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The Brazilian army recently developed the military physical training “Cross Operational” (CO), composed of four different difficulty levels. The moderate/high intensity character of CO can damage the muscle fibers, characterized by alterations at the cellular level, which can be evaluated indirectly by intramuscular proteins showing up in the bloodstream. In this context, the analysis of muscle damage biomarkers can be used as part of an initial injury prevention tool and in the evaluation of training progression and post-exercise recovery. This study aimed to observe the acute effect of CO on indirect markers of muscle damage in military personnel of the Brazilian army. Twenty-four male recruits aged between 18 and 26 years participated in the study. The four sessions corresponding to the CO levels were performed in cross-design, with a seven-day washout period, and blood samples were collected at rest, immediately after, 24 and 48 hours after each training session. The measured markers were creatine kinase (CK), myoglobin (Mb) and lactate dehydrogenase (LDH). In all CO levels, CK had a significant increase 24 hours after CO, whereas Mb and LDH incresead immediately after CO. Regarding recovery time, LDH returned to baseline values within 48 hours in all levels, whereas CK and Mb, after 48 hours of recovery, reduced but did not return to baseline in level 4. The serum elevations of the biomarkers studied, like those found in studies with other consolidated physical training methods, suggest that CO can contribute to the improvement of the physical training of military personnel.

Objective: Mitochondrial proteostasis is critical for cellular function. The molecular chaperone HSP60 is essential for cell function and dysregulation of HSP60 expression has been implicated in cancer and diabetes. The few reported patients carrying HSP60 gene variants show neurodevelopmental delay and brain hypomyelination. Hsp60 interacts with more than 260 mitochondrial proteins but the mitochondrial proteins and functions affected by HSP60 deficiency are poorly characterized. Methods: We studied two model systems for HSP60 deficiency: (1) engineered HEK cells carrying an inducible dominant negative HSP60 mutant protein, (2) zebrafish HSP60 knockout larvae. Both systems were analyzed by RNASeq, proteomics, and targeted metabolomics, and several functional assays relevant for the respective model. In addition, skin fibroblasts from patients with disease-associated HSP60 variants were analyzed by proteomics. Results: We show that HSP60 deficiency leads to a differentially downregulated mitochondrial matrix proteome, transcriptional activation of stress responses, and dysregulated cholesterol biosynthesis. This leads to lipid accumulation in zebrafish knockout larvae. Conclusions: Our data provide a compendium of the effects of HSP60 deficiency on the mitochondrial matrix proteome. We show that HSP60 is a master regulator and modulator of mitochondrial functions and metabolic pathways. HSP60 dysfunction also affects cellular metabolism and disrupts the integrated stress response. The effect on cholesterol synthesis explains the effect of HSP60 dysfunction on myelination observed in patients carrying genetic variants of HSP60.

Leaf senescence in plants is the last stage of leaf development and is characterized by a decline in photosynthetic activity, an active degeneration of cellular structures, and the recycling of accumulated nutrients to areas of active growth, such as buds, young leaves, flowers, fruits, and seeds. This process holds economic significance as it can impact yield, influencing the plant’s ability to maintain an active photosynthetic system during prolonged periods, especially during the grain filling stage, which affects plant weight and oil content. It can be associated with different stresses or environmental conditions, manifesting itself widely in the context of climate change and limiting yield, especially in crops of agronomic relevance. In this work, we study the stability of two widely described sunflower (Helianthus annuus L.) genotypes belonging to the INTA Breeding Program against differential N conditions, to verify their yield stability in control conditions and under N supply. Two inbred lines were utilized, namely R453 (early senescence) and B481-6 (late senescence), with contrasting nitrogen availability in the soil but sharing the same ontogeny cycle length. It was observed that, starting from R5.5, the B481-6 genotype not only delayed senescence but also exhibited a positive response to increased nitrogen availability in the soil. This response included an increase in intercepted radiation, resulting in a statistically significant enhancement in grain yield. Conversely, the R453 genotype did not show significant differences under varying nitrogen availability and exhibited a tendency to decrease grain yield when nitrogen availability was increased. The response to nitrogen can vary depending on the specific genotype.

Leaf senescence is a complex trait which becomes crucial for grain filling because photoassimilates are translocated to the seeds. Therefore, a correct sync between leaf senescence and phenological stages is necessary to obtain increasing yields. In this study, we evaluated the performance of five deep machine-learning methods for the evaluation of the phenological stages of sunflowers using images taken with cell phones in the field. From the analysis, we found that the method based on the pre-trained network resnet50 outperformed the other methods, both in terms of accuracy and velocity. Finally, the model generated, Sunpheno, was used to evaluate the phenological stages of two contrasting lines, B481_6 and R453, during senescence. We observed clear differences in phenological stages, confirming the results obtained in previous studies. A database with 5000 images was generated and was classified by an expert. This is important to end the subjectivity involved in decision making regarding the progression of this trait in the field and could be correlated with performance and senescence parameters that are highly associated with yield increase.

