DNA methylation of HOXA10 in eutopic and ectopic endometrium.

STUDY QUESTION: Does the methylation status of the promoter region of the HOXA10 gene differ in eutopic and ectopic endometrium? SUMMARY ANSWER: The eutopic endometrium in women with endometriosis is significantly more methylated when compared with controls. WHAT IS KNOWN ALREADY: Expression of the HOXA10 gene, which is important for successful implantation, is reduced in women affected by endometriosis. STUDY DESIGN, SIZE AND DURATION: A pilot study was carried out including 18 women admitted for surgery for endometriosis-related pain (cases) and 12 women admitted for surgery because of non-endometriotic disease (control). Sample collection and analysis were performed between November 2010 and July 2013. PARTICIPANTS/MATERIALS, SETTING, METHODS: Endometrial tissue (eutopic and ectopic) underwent sodium bisulfite DNA modification, PCR amplification of two regions of the HOXA10 promoter and pyrosequencing analysis. MAIN RESULTS AND THE ROLE OF CHANCE: The eutopic endometrium of women with endometriosis was significantly more methylated compared with endometrium from the control group (sequence 1: 8.68% in cases and 6.25% in the control group: P = 0.037, sequence 2: 11.89% in cases and 9.25% in the control group: P = 0.032). The eutopic endometrium was significantly more methylated than the ectopic tissue in patients with endometriosis (mean difference -3.6 sequence 1: P = 0.001 and -6.0 sequence 2: P = 0.0001). LIMITATIONS, REASONS FOR CAUTION: The study had a limited sample size and the fertility status of the majority of patients in our study was unknown. WIDER IMPLICATIONS OF THE FINDINGS: Our data regarding methylation state of the ectopic tissues contribute to a better etiopathologic understanding of endometriosis. STUDY FUNDING/COMPETING INTERESTS: No external funding was either sought or obtained for this study. The authors have no conflicts of interests to declare.

Amniotic band syndrome: a case report.

Amniotic band syndrome is an uncommon congenital pathological condition that may lead to malformations and foetal-infant death. We report an autoptic case. The patient was a male preterm infant. At 14 weeks of gestation, a routine ultrasonography showed severe craniofacial anomalies and a close contiguity of the foetal head with the amnios. The neonate survived three days, after which an autopsy was carried out. The infant had a frontoparietal meningoencephalocele; a fibrous band was attached to the skin, close to the meningoencephalocele base. Cleft lip and palate, nose deformation and agenesis of the right eye were also present. At the opening of the cranial cavity, occipital hyperostosis was observed. The herniated brain showed anatomical abnormalities that made identification of normal structures difficult. Microscopically, the nervous parenchyma had architectural disorganization and immaturity, and the fibrous band consisted of amniotic membranes. As evident from this case report, amniotic band syndrome may cause severe malformations and foetal-infant death.

Liquid-based endometrial cytology: the Florence and Bari experience.

Several diagnostic procedures are available to investigate the endometrium, i.e. sonography, hysteroscopy, biopsy, endometrial curettage and cytology. Among these, endometrial cytology is less commonly utilized. Although the use of cytology in the diagnosis of endometrial adenocarcinoma has already been proposed due to its low cost and simple execution, a general consensus has not been reached. The improvement of the diagnostic capacity of endometrial cytology following the introduction of a liquid-based method suggests that this test should be routinely used in endometrial diagnosis. The main advantages of this method are the reduction in confounding factors, the distribution of cells on a thin layer and the possibility to obtain more slides from the same sample. The aim of this article is to focus on the methodological procedures and diagnostic criteria in liquid-based endometrial cytology based on the experience in two Italian centres: Department of Pathology, University of Bari and Department of Human Pathology and Oncology, University of Florence. The sampling method used by the Bari authors consists in the collection of liquid for uterine distension during hysteroscopy, while the Florence group used an endometrial brush. The sensitivity and specificity at Bari were 75% and 83%, respectively, and were 94-100% and 95-100% at Florence, respectively. Endometrial cytology provided sufficient diagnostic material significantly more often than biopsy. We thus propose that endometrial cytology can be used in routine diagnosis either alone or in association with other diagnostic procedures in order to improve diagnostic accuracy.

Gestational diabetes insipidus: a morphological study of the placenta.

