Angelos Daniilidis

The evaluation of p16INK4a immunoexpression/immunostaining and human papillomavirus DNA test in cervical liquid-based cytological samples.

Aim: To evaluate the role of p16INK4a immunoexpression and human papillomavirus (HPV) DNA test for the detection of dyskaryotic cells in high-risk women. Materials and Methods: This work was a retrospective diagnostic study conducted in the University Hospital of Thessaloniki from January to December 2008. The subjects were women with current or previous HPV infection and current or previous cervical intraepithelial lesion (with or without treatment) or clinical warts. All liquid-based cytological samples were tested for P16INKa and HPV DNA test. The accuracy parameters used for the outcome included sensitivity, specificity, and positive predictive value. Results: A total of 226 women were included; the mean age was 29 years. Expression of p16INK4a was detected in the cytological samples of 13% of the negative cases, 44% of the cases of atypical squamous cells of undetermined significance, 46% of the cases of low-grade squamous intraepithelial lesion, and 78% of the cases of high-grade squamous intraepithelial lesion. A total of 91 women tested positive for high-risk HPV infection, and 54 of those had p16INK4a-positive staining reaction cells. The concordance between the 2 tests, HPV DNA and p16, was 59% regarding infection-positive cases. Diffuse strong parabasal p16INK4a immunostaining (nuclear score >2) was observed in 17 cases of the abnormal cytological findings (atypical squamous cells of undetermined significance, 2 cases; low-grade squamous intraepithelial lesion, 8 cases; high-grade squamous intraepithelial lesion, 7 cases). Colposcopy-directed biopsies were used as the criterion standard for the detection of cervical intraepithelial neoplasia in 91 women. The sensitivity of p16INK4a was 95% and the specificity was 92%, whereas the sensitivity of high-risk HPV was 100% and the specificity was 78%. The positive predictive value of p16INK4a was 71%, whereas that of HPV DNA was 44%. Conclusion: The findings suggest that p16INK4a immunostaining can improve the accuracy of cytological examination and HPV DNA test and may be particularly useful in the triage of low-grade lesions.

Osteoprotegerin as a Marker of Atherosclerosis in Diabetic Patients

Atherosclerosis is the principal cause of cardiovascular disease (CVD) and has many risk factors, among which is diabetes. Osteoprotegerin (OPG) is a soluble glycoprotein, involved in bone metabolism. OPG is also found in other tissues, and studies have shown that it is expressed in vascular smooth muscle cells. OPG has been implicated in various inflammations and also has been linked to diabetes mellitus. Increased serum OPG levels were found in patients with diabetes and poor glycemic control. Furthermore, prepubertal children with type 1 diabetes have significantly increased OPG levels. Receptor activator of nuclear factor kappa-B ligand (RANKL) is not found in the vasculature in normal conditions, but may appear in calcifying areas. OPG and RANKL are important regulators of mineral metabolism in both bone and vascular tissues. Few data are available on the relationship between plasma OPG/RANKL levels and endothelial dysfunction as assessed using noninvasive methods like ultrasound indexes, neither in the general population nor, more specifically, in diabetic patients. The aim of our review study was to investigate, based on the existing data, these interrelationships in order to identify a means of predicting, via noninvasive methods, later development of endothelial dysfunction and vascular complications in diabetic patients.

Acute lung injury in preterm fetuses and neonates: mechanisms and molecular pathways.

Abstract Acute lung injury (ALI) results in high morbidity and mortality among preterm neonates and efforts have therefore been devoted to both antenatal and postnatal prevention of the disease. ALI is the result of an inflammatory response which is triggered by a variety of different mechanisms. It mostly affects the fetal lung and, in particular, causes damage to the integrity of the lung’s alveolar-capillary unit while weakening its cellular linings. Chemotactic activity and inflammatory products, such as proinflammatory cytokines TNF-α, IL-1, IL-6, IL-11, VEGF,TGF-α and TGF-β, provoke serious damage to the capillary endothelium and the alveolar epithelium, resulting in hyaline membrane formation and leakage of protein-rich edema fluid into the alveoli. Chorioamnionitis plays a major part in triggering fetal lung inflammation, while mechanical ventilation, the application of which is frequently necessary in preterm neonates, also causes ALI by inducing proinflammatory cytokines. Many different ventilation-strategies have been developed in order to reduce potential lung injury. Furthermore, tissue injury may occur as a result of injurious oxygen by-products (Reactive Oxygen Species, ROS), secondary to hyperoxia. Knowledge of the inflammatory pathways that connect intra-amniotic inflammation and ALI can lead to the formulation of novel interventional procedures. Future research should concentrate on the pathophysiology of ALI in preterm neonates and οn possible pharmaceutical interventions targeting prevention and/or resolution of ALI.

Awareness of human papillomavirus infection, testing and vaccination in midwives and midwifery students in Greece

Summary Midwives play an important role in the implementation of cervical cancer screening. We assessed the knowledge of human papillomavirus (HPV) infection and of its relationship with cervical cancer in 107 midwives and 29 graduating midwifery students. The majority of midwives (78.5%) were aware that a viral infection causes cervical cancer, whereas only 48.3% of the students knew this (p = 0.003). Only one midwife (0.9%) was not aware of HPV infection compared with 10.3% of the students (p = 0.029). Midwives were also more knowledgeable of the relationship between HPV infection and cervical cancer and of the availability of a vaccine against HPV infection (p = 0.005 and p < 0.0001, respectively). In conclusion, Greek midwives have a satisfactory level of knowledge about cervical cancer and HPV infection, in contrast to midwifery students. It is important to better educate midwifery students in order to facilitate the incorporation of HPV testing and vaccination in clinical practice.

Endocrine, paracrine, and autocrine placental mediators in labor.

Considering that preterm birth accounts for about 6–10% of all births in Western countries and of more than 65% of all perinatal deaths, elucidation of the particularly complicated mechanisms of labor is essential for determination of appropriate and effective therapeutic interventions. Labor in humans results from a complex interplay of fetal and maternal factors, which act upon the uterus to trigger pathways leading gradually to a coordinated cervical ripening and myometrial contractility. Although the exact mechanism of labor still remains uncertain, several components have been identified and described in detail. Based on the major role played by the human placenta in pregnancy and the cascade of labor processes activated via placental mediators exerting endocrine, paracrine, and autocrine actions, this review article has aimed at presenting the role of these mediators in term and preterm labor and the molecular pathways of their actions. some of the aforementioned mediators are involved in myometrial activation and preparation and others in myometrial stimulation leading to delivery. In the early stages of pregnancy, myometrial molecules, like progesterone, nitric oxide, and relaxin, contribute to the retention of pregnancy. At late stages of gestation, fetal hypothalamus maturation signals act on the placenta causing the production of hormones, including CRH, in an endocrine manner; the signals then enhance paracrinically the production of more hormones, such as estrogens and neuropeptides, that contribute to cervical ripening and uterine contractility. These molecules act directly on the myometrium through specific receptors, while cytokines and multiple growth factors are also produced, additionally contributing to labor. In situations leading to preterm labor, as in maternal stress and fetal infection, cytokines trigger placental signaling sooner, thus leading to preterm birth.

Probing the impact of sex steroids and menopause-related sex steroid deprivation on modulation of immune senescence.

Immune senescence denotes the general decline in immune system function, characterized by a reduced immune response and an increased inflammatory state. Menopause is a natural change in a womens life, the menopause-related low estrogen levels affecting many body functions, among them the immune system. Numerous human studies with menopausal women and animal models with surgically induced menopause show a clear impact of sex steroids in immune responses. Female superiority in vaccination response and predisposition to infections are eliminated after menopause, while during menopause inflammatory cytokines such as Tumor Necrosis Factor-α (TNF-α), Interleukins-1β, 6, 8 and 13 (IL-1β, IL-6, IL-8, IL-13) and Monocyte Chemoattractant Protein-1 (MCP-1) are increased, implying a molecular connection of sex steroid loss with immune senescence. Moreover, immune cells modify their number and function after the menopausal transition, this offering another explanation for immune senescence. Until now most of the existing studies have concluded that menopause plays an additional role to aging in immune senescence. While it is clear that we are as yet far from thoroughly understanding the molecular pathways connecting sex steroids and menopause with immune senescence, such knowledge is highly likely to enable future targeted interventions in treatment and prevention of age-related diseases in women.

Predictive value of anti-müllerian hormone, follicle-stimulating hormone and antral follicle count on the outcome of ovarian stimulation in women following GnRH-antagonist protocol for IVF/ET

AimTo compare the efficacy of three methods: the levels of anti-müllerian hormone (AMH), the levels of follicle stimulating hormone (FSH) and the antral follicle count (AFC), for the prediction of the number of retrieved mature oocytes and the number of generated embryos by intracytoplasmic sperm injection (ICSI) in women stimulated with a GnRH-antagonist protocol.Materials and methods105 women were enrolled in the study. At the day 2 of a preceding cycle, AFC was performed and FSH and AMH were measured in serum by immunoenzymatic assays. All women were stimulated with a GnRH-antagonist protocol and ovulation was induced with human chorionic gonadotropin. ICSI was performed in all retrieved mature oocytes. Embryo transfers were performed at days 2–3. According to the oocytes retrieved, patients were categorized as poor (<4), normal (4–12) or high responders (>12).ResultsAFC and the levels of baseline FSH and AMH were significantly different among poor, normal and high responders. The number of oocytes as well as the number of embryos was negatively correlated with baseline FSH and positively correlated with baseline AMH and AFC, whereas AFC showed the strongest correlation. Stepwise regression analysis indicated AFC and baseline AMH as the most significant parameters for the prediction of the number of oocytes; for the prediction of the number of embryos, the most significant parameter was AFC.ConclusionsAFC, baseline AMH and baseline FSH are good predictors for the outcome of ovarian stimulation in GnRH-antagonist cycles. However, AFC appears to have the best predictive value.

