Effect of sexual steroids on boar kinematic sperm subpopulations.

Here, we show the effects of sexual steroids, progesterone, testosterone, or estradiol on motility parameters of boar sperm. Sixteen commercial seminal doses, four each of four adult boars, were analyzed using computer assisted sperm analysis (CASA). Mean values of motility parameters were analyzed by bivariate and multivariate statistics. Principal component analysis (PCA), followed by hierarchical clustering, was applied on data of motility parameters, provided automatically as intervals by the CASA system. Effects of sexual steroids were described in the kinematic subpopulations identified from multivariate statistics. Mean values of motility parameters were not significantly changed after addition of sexual steroids. Multivariate graphics showed that sperm subpopulations were not sensitive to the addition of either testosterone or estradiol, but sperm subpopulations responsive to progesterone were found. Distribution of motility parameters were wide in controls but sharpened at distinct concentrations of progesterone. We conclude that kinematic sperm subpopulations responsive to progesterone are present in boar semen, and these subpopulations are masked in evaluations of mean values of motility parameters. © 2017 International Society for Advancement of Cytometry.

Effects of repeated electroejaculations on kinematic sperm subpopulations and quality markers of Mexican creole goats.

Here we show the effects of repeated electroejaculation (EE) on mean values of motility, mitochondrial functionality, and expression of active caspases on goat sperm obtained by EE. Evaluations were done using CASA and flow cytometry. A strategy for identification of kinematic sperm subpopulations, when individual data of sperm are not provided by the CASA system, is provided. Fifty semen samples, five of each of ten adult creole goats, were obtained by electroejaculation. Mean values of total motility, progressive motility and flow cytometry evaluations were compared among EEs. Relationships among mean values of variables were investigated using Spearman correlation and principal component analysis (PCA). For identification of kinematic sperm subpopulations, PCA followed by hierarchical clustering was applied on data of the intervals provided automatically by the CASA system. Total motility does no change after repeated EE. Mean values of motility parameters and molecular markers were unrelated in multivariate space, but bivariate correlations were found. Values in upper and lower intervals defined clearly the sperm subpopulations, which had motility parameters changing over time. Taken together, our results show that repeated EE does not affect mean values of total motility, that molecular markers are not related with motility parameters, and that it is possible to identify kinematic sperm subpopulations when individual data, of motility parameters, are not provided by the CASA system.

First report of the Taiwan sardinella Sardinella hualiensis (Clupeiformes: Clupeidae) in the Philippines.

A range expansion of the Taiwan sardinella Sardinella hualiensis to the Philippines is reported. The data suggested that the southern translocation of S. hualiensis occurred across oceanographic features that are typically barriers to small, pelagic fish dispersal. With this addition, the Philippines is among the most diverse countries for Sardinella biodiversity in the world, second only to India.

The amelogenin C-terminus is required for enamel development.

The abundant amelogenin proteins are responsible for generating proper enamel thickness and structure, and most amelogenins include a conserved hydrophilic C-terminus. To evaluate the importance of the C-terminus, we generated transgenic mice that express an amelogenin lacking the C-terminal 13 amino acids (CTRNC). MicroCT analysis of TgCTRNC29 teeth (low transgene number) indicated that molar enamel density was similar to that of wild-type mice, but TgCTRNC18 molar enamel (high transgene number) was deficient, indicating that extra transgene copies were associated with a more severe phenotype. When amelogenin-null (KO) and TgCTRNC transgenic mice were mated, density and volume of molar enamel from TgCTRNCKO offspring were not different from those of KO mice, indicating that neither TgCTRNC18 nor TgCTRNC29 rescued enamel's physical characteristics. Because transgenic full-length amelogenin partially rescues both density and volume of KO molar enamel, it was concluded that the amelogenin C-terminus is essential for proper enamel density, volume, and organization.

Estudio de los bordes de resección en la cirugía conservadora del cáncer de mama. Experiencia en el Hospital de Barbastro / No disponible