Gestational diabetes insipidus (GDI) refers to the state of excessive water intake and hypotonic polyuria. Those cases manifesting in pregnancy and referred to as GDI may persist thereafter or may be a transient latent form that resolves after delivery. Microscopic examination of affected subjects has not been previously reported. In the literature, there are various case reports and case series on diabetes insipidus in pregnancy. In this study, we present a case that had transient diabetes insipidus during pregnancy in which the placenta was examined.

O6-Methylguanine-DNA-methyltransferase in recurring anaplastic ependymomas: PCR and immunohistochemistry.

Ependymomas are the third most common brain tumor in children. The post surgical management is controversial. There are no convincing data on an effective role for chemotherapy. O(6)-Methylguanine-DNA-Methyltransferase (MGMT) is a DNA repair protein considered to be a chemosensitivity predictor. Hypermethylation of the MGMT gene promoter is an important cause of MGMT inactivation. We evaluated the MGMT gene promoter methylation and the immunohistochemical MGMT protein expression in 12 recurrent anaplastic ependymomas affecting children. Our purpose was to investigate the molecular rationale of the administration of alkylating agents to children affected by recurrent anaplastic ependymomas. All ependymomas lacked MGMT promoter hypermethylation and 9 (75%) showed high MGMT protein expression (>50% tumoral cells). Differences between different recurrences in the same patient were not observed. These results may indicate MGMT as a factor of chemoresistance to alkylating drugs in anaplastic ependymomas and support the uncertainties regarding the actual benefit of chemotherapy for patients with anaplastic ependymomas.

Placental findings in pregnancy-associated haemolytic uremic syndrome: clinicomorphological study of three cases.

Acute renal failure occurring in pregnancy or postpartum is often associated with preeclampsia, HELLP syndrome or haemolytic uremic syndrome: differential diagnosis may be difficult due to the overlapping symptoms of these syndromes. We report our experience on diagnosis, management and outcome of women with pregnancy associated haemolytic uremic syndrome, focusing on placental features.

Occipital pilomyxoid astrocytoma in a 14-year-old girl--case report.

Pilomyxoid astrocytoma is a recently described tumor. Its most typical morphological characteristics are an angiocentric astrocytic proliferation embedded in a myxoid background. The behavior seems to be unfavorable due to the reported high rate of local recurrence. The earlier studies indicated that pilomyxoid astrocytoma typically affects young children and arises in the hypothalamic/chiasmatic region. We report a case of a 14-year-old patient with a 6-year history of absence seizure. Magnetic resonance imaging showed a right occipital lesion of approximately 3 cm in diameter. The patient underwent the surgical procedure with gross total excision. Histologically, the tumor was mainly composed of a monomorphous population of bipolar elongated piloid cells radially arranged around thin-walled blood vessels in a prominent myxoid background. There were focal hemorrhagic foci but no bona fide evidence of tumor necrosis or mitoses. Rosenthal fibers and eosinophilic granular bodies were not observed. The postoperative course was uneventful. No adjuvant therapy was administered. The patient is alive and well at 18-month follow-up. The case presented provides evidence that pilomyxoid astrocytoma can occur at a later age and can arise in regions different from hypothalamic/chiasmatic.

Liquid-based endometrial cytology: cyto-histological correlation in a population of 917 women.

OBJECTIVE: Liquid-based cytology, because of its capacity to reduce the obscuring factors and to provide thin-layer specimens, represents an opportunity to reevaluate endometrial cytology. In order to assess the utility of the liquid-based method in endometrial diagnosis, we evaluated its accuracy in comparison with histology. METHODS: Nine hundred and seventeen women scheduled for hysteroscopy were enrolled in the study. After providing informed consent, all the women proceeded sequentially to hysteroscopy, endometrial cytology and then biopsy endometrial sampling. RESULTS: Cyto-histological correlations were possible in 519 cases (57%): in 361 (39%) cases the biopsy was inadequate, in 15 (2%) the cytology was inadequate, and in 22 (2%) both were inadequate. At biopsy 25 (3%) women had adenocarcinoma, 5 (1%) had adenomatous atypical hyperplasia and 21 (2%) had simple non atypical hyperplasia. At cytology two adenocarcinomas and one adenomatous atypical hyperplasia were underrated as atypical hyperplasias and as non-atypical hyperplasia; two simple non-atypical hyperplasias were reported as negative; and eight cases were false positive (non-atypical hyperplasia at cytology, negative at biopsy). In our population, the cytology provided sufficient material more often than biopsy (P < 0.04). Sensitivity was estimated at 96%, specificity at 98%, positive predictive value at 86% and negative predictive value at 99%. CONCLUSIONS: We concluded that endometrial cytology may be an efficient diagnostic method. It could be applied to selected patients solely or in association with ultrasonography. The combination of these two noninvasive procedures may improve their diagnostic accuracy and reduce unnecessary hysteroscopies, thereby producing benefits for women and society.

