High-Throughput DNA-Encoded Libraries Affinity Selection Platform for Binder Identification with Solid Support Protein Immobilization.

DNA-encoded libraries (DELs) have demonstrated to be one of the most powerful technologies within the ligand identification toolbox, widely used either in academia or biotech and pharma companies. DEL methodology utilizes affinity selection (AS) as the approach to interrogate the protein of interest for the identification of binders. Here we present a high-throughput, fully automated AS platform developed to fulfill industrial standards and compatible with different assay formats to improve the reproducibility of the AS process for DEL binders identification. This platform is flexible enough to virtually set aside all kinds of DELs and AS methods and conditions using immobilized proteins. It bears the two main immobilization methods to support of the proteins of interest: magnetic beads or resin tip columns. A combination of a broad variety of protocol options with a wide range of different experimental conditions can be set up with a throughput of 96 samples at the same time. In addition, small modifications of the protocols provide the platform with the versatility to run not only the routine DEL screens, but also test covalent libraries, the successful immobilization of the proteins of interest, and many other experiments that may be required. This versatile AS platform for DEL can be a powerful instrument for direct application of the technology in academic and industry settings.

Design, synthesis and validation of a new Crimped Head-Piece for DNA-Encoded libraries generation.

Codification of DNA Encoded Libraries (DELs) is critical for successful ligand identification of molecules that bind a protein of interest (POI). There are different encoding strategies that permit, for instance, the customization of a DEL for testing single or dual pharmacophores (single strand DNA) or for producing and screening large diversity libraries of small molecules (double strand DNA). Both approaches challenges, either from the synthetic and encoding point of view, or from the selection methodology to be utilized for the screening. The Head-Piece contains the DNA sequence that is attached to a chemical compound, allowing the encoding of each molecule with a unique DNA tag. Designing the Head-Piece for a DNA-encoded library involves careful consideration of several key aspects including DNA barcode identity, sequence length and attachment chemistry. Here we describe a double stranded DNA versatile Head-Piece that can be used for the generation of single or dual pharmacophore libraries, but also shows other advanced DEL functionalities, stability and enlarged encoding capacity.

Stochastic Syncing in Sinusoidally Driven Atomic Orbital Memory.

Stochastically fluctuating multiwell systems are a promising route toward physical implementations of energy-based machine learning and neuromorphic hardware. One of the challenges is finding tunable material platforms that exhibit such multiwell behavior and understanding how complex dynamic input signals influence their stochastic response. One such platform is the recently discovered atomic Boltzmann machine, where each stochastic unit is represented by a binary orbital memory state of an individual atom. Here, we investigate the stochastic response of binary orbital memory states to sinusoidal input voltages. Using scanning tunneling microscopy, we investigated orbital memory derived from individual Fe and Co atoms on black phosphorus. We quantify the state residence times as a function of various input parameters such as frequency, amplitude, and offset voltage. The state residence times for both species, when driven by a sinusoidal signal, exhibit synchronization that can be quantitatively modeled by a Poisson process based on the switching rates in the absence of a sinusoidal signal. For individual Fe atoms, we also observe a frequency-dependent response of the state favorability, which can be tuned by the input parameters. In contrast to Fe, there is no significant frequency dependence in the state favorability for individual Co atoms. Based on the Poisson model, the difference in the response of the state favorability can be traced to the difference in the voltage-dependent switching rates of the two different species. This platform provides a tunable way to induce population changes in stochastic systems and provides a foundation toward understanding driven stochastic multiwell systems.

The function of wing bullae in mayflies (Insecta: Ephemeroptera) reveals new insights into the early evolution of Pterygota.

