Modeling methyl-sensitive transcription factor motifs with an expanded epigenetic alphabet.

BACKGROUND: Transcription factors bind DNA in specific sequence contexts. In addition to distinguishing one nucleobase from another, some transcription factors can distinguish between unmodified and modified bases. Current models of transcription factor binding tend not to take DNA modifications into account, while the recent few that do often have limitations. This makes a comprehensive and accurate profiling of transcription factor affinities difficult. RESULTS: Here, we develop methods to identify transcription factor binding sites in modified DNA. Our models expand the standard A/C/G/T DNA alphabet to include cytosine modifications. We develop Cytomod to create modified genomic sequences and we also enhance the MEME Suite, adding the capacity to handle custom alphabets. We adapt the well-established position weight matrix (PWM) model of transcription factor binding affinity to this expanded DNA alphabet. Using these methods, we identify modification-sensitive transcription factor binding motifs. We confirm established binding preferences, such as the preference of ZFP57 and C/EBPß for methylated motifs and the preference of c-Myc for unmethylated E-box motifs. CONCLUSIONS: Using known binding preferences to tune model parameters, we discover novel modified motifs for a wide array of transcription factors. Finally, we validate our binding preference predictions for OCT4 using cleavage under targets and release using nuclease (CUT&RUN) experiments across conventional, methylation-, and hydroxymethylation-enriched sequences. Our approach readily extends to other DNA modifications. As more genome-wide single-base resolution modification data becomes available, we expect that our method will yield insights into altered transcription factor binding affinities across many different modifications.

Risk Factors for Postoperative Infections Following Appendectomy of Complicated Appendicitis: A Meta-analysis and Retrospective Single-institutional Study.

Patients with complicated appendicitis have an increased risk for postoperative infections. Potential risk factors for postoperative infections through a meta-analysis and retrospective chart review are discussed. A meta-analysis consisting of 35 studies analyzing complicated appendicitis treated with an appendectomy noting at least 1 postoperative infection was performed. A retrospective review was then conducted in patients diagnosed with complicated appendicitis after appendectomy. Of 5326 patients in total, 15.4% developed postoperative infections. Laparoscopic surgery and perioperative hyperoxygenation were found to be protective factors for the development of infection. Retrospectively, 53.2% of patients presented with complicated appendicitis. Patients with complicated appendicitis were more likely to be older in age and have an increased length of stay. Patient demographics, operative time, and comorbid status had no effect on postoperative infection or readmission rate. Physicians should strongly consider minimally invasive techniques to treat all cases of complicated appendicitis irrespective of comorbidities, age, sex, or body mass index.

Survival benefits of extensive surgery in patients with papillary thyroid microcarcinoma.

BACKGROUND: Despite the guidelines recommending thyroid lobectomy, many papillary thyroid microcarcinoma (PTMC) patients still undergo total thyroidectomy. PTMC's optimal treatment remains unclear. We aimed to determine whether total thyroidectomy improves outcomes compared to less extensive surgery. METHODS: We analyzed 6064 PTMC adult patients from the Surveillance, Epidemiology, and End Results (SEER) database (2000-2019) who underwent either total thyroidectomy (n â€‹= â€‹3652) or less extensive surgery (n â€‹= â€‹2412). Endpoints were overall survival, cancer-specific survival, and recurrence. RESULTS: Total thyroidectomy patients had a 5.2 â€‹% mortality rate versus 8.1 â€‹% with less extensive surgery. Recurrence occurred in 1 (0.03 â€‹%) total thyroidectomy patient compared to 24 (1.0 â€‹%) less extensive surgery patients (HR 0.07, p â€‹= â€‹0.01). Median survival was 8.1 years for total thyroidectomy versus 8.8 years for less extensive surgery. Overall survival favored total thyroidectomy (p â€‹= â€‹0.001) but cancer-specific survival did not differ. CONCLUSION: Although total thyroidectomy may not improve cancer-specific survival, it lowers recurrence risk and confers an overall survival advantage for PTMC patients. These findings may help guide surgical decisions.

Anemonefishes: A model system for evolutionary genomics.