Could gamma/delta T cells influence the mother's rejection in recurrent aborted embryos?

The possibility that the investigation of aborted material may identify aetiologies not easily detectable from even a careful clinical investigation, suggested a study of the T-cell receptors (TCRs) of decidual-infiltrating T-lymphocytes in recurrent spontaneous miscarriage (RSM). From 33 cases of RSM (>3 previous consecutive miscarriages, range 3-5, mean 3.7), PCR products were analysed by 15% acrylamide gel electrophoresis and visualised under UV illumination after ethidium bromide staining. A broad band obtained suggests the presence of a monoclonal T-lymphocyte proliferation. A PCR not showing bands means that the tissue does not contain reactive T cells. A total of 11 samples (33.3%) revealed the presence of receptor TCRgamma with the presence of a specific band. T-cell receptors in RSM were identified in one-third of cases. These data underline the importance of a maternal immune host response to the embryo and the need to study the immune mechanisms with the hope of modulating therapeutic treatment of recurrent abortion.

Tumor markers in ovarian carcinoma.

This review analyzes in 2 ways the prognostic value of markers found in ovarian carcinomas before chemotherapy. It is known that neoangiogenesis, cyclooxygenase activity, and host responsiveness to chemotherapy can be evaluated by means of specific molecules recognized as tumor markers. However, host response as well as tumor histotype, grade of differentiation, clinical characteristics, and histopathologic characteristics must also be taken into account when selecting a treatment. Analysis must therefore focus on the molecular basis of aggressive disease, on tumor peculiarity, on the efficacy of chemotherapy, and on the host's response to the tumor. Although treatment may be more aggressive in patients with unfavorable prognostic elements, it may be modulated according to the molecular and cellular biology of the tumor and the host's response. When the tumor's molecular characterization contributes to the choice of treatment, prognostic markers may turn into predictive markers.

Feasibility and outcome of laser CO2 conization performed within the 18th week of gestation.

The purpose of this study is to evaluate the feasibility, safety, and potential therapeutic benefit of laser CO(2) conization of the cervix for in situ and minimally invasive carcinoma diagnosed during pregnancy. Twenty-six pregnant patients with biopsy-proven carcinoma in situ/cervical intraepithelial neoplasia III but colposcopically suspicious for invasion underwent laser CO(2) conization during the 18th week of gestation in an outpatient setting under local anesthesia. No major intraoperative or postoperative complications occurred, and cervical cerclage was not required in any case. Two cases (7.7%) of occult FIGO stage IA1 minimally invasive cervical cancers with free surgical margins were diagnosed. Both patients delivered vaginally at term and were free of disease at postpartum follow-up. Median length of gestation was 39.1 weeks with a median birth weight of 3450 g. All 1-min Apgar scores were 8 or greater. Twenty patients (76.9%) delivered vaginally, while six patients underwent cesarean section for indications not related to the prior conization. After a mean postpartum follow-up of 18 months (range 3-42), 92.3% of patients continued to have both cytologic and colposcopic evaluations negative for persistent or recurrent disease. Two cases of persistent intraepithelial disease were successfully managed by reconization. In summary, our data suggest that laser CO(2) conization performed within the 18th week of gestation is safe for both the patient and the fetus, provides reliable histologic diagnosis, and can be curative. Further studies are required to confirm the favorable risk-benefit ratio of laser CO(2) conization in the management of non-reassuring cervical lesions observed in the first half of pregnancy.

Liquid-based endometrial cytology: its possible value in postmenopausal asymptomatic women.