Introduction and Objectives: Endoscopic ultrasound guided portal pressure gradient measurement (EUS-PPGm) would provide useful clinical information in patients with liver diseases. However, there is yet scarcely data on the clinical relevance of this EUS-guided procedure.We report our experience on EUS-PPGm focus on safety, clinical relevant findings, technical drawbacks and how to overcome them, aiming to make this procedure more safe, accurate and available. Materials and Patients: EUS-PPGm was performed with a therapeutic echoendoscope and a dedicated 25G needle in 30 consecutive patients. Assessment of NAFLD 25; idiopathic portal hypertension 3; evaluation for curative therapy in hepatocellular carcinoma (HCC) 2. EUS-guided bilobar liver biopsies (EUS-BLB) were also immediately performed in 26 patients (87%) with a 19G needle Results: EUS-PPGm was obtained in 25/30 patients (83%) being >5 mmHg in 10/22 NALFD patients (45%) without endoscopic and/or ultrasonographic signs of portal hypertension neither liver fibrosis on EUS-BLB. Mean time to obtain EUS-PPGm was 21±2 minutes.EUS-PPGm was not obtained in 5 cases. In 4 cases for excesive use of the elevator and up&down wheel and bending of the needle. In another case for exacerbating breathing movements. The hepatic and portal vein were difficult to punction in one and two cases, respectively having to transverse the vessels and reposition the needle.A self-limited bleeding from the cardias and a mild epigastric pain 2 day after a combined procedure were observed without other adverse events one month later. Conclusions: EUS-guided PPGm, even when combined with EUS-BLB, seems safe providing useful clinical information. Almost half of patients affected with NAFLD have portal hypertension diagnosed precociously in reversible stages providing a useful tool in precision Medicine, especially in the setting of obesity pandemic. There are technical aspects related to the needle and the position of the echoendoscope that should be known to improve the safety, accuracy and availibility of this procedure.

Abstract Introduction Contingency measures due to the COVID-19 pandemic limited access to routine prenatal care for pregnant women, increasing the risk of pregnancy complications due to poor prenatal follow-up, especially in those patients at high obstetric risk. This prompted the implementation and adaptation of telemedicine. Objective We aim to evaluate the maternal and perinatal outcomes of patients who received prenatal care in-person and by telemedicine. Methods We conducted a retrospective observational cohort study of pregnant women who received exclusive in-person and alternate (telemedicine and in-person) care from March to December 20,202, determining each group's maternal and neonatal outcomes. Results A total of 1078 patients were included, 156 in the mixed group and 922 in the in-person group. The patients in the mixed group had a higher number of prenatal controls (8 (6–9) vs 6 (4–8) p

Introdução: os níveis de leptina estão relacionados à quantidade de gordura corporal, ao apetite e ao gasto energético, enquanto o índice de massa corporal (IMC) é o parâmetro antropométrico mais amplamente utilizado na identificação do estado nutricional por sua facilidade de mensuração e baixo custo Objetivo: Analisar as concentrações de leptina (LEP) e sua correlação com a massa corporal total (MCT) e o IMC de adultos com sobrepeso. Materiais e Métodos: pesquisa descritiva transversal com 50 voluntários (27,7 ± 5,10 anos), de ambos os sexos (25 homens e 25 mulheres), praticantes regulares de exercício físico, submetidos à avaliação antropométrica (massa corporal total e estatura) e coleta de amostra sanguínea após jejum de 12h para análise dos níveis séricos de LEP pelo método de radioimunoensaio. Realizou-se estatística descritiva, o teste de normalidade de Shapiro-Wilk, o Teste t de Student para amostras independentes e o cálculo do Coeficiente de Correlação de Pearson com significância de p

