Assessing Target Specificity of the Small Molecule Inhibitor MARIMASTAT to Snake Venom Toxins: A Novel Application of Thermal Proteome Profiling.

New treatments that circumvent the pitfalls of traditional antivenom therapies are critical to address the problem of snakebite globally. Numerous snake venom toxin inhibitors have shown promising cross-species neutralization of medically significant venom toxins in vivo and in vitro. The development of high-throughput approaches for the screening of such inhibitors could accelerate their identification, testing, and implementation and thus holds exciting potential for improving the treatments and outcomes of snakebite envenomation worldwide. Energetics-based proteomic approaches, including thermal proteome profiling and proteome integral solubility alteration (PISA) assays, represent "deep proteomics" methods for high throughput, proteome-wide identification of drug targets and ligands. In the following study, we apply thermal proteome profiling and PISA methods to characterize the interactions between venom toxin proteoforms in Crotalus atrox (Western Diamondback Rattlesnake) and the snake venom metalloprotease (SVMP) inhibitor marimastat. We investigate its venom proteome-wide effects and characterize its interactions with specific SVMP proteoforms, as well as its potential targeting of non-SVMP venom toxin families. We also compare the performance of PISA thermal window and soluble supernatant with insoluble precipitate using two inhibitor concentrations, providing the first demonstration of the utility of a sensitive high-throughput PISA-based approach to assess the direct targets of small molecule inhibitors for snake venom.

Serine protease inhibitor, SerpinA3n, regulates cardiac remodelling after myocardial infarction.

AIMS: Following myocardial infarction (MI), the heart repairs itself via a fibrotic repair response. The degree of fibrosis is determined by the balance between deposition of extracellular matrix (ECM) by activated fibroblasts and breakdown of nascent scar tissue by proteases that are secreted predominantly by inflammatory cells. Excessive proteolytic activity and matrix turnover has been observed in human heart failure, and protease inhibitors in the injured heart regulate matrix breakdown. Serine protease inhibitors (Serpins) represent the largest and the most functionally diverse family of evolutionary conserved protease inhibitors, and levels of the specific Serpin, SerpinA3, have been strongly associated with clinical outcomes in human MI as well as non-ischaemic cardiomyopathies. Yet, the role of Serpins in regulating cardiac remodelling is poorly understood. The aim of this study was to understand the role of Serpins in regulating scar formation after MI. METHODS AND RESULTS: Using a SerpinA3n conditional knockout mice model, we observed the robust expression of Serpins in the infarcted murine heart and demonstrate that genetic deletion of SerpinA3n (mouse homologue of SerpinA3) leads to increased activity of substrate proteases, poorly compacted matrix, and significantly worse post-infarct cardiac function. Single-cell transcriptomics complemented with histology in SerpinA3n-deficient animals demonstrated increased inflammation, adverse myocyte hypertrophy, and expression of pro-hypertrophic genes. Proteomic analysis of scar tissue demonstrated decreased cross-linking of ECM peptides consistent with increased proteolysis in SerpinA3n-deficient animals. CONCLUSION: Our study demonstrates a hitherto unappreciated causal role of Serpins in regulating matrix function and post-infarct cardiac remodelling.

Prevalence of sarcopenic obesity according to different diagnostic methods and cut-off points in candidates for bariatric surgery.

BACKGROUND AND AIMS: Sarcopenic obesity (SO) is defined as a combination of low strength and muscle mass along with excess adiposity. Our study aimed to determine the prevalence of sarcopenic obesity in candidates for bariatric surgery, according to ESPEN/EASO criteria using bioimpedance analysis (BIA) and dual-energy X-ray absorptiometry (DXA). METHODS: Retrospective study of adult patients (18-60 years) candidates for bariatric surgery (BMI ≥40 kg/m2 or ≥35 kg/m2 with associated complications). Adiposity was assessed by the percentage of fat mass by DXA, according to Gallagher's cut-off points. Muscle strength was measured by hand grip according to Sánchez-Torralvo, Dodds, and <-2SD cut-off points of healthy reference population. Muscle mass was determined by DXA (ALM/weight according to Batsis) and by BIA (SMM/weight according to Janssen and according to reference population). In addition, the agreement of the different diagnostic methods of sarcopenic obesity was analyzed. RESULTS: A total of 124 subjects were included in the study, with 71.8 % being women. The overall mean age was 42.6 (SD 8.9) years. SO prevalence was found to be 13 %-22 % applying BIA with the SMM/weight equation according to Janssen, 14 %-23 % utilizing BIA with the SMM/weight equation according to the reference population, and 13 %-23 % employing DXA with the ALM/weight equation following Batsis criteria, depending on the specific hand grip strength cut-off points used. In general, we found good or very good concordances with the different diagnostic methods (with kappa values between 0.6 and 0.97). CONCLUSIONS: The prevalence of SO according to ESPEN/EASO criteria in candidates for bariatric surgery was 13 %-23 % based on the diagnostic method and cut-off points used.

