acetate decreased strongly with IOC, and aromatics and proteins contained in the solid matrix remained in the culture. This study showed the importance of oxygen on the final product. A 2% IOC was found to ensure an optimal balance between LAB development, preservation of functional catabolic diversity and bio-product free of microbial pathogens.

Is Musical Auditory Stimulation Able to Influence the Recovery of Autonomic Modulation After Exercise?

Context: The effects of musical auditory stimulation on the human body have received considerable interest, especially when related to health and wellbeing, so the objective was to verify the influence of classical music exposure on autonomic modulation in recovery after exercise. Methods: 35 healthy young men aged 21.74 ± 2.59 years were evaluated. Volunteers perform 2-day evaluations, 1 day without musical stimulus and 1 with musical stimulation through classical music during exercise and recovery. Physical exercises were performed on a treadmill with an intensity of 6.0 km/hr + 1% of inclination in the first 5 minutes for physical "warm-up", followed by 25 minutes with an intensity equivalent to 60% of the maximum velocity (Vmax), with a same slope according to the Conconi's threshold. Finally, the volunteers remain at rest for 60 minutes for recovery. Heart rate variability (HRV) was analyzed in some moments of recovery by linear (RMSSD, SDNN, LF and HF (milliseconds squared and normalized unit) and LF/HF ratio, RRtri, TINN and the Poincaré plot: SD1, SD2 and SD1/SD2) and nonlinear indexes (DFA Total, DFA Alpha 1 and Symbolic Analysis: Shannon's Entropy, 0V%, 1V%, 2LV% and 2ULV%). Results: No differences were found in the comparison between the values of the HRV indexes between protocols and interaction moments vs protocols. Differences were observed between the moments for most indexes (P < .05). Conclusions: Classical music did not significantly influence the autonomic control of heart rate in the recovery phase after submaximal exercise.

Ethylhexyl methoxycinnamate and butyl methoxydibenzoylmethane: Toxicological effects on marine biota and human concerns.

Ethylhexyl methoxycinnamate (EHMC) (CAS number: 5466-77-3) and butyl methoxydibenzoylmethane (BMDM) (CAS number: 70356-09-1) are important sunscreens. However, frequent application of large amounts of these compounds may reflect serious environmental impact, once it enters the environment through indirect release via wastewater treatment or immediate release during water activities. In this article, we reviewed the toxicological effects of EHMC and BMDM on aquatic ecosystems and the human consequences. According to the literature, EHMC and BMDM have been detected in water samples and sediments worldwide. Consequently, these compounds are also present in several marine organisms like fish, invertebrates, coral reefs, marine mammals, and other species, due to its bioaccumulation potential. Studies show that these chemicals are capable of damaging the aquatic beings in different ways. Further, bioaccumulation studies have shown that EHMC biomagnifies through trophic levels, which makes human seafood consumption a concern because the higher position in the trophic chain, the more elevate levels of ultraviolet (UV) filters are detected, and it is established that EHMC present adverse effects on the human organism. In contrast, there are no studies on the BMDM bioaccumulation and biomagnification potential. Different strategies can be adopted to avoid the damage caused by sunscreens in the environment and human organism. Two of them include the use of natural photoprotectors, such as polyphenols, in association with UV filters in sunscreens and the development of new and safer UV filters. Overall, this review shows the importance of studying the impacts of sunscreens in nature and developing safer sunscreens and formulations to safeguard marine fauna, ecosystems, and humans.

Dried Plasma Spots and Oral Fluid as Alternative Matrices for Therapeutic Drug Monitoring of Busulfan: Analytical Method Development and Clinical Evaluation.