Differences of sex development and maturation (SDM) represent a heterogeneous puzzle of rare conditions with a large genetic component whose management and treatment could be improved by an accurate classification of underlying molecular conditions, and next-generation sequencing (NGS) should represent the most appropriate approach. Therefore, we conducted a survey dedicated to the use and potential outcomes of NGS for SDM disorders diagnosis among the 53 health care providers (HCP) of the European Reference Network for rare endocrine conditions. The response rate was 49% with a total of 26 HCPs from 13 countries. All HCPs, except 1, performed NGS investigations for SDM disorders on 6720 patients, 3764 (56%) with differences of sex development (DSD), including 811 unexplained primary ovarian insufficiency, and 2956 (44%) with congenital hypogonadotropic hypogonadism (CHH). The approaches varied from targeted analysis of custom gene panels (range: 11–490 genes) in 81.5% of cases or whole exome sequencing with the extraction of a virtual panel in the remaining cases. These analyses were performed for diagnostic purposes in 21 HCPs, supported by the National Health Systems in 16 cases. The likelihood of finding a variant ranged between 7 and 60%, mainly depending upon the number of analysed genes or criteria used for reporting, most HCPs also reporting variants of uncertain significance. These data illustrate the status of genetic diagnosis of DSD and CHH across Europe. In most countries, these analyses are performed for diagnostic purposes, yielding highly variable results, thus suggesting the need for harmonization and general improvements of NGS approaches.

Diagnostic criteria for juvenile myelomonocytic leukemia (JMML) are currently well defined, however in some patients diagnosis still remains a challenge. Flow cytometry is a well established tool for diagnosis and follow-up of hematological malignancies, nevertheless it is not routinely used for JMML diagnosis. Herewith, we characterized the CD34+ hematopoietic precursor cells collected from 31 children with JMML using a combination of standardized EuroFlow antibody panels to assess the ability to discriminate JMML cells from normal/reactive bone marrow cell as controls (n=29) or from cells of children with other hematological diseases mimicking JMML (n=9). CD34+ precursors in JMML showed markedly reduced B-cell and erythroid-committed precursors compared to controls, whereas monocytic and CD7+ lymphoid precursors were significantly expanded. Moreover, aberrant immunophenotypes were consistently present in CD34+ precursors in JMML, while they were virtually absent in controls. Multivariate logistic regression analysis showed that combined assessment of the number of CD34+CD7+ lymphoid precursors and CD34+ aberrant precursors or erythroid precursors had a great potential in discriminating JMMLs versus controls. Importantly our scoring model allowed highly efficient discrimination of truly JMML versus patients with JMML-like diseases. In conclusion, we show for the first time that CD34+ precursors from JMML patients display a unique immunophenotypic profile which might contribute to a fast and accurate diagnosis of JMML worldwide by applying an easy to standardize single eight-color antibody combination.

Abstract Bromodomain containing 1 (BRD1) encodes an epigenetic regulator that controls the expression of genetic networks linked to mental illness. BRD1 is essential for normal brain development and its role in psychopathology has been demonstrated in genetic and preclinical studies. However, the neurobiology that bridges its molecular and neuropathological effects remains poorly explored. Here, using publicly available datasets, we find that BRD1 targets nuclear genes encoding mitochondrial proteins in cell lines and that modulation of BRD1 expression, irrespective of whether it is downregulation or upregulation of one or the other existing BRD1 isoforms (BRD1-L and BRD1-S), leads to distinct shifts in the expression profile of these genes. We further show that the expression of nuclear genes encoding mitochondrial proteins is negatively correlated with the expression of BRD1 mRNA during human brain development. In accordance, we identify the key gate-keeper of mitochondrial metabolism, Peroxisome proliferator-activated receptor (PPAR) among BRD1’s co-transcription factors and provide evidence that BRD1 acts as a co-repressor of PPAR-mediated transcription. Lastly, when using quantitative PCR, mitochondria-targeted fluorescent probes, and the Seahorse XFe96 Analyzer, we demonstrate that modulation of BRD1 expression in cell lines alters mitochondrial physiology (mtDNA content and mitochondrial mass), metabolism (reducing power), and bioenergetics (among others, basal, maximal, and spare respiration) in an expression level- and isoform-dependent manner. Collectively, our data suggest that BRD1 is a transcriptional regulator of nuclear-encoded mitochondrial proteins and that disruption of BRD1’s genomic actions alters mitochondrial functions. This may be the mechanism underlying the cellular and atrophic changes of neurons previously associated with BRD1 deficiency and suggests that mitochondrial dysfunction may be a possible link between genetic variation in BRD1 and psychopathology in humans.

