Subpoblaciones espermáticas en el semen de conejo refrigerado con diferentes diluyentes

The reproductive activity of various species of mammals is influenced by the sperm subpopulations of their ejaculates. These subpopulations differ in the motility and kinetic characteristics of the sperm and have been related and classified according to their capacity for the fertilization process. The objective of this research was to evaluate the effect of the composition of the semen diluent on the sperm subpopulations of the chilled rabbit semen. The semen of 10 rabbits of the Californian breed (three ejaculated per animal) was obtained. Each ejaculate was divided into four aliquots, which were diluted in one of four diluents, whose basic composition consisted of T1: skim milk and sugars; T2: dextrose, sodium citrate and potassium acetate; T3: sodium caseinates, phosphates and sugars; and T4: tris-citric acid and egg yolk. Each aliquot was kept refrigerated at 16 °C for 72 h. Immediately after dilution and every 24 h, the distribution of four sperm subpopulations was evaluated using a computerized system: Type A (rapid progressive spermatozoa), Type B (rapid spermatozoa and medium progressive spermatozoa), Type C (medium spermatozoa and slow spermatozoa) and Type D (static spermatozoa). The data were analyzed using the PROC GLM and Tukey's HSD test of the SAS v. 9.2. It was found that T1 and T3 produced superior results for curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP) and amplitude of lateral head displacement (ALH) of sperm refrigerated for 24 and 48 hours (p<0.05), while T2 and T4 produced an increase in the beat cross frequency (BCF) (p<0.05). It is concluded that the composition of the diluent influences the distribution of the sperm subpopulations of the refrigerated rabbit semen, by generating changes in the progressivity and kinetics of the sperm.La actividad reproductiva de diversas especies de mamíferos se encuentra influenciada por las subpoblaciones espermáticas de sus eyaculados. Dichas subpoblaciones se diferencian en las características de movilidad y cinética de los espermatozoides y han sido relacionadas y catalogadas según su capacidad para el proceso de fertilización. El objetivo de este estudio fue evaluar el efecto de la composición del diluyente de semen sobre las subpoblaciones espermáticas del semen refrigerado de conejo. Se obtuvo el semen de 10 conejos de la raza Californiano (tres eyaculados por animal). Cada eyaculado se dividió en cuatro alícuotas, las cuales se diluyeron en uno de cuatro diluyentes, cuya composición básica consistió en T1: leche descremada y azucares; T2: dextrosa, citrato sódico y acetato potásico; T3: caseinatos de sodio, fosfatos y azúcares; y T4: tris-ácido cítrico y yema de huevo. Cada alícuota se mantuvo en refrigeración a 16 °C durante 72 horas. Inmediatamente después de la dilución y cada 24 horas se evaluó la distribución de cuatro subpoblaciones de espermatozoides mediante un sistema computarizado: Tipo A (espermatozoides rápidos progresivos), Tipo B (espermatozoides rápidos y espermatozoides medios progresivos), Tipo C (espermatozoides medios y espermatozoides lentos), y Tipo D (espermatozoides estáticos). Los datos se analizaron mediante el PROC GLM y la prueba HSD de Tukey del software SAS v. 9.2. Se encontró que T1 y T3 produjeron resultados superiores para la velocidad curvilínea (VCL), la velocidad rectilínea (VSL), la velocidad promedio (VAP) y la amplitud de desplazamiento lateral de la cabeza (ALH) de los espermatozoides refrigerados durante 24 y 48 horas (p<0.05), mientras que T2 y T4 produjeron un incremento en la frecuencia de batido (BCF) (p<0.05). Se concluye que la composición del diluyente influye sobre la distribución de las subpoblaciones espermáticas del semen refrigerado de conejo, al generar cambios en la progresividad y la cinética de los espermatozoides

The red fox (Vulpes vulpes) as a potential natural reservoir of human cryptosporidiosis by Cryptosporidium hominis in Northwest Spain.