BACKGROUND: Busulfan (BU) is an alkylating agent with a narrow therapeutic index and high intraindividual pharmacokinetic variability used in conditioning therapy for hematopoietic stem cell transplantation. Monitoring BU exposure during high-dose conditioning regimens is recommended and positively impacts outcomes. We aimed to develop, validate, and apply a ultra-high-performance liquid chromatography-mass spectrometry (MS)/MS assay to measure BU concentrations in oral fluid and dried plasma spots (DPS) as alternative matrices to plasma. METHODS: We prepared plasma and oral fluid samples by protein precipitation and DPS after liquid extraction. We analyzed extracts using an LC-MS/MS system with an Acquity HSS T3 column in the positive electrospray ionization mode. The method was validated and applied to 79 paired plasma and oral fluid samples from 7 patients on BU conditioning treatment. DPS were prepared by pipetting plasma onto Whatman 903 paper. The correlation between BU in plasma, oral fluid, and DPS samples was evaluated. RESULTS: Run time was 4.0 minutes. The assay was linear at 50-5000 ng mL-1 (r > 0.99), precise (1.9%-5.3% oral fluid and 1.8%-5.9% DPS), and accurate (98.1%-108.9% oral fluid and 93%-103.1% DPS). BU was stable in DPS at 23°C for 24 hours. BU levels in oral fluid (r = 0.927) and DPS (r = 0.982) were significantly correlated with plasma. Despite the good correlation, we found a wide variation between oral fluid and plasma levels. The area under curves (AUCs) calculated with oral fluid concentrations were 79.1%-167.1% of plasma AUCs. Bland-Altman plots found a better agreement for DPS, with AUCs estimated from corrected DPS levels at 83.1%-114.1% of plasma values. CONCLUSIONS: We developed and validated a simple and fast ultra-high-performance liquid chromatography-MS/MS assay to measure BU in oral fluid and DPS. The results do not support the use of oral fluid as a matrix for routine therapeutic drug monitoring of BU. The AUC estimated from BU measurements in DPS was comparable to that in plasma, supporting the use of DPS in BU therapeutic drug monitoring as an alternative matrix, with adequate short-term stability and logistic advantages.

Interrater Reliability Across 7 Established Risk Stratification Protocols in Cardiac Rehabilitation.

OBJECTIVE: To analyze the interrater agreement among physiotherapists in using 7 risk stratification (RS) protocols to evaluate participants of cardiac rehabilitation (CR) and the main factors associated with disagreements that emerged during the RS process. DESIGN: Cross-sectional observational study. SETTING: Outpatient rehabilitation center. PARTICIPANTS: Patients (N=72) enrolled in CR with a diagnosis of cardiovascular disease or cardiovascular risk factors. Mean age was 65.62±12.14 y, and mean body mass index (calculated as weight in kilograms divided by height in meters squared) was 29.18±4.56. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The main outcome was to the agreement between 2 physiotherapists in the patients' RS process, using 7 protocols established in the literature for use in CR: American Association of Cardiovascular and Pulmonary Rehabilitation, American College of Sports Medicine, American Heart Association, Sociedade Brasileira de Cardiologia, Sociedad Española de Cardiología, and Société Française de Cardiologie. In addition, the main disagreement factors were assessed. RESULTS: Interrater agreement was classified as moderate-to-good in the 7 included RS protocols (kappa index between 0.53-0.76). The most important aspects that led to disagreement between physiotherapists were reported in 5 categories. The protocol with the greater agreement index was the American College of Sports Medicine (93.10%; n=67), and the one with the greater disagreement was the American Association of Cardiovascular and Pulmonary Rehabilitation (27.80%; n=20). CONCLUSIONS: Moderate-to-good interrater agreement among physiotherapists in using 7 RS protocols was observed. Major disagreements were the definition of abnormal hemodynamic responses, rhythm disorders, left ventricular dysfunction, and interpretation of the patient's clinical characteristics.

An investigation into whether cardiac risk stratification protocols actually predict complications in cardiac rehabilitation programs?

OBJECTIVES: This study evaluated the capacity of cardiac risk stratification protocols on simple complications that occur during activities of a cardiovascular rehabilitation program. DESIGN: Observational longitudinal cohort study. SETTING: Outpatient clinic of cardiovascular rehabilitation. SUBJECT: Patients diagnosed with cardiovascular disease and/or risk factors. INTERVENTIONS: Not applicable. MAIN MEASURES: The relationship between the cardiac risk classes of seven risk stratification protocols and the occurrence of simple complications (such angina, abnormal changes in blood pressure, arrhythmias, fatigue, muscle pain, pallor) was assessed using the chi-square test, and when statistical significance was observed, sensitivity, specificity and accuracy were determined. RESULTS: About 76 patients were analyzed. The American Association of Cardiovascular and Pulmonary Rehabilitation (AACVPR) protocol showed a statistically significant relationship between simple complications and cardiac risk classes (P-value = 0.046), however the results of sensitivity (0.53), specificity (0.52), and accuracy (0.53) were not significant. The other protocols analyzed were not significant: American College of Sports Medicine (P-value = 0.801), Brazilian Society of Cardiology (P-value = 0.734), American Heart Association (P-value = 0.957), Pashkow (P-value = 0.790), Society French Cardiology (P-value = 0.314), and Spanish Society of Cardiology (P-value = 0.078). CONCLUSION: The AACVPR protocol showed a significant relationship between the risk classes and the occurrence of simple complications, however, the low values obtained for sensitivity, specificity and accuracy show that it is not useful for this purpose. CLINICAL TRIALS REGISTRATION: NCT03446742.

