PGRMC1 participates in late events of bovine granulosa cells mitosis and oocyte meiosis.

Progesterone Receptor Membrane Component 1 (PGRMC1) is expressed in both oocyte and ovarian somatic cells, where it is found in multiple cellular sub-compartments including the mitotic spindle apparatus. PGRMC1 localization in the maturing bovine oocytes mirrors its localization in mitotic cells, suggesting a possible common action in mitosis and meiosis. To test the hypothesis that altering PGRMC1 activity leads to similar defects in mitosis and meiosis, PGRMC1 function was perturbed in cultured bovine granulosa cells (bGC) and maturing oocytes and the effect on mitotic and meiotic progression assessed. RNA interference-mediated PGRMC1 silencing in bGC significantly reduced cell proliferation, with a concomitant increase in the percentage of cells arrested at G2/M phase, which is consistent with an arrested or prolonged M-phase. This observation was confirmed by time-lapse imaging that revealed defects in late karyokinesis. In agreement with a role during late mitotic events, a direct interaction between PGRMC1 and Aurora Kinase B (AURKB) was observed in the central spindle at of dividing cells. Similarly, treatment with the PGRMC1 inhibitor AG205 or PGRMC1 silencing in the oocyte impaired completion of meiosis I. Specifically the ability of the oocyte to extrude the first polar body was significantly impaired while meiotic figures aberration and chromatin scattering within the ooplasm increased. Finally, analysis of PGRMC1 and AURKB localization in AG205-treated oocytes confirmed an altered localization of both proteins when meiotic errors occur. The present findings demonstrate that PGRMC1 participates in late events of both mammalian mitosis and oocyte meiosis, consistent with PGRMC1's localization at the mid-zone and mid-body of the mitotic and meiotic spindle.

Occurrence of D-aspartate in the harderian gland of Podarcis s. sicula and its effect on gland secretion.

High concentrations of free D-aspartate (D-Asp), an amino acid well known for its neuroexcitatory activity, are endogeneously present in the Harderian gland (HG) of the lizard Podarcis s. sicula. This orbital gland consists of two different parts: the medial part, which is prevalently a mucous acinar gland, and the lateral part, which is a serous tubulo-acinar gland. To determine the physiological effect of D-Asp on exocrine secretion in HG, D-Asp (2.0 micromol/g b.w.) was injected intraperitoneally into lizards. We found that highest accumulations of exogenous D-Asp in HGs occurred 15 hr after the injection. Specifically, exogenous D-Asp prevalently stimulated serous secretion from the lateral portion of the gland, where immunohistochemical analysis revealed a major accumulation. Similarly, in the medial part of the gland, highly sulfated mucosubstances were observed after D-Asp injection. Further, in both parts of the HG, the electron microscope revealed euchromatic nuclei, a prominent rough endoplasmic reticulum, as well as numerous secretory granules within the acinar cells. Thus, following D-Asp injection, a 60% increase in HG total protein was detected. In addition, exogenous D-Asp induced changes in the electrophoretic pattern of HG. In conclusion, although further investigations are still needed to clarify the molecular pathway induced by D-Asp in exocrine secretion, this study does indicate that free D-Asp plays a significant role in the secretory activity of this gland.

Endocrine roles of D-aspartic acid in the testis of lizard Podarcis s. sicula.