Anemonefishes are an iconic group of coral reef fish particularly known for their mutualistic relationship with sea anemones. This mutualism is especially intriguing as it likely prompted the rapid diversification of anemonefish. Understanding the genomic architecture underlying this process has indeed become one of the holy grails of evolutionary research in these fishes. Recently, anemonefishes have also been used as a model system to study the molecular basis of highly complex traits such as color patterning, social sex change, larval dispersal and life span. Extensive genomic resources including several high-quality reference genomes, a linkage map, and various genetic tools have indeed enabled the identification of genomic features controlling some of these fascinating attributes, but also provided insights into the molecular mechanisms underlying adaptive responses to changing environments. Here, we review the latest findings and new avenues of research that have led to this group of fish being regarded as a model for evolutionary genomics.

OPA1 disease-causing mutants have domain-specific effects on mitochondrial ultrastructure and fusion.

Inner mitochondrial membrane fusion and cristae shape depend on optic atrophy protein 1, OPA1. Mutations in OPA1 lead to autosomal dominant optic atrophy (ADOA), an important cause of inherited blindness. The Guanosin Triphosphatase (GTPase) and GTPase effector domains (GEDs) of OPA1 are essential for mitochondrial fusion; yet, their specific roles remain elusive. Intriguingly, patients carrying OPA1 GTPase mutations have a higher risk of developing more severe multisystemic symptoms in addition to optic atrophy, suggesting pathogenic contributions for the GTPase and GED domains, respectively. We studied OPA1 GTPase and GED mutations to understand their domain-specific contribution to protein function by analyzing patient-derived cells and gain-of-function paradigms. Mitochondria from OPA1 GTPase (c.870+5G>A and c.889C>T) and GED (c.2713C>T and c.2818+5G>A) mutants display distinct aberrant cristae ultrastructure. While all OPA1 mutants inhibited mitochondrial fusion, some GTPase mutants resulted in elongated mitochondria, suggesting fission inhibition. We show that the GED is dispensable for fusion and OPA1 oligomer formation but necessary for GTPase activity. Finally, splicing defect mutants displayed a posttranslational haploinsufficiency-like phenotype but retained domain-specific dysfunctions. Thus, OPA1 domain-specific mutants result in distinct impairments in mitochondrial dynamics, providing insight into OPA1 function and its contribution to ADOA pathogenesis and severity.

The chromosome-scale genome assembly of the yellowtail clownfish Amphiprion clarkii provides insights into the melanic pigmentation of anemonefish.

Anemonefish are an emerging group of model organisms for studying genetic, ecological, evolutionary, and developmental traits of coral reef fish. The yellowtail clownfish Amphiprion clarkii possesses species-specific characteristics such as inter-species co-habitation, high intra-species color variation, no anemone specificity, and a broad geographic distribution, that can increase our understanding of anemonefish evolutionary history, behavioral strategies, fish-anemone symbiosis, and color pattern evolution. Despite its position as an emerging model species, the genome of A. clarkii is yet to be published. Using PacBio long-read sequencing and Hi-C chromatin capture technology, we generated a high-quality chromosome-scale genome assembly initially comprised of 1,840 contigs with an N50 of 1,203,211 bp. These contigs were successfully anchored into 24 chromosomes of 843,582,782 bp and annotated with 25,050 protein-coding genes encompassing 97.0% of conserved actinopterygian genes, making the quality and completeness of this genome the highest among all published anemonefish genomes to date. Transcriptomic analysis identified tissue-specific gene expression patterns, with the brain and optic lobe having the largest number of expressed genes. Further analyses revealed higher copy numbers of erbb3b (a gene involved in melanocyte development) in A. clarkii compared with other anemonefish, thus suggesting a possible link between erbb3b and the natural melanism polymorphism observed in A. clarkii. The publication of this high-quality genome, along with A. clarkii's many unique traits, position this species as an ideal model organism for addressing scientific questions across a range of disciplines.

A feasible method for llama embryo paraffin embedding.

Maternal recognition of pregnancy in llamas would be related to a still-unknown signal secreted by the embryo to extend the corpus luteum lifespan. In order to study llamas reproductive physiology and early pregnancy aspects, research on embryos between 8 and 12 days post-mating is essential. However, there is a lack of information on llama embryo paraffin embedding, a procedure that exhibits difficulties, mostly for embryo manipulation. This work presents an accessible, easy and rapid protocol for embedding llama blastocysts. Uterine flushings from four superovulated llamas were performed to recover embryos 8 days post-mating. Each embryo was fixed in 1 ml of 10 % neutral buffered formalin for 24 h, at 4 °C, and transferred to 70 % ethanol. Later, they were submitted to a series of baths: 500 µl PBS with 5 % bovine foetal serum solution (PBS+BFS) for 1 min; zona pellucida permeabilization with 500 µl PBS with 0.2 % Triton X-100 for 20 min; 500 µl PBS+BFS for 1 min; dehydration on baths with ascendent grades of ethanol (70 %, 96 %, and 100 % twice) for 10 min each; and bath of butanol for 5 min. Finally, embryos were embedded in a pellet of paraffin and afterwards in a paraffin block. Complete sections from 8 of 13 grade I and II embryos were obtained. In conclusion, by applying this method, good quality sections with preserved embryo morphology suitable for histological, histochemical or immunohistochemical approaches would be possible to acquire.