Environmental Drivers of Microbial Functioning in Mediterranean Forest Soils.

Mediterranean forests own distinct characteristics resulting from climate, soil, and vegetation that affect soil microbial communities' assembly and their associated functions. We initiated a multi-scalar analysis of environmental drivers of soil functioning to (1) identify pertinent factorial scales and (2) determine the relative importance of soil, vegetation, and geoclimate influences in shaping soil microbial functions across the French Mediterranean forests. Soil samples (0-15 cm) were collected from 60 forest sites and soil physicochemical and microbiological properties were assessed across different factorial scales i.e., bioclimates, slope exposures, and forest stands. Patterns in microbial catabolic potential (i.e., extracellular enzymes and microbial respiration) and carbon (C) source utilization (i.e., catabolic-level physiological profiling) were partitioned between vegetation cover, soil characteristics, and geoclimate components. Our results reveal that the catabolic potential of soil microbes was strongly influenced by the forest stands and mainly relied on ammonium and nitrate contents. In contrast, variation in C source utilization was mainly explained by vegetation cover. Soil metabolic capacities of microorganisms and resulting C dynamics were largely constrained by climate parameters, which suggests potentially important consequences for soil C storage. Our study revealed diverse structuration patterns between the catabolic potential and the carbon source utilization of soil microbial communities, and gives insights into the underlying mechanisms of soil microbial functioning in Mediterranean forests.

On the origin of European sheep as revealed by the diversity of the Balkan breeds and by optimizing population-genetic analysis tools.

BACKGROUND: In the Neolithic, domestic sheep migrated into Europe and subsequently spread in westerly and northwesterly directions. Reconstruction of these migrations and subsequent genetic events requires a more detailed characterization of the current phylogeographic differentiation. RESULTS: We collected 50 K single nucleotide polymorphism (SNP) profiles of Balkan sheep that are currently found near the major Neolithic point of entry into Europe, and combined these data with published genotypes from southwest-Asian, Mediterranean, central-European and north-European sheep and from Asian and European mouflons. We detected clines, ancestral components and admixture by using variants of common analysis tools: geography-informative supervised principal component analysis (PCA), breed-specific admixture analysis, across-breed [Formula: see text] profiles and phylogenetic analysis of regional pools of breeds. The regional Balkan sheep populations exhibit considerable genetic overlap, but are clearly distinct from the breeds in surrounding regions. The Asian mouflon did not influence the differentiation of the European domestic sheep and is only distantly related to present-day sheep, including those from Iran where the mouflons were sampled. We demonstrate the occurrence, from southeast to northwest Europe, of a continuously increasing ancestral component of up to 20% contributed by the European mouflon, which is assumed to descend from the original Neolithic domesticates. The overall patterns indicate that the Balkan region and Italy served as post-domestication migration hubs, from which wool sheep reached Spain and north Italy with subsequent migrations northwards. The documented dispersal of Tarentine wool sheep during the Roman period may have been part of this process. Our results also reproduce the documented 18th century admixture of Spanish Merino sheep into several central-European breeds. CONCLUSIONS: Our results contribute to a better understanding of the events that have created the present diversity pattern, which is relevant for the management of the genetic resources represented by the European sheep population.

Determination of cannabinoids in plasma using salting-out-assisted liquid-liquid extraction followed by LC-MS/MS analysis.

The detection of the markers of Cannabis consumption in biological specimens is an important task for drug testing laboratories in varous contexts. A simple assay combining salting-out assisted liquid-liquid extraction sample preparation and LC-MS/MS analysis was applied to the measurement of Δ9 -tetrahydrocannabinol, 11-nor-9-carboxy-Δ9 -tetrahydrocannabinol (THC-COOH), 11-hydroxy-Δ9 -tetrahydrocannabinol, cannabinol and cannabidiol concentrations in 100 µl plasma specimens. The assay had linearity of 1-100 ng ml-1 for THC-COOH and 0.5-50 ng ml-1 for the other tested cannabinoids. Assay validation criteria were fulfilled. Extraction yields (88.7-97.3%) and internal-standard correct matrix effects (-9.6 to +5.4%) were acceptable. The assay was applied to 238 clinical specimens from trauma patients, with 19 samples presenting quantifiable concentrations of at least one of the target compounds. The developed assay is a simple and efficient strategy for simultaneous measurement of Δ9 -tetrahydrocannabinol, THC-COOH, 11-hydroxy-Δ9 -tetrahydrocannabinol, cannabinol and cannabidiol concentrations in plasma specimens.