Oxidized laminin-1 induces increased monocyte attachment and expression of ICAM-1 in endothelial cells

Atherosclerosis has been associated with increased oxidative stress and monocyte recruitment by endothelial cells. Sub‐endothelial basement membrane proteins, such as laminins that play a central role in cell adhesion, are exposed to reactive oxygen species. In the present study monocyte attachment on human umbilical cord vein endothelial cells (HUVEC) that were preattached to oxidized or native laminin, was investigated. Intracellular cell adhesion molecule‐1 (ICAM‐1) expression by HUVEC was estimated by an enzyme‐linked immunosorbent assay. HUVEC attachment to oxidized or native laminin‐1 was examined using the Hemacolor kit. Anti‐alphaL, anti‐alphaM, anti‐alpha2 and anti‐beta2 integrin subunit antibodies were used in order to further investigate the above phenomena. HUVEC that were preattached to oxidized laminin expressed higher levels of ICAM‐1 and monocytes attached at a higher degree to these cells as compared to HUVEC that were preattached to native laminin. Incubation of monocytes with monoclonal antibodies against the alphaM and beta2 integrin subunits equalized the above mentioned differences. Moreover, HUVEC attached to oxidized laminin at a higher degree as compared to native laminin. This difference was equalized after incubation with the antibody against the alpha2 integrin subunit. These results indicate a modified interaction between HUVEC and the basement membranes in cases where laminin is oxidatively modified. This modified interaction results in increased ICAM‐1 expression by endothelial cells and consequently increased monocyte recruitment capacity.

Anterior vaginal wall reconstruction: anterior colporrhaphy reinforced with tension free vaginal tape underneath bladder base

Objective. To assess the effectiveness of the polypropylene tape in preventing cystocele recurrence. Methods. Fifty Caucasian women aged 50‐77 years (mean age 66.6 years), with stage II–IV pelvic organ prolapse, enrolled into the study. Vaginal reconstructive surgery included an anterior colporrhaphy on all patients, posterior colpoperineorrhaphy on 28 patients and hysterectomy on 36 patients. Patients were divided into a study group of 28 women and a control group of 22 women. As reinforcement to the anterior colporrhaphy procedure, in the study group a polypropylene tape (TVT‐O) was placed underneath the bladder base and fixed with polyglactin sutures. Postoperative follow‐up was carried out every 4 months (total 48 months). The assessment of the anatomic result included both clinical evaluation of the operated sites and perineal sonography. Results. The mean postoperative distance of the bladder base to the inferior edge of the symphysis pubis was 1.5 cm (range: 1.0–2.2 cm) in the study group and 2.8 cm (range: 2.0–3.8) in the control group. Postoperatively, there were two cases of stress incontinence and two cases of urge incontinence, one in each group. No case of tape erosion was noted, no dyspareunia and no recurrent cystocele in the study group. Four cases of recurrent cystocele (20%) were reported in the control group. Conclusion. While the preliminary results of our study are encouraging, larger series of patients and longer follow‐up are required to verify the effectiveness of the aforementioned modification.

Indications of diagnostic hysteroscopy, a brief review of the literature

Plenty of authors propose outpatient hysteroscopy as the gold standard diagnostic method for the evaluation of endometrial pathology. This statement has been strengthened in the recent years due to the wide use of smaller diameter hysteroscopic devices, which have made the dilation of the cervix and the use of anesthesia unnecessary. The main purpose of this paper is to summarize the indications of diagnostic hysteroscopy. In this review, we used the most recent publications in MEDLINE and Cochrane Library in order to specify the indications of diagnostic hysteroscopy and the experience that have been obtained till today in the management of certain pathological uterine conditions. The key words we used were diagnostic hysteroscopy, abnormal uterine bleeding, infertility, endometrial cancer. Hysteroscopy provides an accurate method of evaluation and direct visualization of the endometrial cavity and moreover directed biopsy and sampling of suspected lesions. Last years with the continuous development in the hysteroscopy devices, plenty of women benefit surgical hysteroscopy techniques for uterine abnormalities. Hysteroscopy is useful for the diagnosis in patients with abnormal uterine bleeding, with endometrial cancer and in infertile women. Hysteroscopy has the unique advantage of combining a thorough procedure with great diagnostic accuracy. The only disadvantage is that hysteroscopy requires specific teaching and training and has a long learning curve.