Giardia duodenalis and Cryptosporidium spp. are ubiquitous intestinal protozoa that parasitize domestic and wild animals, as well as human beings. Due to their zoonotic potential, the objective of the present study was to determine the presence of these pathogens in the fox population (Vulpes vulpes) located in Northwest Spain. A total of 197 faecal samples from legally hunted foxes were collected in the autonomous region of Galicia. The presence of G. duodenalis and Cryptosporidium spp. was investigated by PCR-based methods amplifying the small subunit ribosomal RNA (ssu rRNA) gene of the parasites. Attempts to genotype obtained positive samples were subsequently conducted at the glutamate dehydrogenase (gdh) and β-giardin (bg) genes of G. duodenalis, and the 60 kDa glycoprotein (gp60) gene of Cryptosporidium. Giardia duodenalis and Cryptosporidium spp. were identified in 19 (9.6%) and 12 (6.1%) of the investigated samples, respectively. However, five Cryptosporidium species were detected at the ssu rRNA locus: C. hominis (33.4%, 4/12), C. canis (25.0%, 3/12), C. parvum (16.7%, 2/12), C. ubiquitum (8.3%, 1/12) and C. suis (8.3%, 1/12). An additional Cryptosporidium-positive sample was identified at the genus level only. Typing and subtyping of Giardia- and Cryptosporidium-positive samples were unsuccessful. The detection of C. hominis in wild foxes indicates the probable overlapping of sylvatic and domestic cycles of this parasite in rural settings. Besides, this finding raises the question of whether red foxes may act as natural reservoirs of C. hominis. The detection of C. parvum and C. suis is suggestive of active transmission events between farm and wild animals, opening up the possibility of transmission to human beings.The red foxes used in this study were provided by the Wildlife Recovery Centres of Galicia, Dirección Xeral de Patrimonio Natural (Xunta de Galicia, Spain) and by Federación Galega de Caza. Molecular analyses conducted in this survey were funded by the Health Institute Carlos III, Spanish Ministry of Economy and Competitiveness under project CP12/03081.S

Efecto de Cuatro Métodos de Separación Seminal sobre la Calidad y la Capacidad Fertilizante in vitro de Espermatozoides Equinos Criopreservados

The aim of this study was to evaluate the effect of four separation methods on seminal quality and in vitro fertilizing capacity of equine cryopreserved spermatozoa. Straws of equine cryopreserved semen were used for sperm separation through the Androcoll, CushionFluid, EquiPure and Percoll methods. A Sperm Class Analyzer (SCA®) system was used to determine total motility, progressive motility, curvilinear velocity, linear velocity and average speed. Also, intact acrosome and sperm vitality were evaluated by fluorescence microscopy. In vitro fertilizing capacity was assessed by in vitro fertilization of bovine oocytes with spermatozoa obtained by each separation method.Cleavage rates were determined after three days of in vitro culture. The results were analyzed using generalized linear models (GLM) and means of the four methods were compared using the Tukey test. The CushionFluid was superior for most parameters of sperm motility as well as for intact acrosome (p<0.05). Higher means of sperm vitality were obtained with EquiPure and Percoll (p<0.05), but no difference were found on cleavage rates between treatments. It was concluded that cryopreserved stallion semen selected by the CushionFluid method had better sperm quality compared to the other three methods; however no difference was evident between them for in vitro fertilizing capacity of spermatozoa.El objetivo del presente estudio fue evaluar el efecto de cuatro métodos de separación seminal sobre la calidad y la capacidad fertilizante in vitro de espermatozoides equinos criopreservados. Se utilizaron pajillas de semen equino criopreservado para la separación espermática a través de los métodos Androcoll, CushionFluid, EquiPure y Percoll. Mediante un sistema analizador de clase (SCA®) se evaluó la motilidad total, motilidad progresiva, velocidad curvilínea, velocidad lineal y velocidad media, y por microscopía de fluorescencia se determinaron los acrosomas intactos y la vitalidad espermática. La evaluación de la capacidad fertilizante in vitro se realizó mediante la fertilización in vitro de oocitos bovinos con espermatozoides obtenidos en cada método de separación. El clivaje se determinó después de tres días de cultivo in vitro. Los resultados se analizaron mediante modelos lineales generalizados (GLM) y las medias para los diferentes métodos se compararon usando la prueba de Tukey. El CushionFluid fue superior para la mayoría de parámetros de movilidad espermática, así como para acrosomas intactos (p<0.05). Se encontraron medias superiores de vitalidad espermática para EquiPure y Percoll (p<0.05), pero no se encontró diferencia entre las medias de clivaje entre tratamientos. Se concluye que el semen equino criopreservado y seleccionado por el método CushionFluid presenta mejores parámetros de calidad espermática respecto a los otros tres métodos; sin embargo, no se evidencia diferencia entre ellos para la capacidad fertilizante in vitro de los espermatozoides

Mathematical model of photovoltaic systems for distributed maximum power point tracking