Assessment of metal/metalloid occurrence in rivers with their accumulation in macrophyte case study with Myriophyllum alterniflorum.

Water quality monitoring with integrative tools is a main issue of concern for environment assessment. Submerged aquatic macrophyte can be a good candidate for the evaluation of contaminant content in rivers. Indeed, owing to their habitat, aquatic macrophytes interact directly with surface water; they can absorb contaminants and thus allow to detect their presence in water. In situ studies were conducted over 28 days in five aquatic environments, affected by different levels of anthropogenic pressure (domestic wastewater plant, industrial activities), during two field campaigns. We have investigated whether the accumulation of some metals and a metalloid (As) in Myriophyllum alterniflorum could be used to detect their occurrence in river waters. Our results demonstrated that long time bioaccumulation was correlated with the contaminant levels in water. However, the water composition and the duration of exposure affected the studied pollutants' absorption. On an operational point of view, the optimal duration of exposure of Myriophyllum alterniflorum to assess the water quality is conditioned by the contaminant contents in waters that can induce different defense mechanisms as the reduction of pollutant absorption and their efflux. In addition, the nutrient concentration influenced the accumulation of pollutants since the higher the nutrient level, the higher the essential metal accumulation was observed.

Revisiting the Pharmacodynamic Uroselectivity of α 1-Adrenergic Receptor Antagonists.

α1-Adrenoceptor (AR) antagonists are widely used for the relief of urinary retention secondary to benign prostatic hyperplasia (BPH). While the five Food and Drug Administration-approved α 1-AR antagonists (terazosin, doxazosin, alfuzosin, tamsulosin, and silodosin) share similar efficacy, they differ in tolerability, with reports of ejaculatory dysfunction. The aim of the present work was to revisit their α 1-AR subtype selectivity as well as of LDT5 (1-(2-methoxyphenyl)-4-[2-(3,4-dimethoxyphenyl) ethyl]piperazine monohydrochloride), a compound previously described as a multitarget antagonist of α 1A-/α 1D-AR and 5-HT1A receptors, and to estimate their affinity for D2, D3, and 5-HT1A receptors, which are putatively involved in ejaculatory dysfunction. Competition binding assays were performed with native (D2, 5-HT1A) or transfected (human α 1A-, α 1B-, α 1Dt-AR, and D3) receptors for determination of the drug's affinities. Tamsulosin and silodosin have the highest affinities for α 1A-AR, but only silodosin is clearly a selective α 1A-AR antagonist, with K i ratios of 25.3 and 50.2 for the α 1D- and α 1B-AR, respectively. Tamsulosin, silodosin, and LDT5 (but not terazosin, doxazosin, and alfuzosin) have high affinity for the 5-HT1A receptor (K i around 5-10 nM), behaving as antagonists. We conclude that the uroselectivity of tamsulosin is not explained by its too-low selectivity for the α 1A- versus α 1B-AR, and that its affinity for D2 and D3 receptors is probably too low for explaining the ejaculatory dysfunction reported for this drug. Present data also support the design of "better-than-LDT5" new multitarget lead compounds with pharmacokinetic selectivity based on poor brain penetration and that could prevent hyperplastic cell proliferation and BPH progression. SIGNIFICANCE STATEMENT: The present work revisits the uroselectivity of the five Food and Drug Administration-approved α1 adrenoceptor antagonists for the treatment of benign prostatic hyperplasia (BPH). Contrary to what has been claimed by some, our results indicate that the uroselectivity of tamsulosin is probably not fully explained by its too-weak selectivity for the α1A versus α1B adrenoceptors. We also show that tamsulosin affinity for D3 and 5-HT1A receptors is probably too low for explaining the ejaculatory dysfunction reported for this drug. Based on our lead compound LDT5, present data support the search for a multitarget antagonist of α1A-α1D and 5-HT1A receptors with poor brain penetration as an alternative for BPH treatment.

Determination of Endogenous Concentrations of Uracil and Dihydrouracil in Dried Saliva Spots by LC-MS/MS: Method Development, Validation, and Clinical Application.

