Gaspare Carta

Role of the infections in recurrent spontaneous abortion

Embryo–fetal infections have been reported to cause recurrent spontaneous abortions (RSAs) at a rate lower than 4%. The possible mechanisms include production of toxic metabolic byproducts, fetal or placental infection, chronic endometrial infection, and chorio-amnionitis. Viruses appear to be the most frequently involved pathogens, since some of them can produce chronic or recurrent maternal infection. In particular, cytomegalovirus during pregnancy can reach the placenta by viremia, following both primary and recurrent infection, or by ascending route from the cervix, mostly following reactivation. Another herpesvirus, herpes simplex virus type 2, less frequently type 1, causes recurrent infections of the genital tract, which can involve the feto-placental unit. Parvoviruses have also been implicated in the development of repeated fetal loss. Among bacterial infections, Chlamydia trachomatis, Ureaplasma urealyticum,and Mycoplasma hominis have been mostly associated with occurrence of RSA. An increased risk of abortion among women with bacterial vaginosis (BV) during early pregnancy was also shown, but questions arise about the role of chronic BV in its occurrence. Although a definitive relationship between recurrently active infections and RSA is still lacking, mostly due to difficulties in demonstrating the pathogenic role of each individual isolated pathogen, diagnosis and therapy of RSA-related infections should be attempted. The diagnosis of infectious agents as a possible cause of RSA might lead to a therapeutic approach with antiviral drugs and antibiotics or using immunoglobulins, which can display both anti-infective neutralizing and immunomodulating properties.

Waiting Time and Pain During Office Hysteroscopy

STUDY OBJECTIVE To find a correlation between the waiting time between counseling about and performance of office hysteroscopy and the perception of pain. DESIGN Observational study (Canadian Task Force classification II-2). SETTING Academic environment. PATIENTS Two hundred eighty-four women undergoing hysteroscopy. INTERVENTIONS Diagnostic hysteroscopy with endometrial biopsy. MEASUREMENTS AND MAIN RESULTS Before examination, patients were asked to complete 2 forms, the STAI-S (State-Trait Anxiety Inventory, State) and STAI-T (State-Trait Anxiety Inventory, Trait) anxiety scales, for evaluation of their usual anxiety state and their state of anxiety during the examination. Patients were asked to quantify on a visual analog scale the pain felt during the examination. A statistically significant positive correlation, even if weak, was demonstrated between pain and waiting time (r = 0.45; p < .01) but not with the values for the anxiety state (r = 0.06; p = .56) and anxiety trait (r = -0.05; p = .66). Pain (≥4) was significantly associated with waiting time (≥60 minutes) (odds ratio [OR], 5.21; 95% confidence interval [CI], 1.29-35.50), age (OR, 1.57; 95% CI, 0.40-5.87) and menopause (OR, 2.81; 95% CI, 1.10-7.40) but not with STAI-S level (≥34) (OR, 0.87; 95% CI, 0.26-3.12) or STAI-T level (≥34) (OR, 0.65; 95% CI, 0.19-2.32). CONCLUSION Office hysteroscopy is associated with a level of anxiety that can affect patient tolerability of the procedure. However, factors such as reducing waiting time may have a positive effect on patient compliance, making hysteroscopy easier and thereby increasing its diagnostic and therapeutic potential.

Suberoylanilide hydroxamic acid partly reverses resistance to paclitaxel in human ovarian cancer cell lines

OBJECTIVES The purpose of this study was to determine whether the addition of the histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) to paclitaxel (PTX) can sensitize PTX-resistant human ovarian cancer cell lines (CABA-PTX and IGROV-PTX) in vitro. METHODS SAHA was studied in combination with paclitaxel in PTX-sensitive and PTX-resistant human ovarian cancer cell lines. Using cell proliferation analysis, immunofluorescence, and flow cytometric assays, we can determine whether the resistance was partly removed when the cells were treated with a combination of SAHA and PTX. Cells were also assayed for cytochrome c release. The levels of acetylated tubulin, β-tubulin, and HDAC6 were quantified by Western blots. RESULTS SAHA in combination with PTX led to a more pronounced inhibition of cell growth compared with PTX alone. In addition, the combined exposure to PTX and SAHA resulted in a marked arrest in the G2/M phase of the cell cycle and in a significant increase in the percentage of apoptotic cells. The expression of acetylated tubulin was dramatically increased by exposure to the combination of PTX and SAHA. These data paralleled the findings of an increased expression of HDAC6 in the presence of PTX in PTX-resistant cell lines. CONCLUSIONS The results of this study suggest the existence of a novel resistance mechanism based upon the upregulation of HDAC6 and that the histone deacetylase inhibitor SAHA holds promise to overcome PTX resistance in ovarian cancer cell lines.

Peroxisome Proliferator-Activated Receptors in Female Reproduction and Fertility

Reproductive functions may be altered by the exposure to a multitude of endogenous and exogenous agents, drug or environmental pollutants, which are known to affect gene transcription through the peroxisome proliferator-activated receptors (PPARs) activation. PPARs act as ligand activated transcription factors and regulate metabolic processes such as lipid and glucose metabolism, energy homeostasis, inflammation, and cell proliferation and differentiation. All PPARs isotypes are expressed along the hypothalamic-pituitary-gonadal axis and are strictly involved in reproductive functions. Since female fertility and energy metabolism are tightly interconnected, the research on female infertility points towards the exploration of potential PPARs activating/antagonizing compounds, mainly belonging to the class of thiazolidinediones (TZDs) and fibrates, as useful agents for the maintenance of metabolic homeostasis in women with ovarian dysfunctions. In the present review, we discuss the recent evidence about PPARs expression in the hypothalamic-pituitary-gonadal axis and their involvement in female reproduction. Finally, the therapeutic potential of their manipulation through several drugs is also discussed.

Post-ovulatory ageing of mouse oocytes affects the distribution of specific spindle-associated proteins and Akt expression levels

The aim of this study has been to determine the effects of in vivo post-ovulatory ageing (POA) on the distribution of spindle-associated proteins, histone H3/H4 post-translational modifications and on v-akt murine thymoma viral oncogene homolog 1 (Akt) expression levels. To this end, oocytes were retrieved 13, 29 and 33h after human chorionic gonadotrophin (hCG) treatment. The presence and distribution at the meiotic spindle of acetylated tubulin, γ-tubulin, polo kinase-1 and Ser473/Thr308 phosphorylated Akt (pAkt) as well as histone H3 and H4 acetylation and phosphorylation levels were assayed via immunofluorescence. Akt expression levels were determined via reverse transcription-polymerase chain reaction and western blotting analyses. Spindles from oocytes recovered 13h and 29h after hCG treatment showed similar levels of acetylated tubulin but ageing induced: (1) translocation of γ-tubulin from spindle poles to microtubules, (2) absence of Thr308- and Ser473-pAkt in 76% and 30% of oocytes, respectively, and (3) a significant reduction in phosphorylation levels of serine 10 on histone 3. At 29h, a significant decrease in Akt mRNA, but not in pAkt or Akt protein levels, was recorded. By contrast, protein content significantly decreased 33h after hCG. We conclude that POA impairs oocyte viability and fertilisability by altering the expression levels and spindle distribution of proteins that are implicated in cell survival and chromosome segregation. Together, these events could play a role in oocyte apoptosis.

Post-earthquake birth-rate evaluation using the brief cope

Background: The study arises from the idea of analyzing the reasons why many women in L’Aquila decided to conceive in the months following the 2009 earthquake. In the months from January to June 2011, there was a +27.24% increase in the number of newborns (+ 91) delivered in the Obstetrics and Gynecology Unit of the San Salvatore Hospital of L’Aquila compared to the same six-month period in 2010. Methods: Between January 2010 and December 2010, 874 women gave birth in L’Aquila. The women living outside of L’Aquila were excluded from the study. The remaining women, namely a sample of 451 individuals, were administered a questionnaire that focused on the level of stress experienced during the earthquake, and subsequently the Brief Cope. Results: After the earthquake there was a +22.64% increase in the number of women who desired a pregnancy. The results of the Brief Cope show that the coping strategies used by the women in L’Aquila were active coping, planning, acceptance and positive reframing. Conclusions: The desire for motherhood was the main reason accounting for the increase in births that occurred after the earthquake. The decision to have a child was the tangible sign of adaptation to the post-traumatic stress.

Ovarian cancer-derived extracellular vesicles affect normal human fibroblast behavior

ABSTRACT It has become clear that non-tumor cells in the microenvironment, especially fibroblasts, actively participate in tumor progression. Fibroblasts conditioned by tumor cells become “activated” and, as such, are identified as CAFs (cancer-associated fibroblasts). These CAFs remodel the tumor stroma to make it more favourable for cancer progression. The aim of this work was to verify whether EVs (extracellular vesicles - whose role as mediators of information between tumor and stromal cells is well known) released from human ovarian cancer cells were able to activate fibroblasts. EVs isolated from SKOV3 (more aggressive) and CABA I (less aggressive) cells were administered to fibroblasts. The consequent activation was supported by morphological and molecular changes in treated fibroblasts; XTT assays, zymographies, wound healing tests and invasion assays also highlighted higher proliferation, motility, invasiveness and enzyme expression. The secretome of these “activated” fibroblasts was, in turn, able to modulate the responses (proliferation, motility and invasion) of fibroblasts, and of tumor and endothelial cells. These findings support the idea that ovarian cancer cells can modulate fibroblast behaviour through the release of EVs, activating them to a CAFs-like state; the latter are able, in turn, to stimulate the surrounding cells. EVs from SKOV3 rather than from CABA I seem to be more efficient in some processes.