In the lizard Podarcis s. sicula, a substantial amount of D-aspartate (D-Asp) is endogenous to the testis and shows cyclic changes of activity connected with sex hormone profiles during the annual reproductive phases. Testicular D-Asp content shows a direct correlation with testosterone titres and a reverse correlation with 17beta-estradiol titres. In vivo experiments, consisting of i.p. injections of 2.0 micromol/g body weight of D-Asp or other amino acids, in lizards collected during the three main phases of the reproductive cycle (pre-reproductive, reproductive and post-reproductive period), revealed that the testis can specifically take up and accumulate D-Asp alone. Moreover, this amino acid influences the synthesis of testosterone and 17beta-estradiol in all phases of the cycle. This phenomenon is particularly evident during the pre- and post-reproductive period, when endogenous testosterone levels observed in both testis and plasma were the lowest and 17beta-estradiol concentrations were the highest. D-Asp rapidly induces a fall in 17beta-estradiol and a rise in testosterone at 3 h post-injection in the testis and at 6 h post-injection in the blood. In vitro experiments show that testicular tissue converted L-Asp into D-Asp through an aspartate racemase. D-Asp synthesis was measured in all phases of the cycle, but was significantly higher during the reproductive period with a peak at pH 6.0. The exogenous D-Asp also induces a significant increase in the mitotic activity of the testis at 3 h (P < 0.05) and at 6 h (P < 0.01). Induction of spermatogenesis by D-Asp is recognized by an intense immunoreactivity of the germinal epithelium (spermatogonia and spermatids) for proliferation cell nuclear antigen (PCNA). The effects of D-Asp on the testis appear to be specific since they were not seen in lizards injected with other D- or L-forms of amino acids with known excitatory effects on neurosecretion. Our results suggest a regulatory role for D-Asp in the steroido-genesis and spermatogenesis of the testis of the lizard Podarcis s. sicula.

Testicular endocrine activity is upregulated by D-aspartic acid in the green frog, Rana esculenta.

This study investigated the involvement of D-aspartic acid (D-Asp) in testicular steroidogenesis of the green frog Rana esculenta and its effect on stimulation of thumb pad morphology and glandular activity, a typical testosterone-dependent secondary sexual characteristic in this amphibian species. In the testis, D-Asp concentrations vary significantly during the reproductive cycle: they are low in pre- and post-reproductive periods, but reach peak levels in the reproductive period (140-236 nmol/g wet tissue). Moreover, the concentrations of D-Asp in the testis through the sexual cycle positively match the testosterone levels in the gonad and the plasma. The racemase activity evaluated during the cycle expresses its peak when D-Asp and testosterone levels are highest, that is, during the reproductive period, confirming the synthesis of D-Asp from L-Asp by an aspartate racemase. Short-term in vivo experiments consisting of a single injection of D-Asp (2.0 micro mol/g body weight) demonstrated that this amino acid accumulates significantly in the testis, and after 3 h its uptake is coupled with a testosterone increase in both testis and plasma. Moreover, within 18 h of amino acid administration, the D-Asp concentration in the testis decreased along with the testosterone titer to prestimulation levels. Other amino acids (L-Asp, D-Glu and L-Glu) used instead of D-Asp were ineffective, confirming that the significant increase in testicular testosterone was a specific feature of this amino acid. In long-term experiments, D-Asp had been administered chronically to frogs caught during the three phases of the reproductive cycle, inducing testosterone increase and 17beta-estradiol decrease in the gonad during the pre- and post-reproductive period, and vice versa during the reproductive period. The stimulatory effect of D-Asp on testosterone production by the testis is consistent with the stimulation of spermatogenesis and the maturation of thumb pads occurring in D-Asp-treated frogs. In these last animals, there was an increase of seminiferous ampoule area and a higher number of spermatids and sperm. Moreover, in spermatogonia I and II and in spermatocytes, a proliferating cell nuclear antigen (PCNA) intense immunopositivity was observed. In addition, the thumb pads of D-Asp-treated frogs compared with controls showed a significantly thicker epithelial lining, a wider area of their glands with taller secretion cells, and more numerous, PAS-positive-rich secretions. Finally, these results provide functional evidence for a biologic role of D-Asp in amphibian male steroidogenesis; therefore, this unusual amino acid could be considered a modulatory agent for reproductive processes.

[Gestational HSV in the work setting]. / HSV gestazionale in ambiente di lavoro.

OBJECTIVE: The purpose of our studies has been that to individualize the in partnership factors of risk to infection Herpes Simplex Virus-Type 2 in pregnant women and to appraise the role of the job in such context. METHODS: A champion of 79 pregnant women with infection from HSV-2 has responded to a questionnaire regarding the appearance of the infection, her own job and the degree of stress, if they used services hygienic public. RESULTS: In this champion of pregnant women 63% they are workers. Of these the 85% they introduced signs of a reinfection. For the housewives in 67% there was a reinfection. 84% of the housewives it uses services hygienic privacies. The infection from HSV has appeared in 28.5% of the been born by women with first infection from HSV genitalis, and in 12.5% of the been born by women with reinfection from HSV. CONCLUSIONS: The women that work are mostly to risk of reinfection, they have more evident troubles. This is consequence of a style of life that contemplates a state of superior stress and a great tiredness from the same and of the appearance of vulvo-vaginal infections sustained by microorganisms transmitted through the hygienic services.

[Folic acid and cervix dysplasia]. / Acido folico e displasia cervicale.

The localized folate deficiency, which is sometimes misdiagnosed as cervical dysplasia, because of morphological similarities between the cytologic features of megaloblastosis seen with folate deficiency and the changes associated with dysplasia, could be a component of the dysplastic process. In this study we attempted the effect of oral folic in women with cervical dysplasia. A total of 154 subjects with grade 1 or 2 CIN were randomly assigned either 10 mg of folic acid or a placebo daily for 6 months. Clinical status, human papillomavirus type 16 infection and blood folate levels were monitored at 2 month intervals. After 6-months no significant differences were observed between supplemented and unsupplemented subjects regarding dysplasia status, biopsy results, or prevalence of human papillomavirus type 16 infection. Folate deficiency the initiation of cervical dysplasia, but folic acid supplements do not alter the course of established disease.

[Incidence of HPV and CIN in HIV positive women]. / Incidenza dell'infezione da HPV e della CIN in donne HIV-positive.

Cervical intraepithelial lesions associated with genital human papilloma virus (HPV) infection occur with increased frequency and severity among women with immunodeficiency. In this study, we considered 24 HIV-seropositive and 12 HIV-seronegative women. Each woman was interviewed and underwent a cytologic and colposcopic evaluation. Then colposcopic and cytologic findings were correlated with histologic and differences between HIV-seropositives and seronegatives were analyzed. Ten (41%) of 24 HIV-seropositive and one (9%) of 12 HIV-seronegative women had human papilloma virus infection. Among seropositives, eight (34%) had cervical intraepithelial (CIN): of those eight, 5 had CIN I, 2 CIN II and 1 CIN III. There (24%) of the 12 HIV-seronegative had CIN: two had CIN I and one CIN II. Six of the HIV-seropositive women were found to have multicentric disease (two or more sites). The objective of this study is to determine the relationship between human immunodeficiency virus (HIV) and human papilloma virus infection, sexual habits, reproducive history, and risk of cervical intraepithelial neoplasia (CIN). The results of this study suggest that cervical intraepithelial neoplasia is a common finding in HIV-infected women. Papanicolaou tests should be effective for detecting cervical disease in this population.

[Current conservative surgery in cervical intraepithelial neoplasia]. / Attualità della terapia chirurgica conservativa nella neoplasia cervicale intraepiteliale.

5 years (1989-1993) experience of cervical conization including 57 cases, has been reviewed. The mean follow-up was 33 (6-60) months. Mean age was 34.6 (range 21-57) years. Mean height and diameter of the cones were 2.5 +/- 0.3 cm and 1.5 +/- 0.4 cm, respectively. In 55 cases there were free margins of the cones. Cytological diagnosis was CIN I for 8 patients, CIN II for 21 patients, CIN III for 22 patients, CIN+HPV for 6 patients. In 40 of the 57 patients suspected of having a cervical intraepithelial neoplasia on cytology and on colposcopy, the histology was confirmative. Bleeding (the most common pre and postoperative complication) was observed in 6 cases (11.7%). In three patients a new conization was necessary and in the other two patients hysterectomy was necessary. No significant differences were noted in the pregnancy outcome following either treatment in five patients.