Coculture with porcine luteal cells during in vitro porcine oocyte maturation affects lipid content, cortical reaction and zona pellucida ultrastructure.

CONTEXT: In pigs, in vitro fertilisation (IVF) is associated with high polyspermy rates, and for this reason, in vitro embryo production (IVP) is still an inefficient biotechnology. Coculture with somatic cells is an alternative to improve suboptimal in vitro maturation (IVM) conditions. AIM: This study was conducted to test a coculture system of porcine luteal cells (PLC) and cumulus-oocyte complexes (COC) to improve oocyte metabolism. METHODS: COC were matured in vitro with PLC. Oocyte lipid content, mitochondrial activity, zona pellucida (ZP) digestibility and pore size, cortical reaction and in vitro embryo development were assessed. KEY RESULTS: Coculture reduced cytoplasmic lipid content in the oocyte cytoplasm without increasing mitochondrial activity. Although ZP digestibility and ZP pore number were not different between culture systems, ZP pores were smaller in the coculture. Coculture impacted the distribution of cortical granules as they were found immediately under the oolemma, and more of them had released their content in the ZP. Coculture with porcine luteal cells during IVM increased monospermic penetration and embryo development after IVF. CONCLUSIONS: The coculture of COC with PLC affects the metabolism of the oocyte and benefits monospermic penetration and embryo development. IMPLICATIONS: The coculture system with PLC could be an alternative for the conventional maturation medium in pigs.

Host phylogeny and environment shape the diversity of salamander skin bacterial communities.

The composition and diversity of animal-associated microbial communities are shaped by multiple ecological and evolutionary processes acting at different spatial and temporal scales. Skin microbiomes are thought to be strongly influenced by the environment due to the direct interaction of the host's skin with the external media. As expected, the diversity of amphibian skin microbiomes is shaped by climate and host sampling habitats, whereas phylogenetic effects appear to be weak. However, the relative strength of phylogenetic and environmental effects on salamander skin microbiomes remains poorly understood. Here, we analysed sequence data from 1164 adult salamanders of 44 species to characterise and compare the diversity and composition of skin bacteria. We assessed the relative contribution of climate, host sampling habitat, and host phylogeny to the observed patterns of bacterial diversity. We found that bacterial alpha diversity was mainly associated with host sampling habitat and climate, but that bacterial beta diversity was more strongly associated with host taxonomy and phylogeny. This phylogenetic effect predominantly occurred at intermediate levels of host divergence (0-50 Mya). Our results support the importance of environmental factors shaping the diversity of salamander skin microbiota, but also support host phylogenetic history as a major factor shaping these bacterial communities.

Efecto perjudicial de la pandemia por SARS-CoV-2 en la sintomatología digestiva en pacientes con síndrome de intestino irritable / Detrimental impact of SARS-CoV-2 pandemic in the symptomatology of patients with irritable bowel syndrome

Irritable Bowel Syndrome (IBS) is a more frequent disorder in the brain-gut axis interaction in the world. COVID-19 has affected the population's mental health, and its impact on clinical severity in patients with IBS is unknown. Objective: To evaluate the effect of psychosocial stress produced by the pandemic on the severity of gastrointestinal symptoms. Methodology: 54 women and three men with IBS were interviewed by telephone. Factors associated with quality of life, comorbidities, IBS subtype, and COVID-19 diagnosis were asked. Calls were developed between June 2020 to January 2021. Results: 75% had Diarrheal IBS (IBS-D), 67% had comorbidities, 47% with busy work, and 70% in person, five patients (9%) were diagnosed with COVID-19. Of the total, 88% referred to change in gastrointestinal symptoms, 56% increased abdominal pain, and 95% bloating. Abdominal pain was negatively associated with quality of life (p < 0.036), and the incomplete evacuation's sensation positively with difficulty sleeping (p < 0.034). Conclusion: In this study, IBS patients interviewed by telephone reported higher abdominal pain and subjective bloating associated with the pandemic by SARS-CoV-2. Keywords: Irritable