Introducción: El objetivo de este estudio es determinar laamplitud del margen obtenido y la tasa de recidiva local en loscarcinomas de mama operados en el hospital de Barbastrocon cirugía conservadora entre 1989 y 2003.Material y métodos: Se han incluido 141 casos, 119 invasoresy 22 intraductales. El margen se ha definido como positivocuando se hallaba afectado por células tumorales, cercanosi era ≤ 1 mm y libre si era > 1 mm. Se evaluó preferentementemediante estudio macroscópico intraoperatorio.Resultados: La media de la edad fue de 57,36 ± 12,32años, del tamaño tumoral 14,7 ± 0,83 mm y del seguimiento86,05 ± 32,62 meses. El margen resultó positivo en 13 casos,6 reintervenidos. Los 7 restantes afectaban a la cara anteriory/o posterior y no se reintervinieron. Treinta y un casostuvieron margen cercano. Han presentado recidiva local ipsilateral3 carcinomas infiltrantes.Conclusiones: El 73% de los casos tienen bordes libres.Nuestra tasa de recidiva local, tras una media de seguimientode 86 meses, es baja (2%). La afectación de las caras anteriory posterior no ha ocasionado recidiva y creemos que deberíanvalorarse de forma diferente al resto de los márgenes(AU)

Objective: The aim of this study was to analyze the tumormargindistance and the local recurrences of breast cancer casestreated at the Hospital of Barbastro with breast-conservingtherapy between 1989 and 2003.Patients and method: 141 cases are included, 119 invasiveand 22 intraductal carcinomas. Margin was defined aspositive when tumor cells where in contact with india ink,close if ≤ 1 mm and free if > 1 mm. A macroscopic intraoperativeassessment of margin was carried out mainly.Results: The mean age was 57,36 ± 12,32, the mean tumorsize 14,7 ± 0,83 mm and the mean follow-up 86,05 ±32,62 months. The final margin was positive in 13 cases witha need for reintervention in 6 of them. The remaining 7, affectinganterior and/or posterior surfaces, were not operated.The distance was ≤ 1 mm in 31 cases. Local recurrences havebeen found in 3 invasive ipsilateral carcinomas.Conclusion: A free margin was found in 73% of cases. Localrecurrence rate is low (2%) after a median follow-up of 86months. Involvement of anterior and posterior surfaces hasnot been cause of recurrences. Thus it seems they should beassessed in different way to the others(AU)

Transgenic mice that express normal and mutated amelogenins.

Amelogenin proteins are secreted by ameloblasts within the enamel organ during tooth development. To better understand the function of the 180-amino-acid amelogenin (M180), and to test the hypothesis that a single proline-to-threonine (P70T) change would lead to an enamel defect similar to amelogenesis imperfecta (AI) in humans, we generated transgenic mice with expression of M180, or M180 with the proline-to-threonine (P70T) mutation, under control of the Amelx gene regulatory regions. M180 teeth had a relatively normal phenotype; however, P70T mineral was abnormally porous, with aprismatic regions similar to those in enamel of male amelogenesis imperfecta patients with an identical mutation. When Amelx null females were mated with P70T transgenic males, offspring developed structures similar to calcifying epithelial odontogenic tumors in humans. The phenotype argues for dominant-negative activity for the P70T amelogenin, and for the robust nature of the process of amelogenesis.

Amelogenin-mediated regulation of osteoclastogenesis, and periodontal cell proliferation and migration.

We previously reported that amelogenin isoforms M180 and leucine-rich amelogenin peptide (LRAP) are expressed in the periodontal region, and that their absence is associated with increased cementum defects in amelogenin-knockout (KO) mice. The aim of the present study was to characterize the functions of these isoforms in osteoclastogenesis and in the proliferation and migration of cementoblast/periodontal ligament cells. The co-cultures of wild-type (WT) osteoclast progenitor and KO cementoblast/periodontal ligament cells displayed more tartrate-resistant acid phosphatase (TRAP)-positive cells than the co-cultures of WT cells. The addition of LRAP to both co-cultures significantly reduced RANKL expression and the TRAP-positive cells. Proliferation and migration rates of the KO cementoblast/periodontal ligament cells were lower than those of WT cells and increased with the addition of either LRAP or P172 (a porcine homolog of mouse M180). Thus, we demonstrate the regulation of osteoclastogenesis by LRAP, and the proliferation and migration of cementoblast/periodontal ligament cells by LRAP and P172.

Tooth enamel defects in mice with a deletion at the Arhgap 6/Amel X locus.

The amelogenin proteins regulate enamel mineral formation in the developing tooth. The human AMELX gene, which encodes the amelogenin proteins, is located within an intron of the Arhgap 6 gene. ARHGAP 6 encodes a Rho GAP, which regulates activity of Rho A, a small G protein involved in intracellular signal transduction. Mice were generated in which the entire ARHGAP 6 gene was deleted by Cre-mediated recombination, which also removed the nested Amel X gene. Enamel from these mice appeared chalky white, and the molars showed excessive wear. The enamel layer was hypoplastic and non-prismatic, whereas other dental tissues had normal morphology. This phenotype is similar to that reported for Amel X null mice, which have a short deletion that removed the region surrounding the translation initiation site, and resembles some forms of X-linked amelogenesis imperfecta in humans. Analysis of the enamel from the Arhgap 6/Amel X-deleted mice verifies that the Amel X gene is nested within the murine Arhgap 6 gene and shows that removal of the entire Amel X gene leads to a phenotype similar to the earlier Amel X null mouse results, in which no amelogenin protein was detected. However, an unusual layer of aprismatic enamel covers the enamel surface, which may be related to the 1.1-Mb deletion, which included Arhgap 6 in these mice.