Comparative karyotype analysis of the red brocket deer (M. americana sensu lato and M. rufa) complex: evidence of drastic chromosomal evolution and implications on speciation process.

Chromosomal rearrangements are often associated with playing a role in the speciation process. However, the underlying mechanism that favors the genetic isolation associated with chromosomal changes remains elusive. In this sense, the genus Mazama is recognized by its high level of karyotype diversity among species with similar morphology. A cryptic species complex has been identified within the genus, with the red brocket deer (Mazama americana and Mazama rufa) being the most impressive example. The chromosome variation was clustered in cytotypes with diploid numbers ranging from 42 to 53 and was correlated with geographical location. We conducted an analysis of chromosome evolution of the red brocket deer complex using comparative chromosome painting and Bacterial Artificial Chromosome (BAC) clones among different cytotypes. The aim was to deepen our understanding of the karyotypic relationships within the red brocket, thereby elucidating the significant chromosome variation among closely related species. This underscores the significance of chromosome changes as a key evolutionary process shaping their genomes. The results revealed the presence of three distinct cytogenetic lineages characterized by significant karyotypic divergence, suggesting the existence of efficient post-zygotic barriers. Tandem fusions constitute the main mechanism driving karyotype evolution, following a few centric fusions, inversion X-autosomal fusions. The BAC mapping has improved our comprehension of the karyotypic relationships within the red brocket deer complex, prompting questions regarding the role of these changes in the speciation process. We propose the red brocket as a model group to investigate how chromosomal changes contribute to isolation and explore the implications of these changes in taxonomy and conservation.

Are New Ultra-Rapid-Acting Insulins Associated with Improved Glycemic Control and Reduced Hypoglycemia in Comparison to Conventional Rapid-Acting Insulins for Individuals with Type 1 and Type 2 Diabetes? A Systematic Review and Meta-Analysis.

Introduction: This study aimed to compare efficacy and safety of ultra-rapid-acting insulin analogs (URAIs; faster aspart [FAsp], ultra-rapid lispro [URLi], and technosphere insulin [TI]) with rapid-acting insulin analogs (RAI) in individuals with type 1 (T1D) or type 2 diabetes (T2D). Methods: Searching for randomized control trial comparing the effects of URAI versus RAI that lasted at least 12 weeks, we initially selected 15 studies for analysis. Three studies involving TI were excluded due to a high degree of heterogeneity. The final meta-analysis included only 12 studies with either FAsp or URLi. Results: Mealtime URAI significantly reduced overall early 1 h postprandial glycemia in individuals with T1D (-20.230 mg/dL [95% confidence interval, 95% CI -24.040 to -16.421]; P < 0.001; I2 = 33.42%) and those with T2D (-9.138 mg/dL [95% CI -12.612 to -5.663]; P < 0.001; I2 = 0%). However, the significant reduction in 2 h postprandial glucose remained only in individuals with T1D (-17.620 mg/dL [95% CI -26.047 to -9.193]; P < 0.001; I2 = 65.88%). These benefits were lost when URAI was administered postmeal. At 24-26 weeks, there was no significant difference in HbA1c between groups, but at 52 weeks, a slight reduction in HbA1c with mealtime URAI was observed (-0.080% [95% CI -0.147 to -0.013]; P = 0.019; I2 = 0%). No difference in weight or the rate of severe or confirmed hypoglycemia was observed. Only individuals with T1D showed a small, but significant increase in early 1-h hypoglycemia with URAI (1.468 [95% CI 1.235 to 1.747]; P < 0.001; I2 = 0%). Conclusion: Mealtime URAI improves 1 and 2 h postprandial glycemic control compared to RAI without increasing hypoglycemia or weight gain.

Cardiovascular and Locomotor Effects of Binary Mixtures of Common "Bath Salts" Constituents: Studies with Methylone, MDPV, and Caffeine in Rats.

Background and Purpose: The use of "Bath Salts" drug preparations has been associated with high rates of toxicity and death. Preparations often contain mixtures of drugs including multiple synthetic cathinones or synthetic cathinones and caffeine; however, little is known about whether interactions among "Bath Salts" constituents contribute to the adverse effects often reported in users. Experimental Approach: This study used adult male Sprague-Dawley rats to characterize the cardiovascular effects, locomotor effects, and pharmacokinetics of methylone, MDPV, and caffeine, administered alone and as binary mixtures. Dose-addition analyses were used to determine the effect levels predicted for a strictly additive interaction for each dose pair. Key Results: Methylone, MDPV, and caffeine increased heart rate and locomotion, with methylone producing the largest increase in heart rate, MDPV producing the largest increase in locomotor activity, and caffeine being the least effective in stimulating heart rate and locomotor activity. MDPV and caffeine increased mean arterial pressure, with caffeine being more effective than MDPV. The nature of the interactions between methylone and MDPV tended toward sub-additivity for all endpoints, whereas interactions between MDPV or methylone and caffeine tended to be additive or sub-additive for cardiovascular endpoints, and additive or supra-additive for increases in locomotion. No pharmacokinetic interactions were observed between individual constituents, but methylone displayed non-linear pharmacokinetics at the largest dose evaluated. Conclusion and Implications: These findings demonstrate that the composition of "Bath Salts" preparations can impact both cardiovascular and locomotor effects and suggest that such interactions among constituent drugs could contribute to the "Bath Salts" toxidrome reported by human users.