The incidence of endometrial adenocarcinoma in asymptomatic women is low. Nevertheless, some of these women might require endometrial surveillance. In this study, we evaluated the accuracy of liquid-based endometrial cytology compared to biopsy in asymptomatic postmenopausal women. Three hundred twenty women scheduled for hysteroscopy were enrolled for this study. After hysteroscopy, patients were submitted to endometrial cytology and to biopsy. Two hundred ninety-three (92%) women had sonographically thickened endometrium (>5 mm), 53 (17%) were on tamoxifen, and 16 (5%) were on hormonal substitutive treatment. The evaluation of the biopsies determined that six (2%) women had adenocarcinoma, one (<1%) had adenomatous atypical hyperplasia, and eight (3%) had simple nonatypical hyperplasia. Endometrial cytology evidenced 5 (2%) neoplastic cases, 2 (<1%) hyperplastic with atypia cases, and 25 (8%) hyperplastic without atypia cases. Two hundred twenty-two biopsies (69%) and 17 (5%) cytologies were inadequate. One adenocarcinoma and one simple nonatypical hyperplasia were underrated by cytology resulting, respectively, as atypical hyperplasia and as negative. Four cases were false positive (simple nonatypical hyperplasias on cytology, negative on biopsy). The sensitivity and specificity were estimated, respectively, at 94% and 95%; the positive and negative predictive value were estimated, respectively, at 80% and 99%. Endometrial cytology provided sufficient material more often than biopsy (P < 0.01). We suggest to introduce liquid-based endometrial cytology in the management of some subpopulations of asymptomatic postmenopausal women. Particularly, the combination of liquid-based endometrial cytology and transvaginal sonography may improve their diagnostic accuracy and reduce unnecessary more invasive and expensive procedures.

[Embryo and fetal pathology in routine diagnostics: what has changed and what needs to be changed]. / L'embrio-patologia e la patologia feto-perinatale nella diagnostica anatomo-patologica: cosa e cambiato e cosa e necessario cambiare.

The Authors have focused on the most important feto-neonatal and placental diseases in order to develop modern diagnostic tools which can meet the needs of clinicians (obstetricians, gynecologists, and neonatologists) for the best possible management of both the mother and the newborn. Although far from being operational instructions, it should be intended as a programmatic document providing a guideline on the issues that have cropped up in eight years of work of the APEFA group, as well as during several residential and practical classes. First of all, a synopsis is provided of the main issues concerning placental diagnosis in the newborn, as well as in case of fetal loss. A reasoned review is then provided of the main diagnostic criteria in placental pathology, in the light of therapeutical measures toward the mother (monitoring of future pregnancies) and the newborn (management of newborns at risk or with infectious disease). Legal issues in case of fetal distress at the end of pregnancy, neonatal damage and peripartum death have also been discussed with particular attention. Early and late miscarriages have also been separately examined, as well as fetal deaths. For each of these categories, a critical analysis is presented of current issues, followed by some considerations on the development of diagnostic methods and technology, and a modern diagnostic process is then outlined. Reference tables are also provided for diagnostic, auxological parameters, as well as on essential procedures. Issues concerning legal abortions and terminations of pregnancies have also been considered, with particular reference to tests and supplemental genetic and ultrasound examinations, diagnostic questions about malformations and forensic medicine assessments that are often involved with these specific categories. Malformations, fetal distress and growth retardation, sudden fetal and neonatal death, as well as embryo-pathology are all briefly dealt with also with synoptic tables. Diagnostic criteria are thus optimized and specially aimed at solving "human reproduction pathology" issues.

Inducible cyclooxygenase (COX-2) in glioblastoma--clinical and immunohistochemical (COX-2-VEGF) correlations.

Cyclooxygenase-2 (COX-2) is the inducible form of the enzyme responsible for the first step in the prostaglandin synthesis. COX-2 upregulation is demonstrated in different tumors. COX-2 products may modulate tumoral growth, apoptosis, metastasis, multidrug resistance and angiogenesis. Moreover, the antitumoral effect of the COX inhibitors has been documented. We studied the immunohistochemical expression and the prognostic value of COX-2 on 43 surgical specimens of glioblastoma-affected patients. Furthermore, we evaluated the correlation between the immunohistochemical expression of COX-2 and vascular endothelial growth factor (VEGF). Of the glioblastomas, 63% resulted as COX-2-positive. Median survival of the patients with COX-2-positive lesions was 10 months; median survival of the patients with COX-2 negative glioblastoma was 21 months (NS). All 4 patients who survived longer than 24 months had COX-2 negative lesions (p = 0.017). Concordance between COX-2 and VEGF was documented in 60% of the cases. Our findings show that glioblastoma can immunohistochemically express COX-2 and that its expression is unrelated with VEGF and significantly less frequent in the long survivors. Nevertheless, the absence of statistical correlation with survival time advises further studies on larger series to ascertain the concrete prognostic value of COX-2 in glioblastoma.

