Biosensor capability of the endometrium is mediated in part, by altered miRNA cargo from conceptus-derived extracellular vesicles.

We tested the hypothesis that the biosensor capability of the endometrium is mediated in part, by the effect of different cargo contained in the extracellular vesicles secreted by the conceptus during the peri-implantation period of pregnancy. We transferred Bos taurus taurus embryos of different origin, in vivo (high developmental potential (IV)), in vitro (intermediate developmental potential (IVF)), or cloned (low developmental potential (NT)), into Bos taurus indicus recipients. Extracellular vesicles (EVs) recovered from Day 16 conceptus-conditioned medium were characterized and their microRNA (miRNA) cargo sequenced alongside RNA sequencing of their respective endometria. There were substantial differences in the endometrial response to in vivo versus in vitro and in vivo versus cloned conceptuses (1153 and 334DEGs respectively) with limited differences between in vitro Vs cloned conceptuses (36 DEGs). The miRNA cargo contained in conceptus-derived EVs was similar between all three groups (426 miRNA in common). Only 8 miRNAs were different between in vivo and cloned conceptuses, while only 6 miRNAs were different between in vivo and in vitro-derived conceptuses. Treatment of endometrial epithelial cells with mimic or inhibitors for miR-128 and miR-1298 changed the proteomic content of target cells (96 and 85, respectively) of which mRNAs are altered in the endometrium in vivo (PLXDC2, COPG1, HSPA12A, MCM5, TBL1XR1, and TTF). In conclusion, we have determined that the biosensor capability of the endometrium is mediated in part, by its response to different EVs miRNA cargo produced by the conceptus during the peri-implantation period of pregnancy.

The central role of pyruvate metabolism on the epigenetic maturation and transcriptional profile of bovine oocytes.

In brief: Pyruvate metabolism is one of the main metabolic pathways during oocyte maturation. This study demonstrates that pyruvate metabolism also regulates the epigenetic and molecular maturation in bovine oocytes. Abstract: Pyruvate, the final product of glycolysis, undergoes conversion into acetyl-CoA within the mitochondria of oocytes, serving as a primary fuel source for the tricarboxylic acid (TCA) cycle. The citrate generated in the TCA cycle can be transported to the cytoplasm and converted back into acetyl-CoA. This acetyl-CoA can either fuel lipid synthesis or act as a substrate for histone acetylation. This study aimed to investigate how pyruvate metabolism influences lysine 9 histone 3 acetylation (H3K9ac) dynamics and RNA transcription in bovine oocytes during in vitro maturation (IVM). Bovine cumulus-oocyte complexes were cultured in vitro for 24 h, considering three experimental groups: Control (IVM medium only), DCA (IVM supplemented with sodium dichloroacetate, a stimulant of pyruvate oxidation into acetyl-CoA), or IA (IVM supplemented with sodium iodoacetate, a glycolysis inhibitor). The results revealed significant alterations in oocyte metabolism in both treatments, promoting the utilization of lipids as an energy source. These changes during IVM affected the dynamics of H3K9ac, subsequently influencing the oocyte's transcriptional activity. In the DCA and IA groups, a total of 148 and 356 differentially expressed genes were identified, respectively, compared to the control group. These findings suggest that modifications in pyruvate metabolism trigger the activation of metabolic pathways, particularly lipid metabolism, changing acetyl-CoA availability and H3K9ac levels, ultimately impacting the mRNA content of in vitro matured bovine oocytes.

Hematological Normality, Serum Biochemistry, and Acute Phase Proteins in Healthy Beef Calves in the Brazilian Savannah.

The Brazilian savannah region, characterized by high average temperatures, well-defined rainy and dry seasons, soil with low productive potential, and high pressure for parasitic diseases, is home to the highest percentage of the beef herd, which is the world's largest commercial beef producer. Therefore, breeds that present rusticity combined with productivity are the focus of research in cattle breeding in the region. Considering their geographic particularities and their effects on the animals' blood parameters, the objective was to study the behavior of hematological variables, serum biochemistry, and acute phase proteins in beef calves at weaning age. Jugular blood samples were collected in a single day from 30 weaning calves (about eight months old and 200 kg of body weight) from the following breeds: Nellore, Senepol, Nellore × Aberdeen Angus cross, Nellore × Senepol cross, and Nellore × Aberdeen Angus × Senepol cross. Hematological data were obtained using an automatic cell counter, serum biochemical measurements were obtained using commercial kits, and the electrophoretogram was obtained using the SDS-page technique. In general, the results were consistent with data already published in similar situations regarding health status, age, and level of metabolic activity. However, differences observed between groups can be explained by differences observed in other concurrent variables like temperament. The pure zebu breed was more reactive than pure taurine. While crossbreds showed intermediate values, and parasitic infestation, the pure taurine breed with higher parasite infestation, while the zebu breed had lower values, which produced effects in some laboratory tests, and generated differences between breeds. In conclusion, the reference intervals available for healthy animals can be routinely used without interference from the geographic region for animals produced without nutritional failures as long as changes are recorded in pathological, infectious, metabolic, or nutritional deficiency situations. However, it is suggested that a study covering a larger number of herds may demonstrate a greater geographic effect on the studied variables.

An Exploratory Data Analysis from Ovine and Bovine RNA-Seq Identifies Pathways and Key Genes Related to Cervical Dilatation.

The present study developed a review and exploration of data in public and already validated repositories. The main objective was to identify the pathways involved in ruminants' cervical dilatation, which are conserved between cattle and sheep in the follicular and luteal phases of the reproductive cycle. In cattle, 1961 genes were more differentially expressed in the follicular phase and 1560 in the luteal phase. An amount of 24 genes were considered exclusively expressed from these. A total of 18 genes were in the follicular phase and 6 genes were in the luteal phase. In sheep, 2126 genes were more differentially expressed in the follicular phase and 2469 genes were more differentially expressed in the luteal phase. Hoxb genes were identified in both species and are correlated with the PI3K/Akt pathway. PI3K/Akt was also found in both cattle and sheep, appearing prominently in the follicular and luteal phases of both species. Our analyses have pointed out that the PI3K/Akt pathway and the Hoxb genes appear in prominence in modulating mechanisms that involve estrus alterations in the cervix. PI3K/Akt appears to be an important pathway in the cervical relaxation process.

Effect of Cow-Calf Supplementation on Gene Expression, Processes, and Pathways Related to Adipogenesis and Lipogenesis in Longissimus thoracis Muscle of F1 Angus × Nellore Cattle at Weaning.

The aim of this study was to identify differentially expressed genes, biological processes, and metabolic pathways related to adipogenesis and lipogenesis in calves receiving different diets during the cow-calf phase. Forty-eight uncastrated F1 Angus × Nellore males were randomly assigned to two treatments from thirty days of age to weaning: no creep feeding (G1) or creep feeding (G2). The creep feed offered contained ground corn (44.8%), soybean meal (40.4%), and mineral core (14.8%), with 22% crude protein and 65% total digestible nutrients in dry matter. After weaning, the animals were feedlot finished for 180 days and fed a single diet containing 12.6% forage and 87.4% corn-based concentrate. Longissimus thoracis muscle samples were collected by biopsy at weaning for transcriptome analysis and at slaughter for the measurement of intramuscular fat content (IMF) and marbling score (MS). Animals of G2 had 17.2% and 14.0% higher IMF and MS, respectively (p < 0.05). We identified 947 differentially expressed genes (log2 fold change 0.5, FDR 5%); of these, 504 were upregulated and 443 were downregulated in G2. Part of the genes upregulated in G2 were related to PPAR signaling (PPARA, SLC27A1, FABP3, and DBI), unsaturated fatty acid synthesis (FADS1, FADS2, SCD, and SCD5), and fatty acid metabolism (FASN, FADS1, FADS2, SCD, and SCD5). Regarding biological processes, the genes upregulated in G2 were related to cholesterol biosynthesis (EBP, CYP51A1, DHCR24, and LSS), unsaturated fatty acid biosynthesis (FADS2, SCD, SCD5, and FADS1), and insulin sensitivity (INSIG1 and LPIN2). Cow-calf supplementation G2 positively affected energy metabolism and lipid biosynthesis, and thus favored the deposition of marbling fat during the postweaning period, which was shown here in an unprecedented way, by analyzing the transcriptome, genes, pathways, and enriched processes due to the use of creep feeding.

Characterization of histone lysine ß-hydroxybutyrylation in bovine tissues, cells, and cumulus-oocyte complexes.

Besides their canonical roles as energy sources, short-chain fatty acids act as metabolic regulators of gene expression through histone posttranslational modifications. Ketone body ß-hydroxybutyrate (BHB) causes a novel epigenetic modification, histone lysine ß-hydroxybutyrylation (Kbhb), which is associated with genes upregulated in starvation-responsive metabolic pathways. Dairy cows increase BHB in early lactation, and the effects of this increase on cellular epigenomes are unknown. We searched for and identified that Kbhb is present in bovine tissues in vivo and confirmed that this epigenetic mark is responsive to BHB in bovine and human fibroblasts cultured in vitro in a dose-dependent manner. Maturation of cumulus-oocyte complexes with high concentrations of BHB did not affect the competence to complete meiotic maturation or to develop until the blastocyst stage. BHB treatment strongly induced H3K9bhb in cumulus cells, but faintly in oocytes. RNA-seq analysis in cumulus cells indicated that BHB treatment altered the expression of 345 genes. The downregulated genes were mainly involved in glycolysis and ribosome assembly pathways, while the upregulated genes were involved in mitochondrial metabolism and oocyte development. The genes and pathways altered by BHB will provide entry points to carry out functional experiments aiming to mitigate metabolic disorders and improve fertility in cattle.

CeTF: an R/Bioconductor package for transcription factor co-expression networks using regulatory impact factors (RIF) and partial correlation and information (PCIT) analysis.

BACKGROUND: Finding meaningful gene-gene interaction and the main Transcription Factors (TFs) in co-expression networks is one of the most important challenges in gene expression data mining. RESULTS: Here, we developed the R package "CeTF" that integrates the Partial Correlation with Information Theory (PCIT) and Regulatory Impact Factors (RIF) algorithms applied to gene expression data from microarray, RNA-seq, or single-cell RNA-seq platforms. This approach allows identifying the transcription factors most likely to regulate a given network in different biological systems - for example, regulation of gene pathways in tumor stromal cells and tumor cells of the same tumor. This pipeline can be easily integrated into the high-throughput analysis. To demonstrate the CeTF package application, we analyzed gastric cancer RNA-seq data obtained from TCGA (The Cancer Genome Atlas) and found the HOXB3 gene as the second most relevant TFs with a high regulatory impact (TFs-HRi) regulating gene pathways in the cell cycle. CONCLUSION: This preliminary finding shows the potential of CeTF to list master regulators of gene networks. CeTF was designed as a user-friendly tool that provides many highly automated functions without requiring the user to perform many complicated processes. It is available on Bioconductor ( http://bioconductor.org/packages/CeTF ) and GitHub ( http://github.com/cbiagii/CeTF ).

RNA sequencing data of different grade astrocytoma cell lines.

Astrocytomas are the most common and aggressive type of primary brain tumors in adults. The World Health Organization (WHO) assorts them into grades, from I to IV, based on histopathological features that reflect their malignancy [1]. Alongside with tumor progression, comes an increased proliferation, genomic instability, infiltration in normal brain tissue and resistance to treatments. The high genomic instability forges tumor cells enhancing key proteins that avoid cells from collapsing and favor therapy resistance [2]. To explore genes and pathways associated with tumor progression phenotypes we analyzed gene expression in a panel of non-tumor and glioma cell lines, namely: ACBRI371, non-tumor human astrocytes; HDPC, fibroblasts derived from dental pulp; Res186, Res259, Res286 and UW467 that include grade I, II and III astrocytoma cell lines derived from pediatric tumors; and T98G, U343MG, U87MG, U138MG and U251MG, all derived from GBM (grade IV). We also profiled gene expression changes caused by exogenously induced replicative stress, performing RNA sequencing with camptothecin (CPT)-treated cells. Here we describe the RNA-sequencing data set acquired, including quality of reads and sequencing consistency, as well as the bioinformatics strategy used to analyze it. We also compared gene expression patterns and pathway enrichment between non-tumor versus lower-grade (LGG), non-tumor versus GBM, LGG versus GBM, and CPT-treated versus non-treated cells. In brief, a total of 6467 genes showed differential expression and 5 pathways were enriched in tumor progression, while 2279 genes and 7 pathways were altered under the replication stress condition. The raw data was deposited in the NCBI BioProject database under the accession number PRJNA631805. Our dataset is valuable for researchers interested in differential gene expression among different astrocytoma grades and in expression changes caused by replicative stress, facilitating studies that seek novel biomarkers of glioma progression and treatment resistance.

Estrous cycle impacts microRNA content in extracellular vesicles that modulate bovine cumulus cell transcripts during in vitro maturation†.

Extracellular vesicles (EVs) are nanoparticles secreted by ovarian follicle cells. Extracellular vesicles are an important form of intercellular communication, since they carry bioactive contents, such as microRNAs (miRNAs), mRNAs, and proteins. MicroRNAs are small noncoding RNA capable of modulating mRNA translation. Thus, EVs can play a role in follicle and oocyte development. However, it is not clear if EV contents vary with the estrous cycle stage. The aim of this study was to investigate the bovine miRNA content in EVs obtained from follicles at different estrous cycle stages, which are associated with different progesterone (P4) levels in the follicular fluid (FF). We collected FF from 3 to 6 mm follicles and evaluated the miRNA profile of the EVs and their effects on cumulus-oocyte complexes during in vitro maturation. We observed that EVs from low P4 group have a higher abundance of miRNAs predicted to modulate pathways, such as MAPK, RNA transport, Hippo, Cell cycle, FoxO, oocyte meiosis, and TGF-beta. Additionally, EVs were taken up by cumulus cells and, thus, affected the RNA global profile 9 h after EV supplementation. Cumulus cells supplemented with EVs from low P4 presented upregulated genes that could modulate biological processes, such as oocyte development, immune responses, and Notch signaling compared with genes of cumulus cells in the EV free media or with EVs from high P4 follicles. In conclusion, our results demonstrate that EV miRNA contents are distinct in follicles exposed to different estrous cycle stage. Supplementation with EVs impacts gene expression and biological processes in cumulus cells.

Follicular dynamics and in vivo embryo production in Santa Inês ewes treated with smaller doses of pFSH.

To evaluate follicular dynamics, there was assessment of superovulatory response and in vivo embryo production in ewes treated with relatively smaller doses of exogenous pFSH than typically used in combination with a dose of eCG at the beginning of the gonadotropin treatment period. Santa Inês ewes (n = 24) were randomly divided into three groups, based on mg dose of pFSH administered: G200 (n = 8), G133 (n = 8) and G100 (n = 8) in eight decreasing doses at 12 -h intervals. All ewes were treated with 300 IU of eCG concomitantly starting with first pFSH administration. Ovulatory follicular dynamics and follicular wall vascularization (FWV) were evaluated using a B-mode and color Doppler ultrasonic machine, respectively. Superovulatory response and embryo production were evaluated 6 days after estrous detection. In the G200 group, the preovulatory follicle size (PFS) were less (P <  0.05), ovulation time later (P <  0.05), and PFS rate greater (P <  0.05); while in the G100 group ovulation rate, and number and percentage of unfertilized eggs were greater (P <  0.05) than in the G133 group (P <  0.05). Number and percentage of viable embryos were greater in the G200 and G100 compared to G133 group (P <  0.05). The dose of 100 mg of FSH was as efficacious as the traditional dose of 200 mg, in combination with a dose of eCG, for superovulatory response and viable embryo production but there was a greater percentage of unfertilized eggs with this treatment.

B-mode ultrasonography and ecobiometric parameters for assessment of embryonic and fetal development in sheep.

The aim in this study was to evaluate the maternal-fetal development during physiological gestation in sheep by means of ultrasonography and to predict the gestational age by mathematical formulae. Thirty Santa Inês sheep were included. Evaluations were performed weekly from the 3rd to 21st gestational week. The Shapiro test was used to determine normality of the data, and Spearman test was used for determining correlations of real or transformed measurements as gestation advanced. If there was a significant difference detected, variable adjustments and gestational weeks were tested using regression models. Significance level was set at 5% and the results presented as mean ± SD. Principal components analysis (PCA) was used for data clustering and graphic analysis. The gestational vesicle, abdominal, thoracic, biparietal, ocular orbit, and placentome diameters; and the lengths of crown-to-rump, nuchal to snout, scapula, humerus, radius, metacarpus, femur, tibia, metatarsal, heart length, heart width and kidneys were measured. Using pulsed Doppler ultrasonography, heart rate (HR) was obtained. The greatest coefficients of determination (r²) were obtained for the biparietal diameter (BPD; 96.5%) and metacarpus length (MCAR; 93%), followed by the abdominal diameter (AD; 91.7%) and heart width (HTW; 90.2%). The PCA analysis indicated graphically that at week 8 to 9, there was no significant change related to technique, whereas, at weeks 19, 20, and 21 a decrease in growth was observed, with the greatest data variability at week 19. Ultrasonographic assessments indicated there were significant correlations between developmental stages of all maternal-fetal structures and the gestational age, which can be used as reference values for gestational age assessments and detection of development abnormalities.

Levantamento e identificação dos aspectos epidemiológicos da raiva canina no município de Cuiabá – MT / Survey and identification of the epidemiological aspects of canine rabies in the city of Cuiabá-MT

Reconhecida desde a antiguidade, a raiva é uma infecção aguda do sistema nervoso central que acomete mamíferos, sendo causada por um RNA vírus da família Rabidoviridae, gênero Lyssavírus. O Brasil, desde a implantação do programa de controle da raiva em áreas urbanas, vem apresentando um declínio de casos de raiva em cães e em seres humanos. Em Cuiabá, as atividades realizadas pelo Centro de Controle de Zoonoses voltadas para o controle da raiva como: vacinação antirrábica animal, bloqueios de foco, captura de animais errantes, entre outros, permitiu o bloqueio da circulação do vírus na espécie canina. Desde 2008, o município de Cuiabá não registra nenhum caso da doença na espécie canina. Com o objetivo de conhecer a distribuição temporal da raiva canina em Cuiabá- MT, assim como identificar os aspectos epidemiológicos dos cães infectados pelo vírus rábico e verificar as medidas de profilaxia para a raiva canina, adotadas pelo Centro de Controle de Zoonose do município de Cuiabá no período de janeiro de 2002 a dezembro de 2008, realizou-se o presente estudo. Para o estudo observacional descritivo foi empregado o banco de dados referente à série histórica do CCZ e do Laboratório de Apoio a Saúde Animal – LASA ambos situados no município de Cuiabá- MT. O período de estudo foi de janeiro de 2002 a dezembro de 2008. Neste período foram examinadas 1021 amostras de cérebro de cães, destas 8,71% (89/1021) foram positivas e 91,29% (932/1021) negativas para a raiva. Os meses de maior incidência da doença foram os meses de janeiro 15,05% (14/93), fevereiro 12,5% (9/72), março 12,5% (9/72), abril 14,14% (9/72) e dezembro 13,69% (10/73). O maior número de cães doentes era macho 59,55% (53/89) com idade acima de dois anos 50,55% (45/89) e não domiciliados 67,41% (60/89). Em relação à imunoprofilaxia, a maioria dos cães 99,63% foi vacinada em campanhas. A cobertura vacinal ficou acima dos 80% exceto em 2006. Entre os animais que apresentaram a enfermidade o maior número, 67,41% (60/89), foi de cães não domiciliados, os chamados cães errantes. O percentual de cães domiciliados infectados pelos VR foi igual a 32,58% (29/89). As campanhas de vacinação, portanto, são fundamentais para a imunização do maior número de animais possível e a sociedade deve se conscientizar sobre a importância da raiva e as medidas que devem ser tomadas para controlá-la.

Known since the ancient times, the rabies is an acute infection of the central nervous system that affects mammals, caused by an RNA virus of the Rhabidoviridae family, Lyssavirus gender. Since the implantation of the urban area rabies control program in Brazil the number of cases in dogs and human have shown a decrease. In Cuiabá, the efforts to control the rabies, such as animal vaccination, focus isolation, capture of nomad animals and others realized by the Zoonosis Control Center has leaded to locking in the viral circulation in the canine species. Since 2008, zero cases of the disease in dogs were reported. In order to know the temporal distribution of canine rabies in Cuiabá – MT and identify the epidemiologic aspects of the rabies’s infected dogs, and verify the canine rabies’s profilatic actions of the Zoonosis Control Center of the city of Cuiabá, during the period of January, 2002 to December , 2008, the present study was performed. The historic data base from Cuiabá - MT was obtained at the Zoonosis Control Center and at the Animal Health Support Laboratory and used to perform the descriptive and observational study. During the performance of the study, 1021 samples of canine’s brains were examined, 8,71% (89/1021) of them were positive and 91,29% (932/1021) of them were negative for the rabies test. The Months with higher incidence were January with 15,05% (14/93), February with 12,5% (9/72), March with 12,5% (9/72), April with 14,14% (9/72) and December with 13,69% (10/73). The major part of the infected dogs were males with 59,55% (53/89) older than two years old 50,55% (45/89) and were nomads animals 67,41% (60/89). 99,63% of the infected dogs were vaccinated in anti rabies vaccination campaigns. The vaccinal cover was higher than 80%, except in 2006. The percentual of the domiciled infected dogs by the rabies virus were 32,58% (29/89). Therefore the vaccination campaigns are essential for the immunization of a higher number of animals, and the society must be awareabout the importance of rabies and about the actions to control it.