BACKGROUND: The aim of this study was to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for the measurement of uracil (U) and dihydrouracil (UH2) concentrations in dried saliva spots (DSSs), for the evaluation of dihydropyrimidine dehydrogenase (DPD) enzyme activity. RESULTS: Nine 18-mm diameter DSS discs were extracted with acetate:isopropyl alcohol (85:15, vol/vol) and analyzed by LC-MS/MS. The assay was linear in the range of 10-1000 ng·mL, with accuracy between 89% and 112% and precision between 5.7% and 13%. The metabolic ratio [UH2]/[U] was stable in DSS for up to 9 days at 45°C. Concentrations of U and UH2, as well as the metabolic ratio, were highly concordant between matrices. Using a metabolic ratio classification cutoff of 1.16 for the identification of slow DPD metabolizers, 98.7% concordance was achieved between SS and saliva. CONCLUSIONS: DSS samples could be a useful alternative for DPD activity screening, particularly in locations with limited access to highly equipped laboratories.

Pre-treatment of a sugarcane bagasse-based substrate prior to saccharification: Effect of coffee pulp and urea on laccase and cellulase activities of Pycnoporus sanguineus.

Production of second-generation bioethanol uses lignocellulose from agricultural by-products such as sugarcane bagasse (SCB). A lignocellulose pre-treatment is required to degrade lignin, ensuring further efficient saccharification. Two experimental designs were set up to define culture conditions of Pycnoporus sanguineus in mesocosms to increase laccase activities and thus delignification. The first experimental design tested the effect of phenolic complementation (via coffee pulp) and the use of urea as a simple nitrogen source and the second defined more precisely the percentages of coffee pulp and urea to enhance delignification. The responses measured were: lignocellulolytic activities, laccase isoform profiles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the chemical transformation of the substrate using solid-state NMR of 13C. Adding 10% of coffee pulp increased laccase activities and fungal biomass (32.5% and 16% respectively), enhanced two constitutive isoforms (Rf 0.23 and 0.27), induced a new isoform (Rf 0.19) and led to a decrease in total aromatics. However, higher concentrations of coffee pulp (25%) decreased laccase and cellulase activities but no decrease in aromaticity was observed, potentially due to the toxic effect of phenols from coffee pulp. Moreover, laccase production was still inhibited even for lower concentrations of urea (0-5%). Our findings revealed that an agricultural by-product like coffee pulp can enhance laccase activity -though to a threshold- and that urea limited this process, indicating that other N-sources should be tested for the biological delignification of SCB.

Relationship of the Aggregation of Cardiovascular Risk Factors in the Parasympathetic Modulation of Young People with Type 1 Diabetes.

Background and objectives: In healthy individuals, autonomic alterations are associated with the aggregation of cardiovascular risk factors. However, in individuals with type 1 diabetes, who are known to present autonomic alterations, mainly characterized by a reduction in parasympathetic modulation, these associations have not yet been investigated. We assess whether the aggregation of cardiovascular risk factors influences parasympathetic indices of heart rate variability in young people with type 1 diabetes. Materials and methods: This cross-sectional study included 39 individuals with type 1 diabetes (22.54 ± 4.31), evaluated in relation to the risk factors: blood pressure, fat percentage, and resting heart rate. For heart rate variability analysis, heart rate was recorded beat-to-beat using a cardio frequency meter (PolarS810i) for 30 min with the volunteers in dorsal decubitus. The parasympathetic heart rate variability indices were calculated: rMSSD, pNN50, high frequency (HF) n.u (normalized units), SD1, 2LV, and 2ULV. Data collection was carried out in 2014 and analyzed in 2017. Results: Individuals with two aggregate risk factors present a reduction in the values of the indices that reflect parasympathetic autonomic modulation compared to individuals without the risk factors analyzed, regardless of sex and age. Conclusion: In young people with type 1 diabetes, the aggregation of cardiovascular risk factors is associated with parasympathetic autonomic impairment.

An siRNA-based screen in C2C12 myoblasts identifies novel genes involved in myogenic differentiation.

Myogenesis is a highly regulated multi-step process involving myoblast proliferation and differentiation. Although studies over the last decades have identified several factors governing these distinct major phases, many of them are not yet known. In order to identify novel genes, we took advantage of the C2C12 myoblastic line to establish a functional siRNA screen combined with quantitative-imaging analysis of a large amount of differentiated myoblasts. We knocked down 100 preselected mouse genes without a previously characterized role in muscle. Using image analysis, we tracked gene-silencing phenotypes by quantitative assessment of cellular density, myotube quantity, myotube morphology and fusion index. Our results have revealed six genes involved in both stages of C2C12 myogenesis and 13 genes specific to the differentiation stage. These findings prove that our RNAi-based screen could be an efficient tool to detect clear and subtle phenotypes allowing the identification of new myogenic regulators in mammals.