Abstract Background Leaf senescence delay impacts positively in grain yield by maintaining the photosynthetic area during the reproductive stage and during grain filling. Therefore a comprehensive understanding of the gene families associated with leaf senescence is essential. NAC transcription factors (TF) form a large plant-specific gene family involved in regulating development, senescence, and responses to biotic and abiotic stresses. The main goal of this work was to identify sunflower NAC TF (HaNAC) and their association with senescence, studying their orthologous to understand possible functional relationships between genes of different species. Results To clarify the orthologous relationships, we used an in-depth comparative study of four divergent taxa, in dicots and monocots, with completely sequenced genomes (Arabidopsis thaliana, Vitis vinifera, Musa acuminata and Oryza sativa). These orthologous groups provide a curated resource for large scale protein sequence annotation of NAC TF. From the 151 HaNAC genes detected in the latest version of the sunflower genome, 50 genes were associated with senescence traits. These genes showed significant differential expression in two contrasting lines according to an RNAseq assay. An assessment of overexpressing the Arabidopsis line for HaNAC001 (a gene of the same orthologous group of Arabidopsis thaliana ORE1) revealed that this line displayed a significantly higher number of senescent leaves and a pronounced change in development rate. Conclusions This finding suggests HaNAC001 as an interesting candidate to explore the molecular regulation of senescence in sunflower.

Advanced glycation end products (AGEs) arising from dietary intake have been associated with numerous chronic diseases including cardiovascular diseases. The interaction between platelets and AGEs has been proposed to play a role in the etiology of cardiovascular diseases. However, the effects of the interaction between platelets and Maillard reaction products generated from glyoxal (Gly) or methylglyoxal (MG) are poorly understood. In this work, the effects of AGEs generated by the reaction between Gly or MG with Lys or bovine serum albumin (BSA) on platelet activation and aggregation were assessed. AGEs were generated incubating Gly or MG with Lys or BSA during 5 hours or 14 days, respectively. AGEs generation were characterized by kinetic studies and by amino acid analysis. Human platelet-rich plasma (PRP) was incubated with different concentrations of AGEs from Lys-MG or Lys-Gly and BSA-MG or BSA-Gly. Platelet activation was determined quantifying the expression of CD62 (P-selectin) in PRP exposed to different AGEs concentrations. It was found that Lys-MG and Lys-Gly induced an increase in P-selectin expression (p

Toto represents a pleiotropic neuropeptide exerting not only inhibitory effects on reproduction, but it is also related to stress and growth. Research has been carried out to develop encapsulation technologies for GnIH, which permit their dietary incorporation into the feed. In this work, microparticles of GnIH were generated using supercritical antisolvent process (SAS) and subsequently the process of encapsulation of the compound with a biodegradable polymer, Pluronic F-127, was studied. The use of supercritical fluids and particularly SAS process to prepare microparticles removes the problems of the conventional techniques such as thermal degradation, excessive use of solvent, residual solvent concentration, and principally the challenge of controlling particle size throughout processing. In this preliminary work, the influence of the pressure and ratio hormone/polymer on the particle size, morphology, and elemental composition of these particles have been investigated. Most of the experiments led to successful precipitation of a white powder of particles in the micrometer range. The initial morphology of the GnIH was modified, resulting in a generally spherical particle form. The smallest particle size (0.16 µm) and the highest amount of precipitated GnIH in composites was obtained using 100 bar and 1/9 ratio, which were considered the best conditions in this preliminary study.