Modulating Intrafollicular Hormonal Milieu in Controlled Ovarian Stimulation: Insights From PPAR Expression in Human Granulosa Cells

Controlled ovarian stimulation (COS) leading to ovulation of multiple follicles is a crucial aspect of biomedical infertility care. Nevertheless, biomarkers useful for COS management are still lacking. Peroxisome proliferator‐activated receptors (PPARs) are nuclear hormone receptors relevant to steroid metabolism in granulosa cells (GCs). We investigated whether PPARs and their steroidogenic targets were differentially expressed in GCs differentiated under different recombinant or urinary gonadotropin preparations. GCs from women subjected to COS with r‐hFSH, r‐hFSH/r‐hLH, or hMG‐HP were processed to assess expression of PPARα, PPARβ/δ, PPARγ, and steroidogenic enzymes under PPAR modulation. As an evidence of their activation, all PPAR isotypes with their coactivators, the retinoic‐X‐receptors (RXRs), localized in the nucleus. When GCs from r‐hFSH/r‐hLH group were compared with r‐hFSH, a significant reduction of PPARα protein was observed. By contrast, an increase of PPARβ/δ at both protein and mRNA levels along with that of PPARγ protein were detected. The steroidogenic enzymes 17βHSD IV, 3βHSD II, and HMG‐CoA red were downregulated in the r‐hFSH/r‐hLH group in comparison to r‐hFSH unlike CYP19A1 that remained unchanged. In GCs from urinary FSH‐LH stimulation (hMG‐HP), PPARα was more expressed in comparison with r‐hFSH/r‐hLH group. Likewise, 3βHSD II and 17βHSD IV were increased suggesting that hMG‐HP partially mimicked r‐hFSH/r‐hLH effects. In summary, transcript analysis associated to protein investigation revealed differential effects of COS protocols on PPARs and their steroidogenic targets in relation to LH and gonadotropin source. These observations candidate PPARs as new biomarkers of follicle competence opening new hypotheses on COS effects on ovarian physiology. J. Cell. Physiol. 231: 908–914, 2016.

Changes in muscularis propria of anterior vaginal wall in women with pelvic organ prolapse.

The objective of this study was to evaluate the morphological and immunohistochemical alterations of tissue removed from the upper third of anterior vaginal wall in a sample group of the female population presenting homogenous risk factors associated with pelvic organ prolapse (POP). The case study consisted of 14 patients with POP and there were 10 patients in the control group. Patient selection was carried on the basis of specific criteria and all of the patients involved in the study presented one or more of the recognized POP risk factors. Samples were taken from POP patients during vaginal plastic surgery following colpohysterectomy, and from control patients during closure of the posterior fornix following hysterectomy. Samples were processed for histological and immunohistochemical analyses for Collagen I and Collagen III, α-Smooth Muscle Actin (α-SMA), Platelet-Derived-Growth-Factor (PDGF), matrix metalloproteinase 3 (MMP3), tissue inhibitors metalloproteinase 1 (TIMP1), Caspase3. Immunofluorescence analyses for Collagen I and III and PDGF were also carried out. In prolapsed specimens our results show a disorganization of smooth muscle cells that appeared to have been displaced by an increased collagen III deposition resulting in rearrangement of the muscularis propria architecture. These findings suggest that the increase in the expression of collagen fibers in muscularis could probably be due to a phenotypic switch resulting in the dedifferentiation of smooth muscle cells into myofibroblasts. These alterations could be responsible for the compromising of the dynamic functionality of the pelvic floor.

Repeated ovarian stimulation does not affect the expression level of proteins involved in cell cycle control in mouse ovaries and fallopian tubes

PurposeTo understand if repeated cycles (2–4 rounds) of gonadotropin stimulation could affect intracellular localization/content of proteins controlling cell cycle progression in mouse fallopian tubes (FT) and ovaries.MethodsFT and ovaries of estrous mice (control) and of stimulated mice were analyzed to detect Oct-3/4, Sox-2, p53, β-catenin, pAKT and cyclin D1 localization/content. Spindles and chromosome alignment were analyzed in ovulated oocytes.ResultsAfter round 4, FT and ovaries of control and stimulated groups showed no differences in Oct-3/4, Sox-2 and β-catenin localization nor in Oct-3/4, Sox-2, p53, β-catenin and pAKT contents. Cyclin D1 level increased significantly in FT of treated mice. Oocytes number decreased meanwhile frequency of abnormal meiotic spindles increased with treatments.ConclusionsRepetitive stimulations affected oocyte spindle morphology but did not induce changes in a set of proteins involved in cell cycle progression, usually altered in ovarian cancer. The significant increase of cyclin D1 in the FT requires further investigation.