A chromosome-scale genome assembly of the false clownfish, Amphiprion ocellaris.

The false clownfish Amphiprion ocellaris is a popular fish species and an emerging model organism for studying the ecology, evolution, adaptation, and developmental biology of reef fishes. Despite this, high-quality genomic resources for this species are scarce, hindering advanced genomic analyses. Leveraging the power of PacBio long-read sequencing and Hi-C chromosome conformation capture techniques, we constructed a high-quality chromosome-scale genome assembly for the clownfish A. ocellaris. The initial genome assembly comprised of 1,551 contigs of 861.42 Mb, with an N50 of 863.85 kb. Hi-C scaffolding of the genome resulted in 24 chromosomes containing 856.61 Mb. The genome was annotated with 26,797 protein-coding genes and had 96.62% completeness of conserved actinopterygian genes, making this genome the most complete and high quality among published anemonefish genomes. Transcriptomic analysis identified tissue-specific gene expression patterns, with the brain and optic lobe having the largest number of expressed genes. Further, comparative genomic analysis revealed 91 genome elements conserved only in A. ocellaris and its sister species Amphiprion percula, and not in other anemonefish species. These elements are close to genes that are involved in various nervous system functions and exhibited distinct expression patterns in brain tissue, potentially highlighting the genetic toolkits involved in lineage-specific divergence and behaviors of the clownfish branch. Overall, our study provides the highest quality A. ocellaris genome assembly and annotation to date, whilst also providing a valuable resource for understanding the ecology and evolution of reef fishes.

Unveiling RCOR1 as a rheostat at transcriptionally permissive chromatin.

RCOR1 is a known transcription repressor that recruits and positions LSD1 and HDAC1/2 on chromatin to erase histone methylation and acetylation. However, there is currently an incomplete understanding of RCOR1's range of localization and function. Here, we probe RCOR1's distribution on a genome-wide scale and unexpectedly find that RCOR1 is predominantly associated with transcriptionally active genes. Biochemical analysis reveals that RCOR1 associates with RNA Polymerase II (POL-II) during transcription and deacetylates its carboxy-terminal domain (CTD) at lysine 7. We provide evidence that this non-canonical RCOR1 activity is linked to dampening of POL-II productive elongation at actively transcribing genes. Thus, RCOR1 represses transcription in two ways-first, via a canonical mechanism by erasing transcriptionally permissive histone modifications through associating with HDACs and, second, via a non-canonical mechanism that deacetylates RNA POL-II's CTD to inhibit productive elongation. We conclude that RCOR1 is a transcription rheostat.

Needs for a Curricular Change in Primary and Secondary Education From the One Health Perspective: A Pilot Study on Pneumonia in Schools.

This is the first pilot study on alternative conceptions and obstacles pertaining to pneumonia in adolescents of different school vulnerability indexes. Countries with low socioeconomic levels are disproportionately affected, with Latin America and the Caribbean (LAC) being the second-most affected area in the world, after sub-Saharan Africa. In spite of this fact, pneumonia is not included as an important component within the contents of the microbiology curriculum unit in the natural science school program. Therefore, we wanted to study how students knew about this topic by putting One Health into action by building and validating qualitative and quantitative questionnaires, put together by different experts in pedagogy, didactics, microbiology, and veterinary to find out what students knew about pneumonia and their misconceptions about it. A total of 148 students (in 8th and 9th grade) participated in this survey. The results reveal that no statistically significant differences between the different scholar grades (p = 0.3360 Pearson chi∧2) or genders (p = 0.8000 Fisher's exact test) presented higher or lower School Vulnerability Index (SVI). Regardless of the social stratum or the level of vulnerability of the students, they have heard about this disease primarily through their family/relatives, maintaining a superficial notion of the disease, learning wrong ideas about microorganisms and treatments that can contribute to the risk to public health.