Introduction: The objective was to verify the relationship between anthropometric parameters and biomarkers associated with cardiometabolic diseases in military personnel. Methods: This is an analytical cross-sectional study, which involved 26 male Brazilian Army (EB) soldiers, with a mean age of 32.7 ± 2.12, physically active and from various EB military organizations. Serological clinical biomarkers were evaluated: glucose (GLUC), insulin (INSUL), triglyceride (TRIG), total cholesterol (TC) and high-density lipoprotein (HDL-c) and anthropometric variables obtained with a dual energy X-ray absorption densitometer (DXA) and body circumferences. The Shapiro-Wilk test and the Pearson correlation test were applied using the software Statistics® version 12.0. Results: Significant negative correlations between GLUC and lean mass (LM) (r = -0.46; p = 0.031) and fat-free mass (FFM) (r = -0.46; p = 0.032) and positive with the percentage of fat (%F) (r = 0.43; p = 0.043). Insulin (INSUL) showed positive correlations with fat mass (FM) (r = 0.52; p = 0.012); visceral adipose tissue (VAT) (r = 0.48; p = 0.024), waist circumference (WC) (r = 0.53; p = 0.01) and body mass index (BMI) (r = 0, 54; p = 0.009). The index of the model for assessing insulin homeostasis (HOMA-IR) showed positive correlations with %F (r = 0.44; p = 0.04), FM (r = 0.55; p = 0.007), VAT ( r = 0.52; p = 0.014), WC (r = 0.54; p = 0.01) and with the BMI (r = 0.52; p = 0.014). Conclusion: There was a positive association between variables representing insulin resistance and those related to body fat. In addition to negative correlations between GLUC and variables related to muscle mass. Resumen. Introducción. El objetivo fue verificar la relación entre parámetros antropométricos y biomarcadores asociados a enfermedades cardiometabólicas en personal militar. Métodos: Se trata de un estudio analítico de corte transversal, que involucró a 26 hombres soldados del Ejército Brasileño (EB), con una edad media de 32,7 ± 2,12 años, físicamente activos y de diversas organizaciones militares de la EB. Se evaluaron biomarcadores clínicos serológicos: glucosa (GLIC), insulina (INSUL), triglicéridos (TRIG), colesterol total (CT) y lipoproteínas de alta densidad (HDL-c) y variables antropométricas obtenidas con densitómetro de absorción de rayos X de energía dual (DXA) y circunferencias corporales. La prueba de Shapiro-Wilk y la prueba de correlación de Pearson se aplicaron utilizando el software Statistics® versión 12.0. Resultados: Correlaciones negativas significativas entre GLIC y masa magra (LM) (r = -0.46; p = 0.031) y masa libre de grasa (FFM) (r = -0.46; p = 0.032) y positiva con el porcentaje de grasa (% F) (r = 0,43; p = 0,043). La insulina (INSUL) mostró correlaciones positivas con la masa grasa (MG) (r = 0,52; p = 0,012); tejido adiposo visceral (IVA) (r = 0,48; p = 0,024), circunferencia de cintura (CC) (r = 0,53; p = 0,01) e índice de masa corporal (IMC) (r = 0, 54; p = 0,009). El índice del modelo para evaluar la homeostasis de la insulina (HOMA-IR) mostró correlaciones positivas con% F (r = 0.44; p = 0.04), MG (r = 0.55; p = 0.007), TAV (r = 0.52; p = 0.014), CC (r = 0,54; p = 0,01) y con el IMC (r = 0,52; p = 0,014). Conclusión: Hubo una asociación positiva entre las variables que representan la resistencia a la insulina y las relacionadas con la grasa corporal. Además de correlaciones negativas entre GLIC y variables relacionadas con la masa muscular.

A major concern for olive cultivation in many extra-Mediterranean regions is the adaptation of recently introduced cultivars to environmental conditions different from those prevailing in the original area, such as the Mediterranean basin. Some of these cultivars can easily adapt their physiological and biochemical parameters in new agro-environments, whereas others show unbalanced values of oleic acid content. The objective of this study was to evaluate the effects of the thermal regime during oil synthesis on the expression of fatty acid desaturase genes and on the unsaturated fatty acid contents at the field level. Two cultivars (Arbequina and Coratina) were included in the analysis over a wide latitudinal gradient in Argentina. The results suggest that the thermal regime exerts a regulatory effect at the transcriptional level on both OeSAD2 and OeFAD2-2 genes and that this regulation is cultivar-dependent. It was also observed that the accumulated thermal time affects gene expression and the contents of oleic and linoleic acids in cv. Arbequina more than in Coratina. The fatty acid composition of cv. Arbequina is more influenced by the temperature regime than Coratina, suggesting its greater plasticity. Overall, findings from this study may drive future strategies for olive spreading towards areas with different or extreme thermal regimes serve as guidance for the evaluation olive varietal patrimony.

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As the deleterious impacts of conventional food systems on areas including public health, environmental sustainability, and farmers’ livelihoods are progressively unveiled, citizen-led initiatives have ubiquitously sprouted, collectively building what is now known as the alternative food system. Despite recent academic interest in the role of alternative food initiatives in countering a narrow view of democracy based on market-based purchasing power, little attention has been paid to a specific democratizing feature that allows for collective expression beyond consumption, that of collective agency. This article argues that it is precisely by focusing on collective agency as the driving force for food systems’ change that we can recognize the diverse contributions of social innovations to the democratization of food systems. By engaging with the reasonings of consumer sovereignty proponents, building on academic literature on the concept of collective agency, and drawing from empirical work with over a hundred local social innovations of the global North, this article proposes an agency typology that allows for parsing out its different dimensions, highlighting social innovations’ key role as agency enablers and agents of change in the democratization of food systems.