Heparan sulfates and the decrease of N-glycans promote early adipogenic differentiation rather than myogenesis of murine myogenic progenitor cells.

In vitro, extracted muscle satellite cells, called myogenic progenitor cells, can differentiate either in myotubes or preadipocytes, depending on environmental factors and the medium. Transcriptomic analyses on glycosylation genes during satellite cells differentiation into myotubes showed that 31 genes present a significant variation of expression at the early stages of murine myogenic progenitor cells (MPC) differentiation. In the present study, we analyzed the expression of 383 glycosylation related genes during murine MPC differentiation into preadipocytes and compared the data to those previously obtained during their differentiation into myotubes. Fifty-six glycosylation related genes are specifically modified in their expression during early adipogenesis. The variations correspond mainly to: a decrease of N-glycans, and of alpha (2,3) and (2,6) linked sialic acids, and to a high level of heparan sulfates. A high amount of TGF-ß1 in extracellular media during early adipogenesis was also observed. It seems that the increases of heparan sulfates and TGF-ß1 favor pre-adipogenic differentition of MPC and possibly prevent their myogenic differentiation.

Effects of 9 Months of Martial Arts Training on Cardiac Autonomic Modulation in Healthy Children and Adolescents.

PURPOSE: The aim of the study was to evaluate the cardiac autonomic modulation after 9 months of martial arts practice in healthy children and adolescents. METHOD: The study included 59 children and adolescents who were divided into 3 groups: judo, Muay Thai, and control. Heart rate variability was measured by a heart rate monitor, model Polar RS800CX. The intervention occurred twice a week on nonconsecutive days, lasting 60 minutes each session. A 1-way analysis of variance was used to compare participants at baseline. The comparisons between groups at baseline and after the intervention were carried out by a 2-way analysis of variance for repeated measures. RESULTS: After 9 months of intervention, significant increases were observed for root mean square successive differences, with higher values post compared with baseline (19.5%; P = .04). For SD1, an interaction effect was observed, with increased posttraining values compared with baseline (24.1%; P = .04) for the judo group. Qualitative analysis of the Poincaré plot showed greater dispersion of RR intervals, mainly beat to beat, after the judo intervention compared with the baseline. The Muay Thai and control groups presented no improvement. CONCLUSION: After 9 months of intervention, there were increases in cardiac autonomic modulation of children and adolescents participating in judo training. The practice of martial arts, such as judo, can be encouraged from an early age to improve cardiovascular system functioning, possibly providing protection against cardiovascular problems.

Laccase induction by synthetic dyes in Pycnoporus sanguineus and their possible use for sugar cane bagasse delignification.

The use of synthetic dyes for laccase induction in vivo has been scarcely explored. We characterized the effect of adding different synthetic dyes to liquid cultures of Pycnoporus sanguineus on laccase production. We found that carminic acid (CA) can induce 722 % and alizarin yellow 317 % more laccase than control does, and they promoted better fungal biomass development in liquid cultures. Aniline blue and crystal violet did not show such positive effect. CA and alizarin yellow were degraded up to 95 % during P. sanguineus culturing (12 days). With this basis, CA was selected as the best inducer and used to evaluate the induction of laccase on solid-state fermentation (SSF), using sugarcane bagasse (SCB) as substrate, in an attempt to reach selective delignification. We found that laccase induction occurred in SSF, and a slight inhibition of cellulase production was observed when CA was added to the substrate; also, a transformation of SCB under SSF was followed by the 13C cross polarization magic angle spinning (CPMAS) solid-state nuclear magnetic resonance (NMR). Results showed that P. sanguineus can selectively delignify SCB, decreasing aromatic C compounds by 32.67 % in 16 days; O-alkyl C region (polysaccharides) was degraded less than 2 %; delignification values were not correlated with laccase activities. Cellulose-crystallinity index was increased by 27.24 % in absence of CA and 15.94 % when 0.01 mM of CA was added to SCB; this dye also inhibits the production of fungal biomass in SSF (measured as alkyl C gain). We conclude that CA is a good inducer of laccase in liquid media, and that P. sanguineus is a fungus with high potential for biomass delignification.