New Insights From Transcriptomic Data Reveal Differential Effects of CO2 Acidification Stress on Photosynthesis of an Endosymbiotic Dinoflagellate in hospite.

Ocean acidification (OA) has both detrimental as well as beneficial effects on marine life; it negatively affects calcifiers while enhancing the productivity of photosynthetic organisms. To date, many studies have focused on the impacts of OA on calcification in reef-building corals, a process particularly susceptible to acidification. However, little is known about the effects of OA on their photosynthetic algal partners, with some studies suggesting potential benefits for symbiont productivity. Here, we investigated the transcriptomic response of the endosymbiont Symbiodinium microadriaticum (CCMP2467) in the Red Sea coral Stylophora pistillata subjected to different long-term (2 years) OA treatments (pH 8.0, 7.8, 7.4, 7.2). Transcriptomic analyses revealed that symbionts from corals under lower pH treatments responded to acidification by increasing the expression of genes related to photosynthesis and carbon-concentrating mechanisms. These processes were mostly up-regulated and associated metabolic pathways were significantly enriched, suggesting an overall positive effect of OA on the expression of photosynthesis-related genes. To test this conclusion on a physiological level, we analyzed the symbiont's photochemical performance across treatments. However, in contrast to the beneficial effects suggested by the observed gene expression changes, we found significant impairment of photosynthesis with increasing pCO2. Collectively, our data suggest that over-expression of photosynthesis-related genes is not a beneficial effect of OA but rather an acclimation response of the holobiont to different water chemistries. Our study highlights the complex effects of ocean acidification on these symbiotic organisms and the role of the host in determining symbiont productivity and performance.

Easy Access to Energy Fluctuations in Nonequilibrium Quantum Many-Body Systems.

We combine theoretical and experimental efforts to propose a method for studying energy fluctuations, in particular, to obtain the related bistochastic matrix of transition probabilities by means of simple measurements at the end of a protocol that drives a many-body quantum system out of equilibrium. This scheme is integrated with numerical optimizations in order to ensure a proper analysis of the experimental data, leading to physical probabilities. The method is experimentally evaluated employing a two interacting spin-1/2 system in a nuclear magnetic resonance setup. We show how to recover the transition probabilities using only local measures, which enables an experimental verification of the detailed fluctuation theorem in a many-body system driven out of equilibrium.

El trabajo emocional en la asesoría de negocios a emprendimientos y microempresas chilenas / Emotional labor in business consulting advisors for Chilean entrepreneurs and microenterprises

Resumen El trabajo emocional es un proceso multidimensional que adquiere relevancia debido a su creciente prevalencia y las serias consecuencias que tiene para trabajadores y organizaciones. La presente investigación busca describir las experiencias de trabajo emocional en trabajadores pertenecientes a centros de negocios que asesoran a emprendimientos y microempresas chilenas, cómo estas afectan su trabajo y las estrategias adoptadas para gestionarlo. Para esto, se organizaron cuatro grupos focales en cuatro centros de negocios con financiamiento público. Los análisis revelaron que el trabajo de los asesores/as requiere demandas de apoyo y contención emocional que están por fuera de su rol y competencias profesionales, lo cual tiene consecuencias negativas para su trabajo y su salud mental. Estas exigencias les hicieron desplegar una serie de estrategias individuales y grupales para disminuir los efectos negativos del trabajo emocional. Dentro de las estrategias individuales se observaron: establecer límites con los clientes, regular expectativas sobre las asesorías, desarrollar un endurecimiento emocional, practicar el autocuidado a través de la autoexploración emocional y, en algunos casos, buscar ayuda profesional de psicólogos/as. Como estrategia grupal, se observó el apoyo y la contención de los miembros de los equipos, los cuales, a su vez, facilitaban la derivación de clientes con situaciones complejas a asesores/as con más experiencia. Estos resultados muestran el carácter invisibilizado del trabajo emocional en la asesoría de negocios a emprendimientos y microempresas, como también la importancia de reconocerlo y gestionarlo como parte de los riesgos laborales para prevenir sus efectos en trabajadores y en las organizaciones.