The synthetic fatty acid 2-hydroxyoleic acid (2OHOA) has been extensively investigated as a cancer therapy mainly based on its regulation of membrane lipid composition and structure, activating various cell fate pathways. We discovered, additionally, that 2OHOA can uncouple oxidative phosphorylation, but this has never been demonstrated mechanistically. Here, we explored the effect of 2OHOA on mitochondria isolated by ultracentrifugation from U118MG glioblastoma cells. Mitochondria were analyzed by shotgun lipidomics, molecular dynamic simulations, spectrophotometric assays for determining respiratory complex activity, mass spectrometry for assessing beta oxidation and Seahorse technology for bioenergetic profiling. We showed that the main impact of 2OHOA on mitochondrial lipids is their hydroxylation, demonstrated by simulations to decrease co-enzyme Q diffusion in the liquid disordered membranes embedding respiratory complexes. This decreased co-enzyme Q diffusion can explain the inhibition of disjointly measured complexes I–III activity. However, it doesn’t explain how 2OHOA increases complex IV and state 3 respiration in intact mitochondria. This increased respiration probably allows mitochondrial oxidative phosphorylation to maintain ATP production against the 2OHOA-mediated inhibition of glycolytic ATP production. This work correlates 2OHOA function with its modulation of mitochondrial lipid composition, reflecting both 2OHOA anticancer activity and adaptation to it by enhancement of state 3 respiration.

Cercospora kikuchii (Tak. Matsumoto & Tomoy.) M.W. Gardner 1927 is an ascomycete fungal pathogen that causes Cercospora leaf blight and purple seed stain on soybean. Here, we report the first draft genome sequence and assembly of this pathogen. The C. kikuchii strain ARG_18_001 was isolated from soybean purple seed collected from San Pedro, Buenos Aires, Argentina, during the 2018 harvest. The genome was sequenced using a 2 × 150 bp paired-end method by Illumina NovaSeq 6000. The C. kikuchii protein-coding genes were predicted using FunGAP (Fungal Genome Annotation Pipeline). The draft genome assembly was 33.1 Mb in size with a GC-content of 53%. The gene prediction resulted in 14,856 gene models/14,721 protein coding genes. Genomic data of C. kikuchii presented here will be a useful resource for future studies of this pathosystem. The data can be accessed at GenBank under the accession number VTAY00000000 https://www.ncbi.nlm.nih.gov/nuccore/VTAY00000000. Keywords: Cercospora kikuchii, Draft genome, Next generation sequencing (NGS), Cercospora leaf blight (CLB), Purple seed stain (PSS), Agriculture, Bioinformatics, Fungal pathogens

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a heterogeneous, debilitating, and complex disease. Along with disabling fatigue, ME/CFS presents an array of other core symptoms, including autonomic nervous system (ANS) dysfunction, sustained inflammation, altered energy metabolism, and mitochondrial dysfunction. Here, we evaluated patients’ symptomatology and the mitochondrial metabolic parameters in peripheral blood mononuclear cells (PBMCs) and plasma from a clinically well-characterised cohort of six ME/CFS patients compared to age- and gender-matched controls. We performed a comprehensive cellular assessment using bioenergetics (extracellular flux analysis) and protein profiles (quantitative mass spectrometry-based proteomics) together with self-reported symptom measures of fatigue, ANS dysfunction, and overall physical and mental well-being. This ME/CFS cohort presented with severe fatigue, which correlated with the severity of ANS dysfunction and overall physical well-being. PBMCs from ME/CFS patients showed significantly lower mitochondrial coupling efficiency. They exhibited proteome alterations, including altered mitochondrial metabolism, centred on pyruvate dehydrogenase and coenzyme A metabolism, leading to a decreased capacity to provide adequate intracellular ATP levels. Overall, these results indicate that PBMCs from ME/CFS patients have a decreased ability to fulfill their cellular energy demands.

Leaf senescence is a complex mechanism controlled by multiple genetic and environmental variables. Different crops present a delay in leaf senescence with an important impact on grain yield trough the maintenance of the photosynthetic leaf area during the reproductive stage. Additionally, because of the temporal gap between the onset and phenotypic detection of the senescence process, candidate genes are key tools to enable the early detection of this process. In this sense and given the importance of some transcription factors as hub genes in senescence pathways, we present a comprehensive review on senescence-associated transcription factors, in model plant species and in agronomic relevant crops. This review will contribute to the knowledge of leaf senescence process in crops, thus providing a valuable tool to assist molecular crop breeding.