Serotoninergic system blockage in the prepubertal rat inhibits spermatogenesis development.

The stimulatory and inhibitory role of serotonin in gonadotropin secretion and in the onset of puberty in the male rat has been previously described, but its role in the establishment of spermatogenesis is not known. The aim of this study was to investigate the effects of serotoninergic inhibition by p-chloroamphetamine (pCA) on the prepubertal-to-adult stage of the rat reproductive system. Hypothalamic serotonin, gonadotropins and sex steroid hormone concentrations were measured, and a histopathological analysis of seminiferous epithelium was carried out on animals treated with pCA from day 30 and killed at 45 or 65 days of age. The pCA treatment significantly reduced the hypothalamic levels of serotonin and its metabolite (5-hydroxyindole-3-acetic acid). This inhibition did not affect the sex steroid hormone or LH concentrations, but rather it induced an increase in FSH concentration in animals of both ages. Spermatogenesis was impaired by pCA treatment. Disruption of seminiferous epithelium and the death of numerous germ cells were observed. Sperm produced by pCA-treated animals was of poor quality and appeared in small quantities. Apparently, serotonin depletion did not affect communication between the hypothalamus and the pituitary, but the FSH increase could have been related to alterations in the seminiferous epithelium effects. The seminiferous epithelium cycle was altered in rats killed at both 45 and 65 days of age, because at each age of killing the distribution of spermatogenesis stages was different. Germ cell apoptosis did not appear to be related to changes in the FSH concentrations, but other factors produced during spermatogenesis could have been involved in this induction. This study showed that serotonin was necessary for the development of normal spermatogenesis in prepubertal rats.

Vanadium induced ultrastructural changes and apoptosis in male germ cells.

Vanadium is a transition metal that is emitted to the atmosphere during combustion of fossil fuels. In the environment, vanadium occurs in the (V) oxidized form, but in the body it is found exclusively in the (IV) oxidized form. Vanadium tetraoxide is an inorganic chemical species in the (IV) oxidized form that has been shown to induce toxic effects in vitro and in vivo. The reproductive toxicity of vanadium in males was studied through monitoring germ cell apoptosis during spermatogenesis. We analyzed ultrastructural damage, and testosterone and progesterone concentrations following vanadium tetraoxide administered to male mice for 60 days. Spermatogenesis stages I-III and X-XII frequently showed apoptotic germ cells in control and treated animals; vanadium tetraoxide treatment induced an increase in the number of germ cell apoptosis in stages I-III and XII at 9.4 and 18.8 mg/kg, respectively. Although spermatogenesis is regulated by testosterone, in our study this hormone level was not modified by vanadium administration; thus, germ cell death was not related with testosterone concentration. At the ultrastructural level, we observed inclusion structures that varied as to location and content in the Sertoli and germ cells.

[Increasing decayed, missing and filled teeth index among school children in Loja, Spain]. / Incremento del índice de dientes permanentes cariados, perdidos por caries y obturados, entre escolares de Loja, España.

OBJECTIVE: To identify the clinical, socioeconomic, dietary, and hygienic predictors of the increasing Decayed, Missing, and Filled Teeth (DMFT) index, in a cohort of school children in Loja, Granada, Spain, during a three-year period (1994-1997). MATERIAL AND METHODS: A retrospective cohort study was conducted in 812 students to measure the DMFT index, using the methods recommended by the World Health Organization. Statistical analysis was conducted using logistic regression models. RESULTS: During the study period, the DMFT index increased from 1.86 to 4.5 for the study cohort as a whole, and from 3.71 to 4.47 among twelve-year olds. The DMFT at mid-period was associated to an increase in the final DMFT. The relative risk for attending school no. 3 was 0.69 (95% CI 0.60-0.8) vs. school no. 4, and 0.78 (95% CI 0.67-0.90) vs. school no. 5. CONCLUSIONS: The final model showed that the DMFT at mid-period was a risk factor for an increase in the DMFT index and that attending school number 3 was a protective factor.