Abstract Emotional labor is a multidimensional concept that includes the interactions between emotional requirements of the jobs, how emotions are expressed and the regulation strategies that it entails. In the last decades it has increased its importance due to its rising prevalence and its severe consequences for employee's health and organizational performance. Within this context, the present study seeks to address the different components of emotional labor and its effects on business consulting advisors for Chilean entrepreneurs and microenterprises, working in public-funded small business development centers. In doing so, four focus groups were carried out in four small business development centers located in Santiago of Chile. Analyses revealed that mentoring entrepreneurs and micro-enterprises is more complex than it appears, showing that advisors must provide support and comfort to their clients to succeed in their job goals. These demands go beyond their work role and exceed their competencies, which negatively impacts job performance and mental health of business advisors. In order to cope with the job demands, business advisors display several individual and collective strategies. As part of the individual strategies, the following was found: establishing boundaries with clients, regulating clients' expectations of the mentoring sessions, developing "emotional hardening", practicing self-care throughout emotional self-exploration, and in some cases, looking for psychological counseling. Interestingly, the term "emotional hardening" matches the definition of the superficial strategy formulated by Alicia Grandey, in which employees simulate or perform the emotion required by a job without changing their inner selves. Vast evidence has demonstrated that the use of this strategy is associated with serious health consequences for employees, therefore, this finding may be considered as a warning sign for the small business development centers. As a collective strategy, business advisors rely on their work teams. Specifically, advisors look for support and comfort from their co-workers and when confronted with complex clients the team referrers them to more experienced advisors. In addition, analyses showed other aspects that hinder the work of business advisors, such as the high amount and diversity of job tasks that are asked from business advisors, combined with several deadlines coming from government institutions and the centers. Furthermore, woman business advisors expressed they face more obstacles at work compared to their male co-workers, most of these obstacles are related to a gender-based discrimination coming from some clients, who doubt their expertise and competences as female advisors, and prefer to deal with male advisors. Overall, results aligned with the literature on emotional labor, showing that emotional labor is characterized by being a secondary and veiled issue in mentoring entrepreneurs and micro-enterprises. Furthermore, using the strategies revealed that emotional labor involves visible processes but also intrapsychic processes that led to a decrease in employees' resources and energy. Considering the effects of emotion labor in the context of business advisory, our results may help to strengthen the main Chilean strategy, and a well-known international program, to promote the development of entrepreneurs and microenterprises. This is even more relevant in the context of the social and employment crisis that Chile and most of the countries undergo, where entrepreneurship (formal and informal) has emerged as the main alternative to cope with unemployment. In conclusion, despite the long tradition of research on emotional labor and its serious consequences, it remains an invisible and secondary process within organizations. This research highlights the importance of identifying and managing emotions as a nuclear part of the job of business consulting advisors for Chilean entrepreneurs and microenterprises.

Temperature transcends partner specificity in the symbiosis establishment of a cnidarian.

Coral reef research has predominantly focused on the effect of temperature on the breakdown of coral-dinoflagellate symbioses. However, less is known about how increasing temperature affects the establishment of new coral-dinoflagellate associations. Inter-partner specificity and environment-dependent colonization are two constraints proposed to limit the acquisition of more heat tolerant symbionts. Here, we investigated the symbiotic dynamics of various photosymbionts in different host genotypes under "optimal" and elevated temperature conditions. To do this, we inoculated symbiont-free polyps of the sea anemone Exaiptasia pallida originating from Hawaii (H2), North Carolina (CC7), and the Red Sea (RS) with the same mixture of native symbiont strains (Breviolum minutum, Symbiodinium linucheae, S. microadriaticum, and a Breviolum type from the Red Sea) at 25 and 32 °C, and assessed their ITS2 composition, colonization rates, and PSII photochemical efficiency (Fv/Fm). Symbiont communities across thermal conditions differed significantly for all hosts, suggesting that temperature rather than partner specificity had a stronger effect on symbiosis establishment. Overall, we detected higher abundances of more heat resistant Symbiodiniaceae types in the 32 °C treatments. Our data further showed that PSII photophysiology under elevated temperature improved with thermal pre-exposure (i.e., higher Fv/Fm), yet, this effect depended on host genotype and was influenced by active feeding as photochemical efficiency dropped in response to food deprivation. These findings highlight the role of temperature and partner fidelity in the establishment and performance of symbiosis and demonstrate the importance of heterotrophy for symbiotic cnidarians to endure and recover from stress.

Immune-related IncRNA LINC00944 responds to variations in ADAR1 levels and it is associated with breast cancer prognosis.

AIMS: Breast cancer is one of the leading causes of woman deaths worldwide, being a major public health problem. It has been reported that the expression of the RNA-editing enzyme Adenosine Deaminase Acting on RNAs 1 (ADAR1) is upregulated in breast cancer, predicting poor prognosis in patients. A few reports in literature examine ADAR1 and long non-coding RNAs (lncRNAs) interplay in cancer and suggest key roles in cancer-related pathways. This study aimed to investigate whether ADAR1 could alter the expression levels of lncRNAs and explore how those changes are related to breast cancer biology. MAIN METHODS: ADAR1 overexpression and knockdown studies were performed in breast cancer cell lines to analyze the effects over lncRNAs expression. Guilt-by-Association correlation analysis of the TCGA-BRCA cohort was performed to predict the function of the lncRNA LINC00944. KEY FINDINGS: Here, we show that LINC00944 is responsive to ADAR1 up- and downregulation in breast cancer cells. We found that LINC00944 expression has a strong relationship with immune signaling pathways. Further assessment of the TCGA-BRCA cohort showed that LINC00944 expression was positively correlated to tumor-infiltrating T lymphocytes and pro-apoptotic markers. Moreover, we found that LINC00944 expression was correlated to the age at diagnosis, tumor size, and estrogen and progesterone receptor expression. Finally, we show that low expression of LINC00944 is correlated to poor prognosis in breast cancer patients. SIGNIFICANCE: Our study provides further evidence of the effect of ADAR1 over lncRNA expression levels, and on the participation of LINC00944 in breast cancer, suggesting to further investigate its potential role as prognostic biomarker.

Histopathological characterization of the reproductive organs of heifers experimentally infected with Campylobacter fetus venerealis / Caracterización histopatológica de los órganos reproductivos de vaquillonas experimentalmente infectadas con Campylobacter fetus venerealis / Caracterização histopatológica em órgãos reprodutores de novilhas infetadas no modo experimental com Campylobacter fetus venerealis

Abstract Background: Bovine campylobacteriosis is a venereal disease due to infection with Campylobacter fetus venerealis. It causes mainly reproductive failures that lead to considerable economic losses. Objective: To perform a histopathological description of the mucosa from reproductive organs of heifers experimentally infected with Campylobacter fetus venerealis. Methods: Twelve 15-18-months-old Aberdeen Angus heifers were treated for estrous synchronization and exposed to natural breeding. They were then randomly divided into two groups: group A (n=9) was inoculated with C. fetus venerealis; group B (n=3, control) was inoculated with a placebo. Ultrasonography was performed at days 29, 38, and 42 post-breeding, and plasmatic progesterone levels were quantified using ELISA to confirm pregnancies. Animals in group A with plasma progesterone levels below 1 ng/mL and/or diagnosed as non-pregnant were further divided into three subgroups: A1 (n=4), euthanized at day 30 post-breeding; A2 (n=3), euthanized at day 40 post-breeding and A3 (n=2), euthanized at day 55 post-breeding. Heifers from group B, all diagnosed as pregnant, were euthanized each at day 30, 40, and 55 days post-breeding as well. Histological sections from every group were taken from oviducts, uterus, and vagina. Results: Lymphocytic inflammation was the most common lesion in all infected heifers. Trophoblast cells were found in the non-pregnant heifers euthanized at days 40, and 55 post-breeding. The inflammatory process with the presence of lymphoid cells probably altered the balance in the activity of maternal lymphoid cells, as well as gene expression of the trophoblast, finally affecting the embryo survival. Conclusion: This work contributes to the understanding of the histopathological process involved in post-mating infection of Campylobacter fetus bovine.