BACKGROUND: Mayflies are basal winged insects of crucial importance for the understanding of the early evolution of Pterygota. Unlike all other insects, they have two successive winged stages, the subimago and the imago. Their forewings feature so-called bullae, which are desclerotized spots in the anterior main veins. Up to now, they have been considered to play a major role in wing bending during flight. RESULTS: We investigated bullae by multiple methods to reveal their structure and arrangement and to gain new information on the evolution of insect flight. Bullae are mostly present in the anterior negative wing veins, disrupting the otherwise rigid veins. High-speed videography reveals that mayfly wings do not bend during flight. Likewise, different arrangements of bullae in different species do not correlate with different modes of flying. Observations on the moulting of subimagines unravel that they are essential for wing bending during the extraction of the imaginal wing from the subimaginal cuticle. Bullae define predetermined bending lines, which, together with a highly flexible wing membrane enriched with resilin, permit wing bending during subimaginal moulting. Bullae are only absent in those species that remain in the subimaginal stage or that use modified modes of moulting. Bullae are also visible in fossil mayflies and can be traced back to stemgroup mayflies of the Early Permian, the 270 million years old Protereismatidae, which most probably had bullae in both fore- and hind wings. CONCLUSIONS: Bullae in mayfly wings do not play a role in flight as previously thought, but are crucial for wing bending during subimaginal moulting. Thus, the presence of bullae is a reliable morphological marker for a subimaginal life stage, confirming the existence of the subimago already in Permian Protereismatidae. A thorough search for bullae in fossils of other pterygote lineages may reveal wheather they also had subimagines and at what point in evolution this life stage was lost. In mayflies, however, the subimago may have been retained due to selective advantages in connection with the transition from aquatic to terrestrial life or due to morphological requirements for a specialized mating flight.

The standardized extract of Centella asiatica L. Urb attenuates the convulsant effect induced by lithium/pilocarpine without affecting biochemical and haematological parameters in rats.

BACKGROUND: Status epilepticus (SE) is a type of epileptic activity characterized by a failure of the inhibitory mechanisms that limit seizures, which are mainly regulated by the GABAergic system. This imbalance increases glutamatergic neurotransmission and consequently produces epileptic activity. It is also associated with oxidative stress due to an imbalance between reactive oxygen species (ROS) and antioxidant defences. Unfortunately, long-term treatment with anti-epileptic drugs (AEDs) may produce hepatotoxicity, nephrotoxicity, and haematological alterations. In this way, some secondary metabolites of plants have been used to ameliorate the deterioration of nervous system disorders through their antioxidant properties, in addition to their anticonvulsant effects. An example is Centella asiatica, a plant noted to have a reputed neuroprotective effect related to its antioxidant activity. However, similar to conventional drugs, natural molecules may produce side effects when consumed in high doses, which could occur with Centella asiatica. Therefore, we aimed to evaluate the effect of a standardized extract of Centella asiatica L. Urb with tested anticonvulsant activity on biochemical and haematological parameters in rats subjected to lithium/pilocarpine-induced seizures. METHODS: Twenty-eight adult male Wistar rats were randomly divided into four groups (n = 7 each): vehicle (purified water), Centella asiatica (200 and 400 mg/kg), and carbamazepine (CBZ) (300 mg/kg) as a pharmacological control of anticonvulsant activity. Treatments were administered orally every 24 h for 35 consecutive days. On Day 36, SE was induced using the lithium/pilocarpine model (3 mEq/kg, i.p. and 30 mg/kg s.c., respectively), and the behavioural and biochemical effects were evaluated. RESULTS: Centella asiatica 400 mg/kg increased the latency to the first generalized seizure and SE onset and significantly reduced the time to the first generalized seizure compared to values in the vehicle group. Biochemical parameters, i.e., haematic cytometry, blood chemistry, and liver function tests, showed no significant differences among the different treatments. CONCLUSION: The dose of Centella asiatica that produces anticonvulsant activity in the lithium/pilocarpine model devoid of hepatotoxicity, nephrotoxicity, and alterations in haematological parameters suggests that the standardized extract of this plant could be of utility in the development of new safe therapies for the treatment of convulsions associated with epilepsy.

[Risk factors for metabolic dysfunction-associated fatty liver disease in the Hispanic-Mexican population.] / Factores de riesgo de la enfermedad hepática grasa asociada a disfunción metabólica en población hispano-mexicana.

OBJECTIVE: Metabolic dysfunction-associated fatty liver disease (MAFLD) is a poor attended disease, which has gained attention due the elevated number of cases in countries as Mexico, where the incidence is the number 4th globally. MAFLD develops in obese or overweighted individuals and is characterized by triglycerides accumulation in the liver, this condition can develop to hepatocellular carcinoma. It has been observed that MAFLD depends on the genetics and lifestyle. Due to the high prevalence of this disease among Hispanic population, we focused on this study in the characteristics and prevalence of MAFLD in Mexican patients. METHODS: In this study were included 572 overweighted and obese patients, who underwent a screening analysis using the fatty liver index (IHG), clinical parameters were analysed, demographic and comorbidities. Frequency of variables were obtained, and the data were analysed by Chi-square test or Fisher test, odd ratio (OR) and binary logistic regression. RESULTS: A MALFD prevalence of 37% were obtained, where the history of familiar obesity, paracetamol usage, carbohydrate and fat intake are shown to be risk factors. It was found that high blood pressure, central obesity and hypertriglyceridemia were also associated to the MAFLD development. On the other hand, physical exercise was a protector factor. CONCLUSIONS: Our results show the necessity to study the MAFLD causalities in Mexican patients, focused on the paracetamol intake.