Expression and amplification of HER-2/neu oncogene in uterine carcinosarcomas: a marker for potential molecularly targeted treatment?

Surgery is the treatment of choice for uterine carcinosarcomas; nevertheless, the poor effect of chemotherapy and radiotherapy represents an insidious problem for patients with metastatic or unresectable disease, and indeed, new therapeutic approaches are clearly required to improve survival of uterine carcinosarcoma patients. The HER-2 oncogene, located on chromosome 17, encodes for a tyrosine kinase growth factor receptor. We analyzed HER-2/neu overexpression by immunohistochemistry in 28 uterine carcinosarcomas. HER-2/neu amplification with fluorescence in situ hybridization (FISH) was tested in positive cases. The expression of HER-2/neu was correlated with disease-free interval and survival (Kaplan-Meier estimates). We observed HER-2/neu overexpression in nine cases (32.1%) and HER-2/neu amplification in all the four HER-2/neu 3+ score positive cases tested by FISH. HER-2/neu expression was not correlated with clinical outcome. Patients with disease limited to the uterus (stages I-II) displayed a significantly better disease-free survival (P= 0.004) and actuarial survival (P= 0.01). Demonstration of HER-2/neu overexpression and amplification in uterine carcinosarcoma may represent the first rationale step for further investigations. Hence, the results of this analysis may support the challenge of a new therapeutic approach, which could test the role of anti-HER-2 (trastuzumab) in patients with advanced or metastatic uterine carcinosarcoma.

Microvessel density in ovarian carcinoma: computer image analysis in patients with shorter and longer survival.

We previously reported that tumor microvessel density (MVD) may have prognostic significance in ovarian carcinoma. The aim of this study was to compare the intratumoral microvessels using a computer-aided image analysis system between FIGO stage IIIC, serous, G3, ovarian carcinomas obtained from living patients who had no evident disease 5 years after primary treatment and ovarian carcinomas, matched for stage, histopathology, grade of differentiation, and treatment, obtained from patients who had died of progression of disease no later than 1 year after primary treatment. We observed that MVD is statistically correlated, according to the logistic regression in univariate and multivariate ways, with the survival (P= 0.03 and P= 0.05, respectively) and with the progression of the disease during first-line chemotherapy (P= 0.009 and P= 0.012, respectively). In the past years, the modulation of first-line chemotherapeutic treatment has been a question of discussion, because the oncologist observes extremely unpredictable behaviors with surprisingly long survivals and also short survivals. Pathologists may give clinicians some additional prognostic information useful in the management of ovarian carcinoma patients. The results of this study support the hypothesis that the evaluation of MVD with computer image analysis can help clinicians in the choice of the tailored treatment of the single case.

hERG1 channels are overexpressed in glioblastoma multiforme and modulate VEGF secretion in glioblastoma cell lines.

Recent studies have led to considerable advancement in our understanding of the molecular mechanisms that underlie the relentless cell growth and invasiveness of human gliomas. Partial understanding of these mechanisms has (1) improved the classification for gliomas, by identifying prognostic subgroups, and (2) pointed to novel potential therapeutic targets. Some classes of ion channels have turned out to be involved in the pathogenesis and malignancy of gliomas. We studied the expression and properties of K(+) channels in primary cultures obtained from surgical specimens: human ether a gò-gò related (hERG)1 voltage-dependent K(+) channels, which have been found to be overexpressed in various human cancers, and human ether a gò-gò-like 2 channels, that share many of hERG1's biophysical features. The expression pattern of these two channels was compared to that of the classical inward rectifying K(+) channels, IRK, that are widely expressed in astrocytic cells and classically considered a marker of astrocytic differentiation. In our study, hERG1 was found to be specifically overexpressed in high-grade astrocytomas, that is, glioblastoma multiforme (GBM). In addition, we present evidence that, in GBM cell lines, hERG1 channel activity actively contributes to malignancy by promoting vascular endothelial growth factor secretion, thus stimulating the neoangiogenesis typical of high-grade gliomas. Our data provide important confirmation for studies proposing the hERG1 channel as a molecular marker of tumour progression and a possible target for novel anticancer therapies.