Sensory nerve release of CGRP increases tumor growth in HNSCC by suppressing TILs.

BACKGROUND: Perineural invasion (PNI) and nerve density within the tumor microenvironment (TME) have long been associated with worse outcomes in head and neck squamous cell carcinoma (HNSCC). This prompted an investigation into how nerves within the tumor microenvironment affect the adaptive immune system and tumor growth. METHODS: We used RNA sequencing analysis of human tumor tissue from a recent HNSCC clinical trial, proteomics of human nerves from HNSCC patients, and syngeneic orthotopic murine models of HPV-unrelated HNSCC to investigate how sensory nerves modulate the adaptive immune system. FINDINGS: Calcitonin gene-related peptide (CGRP) directly inhibited CD8 T cell activity in vitro, and blocking sensory nerve function surgically, pharmacologically, or genetically increased CD8 and CD4 T cell activity in vivo. CONCLUSIONS: Our data support sensory nerves playing a role in accelerating tumor growth by directly acting on the adaptive immune system to decrease Th1 CD4 T cells and activated CD8 T cells in the TME. These data support further investigation into the role of sensory nerves in the TME of HNSCC and points toward the possible treatment efficacy of blocking sensory nerve function or specifically inhibiting CGRP release or activity within the TME to improve outcomes. FUNDING: 1R01DE028282-01, 1R01DE028529-01, 1P50CA261605-01 (to S.D.K.), 1R01CA284651-01 (to S.D.K.), and F31 DE029997 (to L.B.D.).

Factores asociados a la mala adherencia al tratamiento antirretroviral en pacientes VIH positivo

Introducción: La infección por el virus de inmunodeficiencia humana representa un problema de salud pública en el mundo. La adherencia a la terapia antirretroviral es necesaria para alcanzar un estado de supresión viral máxima y disminuir la tasa de infecciones oportunistas. Sin embargo, diversos son los factores que pueden influir en la adherencia terapéutica. Objetivo: Identificar los factores de riesgo asociados a la mala adherencia a la terapia antirretroviral en pacientes con el virus de inmunodeficiencia humana. Métodos: Se empleó un estudio con diseño observacional, analítico, de tipo casos y controles. El tamaño muestral fue de 276, a partir del cual 138 fueron casos y 138 controles. Los datos recolectados fueron analizados utilizando métodos de estadística descriptiva y estadística inferencial. Resultados: El análisis multivariado determinó una asociación estadísticamente significativa entre la variable adherencia al tratamiento antirretroviral y las variables depresión (odds ratio OR ajustado= 2,15), esquema de terapia antirretroviral alternativo (OR ajustado= 2,40), efectos secundarios (OR ajustado= 4,24) y cambio en la adherencia durante el estado de emergencia (OR ajustado= 5,67), independientemente de las otras variables intervinientes. Conclusiones: La depresión, el uso del esquema de terapia antirretroviral alternativo, la presencia de efectos secundarios a terapia antirretroviral y el cambio en la adherencia durante el estado de emergencia aumentan el riesgo de mala adherencia a la terapia antirretroviral.

Introduction: Human immunodeficiency virus infection represents a public health public problem in the world. Adherence to antiretroviral therapy is necessary to achieve a state of maximum viral suppression and decrease the rate of opportunistic infections. However, there are several factors that can influence therapeutic adherence. Objective: To identify the risk factors associated with poor adherence to antiretroviral therapy in patients with the human immunodeficiency virus. Methods: A study with an observational, analytical, case-control type design was used. The sample size was 276, from which 138 were cases and 138 controls. The collected data was analyzed using methods of descriptive statistics and inferential statistics. Results: The multivariate analysis determined a statistically significant association between the variable adherence to antiretroviral treatment and the variables depression (adjusted Odds Ratio OR = 2.15), alternative antiretroviral therapy scheme (adjusted OR= 2.40), side effects (adjusted OR= 4.24), and change in adherence during the state of emergency (adjusted OR= 5.67) independently of the other intervening variables. Conclusions: Depression, the use of alternative antiretroviral therapy regimens, the presence of side effects to antiretroviral therapy, and the change in adherence during the state of emergency increase the risk of poor adherence to antiretroviral therapy.