We here report molecular investigations of a missense mutation in the HSPE1 gene encoding the HSP10 subunit of the HSP60/ HSP10 chaperonin complex that assists protein folding in the mitochondrial matrix. The mutation was identified in an infant who came to clinical attention due to infantile spasms at three months of age. Clinical exome sequencing revealed heterozygosity for a HSPE1 NM_002157.2:c.217C>T de novo mutation causing replacement of leucine with phenylalanine at position 73 of the HSP10 protein. This variation has never been observed in public exome sequencing databases or the literature.To evaluate whether the mutation may be disease-associated we investigated its effects by in vitro and ex vivo studies. Our in vitro studies indicated that the purified mutant protein was functional, yet its thermal stability, spontaneous refolding propensity, and resistance to proteolytic treatment were profoundly impaired. Mass spectrometric analysis of patient fibroblasts revealed barely detectable levels of HSP10-p.Leu73Phe protein resulting in an almost 2-fold decrease of the ratio of HSP10 to HSP60 subunits. Amounts of the mitochondrial superoxide dismutase SOD2, a protein whose folding is known to strongly depend on the HSP60/HSP10 complex, were decreased to approximately 20% in patient fibroblasts in spite of unchanged SOD2 transcript levels. As a likely consequence, mitochondrial superoxide levels were increased about 2-fold. Although we cannot exclude other causative or contributing factors, our experimental data support the notion that the HSP10-p.Leu73Phe mutation could be the cause or a strong contributing factor for the disorder in the described patient.

Heat shock protein 60 (HSP60) forms together with heat shock protein 10 (HSP10) double-barrel chaperonin complexes that are essential for folding to the native state of proteins in the mitochondrial matrix space. Two extremely rare monogenic disorders have been described that are caused by missense mutations in the HSPD1 gene that encodes the HSP60 subunit of the HSP60/HSP10 chaperonin complex. Investigations of the molecular mechanisms underlying these disorders have revealed that different degrees of reduced HSP60 function produce distinct neurological phenotypes. While mutations with deleterious or strong dominant negative effects are not compatible with life, HSPD1 gene variations found in the human population impair HSP60 function and depending on the mechanism and degree of HSP60 dys- and malfunction cause different phenotypes. We here summarize the knowledge on the effects of disturbances of the function of the HSP60/HSP10 chaperonin complex by disease-associated mutations.

O objetivo desse estudo foi comparar a nova formação de placa dental e a quantidade de fluido gengival secretado apos a remoção profissional de placa dental supragengival —deplacagem- utilizando pasta profilática e clorexidina gel 1%. Teve como característica ser um estudo cruzado e com examinador cego para o tipo de tratamento realizado. A formação de placa dental foi avaliada pelo índice de placa de QUIGLEY e HEIN, modificado por TURESKY (1970), a cada 24 horas por um período de 96 horas na ausência de higiene bucal. O fluido gengival secretado foi coletado inicialmente e apos quatro dias de act mulo de placa dental. Os resultados foram avaliados por medianas, para o índice de placa, e medias para o fluido gengival secretado, sendo os resultados comparados pelo teste T de student e ANOVA (p > 0,05) respectivamente. Através da analise dos resultados pode-se observar que houve um aumento gradual na formação de placa dental nos dois grupos. A utilização de clorexidina gel 1% mostrou diferença estatística com maior número de superfícies livres de placa em 48 horas e com menor formação de placa dental em 72 horas quando comparada com a pasta profilática em situação semelhante. A quantidade de fluido gengival secretado não mostrou ser diferente alai:6 96 horas de acúmulo de placa dental, no inicio e final do estudo, com ambos os tratamentos. Concluiu-se que houve uma menor formação de placa apos o controle profissional supragengival associado com uma (mica aplicação de clorexidina gel 1% para um período de 72 horas.

Cultivated sunflower (Helianthus annuus L.), an important source of edible vegetable oil, shows rapid onset of senescence, which limits production by reducing photosynthetic capacity under specific growing conditions. Carbon for grain filling depends strongly on light interception by green leaf area, which diminishes during grain filling due to leaf senescence. Transcription factors (TFs) regulate the progression of leaf senescence in plants and have been well explored in model systems, but information for many agronomic crops remains limited. Here, we characterize the expression profiles of a set of putative senescence associated genes (SAGs) identified by a candidate gene approach and sunflower microarray expression studies. We examined a time course of sunflower leaves undergoing natural senescence and used quantitative PCR (qPCR) to measure the expression of 11 candidate genes representing the NAC, WRKY, MYB and NF-Y TF families. In addition, we measured physiological parameters such as chlorophyll, total soluble sugars and nitrogen content. The expression of Ha-NAC01, Ha-NAC03, Ha-NAC04, Ha-NAC05 and Ha-MYB01 TFs increased before the remobilization rate increased and therefore, before the appearance of the first physiological symptoms of senescence, whereas Ha-NAC02 expression decreased. In addition, we also examined the trifurcate feed-forward pathway (involving ORE1, miR164, and ethylene insensitive 2) previously reported for Arabidopsis. We measured transcription of Ha-NAC01 (the sunflower homolog of ORE1) and Ha-EIN2, along with the levels of miR164, in two leaves from different stem positions, and identified differences in transcription between basal and upper leaves. Interestingly, Ha-NAC01 and Ha-EIN2 transcription profiles showed an earlier up-regulation in upper leaves of plants close to maturity, compared with basal leaves of plants at pre-anthesis stages. These results suggest that the H. annuus TFs characterized in this work could play important roles as potential triggers of leaf senescence and thus can be considered putative candidate genes for senescence in sunflower.

