There is a paucity of information on the relationships of testicular morphology, echotextural attributes, and blood flow dynamics with pubertal development of rams raised in a subtropical climate. Forty-five Dorper rams (24 rams aged 8-11 months and 21 rams aged 12-24 months) were examined using a portable ultrasound scanner connected to a 7.5-MHz transducer. Computer-assisted analyses of testicular ultrasonograms utilized commercially available Image ProPlus analytical software. Spectral Doppler scans of testicular arteries were performed immediately after scrotal (B-mode) ultrasonography to determine peak systolic velocity (PSV), end-diastolic velocity (EDV), resistive index (RI = [PSV-EDV]/PSV), and pulsatility index (PI = [SPV-EDV]/mean velocity) of the blood vessels. The length of the testes (9.7 ± 0.3 compared with 9.0 ± 0.2 cm) and scrotal circumference (33.3 ± 0.5 compared with 31.8 ± 0.4 cm) were greater (p < 0.05) but testicular depth (4.9 ± 0.08 compared with 4.5 ± 0.1 cm) was less (p < 0.05) in sexually mature compared with peripubertal rams. There were no differences (p > 0.05) between the two age groups of Dorper rams in blood flow indices of testicular arteries. Mean numerical pixel values (100.5 ± 4.1 compared with 89.2 ± 4.8) and pixel heterogeneity (25.6 ± 0.6 compared with 23.6 ± 0.5) of testicular parenchyma were greater (p < 0.05) in peripubertal than in postpubertal rams. Semen volume was negatively correlated with PI of testicular arteries (r = -0.57, p = 0.04). In summary, the attainment of sexual maturity in the rams of the present study was associated with significant changes in testicular length and depth, scrotal circumference, and parenchymal echogenicity/hetrogeneity but not in testicular volume and blood perfusion rates. Testicular artery PI can be used to predict the volume of ejaculate in rams.

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.

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Pesq. Vet. Bras. 33(Supl.1):25-31, dezembro 2013 25 RESUMO.-O conhecimento da dinâmica das alterações nos parâmetros hematológicos e na cinética das proteínas de fase aguda em animais saudáveis nas primeiras semanas de vida é essencial para a interpretação correta dessas avaliações em situações de morbidez e para diferenciar animais sadios e enfermos de forma confiável. Com o intuito de avaliar a cinética desses parâmetros no primeiro mês de vida de bezerros de corte sadios, filhos de vacas primíparas ou pluríparas, amostras de sangue foram coletadas antes da ingestão de colostro e 1, 2, 7, 15 e 30 dias após o nascimento. Os parâmetros eritrocitários foram influenciados pelo número de partos das vacas e o leucograma mostrou alterações características de influência do cortisol fetal liberado por ocasião do nascimento. O teor sérico de proteína total aumentou significativamente após a ingestão do colostro. As concentrações de ceruloplasmina, haptoglobina e proteínas de pesos moleculares 33 kDa e 23 kDa aumentaram significativamente no primeiro dia de vida, seja pela resposta ao nascimento ou pela ingestão do colostro, enquanto os teores de transferrina, albumina e α 1 -glicoproteína ácida mantiveram-se relativamente estáveis nos primeiros dias de vida, aumentando gradualmente até os 30 dias de idade. The knowledge of the dynamic in changes of hematologic parameters and the acute phase proteins kinetics in healthy animals in the first weeks of life is essential for the accurate interpretation of these evaluations in morbidity situations, and to reliably differentiate healthy from sick animals. The aim of the study was to evaluate the dynamic of these parameters in the first month of life of healthy beef calves, born from primiparous or multiparous cows, and so blood samples were collected before colostrum intake and 1, 2, 7, 15 and 30 days thereafter to evaluate hemogram and serum proteinogram. The red cell parameters were influenced by the parturition number of cows, and the leukogram showed characteristic changes of the release of fetal cortisol at birth. The total protein level significantly increased after colostrum intake. The concentrations of acute phase proteins ceruloplasmin, haptoglobin and proteins of molecular weight 33 kDa and 23 kDa significantly increased in the first day of life, influenced either by response to birth or colostrum intake, while the levels of transferrin, albumin and α 1 -acid glycoprotein were relatively stable in the first days, increasing gradually until 30 days of life.INDEX TERMS: Cattle, newborn, hematology, acute phase proteins, SDS-PAGE.

The aim of the present study was to determine and compare ultrasonographic characteristics of accessory sex glands and spectral Doppler indices of the internal iliac arteries in peri- and post-pubertal rams raised in a subtropical climate. Forty-five Dorper rams were examined (24 rams aged 8-11 months and 21 rams aged 12-24 months). Digital images of all accessory sex glands were subjected to morphometric and echotextural analyses, the latter using commercially available image analytical software Image ProPlus. Physical and morphological characteristics of semen and serum concentrations of testosterone were also determined. The dimensions of the prostate gland (12.9±1.2 compared with 14.2±2.7mm; mean±standard deviation) and bulbourethral glands (13.7±1.3 compared with 14.7±1.8mm) were greater (P=0.04) in sexually mature compared with peri-pubertal rams. Pixel intensity of vesicular (181.5±20.8 compared with 164.8±26.8, P=0.02) and bulbourethral gland parenchyma (166.9±16.9 compared with 141.8±29.1, P=0.001) was greater in peri-pubertal compared with sexually mature rams. Semen could be collected by ejaculation into the artificial vagina from 38% (8/21) of post-pubertal rams and 21% (5/24) of peri-pubertal animals (P=0.03). Semen volume was positively correlated with peak systolic velocity (PSV) and end-diastolic velocity (EDV) in the internal iliac arteries (r=0.79, P=0.001 and r=0.67, P=0.01, respectively), while spermatic vigor and progressive motility were inversely related to circulating concentrations of testosterone (r=-0.69, P=0.009 and r=-0.61, P=0.03, respectively). In summary, the attainment of sexual maturity in the rams of the present study was associated with an enlargement of the prostate and bulbourethral glands, and with the shift in echotextural attributes of vesicular and bulbourethral glands. Circulating testosterone concentrations and Doppler blood flow indices of the ram's internal iliac arteries are significant predictors of sperm progressive motility, vigor and the amount of ejaculate.

Mitochondrial function, largely regulated by the dynamics of this organelle, is inextricably linked to the oocyte health. In comparison with most somatic cells, mitochondria in oocytes are smaller and rounder in appearance, suggesting limited fusion. The functional implications of this distinct morphology, and how changes in the mitochondrial shape translate to mitochondrial function in oogenesis is little understood. We, therefore, asked whether the pro‐fusion proteins mitofusins 1 (MFN1) and 2 (MFN2) are required for the oocyte development. Here we show that oocyte‐specific deletion of Mfn1, but not Mfn2, prevents the oocyte growth and ovulation due to a block in folliculogenesis. We pinpoint the loss of oocyte growth and ovulation to impaired PI3K‐Akt signaling and disrupted oocyte‐somatic cell communication. In support, the double loss of Mfn1 and Mfn2 partially rescues the impaired PI3K‐Akt signaling and defects in oocyte development secondary to the single loss of Mfn1. Together, this work demonstrates that the mitochondrial function influences the cellular signaling during the oocyte development, and highlights the importance of distinct, nonredundant roles of MFN1 and MFN2 in oogenesis.

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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).