Transcervical artificial insemination in the brown brocket deer (Subulo gouazoubira): a promising method for assisted reproduction in deer.

The present study aimed to test the efficiency of transcervical artificial insemination techniques with cervical immobilization (TCAI-CI) or cervical traction (TCAI-CT), associated or not with the use of oxytocin (OT) as a protocol for cervical dilation, in the brown brocket deer (Subulo gouazoubira). The study was carried out in a crossover design using four adult females in two replicates with an interval of 60 days. Estrus was synchronized with oral melengestrol acetate (MGA) associated with estradiol benzoate and sodium cloprostenol. TCAI techniques were performed from 18 to 24 h after estrus onset. All females received either an i.v. application of 50 IU of OT (G-OT, n = 4) or 1 mL of saline solution (G-Control, n = 4) 20 min before the TCAI procedure. The TCAIs were performed using frozen-thawed semen motility 40%, vigor 3, acrosome integrity 87%, membrane integrity of 95% and 13% of total post-thaw defects from the same batch. Behavioral estrus was observed in 100% of the females, in both replicates. It was achieved a 50% (4/8) success of cervical transposition with semen deposition in the uterine. Regarding inseminations, most of them (87.5%) were performed using the TCAI-CT technique, and the overall conception rate was 50%. Cervical transposition times (< 1 min) and TCAI procedures (~ 17 min) were considered satisfactory. Thus, the performance of the TCAI-CI and TCAI-CT techniques was successful, regardless of using OT as a cervical dilation protocol. This procedure is proposed as a method of choice for artificial insemination with greater applicability in different conservation centers, compared to more advanced reproductive biotechniques, and with a favorable impact on the conservation of deer species.

Col1a2-Deleted Mice Have Defective Type I Collagen and Secondary Reactive Cardiac Fibrosis with Altered Hypertrophic Dynamics.

RATIONALE: The adult cardiac extracellular matrix (ECM) is largely comprised of type I collagen. In addition to serving as the primary structural support component of the cardiac ECM, type I collagen also provides an organizational platform for other ECM proteins, matricellular proteins, and signaling components that impact cellular stress sensing in vivo. OBJECTIVE: Here we investigated how the content and integrity of type I collagen affect cardiac structure function and response to injury. METHODS AND RESULTS: We generated and characterized Col1a2-/- mice using standard gene targeting. Col1a2-/- mice were viable, although by young adulthood their hearts showed alterations in ECM mechanical properties, as well as an unanticipated activation of cardiac fibroblasts and induction of a progressive fibrotic response. This included augmented TGFß activity, increases in fibroblast number, and progressive cardiac hypertrophy, with reduced functional performance by 9 months of age. Col1a2-loxP-targeted mice were also generated and crossed with the tamoxifen-inducible Postn-MerCreMer mice to delete the Col1a2 gene in myofibroblasts with pressure overload injury. Interestingly, while germline Col1a2-/- mice showed gradual pathologic hypertrophy and fibrosis with aging, the acute deletion of Col1a2 from activated adult myofibroblasts showed a loss of total collagen deposition with acute cardiac injury and an acute reduction in pressure overload-induce cardiac hypertrophy. However, this reduction in hypertrophy due to myofibroblast-specific Col1a2 deletion was lost after 2 and 6 weeks of pressure overload, as fibrotic deposition accumulated. CONCLUSIONS: Defective type I collagen in the heart alters the structural integrity of the ECM and leads to cardiomyopathy in adulthood, with fibroblast expansion, activation, and alternate fibrotic ECM deposition. However, acute inhibition of type I collagen production can have an anti-fibrotic and anti-hypertrophic effect.

Evaluation of minimally invasive estrus synchronization protocols in brown brocket deer (Subulo gouazoubira).

This study aimed to evaluate minimally invasive protocols for estrus synchronization in the brown brocket deer (Subulo gouazoubira). Females were submitted to Latin square design, in different treatments. All females received 0.25 mg of estradiol benzoate on the first day of treatment, concomitant with one of the following sources of progesterone: (1) DIP: an intravaginal progesterone releasing device for eight days, (2) MGA1x: once a day (in the morning) oral dose of 1 mg melengestrol acetate for eight days, (3) MGA2x: twice a day (morning and afternoon) oral doses of 0.5 mg of MGA for eight days, (4) P4LA: a single i.m. administration of 75 mg of long-acting progesterone (P4LA). Eight days after the beginning of each treatment, females received an i.m. administration of 265 µg of prostaglandin (PGF2α; cloprostenol). Treatment efficacy was evaluated by manifestation of behavioral estrus after treatment and concentration of fecal progesterone metabolites (FPM). The time to onset of estrus in treatment P4LA was significantly longer (180 ± 38.9 h) compared to DIP (63 ± 6.6 h), MGA1x (53 ± 14.4 h) and MGA2x (41 ± 10.1 h) (P = 0.008). According to individual baseline FPM and FPM concentration during the days after estrus, the corpus luteum formation was suggested in all females which responded to the treatments (93.75 %). Low synchrony, longer interval between PGF2α administration and onset of estrus suggest that the P4LA dose (75 mg) is too high and not effective for S. gouazoubira. DIP, MGA 1x and MGA 2x, were effective in estrus synchronization.