BACKGROUND: The diagnosis of malignant hematologic diseases has become increasingly complex during the last decade. It is based on the interpretation of results from different laboratory analyses, which range from microscopy to gene expression profiling. Recently, a method for the analysis of RNA phenotypes has been developed, the nCounter technology (Nanostring® Technologies), which allows for simultaneous quantification of hundreds of RNA molecules in biological samples. We evaluated this technique in a Swiss multi-center study on eighty-six samples from acute leukemia patients. METHODS: mRNA and protein profiles were established for normal peripheral blood and bone marrow samples. Signal intensities of the various tested antigens with surface expression were similar to those found in previously performed Affymetrix microarray analyses. Acute leukemia samples were analyzed for a set of twenty-two validated antigens and the Pearson Correlation Coefficient for nCounter and flow cytometry results was calculated. RESULTS: Highly significant values between 0.40 and 0.97 were found for the twenty-two antigens tested. A second correlation analysis performed on a per sample basis resulted in concordant results between flow cytometry and nCounter in 44-100% of the antigens tested (mean = 76%), depending on the number of blasts present in a sample, the homogeneity of the blast population, and the type of leukemia (AML or ALL). CONCLUSIONS: The nCounter technology allows for fast and easy depiction of a mRNA profile from hematologic samples. This technology has the potential to become a valuable tool for the diagnosis of acute leukemias, in addition to multi-color flow cytometry.

Oligonucleotide-based microarrays with accurate gene coverage represent a key strategy for transcriptional studies in orphan species such as sunflower, H. annuus L., which lacks full genome sequences. The goal of this study was the development and functional annotation of a comprehensive sunflower unigene collection and the design and validation of a custom sunflower oligonucleotide-based microarray. A large scale EST (>130,000 ESTs) curation, assembly and sequence annotation was performed using Blast2GO (www.blast2go.de). The EST assembly comprises 41,013 putative transcripts (12,924 contigs and 28,089 singletons). The resulting Sunflower Unigen Resource (SUR version 1.0) was used to design an oligonucleotide-based Agilent microarray for cultivated sunflower. This microarray includes a total of 42,326 features: 1,417 Agilent controls, 74 control probes for sunflower replicated 10 times (740 controls) and 40,169 different non-control probes. Microarray performance was validated using a model experiment examining the induction of senescence by water deficit. Pre-processing and differential expression analysis of Agilent microarrays was performed using the Bioconductor limma package. The analyses based on p-values calculated by eBayes (p

Background Tatton- Brown- Rahman syndrome (TBRS) is a rare disorder, caused by DNMT3A heterozygous pathogenic variants, and first described in 2014. TBRS is characterised by overgrowth, intellectual disability, facial dysmorphism, hypotonia and musculoskeletal features, as well as neurological and psychiatric features. Cardiac manifestations have also been reported, mainly congenital malformations such as atrial septal defect, ventricular septal defect and cardiac valvular disease. Aortic dilatation has rarely been described. Methods Here we have undertaken a detailed clinical and molecular description of eight previously unreported individuals, who had TBRS and arterial dilatation and/ or dissection, mainly thoracic aortic aneurysm (TAA). We have also reviewed the seven previously published cases of TAA in individuals with TBRS to try to better delineate the vascular phenotype and to determine specific followup for this condition. Results We include eight new patients with TBRS who presented with arterial aneurysms mainly involving aorta. Three of these patients presented with dissection that required critical surgery. Conclusions Arterial aneurysms and dissections are a potentially lethal, age- dependent manifestation. The prevalence of aortic disease in individuals with TBRS is far in excess of that expected in the general population. This cohort, together with individuals previously published, illustrates the importance to consider dilatation/dissection, mainly in aorta but also in other arteries. Arterial vascular weakness may therefore also be a cardinal feature of TBRS and vascular surveillance is recommended. [ABSTRACT FROM AUTHOR]