Resumen Antecedentes: La campilobacteriosis bovina es una enfermedad venérea causada por el Campylobacter fetus venerealis, que produce principalmente fallas reproductivas ocasionando grandes pérdidas económicas Objetivo: Describir las características histopatológicas de la mucosa de órganos reproductores de vaquillonas infectadas experimentalmente con Campylobacter fetus venerealis. Métodos: Doce vaquillonas Aberdeen Angus (15 a 18 meses de edad) con celo sincronizado, recibieron servicio natural, e inmediatamente se dividieron aleatoriamente en dos grupos: A (n=9), inoculadas con Campylobacter fetus venerealis; B (n=3; control), inoculadas con placebo. El diagnóstico de preñez se realizó por ultrasonografía a los 29, 38 y 42 días post-servicio; los niveles plasmáticos de progesterona fueron determinados por ELISA. Las vaquillonas del grupo A con niveles de progesterona plasmáticos menores a 1 ng/mL y/o diagnosticadas no preñadas, fueron consideradas para eutanasia y divididas en tres subgrupos: A1-eutanasia día 30 (n=4); A2-día 40 (n=3); y A3-día 55 (n=2) post-servicio. Las vaquillonas del grupo B, diagnosticadas preñadas, fueron eutanasiadas a los 30, 40 y 55 días. Se tomaron muestras de oviductos, útero y vagina. Resultados: Se observó inflamación linfocitaria en la totalidad de muestras del grupo A. Células trofoblásticas fueron encontradas en muestras correspondientes a los grupos A2 y A3. Probablemente, el proceso inflamatorio alteró el equilibrio de las células linfoides maternas y la expresión génica del trofoblasto, afectando la supervivencia embrionaria. Conclusión: Este trabajo contribuye a la comprensión del proceso histopatológico involucrado en la infección poscoital por Campylobacter fetus bovino.

Resumo Antecedentes: A campilobacteriose bovina é uma doença venérea originada pelo Campylobacter fetus venerealis, quem produz principalmente falha reprodutiva e porém grandes perdas económicas. Objetivo: Descrever as características histopatológicas da mucosa dos órgãos reprodutores de novilhas infetadas no modo experimental com Campylobacter fetus venerealis. Métodos: Doze novilhas Aberdeen Angus de 15 até 18 meses com cio sincronizado, receberam serviço natural. Logo após, foram aleatóreamente separados em grupos: A (n=9) inoculados com Campylobacter fetus venerealis e grupo B (n=3; controle) inoculadas com um placebo. O diagnóstico da gestação foi realizado por ultrasom nos dias 29, 38 y 42 pós-serviço. Os níveis plasmáticos da progesterona foram determinados por ELISA. As novilhas do grupo A, com níveis plasmáticos de progesterona menores a 1 ng/mL e/ou diagnosticadas não grávidas, foram consideradas para eutanásia e foram divididas em três subgrupos: A1-eutanásia aos 30 dias pós- serviço (n=4); A2-dia 40 (n=3); A3-dia 55 (n=2). Foram realizada eutanásia ás novilhas do grupo B diagnosticadas prenhadas, aos 30, 40 e 55 dias e a amostragem de ovidutos, útero e vagina. Resultados: A presença de inflamação linfocitária foi observada na totalidade das amostras do grupo A. Foram achadas células trofoblásticas nas amostras correspondente aos grupos A2 e A3. Provavelmente, pelo processo inflamatório tenha sido alterado o equilíbrio das células linfoides maternas, assim também como a expressão gênica do trofoblasto, afetando a supervivência embrionária. Conclusão: Este trabalho contribue á compreensão do processo histopatologico na infecção com Campylobacter fetus bovino pós-acasalamento.

Unfamiliar partnerships limit cnidarian holobiont acclimation to warming.

Enhancing the resilience of corals to rising temperatures is now a matter of urgency, leading to growing efforts to explore the use of heat tolerant symbiont species to improve their thermal resilience. The notion that adaptive traits can be retained by transferring the symbionts alone, however, challenges the holobiont concept, a fundamental paradigm in coral research. Holobiont traits are products of a specific community (holobiont) and all its co-evolutionary and local adaptations, which might limit the retention or transference of holobiont traits by exchanging only one partner. Here we evaluate how interchanging partners affect the short- and long-term performance of holobionts under heat stress using clonal lineages of the cnidarian model system Aiptasia (host and Symbiodiniaceae strains) originating from distinct thermal environments. Our results show that holobionts from more thermally variable environments have higher plasticity to heat stress, but this resilience could not be transferred to other host genotypes through the exchange of symbionts. Importantly, our findings highlight the role of the host in determining holobiont productivity in response to thermal stress and indicate that local adaptations of holobionts will likely limit the efficacy of interchanging unfamiliar compartments to enhance thermal tolerance.

Effect of Mycobacterium cell wall fraction on endometrial histomorphometry of mares resistant and susceptible to persistent breeding-induced endometritis.