OBJETIVO: La enfermedad hepática grasa asociada a disfunción metabólica (MAFLD) es una enfermedad poco considerada, que ha recibido atención debido al número de casos en países como México, donde ocupa el 4º lugar mundial de incidencia. La MAFLD se desarrolla en personas con sobrepeso u obesidad y se caracteriza por la acumulación de triglicéridos en el hígado, donde puede evolucionar hacia carcinoma hepatocelular. Se ha observado que la MAFLD depende de la genética y del estilo de vida. Tomando en cuenta la alta prevalencia de MAFLD en la población hispana, nos enfocamos en este trabajo en estudiar la prevalencia y características relacionadas con esta enfermedad en pacientes mexicanos. METODOS: En este estudio se incluyeron 572 pacientes con sobrepeso u obesidad, a los cuales se les realizó un análisis de cribado mediante el índice de hígado graso (IHG), se analizaron parámetros clínicos, demográficos y comorbilidades. Se obtuvieron frecuencias de las variables y se analizaron los datos mediante chi cuadrado o exacta de Fisher, razón de momios (OR) y regresión logística binaria. RESULTADOS: Se obtuvo una prevalencia del 37% de MAFLD, donde la historia familiar de obesidad, el uso de paracetamol, así como el consumo de carbohidratos y grasas fueron factores de riesgo para su desarrollo. Se encontró que la hipertensión arterial, la obesidad visceral y la hipertrigliceridemia también estaban asociados al desarrollo de la MAFLD. Por otro lado, el ejercicio fue un factor protector. CONCLUSIONES: Nuestros resultados ponen de manifiesto la necesidad de realizar estudios relacionados con las causalidades de la MAFLD en los pacientes mexicanos, principalmente en el uso del paracetamol.

Efficacy of nutrition therapy with food rich in methionine for treating nonalcoholic fatty liver / Eficacia de la terapia nutricional con alimentos ricos en metionina para el tratamiento de hígado graso no alcohólico

Non-Alcoholic Fatty Liver disease (NAFLD) can lead to Non Alcoholic steatohepatitis (NASH), cirrhosis, and liver cancer. The treatment for NAFLD involves modification of caloric intake and physical activity. NAFLD has a pro-oxidant nature; therefore, it is logical to suppose that the antioxidant methionine can be used as a treatment for this disease. Aim. This study aimed to evaluate the effect of high-methionine dietary therapy on patients with NAFLD. Materials and methods. A randomized clinical study was conducted over three months. In this study, 121 NAFLD patients participated, and the age of the participants was ≥ 20 years (experimental group included 56 and control group 65), all of whom were randomized and matched by sex, recluted from the ISSSTE hospital in Xalapa, Mexico. The patients were instructed to consume food to cover the recommended methionine daily doses, and the daily amount consumed was calculated. Methionine effect was measured as NAFLD regression and quality of life improvement. Results. Nutritional therapy induced NAFLD regression and diminished central fat accumulation, blood pressure, and the fatty liver index. Some parameters, such as liver enzymes, did not changed. The quality of life of patients improved after treatment. Conclusions. In this study, we show a hepatoprotective effect induced only in three months of chances in the diet, thus, a longer diet may generate more relevant benefits in the resistant parameters of our study(AU)

La enfermedad del hígado graso no alcohólico (NAFLD) puede conducir a la esteatohepatitis no alcohólica (NASH), la cirrosis y el cáncer de hígado. El tratamiento para NAFLD es la modificación de la ingesta calórica y la actividad física. Debido a que NAFLD tiene una naturaleza pro-oxidante; es lógico suponer que el antioxidante metionina puede utilizarse en el tratamiento de esta enfermedad. Objetivo. el presente trabajo evaluó el papel de la terapia nutricional con alimentos ricos en metioninaen pacientes con NAFLD. Materiales y Métodos. Se realizó un ensayo clínico aleatorizado durante tres meses. Participaron en el estudio 121 pacientes con NAFLD con edad ≥ 20 años (56 en el grupo experimental y 65 en el control), todos aleatorizados y pareados por sexo, reclutados de la Clínica Hospital ISSTE en la ciudad de Xalapa, México, en el año 2015. Se instruyó a los pacientes en consumir los alimentos hasta completar la dosis diaria recomendada de metioninay se calculó la cantidad diaria consumida. Su efecto se midió como la regresión de NAFLD y la mejora de la calidad de vida. Resultados. La terapia nutricional retrocedió NAFLD; disminuyó la acumulación de grasa central, la presión arterial y el índice de hígado graso. Algunos parámetros, como las enzimas de la función hepática, no se modificaron con el tratamiento. Otro parámetro fue la mejora de la calidad de vida de los pacientes tratados. Conclusiones. En este trabajo mostramos un impacto hepatoprotector producido con tan solo tres meses de cambios en la dieta, por lo que una dieta más prolongada podría generar beneficios aún más significativos en los parámetros resistentes en nuestro protocolo(AU)