El Síndrome de Intestino Irritable (SII) es uno de los trastornos en la interacción cerebrointestino más frecuentes en el mundo. La pandemia COVID-19 ha afectado la salud mental de la población, siendo desconocido su impacto en la severidad clínica en pacientes con SII. Objetivo: Evaluar el efecto del estrés psicosocial producido por la pandemia en la severidad de síntomas gastrointestinales de pacientes con SII. Metodología: 54 mujeres y 3 hombres con SII fueron entrevistados vía telefónica. Se preguntó por factores asociados a calidad de vida, comorbilidades, subtipo de SII y diagnóstico de COVID-19. Las llamadas se realizaron entre junio de 2020 hasta enero de 2021. Resultados: Un 75% presentó SII Diarreico (SII-D), el 67% comorbilidades, el 47% con trabajo activo y 70% presencial, 5 pacientes (9%) diagnosticados COVID-19. Del total, 88% refirió cambio en síntomas gastrointestinales, 56% aumentó el dolor abdominal y 95% la distensión abdominal. El dolor abdominal se asoció negativamente con la calidad de vida (p < 0,036), y la sensación de evacuación incompleta positivamente con la dificultad para dormir (p < 0,034). Conclusión: En este estudio, los pacientes con SII entrevistados vía telefónica reportaron mayor dolor y distensión abdominal subjetiva asociado a la pandemia por SARS-CoV-2.

Expression of GnRH receptor and 3ßHSD during meiosis and foliculogénesis in Columba livia (Aves: Columbiformes): Histological and immunohistochemical analysis.

During the ovarian ontogeny in birds, five fundamental events can be recognized: migration and colonization of the primordial germ cells, differentiation and proliferation of oogonies, an organization of germinal nests, beginning of the meiotic process and folliculogenesis. The knowledge of these events is fundamental for the interpretation of the processes involved in the differentiation of female gametes. However, there are only references for some model species such as Gallus gallus domesticus and Coturnix coturnix. In a previous study, the histological structure of embryonic ovaries of Columba livia was revealed. Therefore, the objective of this work is to characterize the processes of meiosis and folliculogenesis C. livia from the analysis of the expression of the GnRH receptor, the 3ßHSD enzyme and the cell proliferation protein PCNA in embryonic and postnatal ovaries. Therefore, the expression of GnRHR, 3ßHSD, and PCNA was revealed in histological testicular and ovarian preparations in embryos (stages 41-43) and neonates (2, 5, 7, 10 and 75 days post-hatching). The present study demonstrates that the fate of germline cells is dictated by their location during gonadal development. Thus, the germline cells located in the cortex of the left gonad enter meiosis, while those in the right gonad and those in the medulla of the left ovary fail to go into meiosis. This indicates that somatic signals, instead of an autonomous cellular mechanism, regulate the entry of the germline cells into meiosis in the C. livia embryo. Future studies will be focused on the analysis of proteins associated with meiotic events and folliculogenesis in embryonic and neonatal ovaries of C. livia, to evaluate the regulation of meiosis in vitro.

Differential expression of E- and N- cadherins during gonadal histogenesis in Columba livia (Aves: Columbiformes).

The spatial organization of cells during tissue differentiation is a crucial process in the morphogenesis of vertebrates. This process involves the movement, separation, and connection of cells. It is essential to elucidate the molecular mechanisms involved in these processes for the understanding of animal morphogenesis. Cell-cell adhesion molecules, called cadherins, are involved in the selective adhesion of cells. In the case of birds, the expression of these molecules in various organ systems during embryonic development has been reported in Gallus gallus domesticus. In this work, we present the immunohistochemical analysis of the differential expression of E and N-cadherin binding molecules in Columba livia embryos at various stages of gonadal morphogenesis. The expression of E and N-cadherin in embryos corresponding to the stages 41, 43 and in neonates of 2, 5, 7 and 75 post-hatching days were assessed by immunohistochemistry. Results revealed the expression of N-cadherin in the plasma membrane and the perinuclear zone of germline cells in ovaries and testes. However, the expression of E-cadherin was noticed with similar immunoreactivity pattern, in Sertoli cells and in the cells of the follicular nests. The differential expression of follicular cells and Sertoli cells positive for E-cadherin and germline cell N-cadherin positive cells were evidenced in the present work at the cell-cell interaction level. Future studies will focus on determining the expression of E and N-cadherin molecules during the migration of the primordial germ cells and the colonization of the genital ridge.

Anatomical, histological and immunohistochemical study of testicular development in Columba livia (Aves: Columbiformes).

In this work, testicular ontogeny is analyzed at the anatomical, histological and immunohistochemical levels; the latter through the detection of GnRHR and PCNA in the testicles of embryos, neonates and juveniles of Columba livia. We analyzed 150 embryos, 25 neonates and 5 juveniles by means of observations under a stereoscopic magnifying glass and scanning electron microscope (SEM). The histological analysis was performed using hematoxylin-eosin staining techniques and the PAS reaction. For the immunohistochemical analysis, the expression of GnRHR and PCNA in embryos corresponding to stages 41, 43 and in neonates of 2, 5, 7 and 75 days post-hatch was revealed in testicular histological preparations. That gonadal outline is evident in stage 18. In stage 29, the testes are constituted of a medulla in which the PGCs are surrounded by the Sertoli cells, constituting the seminiferous tubules. From stage 37 a greater organization of the tubules is visualized and at the time of hatching the testicle is constituted of the closed seminiferous tubules, formed of the PGCs and Sertoli cells. The Leydig cells are evident outside the tubules. In the juvenile stages, the differentiation of germline cells and the organization of small vessels that irrigate the developing testicle begin to be visible. In the analyzed stages, the immunodetection of the GnRHR receptor and PCNA revealed specific marking in the plasma membrane and in the perinuclear zone for GnRHR and in the nucleus of the germline cells in juvenile testicles for PCNA. These results can be used as a basis for further study of endocrine regulation events during testicular ontogeny in avian species.

Apoptosis of bovine granulosa cells: Intracellular pathways and differentiation.

Follicular atresia in granulosa and theca cells occurs by apoptosis through weak hormonal stimulation. We have previously proposed an in vitro model to study this process by inducing apoptosis in BGC-1, a bovine granulosa cell line, and in primary cultures from ovaries with or without corpus luteum (CPGB+ and CPGB-, respectively), with different doses of gonadotropin releasing hormone (GnRH) analogs (leuprolide acetate (LA) as agonist and antide as antagonist). BGC-1 represent immature granulosa cells, whereas CPGB represent different degrees of luteinization. Our aim was to evaluate the intracellular pathways involved in the GnRH regulation of apoptosis in BGC-1. Treatment with LA 100nM but not with antide led to an increase in BAX over BCL-2 expression, showing antagonism of antide. All treatments inhibited phospholipase-D (PLD) activity compared to control, implying agonist behavior of antide. Progesterone in vitro production and 3ß-hydroxysteroid dehydrogenase (3ß-HSD) expression revealed different degrees of luteinization: BGC-1 were immature, whereas CPGB+ were less differentiated than CPGB-. We concluded that LA-induced apoptosis in BGC-1 occurs by activation of the mitochondrial pathway and by inhibition of PLD activity and that antide might work both as an antagonist of the intrinsic pathway and as an agonist of the extrinsic protection pathway by inhibiting PLD activity.

Tracking climate change in a dispersal-limited species: reduced spatial and genetic connectivity in a montane salamander.

Tropical montane taxa are often locally adapted to very specific climatic conditions, contributing to their lower dispersal potential across complex landscapes. Climate and landscape features in montane regions affect population genetic structure in predictable ways, yet few empirical studies quantify the effects of both factors in shaping genetic structure of montane-adapted taxa. Here, we considered temporal and spatial variability in climate to explain contemporary genetic differentiation between populations of the montane salamander, Pseudoeurycea leprosa. Specifically, we used ecological niche modelling (ENM) and measured spatial connectivity and gene flow (using both mtDNA and microsatellite markers) across extant populations of P. leprosa in the Trans-Mexican Volcanic Belt (TVB). Our results indicate significant spatial and genetic isolation among populations, but we cannot distinguish between isolation by distance over time or current landscape barriers as mechanisms shaping population genetic divergences. Combining ecological niche modelling, spatial connectivity analyses, and historical and contemporary genetic signatures from different classes of genetic markers allows for inference of historical evolutionary processes and predictions of the impacts future climate change will have on the genetic diversity of montane taxa with low dispersal rates. Pseudoeurycea leprosa is one montane species among many endemic to this region and thus is a case study for the continued persistence of spatially and genetically isolated populations in the highly biodiverse TVB of central Mexico.

A new genus and species of Brachycoeliidae (Digenea) from Chiropterotriton sp. (Caudata: Plethodontidae) in Mexico and its phylogenetic position within the Plagiorchiida based on partial sequences of the 28S ribosomal RNA gene.

Parabrachycoelium longicaecum n. gen., n. sp. (Digenea: Brachycoeliidae) is described from the intestine of a plethodontid salamander Chiropterotriton sp. Hosts were collected in bromeliads at the cloud forest of Tlaquilpa, Veracruz, Mexico. Members of the Brachycoeliidae Looss, 1899 (sensu Yamaguti, 1971) are characterized by having a spined tegument; ceca usually short, not passing level of gonads, but longer in some species; gonads posterior to, or in region of, acetabulum, with ovary anterior to testes; a well developed cirrus pouch containing a bipartite seminal vesicle; and uterus occupying entire hind-body posterior to testes. However, this combination of morphological traits prevents the inclusion of the new taxon in any of the genera in that family; a new genus was, therefore, erected to accommodate the new species. The new taxon is readily distinguished from members belonging to Brachycoelium Dujardin, 1845, Mesocoelium Odhner, 1910, and Tremiorchis Mehra and Negi, 1925, by having long ceca extending into the posterior third of the body, slightly surpassing the testes, and vitellaria extending along the body. The new species morphologically resembles Caudouterina rhyacotritoni Martin, 1966, a digenean parasitizing a plethodontid salamander; however, the latter species lacks spines in the tegument and is actually placed within the Allocreadiidae. To demonstrate further the phylogenetic position of the new taxon, we sequenced the D1-D3 regions of 28S rRNA gene and conducted a phylogenetic analysis of available sequences for the order to which brachycoeliids belong (Plagiorchiida). Sequence divergence of the partial 28S rRNA gene confirms its distinction from the aforementioned brachycoeliids, and the phylogenetic position within the Plagiorchiida places the new species as closely related to a clade formed by Brachycoelium + Mesocoelium. Divergence levels and phylogenetic position within the Plagiorchiida verifies the validity of the new genus and its inclusion in Brachycoeliidae.

Extreme morphological and ecological homoplasy in tropical salamanders.

Fossorial salamanders typically have elongate and attenuated heads and bodies, diminutive limbs, hands and feet, and extremely elongate tails. Batrachoseps from California, Lineatriton from eastern México, and Oedipina from southern México to Ecuador, all members of the family Plethodontidae, tribe Bolitoglossini, resemble one another in external morphology, which has evolved independently. Whereas Oedipina and Batrachoseps are elongate because there are more trunk vertebrae, a widespread homoplasy (parallelism) in salamanders, the genus Lineatriton is unique in having evolved convergently by an alternate "giraffe-neck" developmental program. Lineatriton has the same number of trunk vertebrae as related, nonelongated taxa, but individual trunk vertebrae are elongated. A robust phylogenetic hypothesis, based on sequences of three mtDNA genes, finds Lineatriton to be deeply nested within a clade characterized by generalized ecology and morphology. Lineatriton lineolus, the only currently recognized taxon in the genus, shows unanticipated genetic diversity. Surprisingly, geographically separated populations of L. lineolus are not monophyletic, but are sister taxa of different species of the morphologically generalized genus Pseudoeurycea. Lineatriton, long thought to be a unique monospecific lineage, is polyphyletic. Accordingly, the specialized morphology of Lineatriton displays homoplasy at two hierarchical levels: (i) with respect to other elongate lineages in the family (convergence), and (ii) within what is currently recognized as a single taxon (parallelism). These evolutionary events are of adaptive significance because to invade the lowland tropics salamanders must be either arboreal or fossorial; the repeated evolution of elongation and attenuation has led to multiple lowland invasions.

Biodiversity of Costa Rican salamanders: implications of high levels of genetic differentiation and phylogeographic structure for species formation.

Although salamanders are characteristic amphibians in Holarctic temperate habitats, in tropical regions they have diversified evolutionarily only in tropical America. An adaptive radiation centered in Middle America occurred late in the history of a single clade, the supergenus Bolitoglossa (Plethodontidae), and large numbers of species now occur in diverse habitats. Sublineages within this clade decrease in number from the northern to southern parts of Middle America, and in Costa Rica, there are but three. Despite this phylogenetic constraint, Costa Rica has many species; the number of salamander species on one local elevational transect in the Cordillera de Talamanca may be the largest for any such transect in the world. Extraordinary variation in sequences of the mitochondrial gene cytochrome b within a clade of the genus Bolitoglossa in Costa Rica reveals strong phylogeographic structure within a single species, Bolitoglossa pesrubra. Allozymic variation in 19 proteins reveals a pattern largely concordant with the mitochondrial DNA phylogeography. More species exist than are currently recognized. Diversification occurs in restricted geographic areas and involves sharp geographic and elevational differentiation and zonation. In their degree of genetic differentiation at a local scale, these species of the deep tropics exceed the known variation of extratropical salamanders, which also differ in being less restricted in elevational range. Salamanders display "tropicality" in that although speciose, they are usually local in distribution and rare. They display strong ecological and physiological differentiation that may contribute importantly to morphological divergence and species formation.