Este articulo presenta un modelo para sistemas de generación fotovoltaica orientado a evaluar el desempeño de algoritmos distribuidos de búsqueda del punto de máxima potencia. El modelo considera la conexión en serie de n conjuntos módulo-convertidor que pueden ser dimensionados dependiendo de la aplicación en particular. Dicho modelo está compuesto por un conjunto de ecuaciones que pueden ser implementadas en cualquier software de cálculo matemático o en un lenguaje de programación que soluciones este tipo de ecuaciones. El modelo incluye el comportamiento estático del módulo fotovoltaico a partir de una ecuación que estima su relación voltaje-corriente, además, incluye el modelo del convertidor elevador de potencia con pérdidas en el inductor, esto con el fin de aproximar el modelo general a una solución realística de la aplicación. El modelo final es validado mediante simulaciones y comparado con una implementación del mismo en un software de simulación electrónica tradicional.This paper presents a model of photovoltaic generation systems oriented to evaluate the performance of distributed maximum power point tracking algorithms. The model considers the series connection of n module-converter sets, which can be dimensioned depending on the particular application. This model is composed of an equation set that can be implemented in any mathematical analysis software or programming language that allows to solve this kind of equations. The proposed model also includes the static behavior of the photovoltaic module, described by one equation that estimates the current-voltage relation. Additionally, it includes a model of the Boost power electronic converter with inductor lost, which allows an approximation to realistic applications. The final model is validated using simulations, and it is compared with a traditional electronic simulation software implementation

Kretzoiarctos gen. nov., the Oldest Member of the Giant Panda Clade

The phylogenetic position of the giant panda, Ailuropoda melanoleuca (Carnivora: Ursidae: Ailuropodinae), has been one of the most hotly debated topics by mammalian biologists and paleontologists during the last century. Based on molecular data, it is currently recognized as a true ursid, sister-taxon of the remaining extant bears, from which it would have diverged by the Early Miocene. However, from a paleobiogeographic and chronological perspective, the origin of the giant panda lineage has remained elusive due to the scarcity of the available Miocene fossil record. Until recently, the genus Ailurarctos from the Late Miocene of China (ca. 8–7 mya) was recognized as the oldest undoubted member of the Ailuropodinae, suggesting that the panda lineage might have originated from an Ursavus ancestor. The role of the purported ailuropodine Agriarctos, from the Miocene of Europe, in the origins of this clade has been generally dismissed due to the paucity of the available material. Here, we describe a new ailuropodine genus, Kretzoiarctos gen. nov., based on remains from two Middle Miocene (ca. 12–11 Ma) Spanish localities. A cladistic analysis of fossil and extant members of the Ursoidea confirms the inclusion of the new genus into the Ailuropodinae. Moreover, Kretzoiarctos precedes in time the previously-known, Late Miocene members of the giant panda clade from Eurasia (Agriarctos and Ailurarctos). The former can be therefore considered the oldest recorded member of the giant panda lineage, which has significant implications for understanding the origins of this clade from a paleobiogeographic viewpoint

Impact of the Mitochondrial Genetic Background in Complex III Deficiency

BACKGROUND: In recent years clinical evidence has emphasized the importance of the mtDNA genetic background that hosts a primary pathogenic mutation in the clinical expression of mitochondrial disorders, but little experimental confirmation has been provided. We have analyzed the pathogenic role of a novel homoplasmic mutation (m.15533 A>G) in the cytochrome b (MT-CYB) gene in a patient presenting with lactic acidosis, seizures, mild mental delay, and behaviour abnormalities. METHODOLOGY: Spectrophotometric analyses of the respiratory chain enzyme activities were performed in different tissues, the whole muscle mitochondrial DNA of the patient was sequenced, and the novel mutation was confirmed by PCR-RFLP. Transmitochondrial cybrids were constructed to confirm the pathogenicity of the mutation, and assembly/stability studies were carried out in fibroblasts and cybrids by means of mitochondrial translation inhibition in combination with blue native gel electrophoresis. PRINCIPAL FINDINGS: Biochemical analyses revealed a decrease in respiratory chain complex III activity in patient's skeletal muscle, and a combined enzyme defect of complexes III and IV in fibroblasts. Mutant transmitochondrial cybrids restored normal enzyme activities and steady-state protein levels, the mutation was mildly conserved along evolution, and the proband's mother and maternal aunt, both clinically unaffected, also harboured the homoplasmic mutation. These data suggested a nuclear genetic origin of the disease. However, by forcing the de novo functioning of the OXPHOS system, a severe delay in the biogenesis of the respiratory chain complexes was observed in the mutants, which demonstrated a direct functional effect of the mitochondrial genetic background. CONCLUSIONS: Our results point to possible pitfalls in the detection of pathogenic mitochondrial mutations, and highlight the role of the genetic mtDNA background in the development of mitochondrial disorders