A Comparative Study of Metabolic Syndrome Using NCEP-ATP III and IDF Criteria in Children and Its Relationship with Biochemical Indicators in Huatusco, Veracruz, Mexico.

Metabolic syndrome includes a set of metabolic alterations associated with overweight and obesity. The criteria for its diagnosis are heterogeneous, and there have been few studies about prevalence in the pediatric population. The aim of this study was to describe how the estimated prevalence of metabolic syndrome varies by International Diabetes Federation (IDF) vs. National Cholesterol Education Program-Adult Treatment Panel III (NCEP-ATP) criteria. We conducted a cross-sectional study in which anthropometric information, triglyceride, cholesterol, glycemia, and insulin levels, among others, were collected. We compared the group potentially misclassified by IDF with the group classified without metabolic syndrome by NCEP-ATP with respect to weight status and biomarkers. Statistical analysis included linear regression, Mann-Whitney U test, Fisher´s exact test, and odds ratio calculation. The IDF criteria missed the association with obesity, although the undetected group differed significantly from the nonmetabolic syndrome group in terms of IBM and weight. The associated biomarkers were ultrasensitive C-reactive protein (Hs-CRP), alanine aminotransferase (ALT) enzyme, insulin, triglycerides, and high-density lipoprotein (HDL) cholesterol. Waist circumference was the parameter with the strongest association for presenting metabolic syndrome, with an odds ratio of 18.33. The results of this study showed the estimated prevalence of MS varies by criteria, due to cutoff points, and how the high prevalence of MS strongly associated with obesity.

Sordera súbita idiopática: factores pronóstico y resultados del tratamiento de rescate con dexametasona intratimpánica / Idiopathic sudden sensorineural hearing loss: prognostic factors and outcomes of salvage treatment with intratympanic dexamethasone

Introducción y objetivos: La sordera súbita idiopática se define habitualmente, como una pérdida auditiva superior o igual a 30 dB HL, en 3 o más frecuencias consecutivas, con un tiempo de instauración inferior a 72 horas, de causa desconocida. Siendo actualmente motivo de discusión el efecto de la terapia esteroidea en este síndrome, evaluaremos la eficacia del corticoide intratimpánico (CIT) como tratamiento de rescate. Como objetivo secundario, estudiaremos la influencia de determinados factores considerados de riesgo o pronóstico. Pacientes y métodos: Se comparan los resultados de dos grupos de población con edad y pérdida auditiva similar. El primero, de 99 pacientes, recibió exclusivamente tratamiento sistémico con corticoides. En el segundo de 70, con una sordera súbita idiopática refractaria al tratamiento general, instilamos corticoide intratimpánico. Comparamos las diferencias de umbral medio auditivo en el momento del diagnóstico y a los 3 meses, además del grado general de recuperación. Resultados: La aplicación intratimpánica de 3 dosis de dexametasona (8mg/ml) en la sordera súbita idiopática refractaria, no ha producido, en nuestra población, un cambio significativo respecto al tratamiento sistémico. Analizamos también la influencia de los factores pronósticos seleccionados. Discusión: Desconocemos la eficacia del tratamiento con corticoides en la sordera súbita idiopática. El tipo de lesión y las características del paciente podrían ser decisivos en el pronóstico final y en el resultado terapéutico. Conclusiones: El tratamiento de rescate con CIT, no ha aportado mejoría auditiva significativa en la población estudiada. Los factores con peor pronóstico son: la presencia de síntomas vestibulares, la hipoacusia de grado profundo, la caída audiométrica hacia tonos agudos, el inicio del tratamiento después de 2 semanas y fundamentalmente, la falta de mejoría de al menos 10 dB el 4º día de tratamiento. (AU)