Mycobacterium cell wall fraction (MCWF) is a biological component made up of molecules with immunostimulant properties, which is therapeutically used to modulate persistent breeding-induced endometritis (PBIE). The aim of this study was to analyze the effect of this immunomodulator on the endometrial histological structure during the diestrus of PBIE-resistant and -susceptible mares that either received treatment with MCWF or not. The experiment was conducted with 10 resistant mares (RM) and 9 susceptible mares (SM). In the first estrous cycle of the trial, all mares were inseminated with dead semen as an inflammatory stimulus (Group A); at the next cycle, all mares were inseminated with dead semen and treated with a MCWF commercial immunomodulator (Group B). In both groups, endometrial biopsies were taken on day 7 post-ovulation (diestrus). Endometrial biopsies of untreated-RM (UTRM, n = 6), untreated-SM (UTSM, n = 7) MCWF-treated-RM (TRM, n = 6) and MCWF-treated-SM (TSM, n = 6) were evaluated. They were randomly chosen as representative mares of Group A and B, respectively. The height of lining and glandular epithelia, glandular diameter, glandular density and glandular area were evaluated. The histological structure revealed lymphocytic infiltration and dilated, tortuous glands with some glandular nests, particularly in UTSM. The histomorphometrical results showed no differences (ρ > 0.05) between the analyzed groups. This would indicate that post-service treatment with the MCWF immunomodulator does not modify the endometrial histoarchitecture but, apparently, its action would be mainly based on the stimulation of the cellular and humoral immune responses.

Long-Term Temperature Stress in the Coral Model Aiptasia Supports the "Anna Karenina Principle" for Bacterial Microbiomes.

The understanding of host-microbial partnerships has become a hot topic during the last decade as it has been shown that associated microbiota play critical roles in the host physiological functions and susceptibility to diseases. Moreover, the microbiome may contribute to host resilience to environmental stressors. The sea anemone Aiptasia is a good laboratory model system to study corals and their microbial symbiosis. In this regard, studying its bacterial microbiota provides a better understanding of cnidarian metaorganisms as a whole. Here, we investigated the bacterial communities of different Aiptasia host-symbiont combinations under long-term heat stress in laboratory conditions. Following a 16S rRNA gene sequencing approach we were able to detect significant differences in the bacterial composition and structure of Aiptasia reared at different temperatures. A higher number of taxa (i.e., species richness), and consequently increased α-diversity and ß-dispersion, were observed in the microbiomes of heat-stressed individuals across all host strains and experimental batches. Our findings are in line with the recently proposed Anna Karenina principle (AKP) for animal microbiomes, which states that dysbiotic or stressed organisms have a more variable and unstable microbiome than healthy ones. Microbial interactions affect the fitness and survival of their hosts, thus exploring the AKP effect on animal microbiomes is important to understand host resilience. Our data contributes to the current knowledge of the Aiptasia holobiont and to the growing field of study of host-associated microbiomes.

Complex Role for E-Prostanoid 4 Receptors in Hypertension.

Background Prostaglandin E2 ( PGE 2) is a major prostanoid with multiple actions that potentially affect blood pressure ( BP ). PGE 2 acts through 4 distinct E-prostanoid ( EP ) receptor isoforms: EP 1 to EP 4. The EP 4 receptor ( EP 4R) promotes PGE 2-dependent vasodilation, but its role in the pathogenesis of hypertension is not clear. Methods and Results To address this issue, we studied mice after temporal- and cell-specific deletion of EP 4R. First, using a mouse line with loss of EP 4 expression induced universally after birth, we confirm that EP 4R mediates a major portion of the acute vasodilatory effects of infused PGE 2. In addition, EP 4 contributes to control of resting BP , which was increased by 5±1 mm Hg in animals with generalized deficiency of this receptor. We also show that EP 4 is critical for limiting elevations in BP caused by high salt feeding and long-term infusion of angiotensin II . To more precisely identify the mechanism for these actions, we generated mice in which EP 4R loss is induced after birth and is limited to smooth muscle. In these mice, acute PGE 2-dependent vasodilation was attenuated, indicating that this response is mediated by EP 4R in vascular smooth muscle cells. However, absence of EP 4R only in this vascular compartment had a paradoxical effect of lowering resting BP , whereas the protective effect of EP 4R on limiting angiotensin II-dependent hypertension was unaffected. Conclusions Taken together, our findings support a complex role for EP 4R in regulation of BP and in hypertension, which appears to involve actions of the EP 4R in tissues beyond vascular smooth muscle cells.