Revalidation of Passalites Gloger, 1841 for the Amazon brown brocket deer P.nemorivagus (Cuvier, 1817) (Mammalia, Artiodactyla, Cervidae).

Mazamanemorivaga (Cuvier, 1817) is a gray brocket deer that inhabits the Amazon region. An assessment of previous studies revealed inconsistencies in its current taxonomic classification, suggesting the need for an update in its genus classification. A taxonomic repositioning of this species is proposed through the collection of a specimen from its type locality (French Guiana) with subsequent morphological (coloring pattern, body measurements, and craniometry), cytogenetics (G Band, C Band, conventional Giemsa, Ag-NOR staining, and BAC probe mapping), and molecular phylogenetic analysis (mitochondrial genes Cyt B of 920 bp, COI I of 658 bp, D-loop 610 bp), and comparisons with other specimens of the same taxon, as well as other Neotropical deer species. The morphological and cytogenetic differences between this and other Neotropical Cervidae confirm the taxon as a unique and valid species. The phylogenetic analysis evidenced the basal position of the M.nemorivaga specimens within the Blastocerina clade. This shows early diversification and wide divergence from the other species, suggesting that the taxon should be transferred to a different genus. A taxonomic update of the genus name is proposed through the validation of Passalites Gloger, 1841, with Passalitesnemorivagus (Cuvier, 1817) as the type species. Future research should focus on evaluating the potential existence of other species within the genus Passalites, as suggested in the literature.

Valoración de la baja masa y fuerza muscular en una población de control / Assessment of low muscle mass and strength in a control population

Introducción y objetivos: la baja masa muscular (BMM) condiciona el estado nutricional de un individuo y tiene implicaciones en la calidad y el pronóstico de vida. El objetivo de este trabajo fue evaluar la composición corporal y determinar los valores de normalidad en el diagnóstico de BMM en un grupo de control de individuos sanos. Material y método: estudio transversal de voluntarios sanos con edades entre 18 y 45 años, con un índice de masa corporal (IMC) < 30 kg/m2. Se realizó un estudio descriptivo incluyendo variables demográficas, clínicas, antropométricas y de composición corporal (mediante bioimpedancia, TANITA MC 780 MA; TANITA, Tokio, Japón), con estratificación por edad, sexo e IMC. Se determinaron los valores correspondientes a -1/-2 desviaciones estándar (DE) para considerar una baja masa/función muscular. Resultados: se incluyeron 67 pacientes, el 71,60 % mujeres, con una mediana de edad de 28,29 (RIC: 4,05) años. Los varones presentan mayor peso, IMC, masa libre de grasa (MLG), masa muscular (MM), masa muscular apendicular (MMA), índice de masa muscular apendicular (IMMA) y dinamometría con respecto a las mujeres. Se determinaron los valores de -1/-2 DE de los distintos parámetros musculares en función del sexo. Conclusiones: este estudio determina los valores normales de BMM en una población sana y joven de nuestro medio, y los índices más empleados para expresarla, lo que permitirá diagnosticar la BMM en situaciones patológicas empleando el valor correspondiente a -2 DE. (AU)

Background and aims: low muscle mass (LMM) conditions the nutritional status of an individual and has implications for quality of life and prognosis. The aim of this study was to evaluate body composition and determine normal values in the diagnosis of LMM in a control group of healthy individuals. Methods: a cross-sectional study of healthy volunteers aged 18 to 45 years with body mass index (BMI) < 30 kg/m2, from January 2021 to October 2021. A descriptive study was performed including demographic, clinical, anthropometric, and body composition variables (by bioimpedance, TANITA MC 780 MA; TANITA, Tokyo, Japan), stratified by age, sex and BMI. Values corresponding to -1/-2 standard deviations (SD) were determined to consider low muscle mass/function. Results: we included 67 patients, 71,60 % women, with a median age of 28.29 (IQR 4.05) years. Males presented higher weight, BMI, fat-free mass (FFM), muscle mass (MM), appendicular lean mass (ALM), appendicular lean mass index (ALMI), and dynamometry values when compared to females. The -1/-2 SD values of the various muscle parameters were determined according to sex. Conclusion: this study determined normal LMM values in healthy and young people, and the most commonly used indexes to express it, which will allow the diagnosis of LMM in disease-related situations using the corresponding -2 DS value. (AU)

[Assessment of low muscle mass and strength in a control population]. / Valoración de la baja masa y fuerza muscular en una población de control.