Structure and non-structure of centrosomal proteins

Here we perform a large-scale study of the structural properties and the expression of proteins that constitute the human Centrosome. Centrosomal proteins tend to be larger than generic human proteins (control set), since their genes contain in average more exons (20.3 versus 14.6). They are rich in predicted disordered regions, which cover 57% of their length, compared to 39% in the general human proteome. They also contain several regions that are dually predicted to be disordered and coiled-coil at the same time: 55 proteins (15%) contain disordered and coiled-coil fragments that cover more than 20% of their length. Helices prevail over strands in regions homologous to known structures (47% predicted helical residues against 17% predicted as strands), and even more in the whole centrosomal proteome (52% against 7%), while for control human proteins 34.5% of the residues are predicted as helical and 12.8% are predicted as strands. This difference is mainly due to residues predicted as disordered and helical (30% in centrosomal and 9.4% in control proteins), which may correspond to alpha-helix forming molecular recognition features (α-MoRFs). We performed expression assays for 120 full-length centrosomal proteins and 72 domain constructs that we have predicted to be globular. These full-length proteins are often insoluble: Only 39 out of 120 expressed proteins (32%) and 19 out of 72 domains (26%) were soluble. We built or retrieved structural models for 277 out of 361 human proteins whose centrosomal localization has been experimentally verified. We could not find any suitable structural template with more than 20% sequence identity for 84 centrosomal proteins (23%), for which around 74% of the residues are predicted to be disordered or coiled-coils. The three-dimensional models that we built are available at http://ub.cbm.uam.es/centrosome/models/index.php

Identification of Plasmodium vivax Proteins with Potential Role in Invasion Using Sequence Redundancy Reduction and Profile Hidden Markov Models

BACKGROUND: This study describes a bioinformatics approach designed to identify Plasmodium vivax proteins potentially involved in reticulocyte invasion. Specifically, different protein training sets were built and tuned based on different biological parameters, such as experimental evidence of secretion and/or involvement in invasion-related processes. A profile-based sequence method supported by hidden Markov models (HMMs) was then used to build classifiers to search for biologically-related proteins. The transcriptional profile of the P. vivax intra-erythrocyte developmental cycle was then screened using these classifiers. RESULTS: A bioinformatics methodology for identifying potentially secreted P. vivax proteins was designed using sequence redundancy reduction and probabilistic profiles. This methodology led to identifying a set of 45 proteins that are potentially secreted during the P. vivax intra-erythrocyte development cycle and could be involved in cell invasion. Thirteen of the 45 proteins have already been described as vaccine candidates; there is experimental evidence of protein expression for 7 of the 32 remaining ones, while no previous studies of expression, function or immunology have been carried out for the additional 25. CONCLUSIONS: The results support the idea that probabilistic techniques like profile HMMs improve similarity searches. Also, different adjustments such as sequence redundancy reduction using Pisces or Cd-Hit allowed data clustering based on rational reproducible measurements. This kind of approach for selecting proteins with specific functions is highly important for supporting large-scale analyses that could aid in the identification of genes encoding potential new target antigens for vaccine development and drug design. The present study has led to targeting 32 proteins for further testing regarding their ability to induce protective immune responses against P. vivax malaria

Estimating the global conservation status of more than 15,000 Amazonian tree species

Estimates of extinction risk for Amazonian plant and animal species are rare and not often incorporated into land-use policy and conservation planning. We overlay spatial distribution models with historical and projected deforestation to show that at least 36% and up to 57% of all Amazonian tree species are likely to qualify as globally threatened under International Union for Conservation of Nature (IUCN) Red List criteria. If confirmed, these results would increase the number of threatened plant species on Earth by 22%. We show that the trends observed in Amazonia apply to trees throughout the tropics, and we predict thatmost of the world’s >40,000 tropical tree species now qualify as globally threatened. A gap analysis suggests that existing Amazonian protected areas and indigenous territories will protect viable populations of most threatened species if these areas suffer no further degradation, highlighting the key roles that protected areas, indigenous peoples, and improved governance can play in preventing large-scale extinctions in the tropics in this century

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality