Identification of genomic regions associated with total and progressive sperm motility in Italian Holstein bulls.

Sperm motility is directly related to the ability of sperm to move through the female reproductive tract to reach the ovum. Sperm motility is a complex trait that is influenced by environmental and genetic factors and is associated with male fertility, oocyte penetration rate, and reproductive success of cattle. In this study we carried out a GWAS in Italian Holstein bulls to identify candidate regions and genes associated with variations in progressive and total motility (PM and TM, respectively). After quality control, the final data set consisted of 5,960 records from 949 bulls having semen collected in 10 artificial insemination stations and genotyped at 412,737 SNPs (call rate >95%; minor allele frequency >5%). (Co)variance components were estimated using single trait mixed models, and associations between SNPs and phenotypes were assessed using a genomic BLUP approach. Ten windows that explained the greatest percentage of genetic variance were located on Bos taurus autosomes 1, 2, 4, 6, 7, 23, and 26 for TM and Bos taurus autosomes 1, 2, 4, 6, 8, 16, 23, and 26 for PM. A total of 150 genes for TM and 72 genes for PM were identified within these genomic regions. Gene Ontology enrichment analyses identified significant Gene Ontology terms involved with energy homeostasis, membrane functions, sperm-egg interactions, protection against oxidative stress, olfactory receptors, and immune system. There was significant enrichment of quantitative trait loci for fertility, calving ease, immune response, feed intake, and carcass weight within the candidate windows. These results contribute to understanding the architecture of the genetic control of sperm motility and may aid in the development of strategies to identify subfertile bulls and improve reproductive success.

Adjuvant pretreatment with alum protects neonatal mice in sepsis through myeloid cell activation.

The high mortality in neonatal sepsis has been related to both quantitative and qualitative differences in host protective immunity. Pretreatment strategies to prevent sepsis have received inadequate consideration, especially in the premature neonate, where outcomes from sepsis are so dismal. Aluminium salts-based adjuvants (alum) are used currently in many paediatric vaccines, but their use as an innate immune stimulant alone has not been well studied. We asked whether pretreatment with alum adjuvant alone could improve outcome and host innate immunity in neonatal mice given polymicrobial sepsis. Subcutaneous alum pretreatment improves survival to polymicrobial sepsis in both wild-type and T and B cell-deficient neonatal mice, but not in caspase-1/11 null mice. Moreover, alum increases peritoneal macrophage and neutrophil phagocytosis, and decreases bacterial colonization in the peritoneum. Bone marrow-derived neutrophils from alum-pretreated neonates produce more neutrophil extracellular traps (NETs) and exhibit increased expression of neutrophil elastase (NE) after in-vitro stimulation with phorbol esters. In addition, alum pretreatment increases bone marrow and splenic haematopoietic stem cell expansion following sepsis. Pretreatment of neonatal mice with an alum-based adjuvant can stimulate multiple innate immune cell functions and improve survival. These novel findings suggest a therapeutic pathway for the use of existing alum-based adjuvants for preventing sepsis in premature infants.

Un caso de desorden del desarrollo sexual en un canino mestizo / A case of a disorder sexual development in a hybrid dog

Los desórdenes del desarrollo sexual se pueden presentar en diferentes animales domésticos, aunque no son muy comunes. Se clasifican en tres grupos de anormalidades: del desarrollo cromosómico, del desarrollo gonadal y del sexo fenotípico, y presentan diferentes subcategorías. El objetivo de este trabajo fue describir las características fenotípicas, cromosómicas y anatómicas de los órganos reproductores de un paciente canino, aparentemente hembra, de 3 años de edad, con desorden del desarrollo sexual, que presentaba una protuberancia a nivel vaginal con sangrado y presencia de pus; en la anamnesis reportan comportamiento de macho. Se realizó un examen general por sistemas, ecografía abdominal ventral, radiografía latero-lateral, cuadro hemático y cariotipo con bandas R-replicativas. Tras la evaluación se encontró un clítoris agrandado (pseudopene) con orificio uretral que mostró resistencia a la colocación de una sonda. La radiografía mostró una estructura similar al hueso del pene y la ecografía reveló una estructura compatible con el cuello del útero en una hembra y una estructura lateral parecida al tejido gonadal. El cariotipo fue típico de un macho, compatible con un macho seudohermafrodita, lo cual permite clasificar al individuo como XY, con un desorden del desarrollo sexual fenotípico (78, XY) según la nueva clasificación. Con las herramientas diagnósticas disponibles en Colombia es posible realizar un diagnóstico diferencial adecuado. Sin embargo, falta disponibilidad de pruebas diagnósticas específicas como FISH y mediciones serológicas.

The disorders of sex development can occur in different domestic animals, but they are not very common. They are classified as sex chromosomic disorders, gonadal sex development disorders and phenotypic sex disorders and have different subcategories. The aim of this study was to describe the phenotypic, chromosomal and anatomical traits of the reproductive organs of a canine patient 3-year-old, apparently female with disorder of sexual development, which presented a protuberance into the vagina with bleeding and pus, which anamnesis male behavior report. A general examination was performed by systems, ventral abdominal ultrasound, latero-lateral radiography, blood count and karyotype whit R-replicative bands. After the evaluation found an enlarged clitoris (pseudopene) with urethral opening that showed resistance to placing a catheter. Radiography showed a structure similar to penis bone and the ultrasonography a structure consistent with the cervix in a female and a structure similar to gonadal tissue in the side. The karyotype was typical of a male, compatible with a male pseudo-hermaphrodite, which classifies the individual as XY with a phenotypic disorder of sex development (78, XY) according to the new classification. With the diagnostic tools found in Colombia is possible to make an appropriate differential diagnosis. But nevertheless, lack of availability of specific diagnostic tests such as FISH and serological measurements.

Effect of two single nucleotide polymorphisms on milk yield and composition.

The bovine prolactin (PRL) and bovine growth hormone (bGH) genes exhibit several polymorphisms. Some of them can be detected by molecular techniques using restriction endonucleases, such as RsaI for the PRL gene and MspI for the bGH gene. We examined the relationship between the PRL-RsaI and bGH-MspI polymorphisms and some economically important characteristics of Holstein cows. Research was conducted on 315 Holstein cows from 5 municipalities in the Department of Antioquia, Colombia. Individuals were genotyped using PCR/RFLP. The statistical analysis was carried out using generalized linear models and a regression analysis. Polymorphism of the bGH gene was found to have a significant association with the percentage of protein in milk and milk yield. Genotype (-/-) was favorable for dairy yield, while genotype (+/+) was favorable for protein percentage. The PRL gene showed no significant association with any of the evaluated characteristics. The bGH gene appears to be a candidate for the implementation of marker-assisted selection programs. To determine the effect of the prolactin gene, research should be conducted with a larger sample size and a group of animals with more balanced genotypes.

The epithelial cell rests of Malassez--a role in periodontal regeneration?

This article reviews general aspects about the epithelial cell rests of Malassez (ERM). The historical and general morphological features of the ERM are briefly described. The embryological derivation of the ERM is presented as an important consideration in understanding the events associated with their origin and possible functional roles within the periodontal ligament. The ultrastructural description of the ERM is also included to complement the morphological characteristics which distinguish these cells as the unique epithelial element of the periodontal ligament. The unique ability of these cells to synthesize and secrete a number of proteins usually associated with cells of mesenchymal origin, rather than ectodermal origin, is discussed in light of their role in cementum repair and regeneration. Such considerations lead to our hypothesis that one of the functional roles of the ERM may lie not only their role in maintaining and contributing to the normal periodontal cellular elements and function but also contributing, in a significant manner, to periodontal regeneration.

Enhanced proliferation, attachment and osteopontin expression by porcine periodontal cells exposed to Emdogain.