Introduction and objectives: Idiopathic Sudden Sensorineural Hearing Loss (ISSNHL) is commonly defined as greater than 30 dB of hearing loss in at least 3 audiometric frequencies occurring over 3 days or less, of unknown cause. Since the effect of steroid therapy in this syndrome is currently under discussion, we will evaluate the efficacy of intratympanic corticosteroid (ITC) as rescue treatment. As a secondary objective, we will study the influence of certain factors considered risk or prognostic. Patients and methods: The results of two population groups with similar age and hearing loss are compared. The first of 99 patients received exclusively systemic treatment with corticosteroids. In the second group of 70, with ISSNHL refractory to general treatment, we instilled intratympanic corticosteroids. We compared the differences in mean hearing threshold at diagnosis and at 3 months, as well as the general degree of recovery. Results: Intratympanic application of 3 doses of dexamethasone (8mg/ml) in refractory ISSNHL did not produce, in our population, a significant change with respect to systemic treatment. We also analyzed the influence of selected prognostic factors. Discussion: We do not know the efficacy of corticosteroid treatment in idiopathic sudden deafness. The type of lesion and patient characteristics could be decisive in the final prognosis and therapeutic outcome. Conclusions: Rescue treatment with ITC has not provided significant hearing improvement in the population studied. The factors with the worst prognosis are the presence of vestibular symptoms, profound hearing loss, high-frequency hearing loss in audiometry, the beginning of treatment after 2 weeks and fundamentally, the lack of improvement of at least 10 dB on the 4th day of treatment. (AU)

Telemonitoring System Oriented towards High-Risk Pregnant Women.

A high-risk pregnancy is one in which pathological problems or abnormal conditions are latent during pregnancy and childbirth, increasing dangers to the mother's or the infant's health. Based on international standards and studies, most of the harms and risks to both the mother and the infant can be detected, treated, and prevented through proper pregnancy monitoring, as well as through appropriate and timely diagnosis. In this paper, we present the analysis, design, development, and usability assessment of a telemonitoring system focused on the remote monitoring and control of pregnancy in women suffering from hypertension, diabetes, or high-risk pregnancy. Our system is composed of two mobile web applications. One of these is designed for the medical area, allowing remote monitoring of the patient's pregnancy, and the other one is directed towards the patient, who enters the alarm symptom data, hypertension data, diabetes data, and clinical analyses, allowing the detection of a risk situation on time. Furthermore, we performed a usability assessment of our system based on a laboratory study with seven doctors and seven patients to evaluate the users' satisfaction. Our telemonitoring system shows a satisfactory/favorable opinion from the users' perspectives based on the obtained results.

Linear breakwater reefs of the greater Caribbean: Classification, distribution & morphology.

Geomorphic differences among Caribbean reefs have long been noted. These differences are considered to reflect the presence of reefs in different stages of development, following an incomplete recovery from rapid deglacial sea-level rise. But the possibility that these reflect real developmental differences caused by variation in wind, wave, and climate regime, has never been fully considered. Here, for the first time, we quantify the geomorphology and distribution of Greater Caribbean reefs using satellite images in Google Earth and public-domain bathymetry. To do this, we first standardise their classification based on shallow geomorphology, substrate depth, and physiographic setting, and then count and categorise the total number of reefs. These data show a total of 1023 linear breakwater reefs with a combined length of 2237 km. Of this total length, 80% are fringing reefs, 16% are barriers and 4% are faros and atolls. In terms of categories, there are 16 reef subtypes present, but only 9 are common. Their distribution, however, is not uniform. In particular, flat-subtypes form 60% of breakwater reefs in southern regions, but are less common in northern regions where crest-subtypes dominate (80%). To distinguish the geomorphology of these common reef subtypes, we collect size- and length-related morphometric data from their main reef zones. These data reveal that flat and crest subtypes also have morphometric differences: flat subtypes have well-constrained morphologies with statistically consistent unimodal morphometrics characterised by large back-reef zones, smaller and steeper reef fronts, and more sinuous and persistent crestlines. Crest subtypes, by contrast, have multimodal morphometrics suggesting less consistent morphologies (or unresolved subtypes), and are characterised by crestlines with lower sinuosity, more variable back-reef and reef-front areas, and slopes. These differences in geomorphology and distribution imply that flat- and crest-subtypes are neither successional stages of a single reef type, nor a genetically related sequence of types, but distinct reefal geoforms with different modes of development. In subsequent work we will explore what controls these differences.