Introduction: Background and aims: low muscle mass (LMM) conditions the nutritional status of an individual and has implications for quality of life and prognosis. The aim of this study was to evaluate body composition and determine normal values in the diagnosis of LMM in a control group of healthy individuals. Methods: a cross-sectional study of healthy volunteers aged 18 to 45 years with body mass index (BMI) < 30 kg/m2. A descriptive study was performed including demographic, clinical, anthropometric, and body composition variables (by bioimpedance, TANITA MC 780 MA; TANITA, Tokyo, Japan), stratified by age, sex and BMI. Values corresponding to -1/-2 standard deviations (SD) were determined to consider low muscle mass/function. Results: we included 67 patients, 71,60 % women, with a median age of 28.29 (IQR 4.05) years. Males presented higher weight, BMI, fat-free mass (FFM), muscle mass (MM), appendicular lean mass (ALM), appendicular lean mass index (ALMI), and dynamometry values when compared to females. The -1/-2 SD values of the various muscle parameters were determined according to sex. Conclusion: this study determined normal LMM values in healthy and young people, and the most commonly used indexes to express it, which will allow the diagnosis of LMM in disease-related situations using the corresponding -2 DS value.

Introducción: Introducción y objetivos: la baja masa muscular (BMM) condiciona el estado nutricional de un individuo y tiene implicaciones en la calidad y el pronóstico de vida. El objetivo de este trabajo fue evaluar la composición corporal y determinar los valores de normalidad en el diagnóstico de BMM en un grupo de control de individuos sanos. Material y método: estudio transversal de voluntarios sanos con edades entre 18 y 45 años, con un índice de masa corporal (IMC) < 30 kg/m2. Se realizó un estudio descriptivo incluyendo variables demográficas, clínicas, antropométricas y de composición corporal (mediante bioimpedancia, TANITA MC 780 MA; TANITA, Tokio, Japón), con estratificación por edad, sexo e IMC. Se determinaron los valores correspondientes a -1/-2 desviaciones estándar (DE) para considerar una baja masa/función muscular. Resultados: se incluyeron 67 pacientes, el 71,60 % mujeres, con una mediana de edad de 28,29 (RIC: 4,05) años. Los varones presentan mayor peso, IMC, masa libre de grasa (MLG), masa muscular (MM), masa muscular apendicular (MMA), índice de masa muscular apendicular (IMMA) y dinamometría con respecto a las mujeres. Se determinaron los valores de -1/-2 DE de los distintos parámetros musculares en función del sexo. Conclusiones: este estudio determina los valores normales de BMM en una población sana y joven de nuestro medio, y los índices más empleados para expresarla, lo que permitirá diagnosticar la BMM en situaciones patológicas empleando el valor correspondiente a -2 DE.

Increased incidence of Graves' disease during the SARS-CoV2 pandemic.

INTRODUCTION: COVID-19 has a wide spectrum of clinical severity and there is evidence that SARS-Cov2 affects several organs and systems. Among the organs affected since the beginning of the pandemic, the relationship between SARS-CoV-2 infection and thyroid involvement has been demonstrated. Novel and highly effective messenger RNA and DNA-based vaccines have been rapidly developed to decrease SARS-CoV-2 morbidity and mortality. Early after mass vaccinations, cases of thyroid dysfunction mainly including episodes of subacute thyroiditis, began to be reported like adverse effects. The objective of this study is to determine the impact of the pandemic, both due to SARS-CoV2 infections and vaccinations, on the incidence of Graves' disease (GD). METHODS: Cross-sectional, observational study comparing incidence of GD in adult population (over 18 years) before (2017-2019) and after (2020-2021) Covid-19 pandemic. Only patients with new cases of GD, no relapsed diseases, were included. SARS-CoV-2 diagnosis was based on nucleic acid amplification tests on nasopharyngeal swabs or measurement of class M and class G antibodies to SARS-CoV-2 by highly specific assays. Data on incidence and vaccination related to SARS-CoV-2 infection were obtained from the public records from Castilla y León autonomous regional government. RESULTS: A total of 180 subjects were diagnosed and treated for GD during the study period. We observed a notable increase in expected GD cases in 2021 compared to 2017-19. The number of GD cases was higher in the second (Q2) quarter. Among 2021 GD cases, 42/66 patients (63.6%) had been vaccinated in the 90 days before symptom onset, but none of them in the first quarter of the year. A total of 97.7% were women with a mean age of 48.9 (SD 15.6) years. On average they were diagnosed 19.9 (SD 17.6) days after receiving the vaccine. A total of 7/42 (16.67%) had another previously diagnosed autoimmune disease and 11/42 (26.19%) were smokers. DISCUSSION: Our results show a notable increase in the incidence of GD during the year 2021, specially in women with a history of smoking. Hyper activation of the immune system induced by SARS-CoV2 and by the recently released SARS-COV-2 vaccines has been highlighted in recent months. To assess whether this observed increase in the incidence of GD is sustained in the coming years or has simply been a precipitous trigger for individuals who were already predisposed to develop the disease, future studies will be needed.

Consensus document on the management of hyponatraemia of the Acqua Group of the Spanish Society of Endocrinology and Nutrition.

INTRODUCTION: Hyponatremia is the most prevalent electrolyte disorder in the outpatient and inpatient settings. Despite this frequency, hyponatremia, including severe hyponatremia, is frequently underestimated and inadequately treated, thus highlighting the need to produce consensus documents and clinical practice guidelines geared towards improving the diagnostic and therapeutic approach to it in a structured fashion. MATERIAL AND METHODS: Members of the Acqua Group of the Spanish Society of Endocrinology and Nutrition (SEEN) met using a networking methodology over a period of 20 months (between October 2019 and August 2021) with the aim of discussing and developing an updated guideline for the management of hyponatraemia. A literature search of the available scientific evidence for each section presented in this document was performed. RESULTS: A document with 8 sections was produced, which sets out to provide updated guidance on the most clinically relevant questions in the management of hyponatraemia. The management of severe hyponatraemia is based on the i.v. administration of a 3% hypertonic solution. For the management of chronic euvolemic hyponatraemia, algorithms for the initiation of treatment with the two pharmacological therapeutic options currently available in Spain are presented: urea and tolvaptan. CONCLUSIONS: This document sets out to simplify the approach to and the treatment of hyponatraemia, making it easier to learn and thus improve the clinical approach to hyponatremia.

Cytogenetic Mapping of Cattle BAC Probes for the Hypothetical Ancestral Karyotype of the Family Cervidae.

Cervids are characterized by their greatest karyotypic diversity among mammals. A great diversity of chromosome numbers in notably similar morphological groups leads to the existence of several complexes of cryptic species and taxonomic uncertainties. Some deer lineages, such as those of Neotropical deer, stand out for a rapid chromosomal reorganization and intraspecific chromosome polymorphisms, which have not been properly explored yet. For that reason, we contribute to the study of deer karyotype diversity and taxonomy by producing and characterizing new molecular cytogenetic markers for the gray brocket deer (Subulo gouazoubira), a deer species that retained the hypothetical ancestral karyotype of Cervidae. We used bacterial artificial chromosome (BAC) clones derived from the cattle genome (Bos taurus) as markers, which were hybridized on S. gouazoubira metaphase chromosomes. In total, we mapped 108 markers, encompassing all gray brocket deer chromosomes, except the Y chromosome. The detailed analysis of fluorescent in situ hybridization results showed 6 fissions and 1 fusion as interchromosomal rearrangements that have separated cattle and gray brocket deer karyotypes. Each group of BAC probes derived from bovine chromosome pairs 1, 2, 5, 6, 8, and 9 showed hybridization signals on 2 different chromosomes, while pairs 28 and 26 are fused in tandem in a single acrocentric chromosome in S. gouazoubira. Furthermore, the BAC markers detected the occurrence of intrachromosomal rearrangements in the S. gouazoubira chromosomes homologous to pair 1 and the X chromosome of cattle. We present a karyotypic map of the 108 new markers, which will be of great importance for future karyotypic evolution studies in cervids and, consequently, help in their conservation and taxonomy resolution.

Establishment of the upstream processing for renewable production of hydrogen using vermicomposting-tea and molasses as substrate.

This study aimed to establish the optimal operational conditions for hydrogen production using vermicomposting-tea and sugarcane molasses as substrate. The experiments were carried out by triplicate in 110 ml serological bottles, a Box-Behnken design of experiments was performed in anaerobic dark conditions. The maximal hydrogen production (HP), hydrogen production rate (HPR), and hydrogen yield (HY) attained were 1021.0 mlL-1, 5.32 mlL-1h-1, and 60.3 mlLH2-1/gTCC, respectively. The statistical model showed that the optimal operational conditions for pH, molasses concentration, and temperature were 6.5; 30 % (v/v) and 25 °C. The bioreactor run showed 17.202 L of hydrogen, 0.58 Lh-1, and 77.2 mlH2gTCC-1 For HP, HPR, and HY. Chemometric analysis for the volatile fatty acids obtained at the fermentation showed that only two principal components are required to explain 90 % of the variance. The representative pathways for hydrogen production were acetic and butyric acids. This study established the operational conditions for the upstream processing amenable to pilot and industrial-scale operations. Our results add value to molasses within the circular economy for hydrogen production using a novel consortium from vermicompost.

Bioelectrical impedance analysis as an alternative to dual-energy x-ray absorptiometry in the assessment of fat mass and appendicular lean mass in patients with obesity.

OBJECTIVE: Obesity is a challenge for bioelectrical impedance analysis (BIA) estimations of skeletal muscle and fat mass (FM), and none of the equations used for appendicular lean mass (ALM) have been developed for people with obesity. By using different equations and proposing a new equation, this study aimed to assess the estimation of FM and ALM using BIA compared with dual-energy x-ray absorptiometry (DXA) as a reference method in a cohort of people with severe obesity. METHODS: This cross-sectional study compared a multifrequency BIA (TANITA MC-780A) versus DXA for body composition assessment in adult patients with severe obesity (body mass index [BMI] of >35 kg/m2). Comparisons between measured (DXA) and predicted (BIA) data for FM and ALM were performed using the original proprietary equations of the device and the equations proposed by Kyle, Sergi, and Yamada. Bland-Altman plots were drawn to evaluate the agreement between DXA and BIA, calculating bias and limits of agreement (LOA). Reliability was analyzed using intraclass correlation coefficient (ICC). Stepwise multiple regression analysis was used to derive a new equation to predict ALM in patients with obesity and was validated in a subsample of our cohort. RESULTS: In this study, 115 patients (72.4% women) with severe obesity (mean BMI of 46.1 [5.2] kg/m2) were included (mean age 43.5 [8.6] y). FMDXA was 61.4 (10.1) kg, FMBIA was 57.9 (10.3) kg, and ICC was 0.925 (P < 0.001). Bias was -3.4 (4.4) kg (-5.2%), and LOA was -14.0, +7.3 kg. Using the proprietary equations, ALMDXA was 21.8 (4.7) kg and ALMBIA was 29.0 (6.8) kg with an ICC 0.868, bias +7.3 (4.0) kg (+34.1%) and LOA -0.5, +15.1. When applying other equations for ALM, the ICC for Sergi, et al. was 0.880, the ICC for Kyle, et al. was 0.891, and the best ICC estimation for Yamada, et al. was 0.914 (P < 0.001). Bias was +2.8 (2.8), +4.1 (2.9), and +2.7 (2.8) kg, respectively. The best-fitting regression equation to predict ALMDXA in our population derived from a development cohort (n = 77) was: ALM = 13.861 + (0.259 x H2/Z) - (0.085 x age) - (3.983 x sex [0 = men; 1 = women]). When applied to our validation cohort (n = 38), the ICC was 0.864, and the bias was the lowest compared with the rest of the equations +0.3 (+0.5) kg (+2.7%) LOA -5.4, +6.0 kg. CONCLUSION: BIA using multifrequency BIA in people with obesity is reliable enough for the estimation of FM, with good correlation and low bias to DXA. Regarding the estimation of ALM, BIA showed a good correlation with DXA, although it overestimated ALM, especially when proprietary equations were used. The use of equations developed using the same device improved the prediction, and our new equation showed a low bias for ALM.

Revalidation of Mazama rufa (Illiger 1815) (Artiodactyla: Cervidae) as a Distinct Species out of the Complex Mazama americana (Erxleben 1777).

The red brocket deer Mazama americana Erxleben, 1777 is considered a polyphyletic complex of cryptic species with wide chromosomal divergence. Evidence indicates that the observed chromosomal divergences result in reproductive isolation. The description of a neotype for M. americana allowed its genetic characterization and represented a comparative basis to resolve the taxonomic uncertainties of the group. Thus, we designated a neotype for the synonym Mazama rufa Illiger, 1815 and tested its recognition as a distinct species from the M. americana complex with the analysis of morphological, cytogenetic and molecular data. We also evaluated its distribution by sampling fecal DNA in the wild. Morphological data from craniometry and body biometry indicated an overlap of quantitative measurements between M. rufa and the entire M. americana complex. The phylogenetic hypothesis obtained through mtDNA confirmed the reciprocal monophyly relationship between M. americana and M. rufa, and both were identified as distinct molecular operational taxonomic units by the General Mixed Yule Coalescent species delimitation analysis. Finally, classic cytogenetic data and fluorescence in situ hybridization with whole chromosome painting probes showed M. rufa with a karyotype of 2n = 52, FN = 56. Comparative analysis indicate that at least fifteen rearrangements separate M. rufa and M. americana (sensu stricto) karyotypes, which confirmed their substantial chromosomal divergence. This divergence should represent an important reproductive barrier and allow its characterization as a distinct and valid species. Genetic analysis of fecal samples demonstrated a wide distribution of M. rufa in the South American continent through the Atlantic Forest, Cerrado and south region of Amazon. Thus, we conclude for the revalidation of M. rufa as a distinct species under the concept of biological isolation, with its karyotype as the main diagnostic character. The present work serves as a basis for the taxonomic review of the M. americana complex, which should be mainly based on cytogenetic characterization and directed towards a better sampling of the Amazon region, the evaluation of available names in the species synonymy and a multi-locus phylogenetic analysis.