Emdogain (EMD) is an enamel matrix derivative extracted from developing porcine teeth with demonstrated periodontal regenerative potential. EMD has been shown to influence a number of properties of periodontal ligament cells including proliferation, cell attachment and matrix synthesis. To date, the effect of EMD on the epithelial cell rests of Malassez (ERM) is unknown. In this study, periodontal ligament fibroblasts, ERM, alveolar bone cells and gingival fibroblasts were obtained from porcine periodontal ligament, alveolar bone and gingiva. This study investigated, in vitro, the effect of EMD at three concentrations on proliferation, cell attachment and expression of mRNA for two mineralised tissue-related proteins (osteopontin and bone sialoprotein). As for other periodontal cells, the ERM proliferative response was enhanced by EMD. Attachment assays revealed a highly significant increase for ERM and gingival fibroblasts after EMD treatment at all concentrations. This study has also shown that EMD stimulated expression of osteopontin mRNA by ERM and alveolar bone cells. The results from this study provide evidence that EMD enhanced cellular events related with proliferation, attachment and osteopontin mRNA expression by porcine periodontal cells, in a manner consistent with its role in periodontal regenerative therapy.

Effect of Emdogain on human periodontal fibroblasts in an in vitro wound-healing model.

OBJECTIVE: The aim of this study was to evaluate the influence of Emdogain (EMD) on cultured gingival fibroblasts, periodontal ligament fibroblasts and dermal fibroblasts, using an in vitro model of wound healing. BACKGROUND: Enamel matrix derivative has been demonstrated to promote periodontal regeneration. However, the precise mechanisms by which this agent acts are still unclear. METHODS: The effect of EMD on proliferation of the cells was studied using subconfluent cultures of gingival fibroblasts and periodontal ligament fibroblasts. The cells were made quiescent overnight and then stimulated with various concentrations of EMD (10, 50, 100 and 150 microg/ml) for 24 h. Negative and positive controls were cells cultured in media containing 0.2% and 10% fetal calf serum (FCS). The DNA synthesis was measured by the cellular uptake of [3H]thymidine. For in vitro wounding the cells were cultured, wounded and stimulated with 0.2% FCS, 10% FCS and EMD at a concentration of 20 microg/ml. The percentage of wound fill after treatment was measured after d 1, 4, 6, 12 and 16. The proliferation of cells was also calculated by the extent of incorporation of crystal violet. RESULTS: The results demonstrated that cells in vitro fill an empty space by a combination of proliferation and cell migration. The most rapid closure of a wound area occurred where both proliferation and migration can occur as was seen when wounded cultures were maintained in 10% FCS or at a concentration of 20 microg/ml EMD which promoted proliferation. CONCLUSIONS: Therefore, EMD appears to exert an influence on cells that is compatible with improved wound healing.

Molecular cloning of a cDNA encoding human SPH-binding factor, a conserved protein that binds to the enhancer-like region of the U6 small nuclear RNA gene promoter.

Many vertebrate small nuclear RNA gene promoters contain an SPH motif in their distal control regions that can confer transcriptional stimulation by RNA polymerase II or RNA polymerase III. Using the human U6 gene SPH motif as a probe, we isolated a cDNA encoding human SPH-binding factor (hSBF) from a HeLa cell expression library. The coding region of hSBF is almost identical to ZNF143, a 626 amino acid, seven zinc finger protein of previously unknown function. Furthermore, the predicted amino acid sequence of hSBF is highly homologous to Xenopus laevis and mouse Staf proteins, that bind to SPH motifs and stimulate transcription of selenocysteine tRNA gene promoters. Recombinant hSBF expressed in vitro or from Escherichia coli bound specifically to the human U6 gene SPH motif as shown by DNase I footprinting and electrophoretic mobility shift assays using various mutant SPH sites as competitors. Antibodies prepared against recombinant hSBF inhibited assembly of native SBF-DNA complexes. Immunodepleted HeLa S100 transcription extract no longer supported elevated levels of transcription by RNA polymerase III from a U6 promoter containing an SPH motif, whereas addition of recombinant hSBF protein to the immunodepleted extract reconstituted stimulated transcription.