Determinación de zonas de ruido ambiental dañino en instalaciones no operativas de bases aéreas militares / Determination of harmful environmental noise zones in non-operating military air base facilities

Resumen Introducción: El ruido es un sonido desagradable que afecta la salud y que es factor de riesgo para hipertensión y cardiopatía isquémica. La aviación genera ruido y contaminación del aire que daña la audición y es un problema a nivel mundial y para la población en general y para quienes trabajan en los aeropuertos, por lo que debe ser analizado. Sobre el tema se han publicado trabajos diversos, más sin embargo son pocos los estudios hechos en zonas aledañas a bases aéreas y brindándose poca atención preventiva a la población que labora dentro de ellas. Objetivo: Determinar la existencia de niveles de ruido ambiental dañino en instalaciones de trabajo no operativas de bases aéreas militares. Material y métodos: Se midió el ruido ambiental en turnos laborales por 7 días, en las bases aéreas de Santa Lucía, y Zapopan, con sonómetro ambiental Extech® Mod. 407732, clase 2, normas ANSI y IEC 651 Type 2 y calibrador 407744 de 94 ó 114 dB., en áreas expuestas al ruido cercanas a la pista de aterrizaje en horas de máxima actividad aérea. Resultados: Se encontró ruido dañino no satisfactorio y peligroso entre los 68 dB y 82.06 dB de nivel equivalente sonoro de ruido (LEQ), fuera de norma para ruido ambiental. Conclusiones: Los niveles de ruido encontrado se salen de los parámetros aceptables de exposición al ruido ambiental en la comunidad, por tanto resulta nocivo y se debe contar con medios de protección y programas preventivos para evitar la pérdida de la audición.

Abstract Introduction: Noise is an unpleasant sound that affects health and is a risk factor for hypertension and ischemic heart disease. Aviation generates noise and air pollution that damages hearing and is a problem worldwide and for the general population and for those who work in airports, so it must be analyzed. Various works have been published on the subject, however, there are few studies carried out in areas surrounding air bases and little preventive care is provided to the population that works within them. Objective: Determine the existence of harmful environmental noise levels in non-operational work installations of military air bases. Material and Methods: Ambient noise was measured in work shifts for 7 days, at the air bases of Santa Lucía and Zapopan, with Extech® Mod. 407732, class 2 environmental sound level meter, ANSI and IEC 651 Type 2 standards and 407766 gauge of 94 or 114 dB, in áreas exposed to noise near the runway at peak hours air activit Results: No satisfactory and dangerous harmful noise was found between 68 dB and 82.06 dB of noise equivalent noise level (LEQ), out of the norm for ambient noise. Conclusions: The noise found does not meet what is established for the levels of exposure of environmental noise in the community, it must have means of protection and preventive programs to prevent hearing loss.

Effects of experimental warming on two tropical Andean aquatic insects.

Temperatures have increased around the globe, affecting many ecosystems, including high-elevation Andean streams where important aquatic insect species coexist. Depending on the magnitude of change, warming could lead to the mortality of sensitive species, and those tolerant to rising water temperatures may exhibit differences in growth rates and development. Taxon-specific optimal temperature ranges for growth determine how high or low temperatures alter an organism's body size. In this study, we observed the effects of different climate change scenarios (following three scenarios of the 2021 IPCC predictions) in two aquatic insect species distributed in high-elevation streams in Ecuador: the mayfly Andesiops peruvianus (Ephemeroptera: Baetidae) and the caddisfly Anomalocosmoecus illiesi (Trichoptera: Limnephilidae). We assessed how increased water temperatures affect larval growth rates and mortality during a 10-day microcosm experiment. Our results showed that Andesiops peruvianus was more thermally sensitive than Anomalocosmoecus illiesi. Mortality was higher (more than 50% of the individuals) in mayflies than in caddisflies, which presented mortality below 12% at +2.5°C and +5°C. Mortality in mayflies was related to lower dissolved oxygen levels in increased temperature chambers. Higher temperatures affected body size and dry mass with a faster growth rate of Andesiops peruvianus larvae at experimentally higher temperatures, suggesting an important response of this hemimetabolous species to stream temperatures. For Anomalocosmoecus illiesi, we did not find significant changes in mortality, body size or growth rate in response to temperature changes during our experiment. In situ outcomes of species survival and growth in Andean streams are difficult to predict. Nevertheless, our results suggest that at only +2.5°C, a water temperature increase affected the two insect taxa differentially, leading to a drastic outcome for one species' larvae while selecting for a more tolerant species. Our study suggests that climate change might produce significant mortality and growth rate effects on ectotherm tropical aquatic insects, especially Andean mayflies, which showed higher sensitivity to increased water temperature scenarios.

Anticonvulsant Effect of Turmeric and Resveratrol in Lithium/Pilocarpine-Induced Status Epilepticus in Wistar Rats.

Epilepsy is a chronic neurological disorder that lacks a cure. The use of plant-derived antioxidant molecules such as those contained in turmeric powder and resveratrol may produce short-term anticonvulsant effects. A total of 42 three-month-old male Wistar rats were divided into six groups (n = 7 in each group): Vehicle (purified water), turmeric (150 and 300 mg/kg, respectively), and resveratrol (30 and 60 mg/kg, respectively), administered per os (p.o.) every 24 h for 35 days. Carbamazepine (300 mg/kg/5 days) was used as a pharmacological control for anticonvulsant activity. At the end of the treatment, status epilepticus was induced using the lithium-pilocarpine model [3 mEq/kg, intraperitoneally (i.p.) and 30 mg/kg subcutaneously (s.c.), respectively]. Seizures were evaluated using the Racine scale. The 300 mg/kg of turmeric and 60 mg/kg of resveratrol groups had an increased latency to the first generalized seizure. The groups treated with 150 and 300 mg/kg of turmeric and 60 mg/kg of resveratrol also had an increased latency to status epilepticus and a decreased number of generalized seizures compared to the vehicle group. The chronic administration of turmeric and resveratrol exerts anticonvulsant effects without producing kidney or liver damage. This suggests that both of these natural products of plant origin could work as adjuvants in the treatment of epilepsy.

Optimizing rat and human blood cells sampling for in silico morphometric analysis.

Measurements of Morphometric Parameters of the Blood Cells (MPBC) are key for the diagnosis of both mental and metabolic diseases. Several manual approaches or computational methodologies are useful to provide reliable clinical diagnosis. The sample processing and data analysis is relevant, however the sample handling on the pre-analytical phase remains scarcely evaluated. The main goal of this study was to favor the preservation of blood smear using a histological resin. This strategy lead us two practical approaches, give a detailed morphometric description of white blood cells and establish reference intervals in male Wistar rats, which are scarcely reported. Blood smears from male Wistar rats (n = 120) and adult men were collected at room temperature. The integrity of Wright-stained cells was evaluated by an in silico image analysis from rat and human blood smear preserved with a toluene-based synthetic resin mounting medium. A single sample of human blood was used as a control of procedure. The reference intervals was established by cell counting. Based on the results of segmentation algorithm followed by an automatic thresholding analysis, the incorporation of resin favor the conservation of cell blood populations, and lead to identify morphologic features such as nucleus/cytoplasmic shape, granules presence and DNA appearance in nucleus of white blood cells. The use of a histological resin could favor a fast and efficient sample handling in silico MPBC measurements both in the species studied as in wild animals.

Identification of Sodium Transients Through NaV1.5 Channels as Regulators of Differentiation in Immortalized Dorsal Root Ganglia Neurons.

Neuronal differentiation is a complex process through which newborn neurons acquire the morphology of mature neurons and become excitable. We employed a combination of functional and transcriptomic approaches to deconvolute and identify key regulators of the differentiation process of a DRG neuron-derived cell line, and we focused our study on the Na V 1.5 ion channel (encoded by Scn5a) as a channel involved in the acquisition of DRG neuronal features. Overexpression of Scn5a enhances the acquisition of neuronal phenotypic features and increases the KCl-elicited hyperexcitability response in a DRG-derived cell line. Moreover, pharmacologic inhibition of the Na V 1.5 channel during differentiation hinders the acquisition of phenotypic features of neuronal cells and the hyperexcitability increase in response to changes in the extracellular medium ionic composition. Taken together, these data highlight the relevance of sodium transients in regulating the neuronal differentiation process in a DRG neuron-derived cell line.

Multi-tissue cytogenetic analysis for the diagnosis of mosaic Down syndrome: A case report.

Less than one percent of individuals with Down syndrome exhibit mosaicism, a biological phenomenon that describes an individual who has two or more genetically distinct cell lines. The percentage of mosaicism in different tissues can impact the presence of clinical findings and hinder cytogenetic diagnosis. We report a case of mosaicism for trisomy 21 diagnosed after multi-tissue cytogenetic analysis of peripheral blood and buccal mucosa, associated with significant intellectual disability, dysmorphic facial features, congenital heart defects, macropenis, and imperforate anus.

Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist.

BACKGROUND: CLEC-2 is a platelet receptor with an important role in thromboinflammation but a minor role in hemostasis. Two endogenous ligands of CLEC-2 have been identified, the transmembrane protein podoplanin and iron-containing porphyrin hemin, which is formed following hemolysis from red blood cells. Other exogenous ligands such as rhodocytin have contributed to our understanding of the role of CLEC-2. OBJECTIVES: To identify novel CLEC-2 small-molecule ligands to aid therapeutic targeting of CLEC-2. METHODS: ALPHA screen technology has been used for the development of a high-throughput screening (HTS) assay recapitulating the podoplanin-CLEC-2 interaction. Light transmission aggregometry was used to evaluate platelet aggregation. Immunoprecipitation and western blot were used to evaluate direct phosphorylation of CLEC-2 and downstream protein phosphorylation. Autodock vina software was used to predict the molecular binding site of katacine and mass spectrometry to determine the polymeric nature of the ligand. RESULTS AND CONCLUSION: We developed a CLEC-2-podoplanin interaction assay in a HTS format and screened 5,016 compounds from a European Union-open screen library. We identified katacine, a mixture of polymers of proanthocyanidins, as a novel ligand for CLEC-2 and showed that it induces platelet aggregation and CLEC-2 phosphorylation via Syk and Src kinases. Platelet aggregation induced by katacine is inhibited by the anti-CLEC-2 monoclonal antibody fragment AYP1 F(ab)'2. Katacine is a novel nonprotein ligand of CLEC-2 that could contribute to a better understanding of CLEC-2 activation in human platelets.

Identification of Novel Regulators of Zalcitabine-Induced Neuropathic Pain.

Neuropathic pain is one of the foremost adverse effects that worsens quality of life for patients undergoing an antiretroviral treatment. Currently, there are no effective analgesics for relieving it; thus, there is an urgent need to develop novel treatments for neuropathic pain. Previously, we described and validated F11 cells as a model of DRG (dorsal root ganglia) neurons. In the current work, we employed F11 cells to identify regulators of antiretroviral-induced neuropathic pain combining functional and transcriptomic analysis. The antiretroviral zalcitabine (ddC) increased the excitability of differentiated F11 cells associated with calcium signaling without morphological changes in the neuronal phenotype, mimicking the observed increase of painful signaling in patients suffering from antiretroviral-induced neuropathic pain. Employing RNA sequencing, we observed that zalcitabine treatment upregulated genes related with oxidative stress and calcium homeostasis. The functional impact of the transcriptomic changes was explored, finding that the exposure to zalcitabine significantly increased intracellular oxidative stress and reduced store-operated calcium entry (SOCE). Because the functional and transcriptomic evidence points toward fundamental changes in calcium signaling and oxidative stress upon zalcitabine exposure, we identified that NAD(P)H quinone dehydrogenase and the sarcoplasmic/endoplasmic reticulum calcium ATPase 3 were involved in zalcitabine-induced hyperexcitability of F11 cells. Overexpression of those genes increases the calcium-elicited hyperexcitability response and reduces SOCE, as well as increases intracellular ROS levels. These data do not only mimic the effects of zalcitabine but also highlight the relevance of oxidative stress and of calcium-mediated signaling in antiretroviral-induced hyperexcitability of sensory neurons, shedding light on new therapeutic targets for antiviral-induced neuropathic pain.

Mayfly taxonomy (Arthropoda: Hexapoda: Ephemeroptera) during the first two decades of the twenty-first century and the concentration of taxonomic publishing.

The twentieth anniversary of the first issue of Zootaxa (De Moraes Freire, 2001) provides an appropriate opportunity to reflect on some trends in global Ephemeroptera taxonomy publishing over the last two decades, with a focus on the description of new species and the outsized role of the journals Zootaxa and ZooKeys, in particular. Detailed reviews of world Ephemeroptera knowledge up to about 2000 were collected in a series of nine papers from a symposium on the subject, published together in the proceedings of the ninth International Conference on Ephemeroptera (Domínguez 2001). Domínguez Dos Santos (2014) provided updates and analysis for South America up to the year 2012. More recent detailed accounts of regional and taxonomic diversity, and other aspects of mayfly biology and ecology, were reviewed by Jacobus et al. (2019), while Ogden et al. (2019) discussed current issues involving higher classification.