Efectos cardiovasculares del acortamiento del intervalo AV mediante estimulación eléctrica del corazón en miocardiopatía dilatada / Cardiovascular effects of AV interval shortening with heart electric stimulation in dilated cardiomyopathy

Patients and methods: Nine patients with dilated cardiomyopathy were studied. Hemodynamic and tissular perfusion values, echocardiographic and radioisotopic ventricular function parameters were measured before and after six hours of AV interval shortening with electrical stimulation of the heart. Results: After electrical stimulation, cardiac output increased from 3.38 ñ 0.8 to 32.87 ñ 0.79 l/min (p < 0.05). Pulmonary capillary pressure decreased from 23.8 ñ 8.9 to 19.8 ñ 9.2 mm Hg (p = NS). There were no significant changes in ventricular function parameters or in systemic and pulmonary pressures. Conclusions: Electrical shortening of AV interval in patients with dilated cardiomyopathy increases cardiac output but does not change ventricular function parameters

El valor de la cintigrafía en el diagnóstico de filtración biliar en el postoperatorio de la colecistectomía laparoscópica / The value of cintigraphy in biliary filtration diagnosis in laparoscopic cholecystectomy postsurgery

Los objetivos de este estudio fueron determinar la utilidad de la cintigrafía en el diagnóstico de filtración biliar en el postoperatorio de la colecistectomía laparoscópica y la incidencia de esta complicación en pacientes asintomáticos. Entre mayo de 1991 y marzo de 1993, 813 pacientes fueron sometidos a colecistectomía laparoscópica. A aquellos en que se sospechó la presencia de una filtración biliar se les practicó ecografía abdominal y cintigrafía biliar con TC 99-DISIDA. Entre enero y junio de 1994, a 43 pacientes sometidos a colecistectomía laparoscópica, cuya evolución postoperatoria fue normal, se les practicó una cintigrafía biliar a las 24 h del postoperatorio. De los 813 casos analizados en el primer período en cuatro de ellos se diagnosticó una colección intraabdominal y en 3 de éstos se demostró una filtración biliar por cintigrafía. La incidencia de filtración biliar sintomática fue de 0,36 porciento. De los pacientes estudiados al azar, la cintigrafía biliar fue normal en todos los casos con colelitiasis simple y en un paciente con colecistitis aguda se detectó una filtración biliar asintomática (2,32 porciento). Se concluye que la filtración biliar es una complicación poco frecuente de la colecistectomía laparoscópica y la cintigrafía biliar es un buen método para el diagnóstico de esta complicación

Evaluación cintigráfica de patología nefrourológica diagnosticada en período antenatal / Scintigraphic assessment of nephrourologic pathology with prenatal diagnosis

Desde el advenimiento de la ecografía obstétrica, el diagnóstico antenatal (DA) de patología nefrourológica (PNU) es relativamente frecuente. El objetivo de este trabajo es evaluar retrospectivamente la cintigrafía renal dinámica (CRD) y estática (CRE) en 30 pacientes (pac) entre 2 días y 13 meses de vida, 19 de los 30 pacientes eran recién nacidos, de los cuales siete fueron estudiados durante la primera semana de vida. Se realizó CRD con Tc99n DTPA ó 13, CRE con Tc99m DMSA en 17 pacientes y ambos estudios en 14 pacientes. Trece de las 60 unidades (UR) (21.7 por ciento) estaban excluídas, uno de las cuales bilateral, secundaria a valvas de la uretra posterior. La causa más común de exclusión fue riñón multicístico. Siete de los doce pacientes con exclusión unilateral presentaron patología del riñon contralateral (EPU). En ninguno de estos siete pacientes existía compromiso significativo de la función renal al momento del cintigrama. En diez UR la CRD mostró un patrón obstructivo, siendo en la mayoría (6UR) secundaria a EPU. En 17 UR la CRD mostró retención pielocaliceal con respuesta normal al diurético, 14 UR eran hidronefrosis transitoria del recién nacido. En 11 niños se realizó seguimiento cintigráfico, siete postquirúrgico, en 5 de estos pacientes se observó una mejoría de la curva de excreción (71,4 por ciento). En tres de los 4 pacientes no operados el seguimiento cintigráfico demostró un deterioro en la función renal y/o en la curva de excreción, lo que motivó una corrección quirúrgica