Molecular characterization of novel Ehrlichia genotypes in Ixodes auritulus from Uruguay

Ehrlichia are small intracellular Gram-negative bacteria transmitted by ticks. These microorganisms cause ehrlichiosis, a complex of life-threatening emerging zoonoses and diseases of global veterinary relevance. The aim of this study was to investigate the presence of Ehrlichia in free-living Ixodes auritulus collected in Uruguay. Ticks were collected from vegetation in five localities from the southeast and northeast of the country between 2014 and 2017. Detection of Ehrlichia DNA was performed in pools of adults or nymphs grouped according to the collection site and date. A total of 1,548 I. auritulus ticks were collected in four of the five locations sampled. Fragments of three loci (16S rRNA, dsb and groEL) were obtained by PCR, and phylogenies inferred using Bayesian inference analysis for each gene independently. DNA of Ehrlichia spp. was found in 15 out of 42 tick pools. Based on the topology of the phylogenetic trees, our sequences represent two novel genotypes for the genus named as Ehrlichia sp. Serrana and Ehrlichia sp. Laguna Negra. Both genotypes were closely related to Ehrlichia sp. Magellanica, a species detected in Ixodes uriae and Magellanic penguins. Considering that all stages of I. auritulus and I. uriae are parasites of birds, their phylogenetic relationships, and common eco-epidemiological profiles, it is reasonable to state that these genotypes of Ehrlichia spp. may represent a natural group likely associated with birds. Our results constitute the first characterization of Ehrlichia spp. in Uruguay. Future studies on birds reported as hosts for I. auritulus are needed to further understand the epidemiological cycles of both Ehrlichia genotypes in the country. Finally, I. auritulus does not feed on humans, so the two Ehrlichia species reported herein might have no implications in human health.EEA RafaelaFil: Félix, María L. Universidad de la República. Facultad de Veterinaria. Laboratorio de Vectores y Enfermedades Transmitidas; UruguayFil: Muñoz-Leal, Sebastian. Universidad de Concepción. Facultad de Ciencias Veterinarias. Departamento de Patología y Medicina Preventiva; ChileFil: Carvalho, Luis A. AgResearch. Grasslands Research Centre; Nueva ZelandaFil: Queirolo, Diego. Universidad de La República. CENUR Noreste; UruguayFil: Remesar Alonso, Susana. Universidade de Santiago de Compostela. Faculty of Veterinary Sciences. Department of Animal Pathology (INVESAGA Group); EspañaFil: Nava, Santiago. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Nava, Santiago. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Armúa-Fernández, María T. Universidad de la República. Facultad de Veterinaria. Laboratorio de Vectores y Enfermedades Transmitidas; UruguayFil: Venzal, José Manuel. Universidad de la República. Facultad de Veterinaria. Laboratorio de Vectores y Enfermedades Transmitidas; Urugua

Variable structure control in natural frame for three-phase grid-connected inverters with LCL filter

This paper presents a variable structure control in natural frame for a three-phase voltage source inverter. The proposed control method is based on modifying the converter model in natural reference frame, preserving the low frequency state space variables dynamics. Using this model in a Kalman filter, the system state-space variables are estimated allowing to design three robust current sliding-mode controllers in natural frame. The main closed-loop features of the proposed method are: 1) robustness against grid inductance variations because the proposed model is independent of the grid inductance, 2) the power losses are reduced since physical damping resistors are avoided, and 3) the control bandwidth can be increased due to the combination of a variable hysteresis controller with a Kalman filter. To complete the control scheme, a theoretical stability analysis is developed. Finally, selected experimental results validate the proposed control strategy and permit illustrating all its appealing features.Postprint (author's final draft

Aislamiento de núcleos para citometría de flujo en plantas con alto contenido de compuestos mucilaginosos: un ejemplo en el género Viola L. (Violaceae)

Flow cytometry analysis has been widely applied in the determination of nuclear DNA content and ploidy level in many organisms. Despite being the most appropriate method for DNA content measurement, flow cytometry also presents some limitations. A fairly common, but little-studied problem is the effect on measurements of the presence of secondary metabolites. A good example is the genus Viola, which is composed of 525-600 species distributed worldwide. These species have proved to be problematic for flow cytometric analyses due to the release of extremely mucilaginous compounds into the nuclear suspension. In this work, the genome size of 13 species of Viola using flow cytometry are presented for the first time. Despite obtaining histograms with high coefficients of variation, we here present an optimized protocol to remove cytoplasmic compounds, particularly mucilaginous ones, from plant nuclei that pave the way for its application to estimate the genome size of other species exhibiting similar problems. Statistical analyses revealed significant differences between sections Viola and Melanium, and within each section (P < 0.001). Furthermore, statistically significant differences were not detected among samples of the same species.El análisis mediante citometría de flujo ha sido aplicado de modo general para determinar el contenido de ADN nuclear y el nivel de ploidía en muchos organismos. A pesar de ser el método más adecuado para medir la cantidad de ADN, esta técnica también presenta algunas limitaciones. Un problema bastante común, aunque poco estudiado, es el efecto de los metabolitos secundarios en los resultados obtenidos. Un ejemplo respecto a la presencia de estos compuestos se encuentra en el género Viola, compuesto por 525-600 especies distribuidas por todo el mundo. Las especies de este género ya han sido previamente descritas como problemáticas en los análisis de citometría de flujo debido a la presencia de compuestos extremadamente mucilaginosos en las suspensiones de núcleos. En el presente trabajo se analiza por primera vez el tamaño genómico de 13 especies del genero Viola mediante el empleo de citometría de flujo. A pesar de los altos valores mostrados en los coeficientes de variación de los histogramas, se presenta un protocolo optimizado para eliminar compuestos citoplasmáticos, y más concretamente mucilaginosos de las suspensiones nucleares, siendo de aplicación en la estimación del tamaño genómico de plantas con problemas similares. Los análisis estadísticos mostraron diferencias significativas entre las secciones Viola y Melanium, así como dentro de cada sección (P < 0,001). Además, no se encontraron diferencias significativas entre aquellas muestras pertenecientes a la misma especie

Human Monkeypox: A Comprehensive Overview of Epidemiology, Pathogenesis, Diagnosis, Treatment, and Prevention Strategies.

Monkeypox virus (MPXV) is an emerging zoonotic virus that belongs to the Orthopoxvirus genus and presents clinical symptoms similar to those of smallpox, such as fever and vesicular-pustular skin lesions. However, the differential diagnosis between smallpox and monkeypox is that smallpox does not cause lymphadenopathy but monkeypox generates swelling in the lymph nodes. Since the eradication of smallpox, MPXV has been identified as the most common Orthopoxvirus to cause human disease. Despite MPXV being endemic to certain regions of Africa, the current MPXV outbreak, which began in early 2022, has spread to numerous countries worldwide, raising global concern. As of the end of May 2023, over 87,545 cases and 141 deaths have been reported, with most cases identified in non-endemic countries, primarily due to human-to-human transmission. To better understand this emerging threat, this review presents an overview of key aspects of MPXV infection, including its animal reservoirs, modes of transmission, animal models, epidemiology, clinical and immunological features, diagnosis, treatments, vaccines, and prevention strategies. The material presented here provides a comprehensive understanding of MPXV as a disease, while emphasizing the significance and unique characteristics of the 2022 outbreak. This offers valuable information that can inform future research and aid in the development of effective interventions.J.M.J.-G. received support from the Grant RYC2021-031227-I funded by the MCIN/AEI/10.13039/501100011033 and the European Union “NextGenerationEU”/PRTR and the Grant Proyecto19 funded by the University of Málaga Plan Propio (Ayuda G). Partial funding for open access charge: Universidad de Málaga

Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus <i>Viola</i> L. (Violaceae)

Cires, E., Cuesta, C., Fernández, M.A., Nava, H.S., Vázquez, V.M. & Fernández, J.A. 2011. Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae). Anales.Iard. Bot. Madrid 68(2): 139-154. Flow cytometry analysis has been widely appljed in the determination of nuclear DNA content and ploidy level in many organisms. Despite being the most appropriate method for DNA content measurement, flow cytometry also presents some limitations. A fairly common, but little-studied problem is the effect on measurements of the presente of secondary metabolites. A good example is the genus Viola, which is composed of 525-600 species distributed worldwide. These species have proved to be problematic for flow cytometric analyses due to the release of extremely mucilaginous compounds into the nuclear suspension. In this work, the genome size of 13 species of Viola using flow cytometry are presented for the first time. Despite obtaining histograms with high coefficients of variation, we here pre-sent an optimized protocol to remove cytoplasmic compounds, particularly mucilaginous ones, from plant nudei that paye the way for its application to estimate the genome size of other species exhibiting similar problems. Statistical analyses revealed significant differences between sections Viola and Melanium, and within each section (P < 0.001). Furthermore, statistically significant differences were not detected among samples of the same species.Cires, E., Cuesta, C., Fernández, M.A., Nava, H.S., Vázquez, V.M. & Fernández, J.A. 2011. Aislamiento de núcleos para citometría de flujo en plantas con alto contenido de compuestos mucilaginosos: un ejemplo en el género Viola L. (Violaceae). Anales.lard. Bot. Madrid 68(2): 139-154 (en inglés). El análisis mediante citometría de flujo ha sido aplicado de modo general para determinar el contenido de ADN nuclear y el nivel de ploidía en muchos organismos. A pesar de ser el método más adecuado para medir la cantidad de ADN, esta técnica también presenta algunas limitaciones. Un problema bastante común, aunque poco estudiado, es el efecto de los metabolitos secundarios en los resultados obtenidos. Un ejemplo respecto a la presencia de estos compuestos se encuentra en el género Viola, compuesto por 525-600 especies distribuidas por todo el mundo. Las especies de este género ya han sido previamente descritas como problemáticas en los análisis de citometría de flujo debido a la presencia de compuestos extremadamente mucilaginosos en las suspensiones de núcleos. En el presente trabajo se analiza por primera vez el tamaño genómico de 13 especies del genero Viola mediante el empleo de citometría de flujo. A pesar de los altos valores mostrados en los coeficientes de variación de los histogramas, se presenta un protocolo optimizado para eli-minar compuestos citoplasmáticos, y más concretamente mucilaginosos de las suspensiones nucleares, siendo de aplicación en la estimación del tamaño genómico de plantas con problemas similares. Los análisis estadísticos mostraron diferencias significativas entre las secciones Viola y Melanium, así como dentro de cada sección (P < 0,001). Además, no se encontraron diferencias significativas entre aquellas muestras pertenecientes a la misma especie

Impact of Tinnitus in Patients with Ménière's Disease

Introduction and objective: Tinnitus, with concurrent episodes of vertigo, neurosensorial hearing loss and aural fullness are the cardinal symptoms of Ménière Disease. Not achieving the importance it deserves in the management of the disease, tinnitus has a relevant impact on the lives of patients, especially in the psychological sphere. Method: Fifteen patients with Ménière's disease were selected according to the Bárány Society diagnostic criteria at the University Hospital of Salamanca. Specific variables were recorded for the analysis of the cardinal symptoms of Ménière's disease. Facts about the age of onset and years of disease evolution were collected and in the same strategy for tinnitus, bilateral or unilateral involvement of the disease and in the latter case, the affected ear. Patients were subjected to audiometric test and tinnitus matching. The main parameters defining tinnitus were assessed: frequency (Hz), intensity (dB), minimum masking level (dB) and residual inhibition (seconds). Tinnitus Handicap Inventory (THI) and Visual Analogue Scale (VAS) were performed to determine tinnitus severity. The assessment of mental symptoms was obtained through the Hospital Anxiety and Depression Scale (HADS). Results: Ménière's disease is diagnosed at the mean age of 47.47 years. The average hearing loss at each frequency due to Ménière's disease is approximately 50 dB. Tinnitus is not prevailing in any of the ears (p&gt;0.05). A significant correlation was found between THI and VAS (r=0.72; p&lt;0.01). Both, THI and VAS, were not related to mean values of hearing loss in either ears. Patients with over two years of disease evolution are associated with higher scores on the THI and HADS depression subscales (r=0.821; p=0.001). Discussion: The intensity of tinnitus, quantified subjectively with THI, is not related to hearing loss in patients with Ménière's disease. The study of the psychological sphere through the HADS demonstrates certain predominance of depressive pathology in patients who present longstanding tinnitus.Introducción y objetivo: El acúfeno conforma junto con los episodios de vértigo, hipoacusia neurosensorial y la sensación de plenitud ótica los síntomas cardinales de la Enfermedad de Ménière. No alcanzando la importancia que le corresponde en el manejo de esta enfermedad, el acúfeno repercute profundamente en la vida de los pacientes, sobre todo en la esfera psicológica. Método: Se seleccionaron 15 pacientes con Enfermedad de Ménière diagnosticados según los criterios de la Sociedad de Bárány en Hospital Universitario de Salamanca. Se registraron variables específicas para el análisis de los síntomas cardinales de la enfermedad de Ménière. Se recogieron datos sobre la edad de aparición y años de evolución de la enfermedad y de la misma manera para el acúfeno, la afectación bilateral o unilateral de la misma y en este último supuesto, el oído afecto. Se sometió a los pacientes a estudio audiométrico junto con acufenometría. Se valoraron los principales parámetros definitorios del acúfeno: frecuencia (Hz), intensidad (dB), umbral de enmascaramiento (dB) e inhibición residual (segundos). Los pacientes realizaron el test Tinnitus Handicap Inventory (THI) y Escala visual analógica (VAS). La valoración de alteraciones en el estado de ánimo se obtuvo a través de la Escala Hospitalaria de Ansiedad y Depresión (HADS). Resultados: La edad media de diagnóstico de la Enfermedad de Ménière es 47,47 años. Las medias de pérdida auditiva en cada frecuencia por la enfermedad de Ménière se sitúan en torno a los 50 dB. El acúfeno no predomina en ninguno de los oídos (p&gt;0,05). Se determinó una relación fuertemente significativa entre THI y VAS (r=0,72; p&lt;0,01). Tanto THI como VAS no se relacionaron con los valores de la media de las pérdidas auditivas de ambos oídos. Los pacientes con más de dos años de evolución de la enfermedad se corresponden estadísticamente con puntuaciones mayores en THI y en la subescala de depresión del HADS (r= 0,821; p=0,001). Discusión: La intensidad del acúfeno, cuantificada de manera subjetiva con THI, no se relaciona con el grado de pérdida auditiva en los pacientes con Enfermedad de Ménière. El estudio de la esfera psicológica a través de la escala HADS demuestra la existencia de trastornos del ánimo en estos pacientes, prevaleciendo la patología depresiva en aquellos que presentan acúfenos de larga evolución

The origin and collapse of rock glaciers during the Bølling-Allerød interstadial: A new study case from the Cantabrian Mountains (Spain)

.During the Late Pleistocene, the main mountain ranges of the Iberian Peninsula were covered by small icefields and cirque and alpine glaciers. The deglaciation triggered paraglacial processes that generated landforms, mostly within the ice-free glacial cirques. In this research we analyse the deglaciation process in the Muxivén Cirque (42°15′N – 6°16′W), in the upper Sil River Basin, which includes some of the largest relict rock glaciers of the Cantabrian Mountains. We addressed this objective by means of accurate geomorphological reconstructions, sedimentological analysis, Schmidt-hammer surface weathering measurements and a dataset of 10 10Be Cosmic-Ray Exposure ages. Results reveal that after ~16 ka, glaciers retreated to the bottom of the cirques at the headwaters of the valley, leaving the walls free of ice and triggering rock avalanches onto the remnants of these glaciers. This paraglacial process supplied debris to a small glacier within Muxivén Cirque, which transformed in two rock glaciers. These debris isolated the ice inside the rock glaciers only for a very short period of time and ended up melting completely before the Younger Dryas. The lower sector of the largest one stabilized at 14.5 ± 1.5 ka, while the upper sector remained active until 13.5 ± 0.8 ka. Previous to the stabilization of the lower sector of the northern rock glacier, at its margin a high-energy debris avalanche occurred at ~14.0 ± 0.9 ka. These data agree with previous research, corroborating the paraglacial origin of most Iberian rock glaciers during the Bølling-Allerød interstadial.S

Hearing loss after intratympanic gentamicin in Ménière's disease. Retrospective study

Introduction and objective: Ménière's disease (MS) is characterized by recurrent vertigo attacks with tinnitus, hearing loss and aural fullness. Among the therapeutic options, intratympanic injection of gentamicin (ITG) will be discussed, among others. The objective is to evaluate hearing loss after ITG in patients with MS. Method: 33 patients of the Otorhinolaryngology and Head and Neck Surgery Service of the Hospital Clínico Universitario de Salamanca will be studied, all of them with definitive MS to whom ITG was administered. Hearing loss will be assessed one month after the ITG, at 6, 12 and 24 months. Results: Of the 33 patients analyzed, 78.8% had unilateral MS; 21.2%, bilateral. 48.5% were women; 51.5%, men. 39.4% had already been treated with betahistine and corticosteroids. 9 patients presented Tumarkin crisis before ITG. The PTA (Pure Tone Average) prior to ITG administration was compared with that obtained at 2 years and hearing decreased from 57.50 dB to 62.58 dB. 9 patients required a second round of ITG, with a hearing loss ranging from 66.72 dB to 68.96 dB. And after this, 3 patients needed a third wave, hearing from 63.75dB to 72.50 dB. Conclusions: The use of ITG in MS causes a fall in PTA of around 5 dB at the expense of an improvement in the frequency and intensity of vertigo attacks./nIntroducción y objetivo: La enfermedad de Ménière (EM) se caracteriza por crisis de vértigo recurrentes acompañadas de acúfenos, pérdida de audición y sensacio?n de plenitud o?tica. Dentro de las opciones terapéuticas se hablará, entre otras, de inyección de gentamicina intratimpánica (ITG). El objetivo es evaluar la pérdida de audición tras la ITG en pacientes con EM. Método: Se estudiaron 33 pacientes con EM definitiva a los cuales se les administró ITG. Se valoró la pérdida de audición al mes de la ITG, a los 6, 12 y 24 meses. Resultados: De los 33 pacientes analizados, el 78,8% presentaban EM unilateral; el 21,2%, bilateral. El 48,5% fueron mujeres; el 51,5%, hombres. El 39,4% ya habían sido tratados con betahistina y corticoides. 9 pacientes presentaron crisis de Tumarkin antes de la ITG. Se comparó el PTA (Pure Tone Average) previo a la administración de ITG con el obtenido a los 2 años y la audición disminuyó de 57,50 dB a 62,58 dB. 9 pacientes precisaron una segunda tanda de ITG, con una pérdida de audición desde 66,72 dB a 68,96 dB. Y tras esto, 3 pacientes necesitaron una tercera tanda, pasando la audición de 63,75dB a 72,50 dB. Conclusiones: La utilización de ITG en la EM y provoca una caída del PTA de alrededor de 5 dB a expensas de una mejoría en la frecuencia e intensidad de las crisis de vértigo./

Las Vallinas (Teverga, Asturias). An Iron Age and roman farmsteas in northern Spain

[ES] Los obstáculos para la prospección a lo largo del paisaje costero del Golfo de Vizcaya, las limitaciones estructurales y materiales de las unidades de asentamiento más pequeñas, y la falta de un control exhaustivo de las alteraciones actuales del terreno, han ocasionado un vacío en el conocimiento sobre la posible presencia de granjas o caseríos en espacios abiertos durante la Edad del Hierro en la región cantábrica. Presentamos por primera vez la existencia de estas granjas, con los hallazgos en Las Vallinas que confirman el uso agrícola del terreno. Al comparar la información de esta alquería con la de castros habitados de la misma época, aportamos una visión más amplia del desarrollo económico de los antiguos Astures desde finales de la Prehistoria hasta el inicio de la ocupación romana. El impacto de la romanización provocó una alteración sin precedentes de las antiguas estructuras sociales y económicas de los Astures en esta zona, dando lugar a una lenta y continua recuperación de la economía rural a partir de finales del siglo I d.C[EN] Obstacles to prospection along the coastal landscape of the Bay of Biscay, structural and material limitations in smaller settlement units, and a lack of comprehensive monitoring of current alterations to the terrain, have all left a dearth of knowledge about the possible presence of open Iron Age farms or hamlets in the Cantabrian region. Presented here for the first time, we demonstrate the existence of these farms with findings verifying agricultural land use at Las Vallinas. In comparing information from this farm with that from inhabited hillforts of the same period, we are also given a wider picture of the economic development of the ancient Astures from the end of prehistory to the beginning of Roman occupation. The impact of Romanization caused an unprecedented upheaval of the old social and economic structures of the Asturs in the area, instigating a slow and steady recovery of a rural economy from the late 1st century AD

Generation of NKX2.5(GFP) Reporter Human iPSCs and Differentiation Into Functional Cardiac Fibroblasts

Direct cardiac reprogramming has emerged as an interesting approach for the treatment and regeneration of damaged hearts through the direct conversion of fibroblasts into cardiomyocytes or cardiovascular progenitors. However, in studies with human cells, the lack of reporter fibroblasts has hindered the screening of factors and consequently, the development of robust direct cardiac reprogramming protocols.In this study, we have generated functional human NKX2.5(GFP) reporter cardiac fibroblasts. We first established a new NKX2.5(GFP) reporter human induced pluripotent stem cell (hiPSC) line using a CRISPR-Cas9-based knock-in approach in order to preserve function which could alter the biology of the cells. The reporter was found to faithfully track NKX2.5 expressing cells in differentiated NKX2.5(GFP) hiPSC and the potential of NKX2.5-GFP + cells to give rise to the expected cardiac lineages, including functional ventricular- and atrial-like cardiomyocytes, was demonstrated. Then NKX2.5(GFP) cardiac fibroblasts were obtained through directed differentiation, and these showed typical fibroblast-like morphology, a specific marker expression profile and, more importantly, functionality similar to patient-derived cardiac fibroblasts. The advantage of using this approach is that it offers an unlimited supply of cellular models for research in cardiac reprogramming, and since NKX2.5 is expressed not only in cardiomyocytes but also in cardiovascular precursors, the detection of both induced cell types would be possible. These reporter lines will be useful tools for human direct cardiac reprogramming research and progress in this field.This work was supported by PID 2019-107150RB-I00/AEI/ 10.13039/501100011033 to XC-V; by the “Ramón y Cajal” State Program, Ministry of Economy and Competitivenes