The value of serological markers in the diagnosis and prognosis of endometriosis: a prospective case-control study.

OBJECTIVE: We analyzed selected well-known and less well-known serum markers that have been proposed for diagnosis and severity assessment of endometriosis, in a case-control study. STUDY DESIGN: This prospective study was carried out in a Clinical Department of Gynecology in Iasi, Romania. Study participants included endometriosis patients, and controls in whom laparoscopy had excluded endometriosis. Each case and control was investigated for serum levels of CA125, TNF, IL-1, IL-6 and IL-8. The data were correlated with clinical symptoms and revised American Fertility Society (rAFS) score and stage, and interpreted by Mann-Whitney U-test and ANOVA regression analysis. RESULTS: Over the course of 1 year, 24 cases of endometriosis and 24 controls of matched age were selected. The rAFS stages were: stage I, 12.5%; stage II, 16.7%; stage III, 58.3%; and stage IV, 12.5%. CA125 levels were over the cut-off of 35 IU/l in 54% of patients (versus 8% of controls), averaging 67.5 (CI95: ±17.5). The sensitivity and specificity were 54% and 91%, respectively, with a p value of <0.001 (statistically significant). For IL-6, 71% of cases and 87% of controls were above the cut-off of 2 pg/ml, with an average of 11.83 ± 7. The sensitivity and specificity were 71% and 12%, respectively, but the difference was not statistically significant, p = 0.071. Other tested serum markers had no discrimination value. A correlation with severity of endometriosis was seen for CA125 (p = 0.03) but not for IL-6, by ANOVA. CONCLUSION: CA125 correlated with endometriosis screening and severity, indicating its superiority as a marker for further, larger studies.

Tumor biomarkers in cervical cancer: focus on Ki-67 proliferation factor and E-cadherin expression.

UNLABELLED: Despite recent advances in the immune mechanisms of cervical cancer (CC) and complex management opportunities, relapse remains still an actual issue. While predictive factors are required, current research is directed towards proliferation and tumor aggressiveness biomarkers as potential negative factors in CC. The main objectives were to assess tumor proliferation and invasiveness biomarkers (Ki-67, E-cadherin) and to identify potential correlation between biomarkers and classic prognostic factors in CC. Radical hysterectomy specimens from 61 consecutive CC were immunohistochemically investigated for Ki-67 and E-cadherin. Nuclear immunostaining for Ki-67 proliferation index was assigned scores 1 to 3, "+" meaning low (10-30%), "++" moderate (30-50%), "+++" high-proliferation rate (>50%); cell membrane E-cadherin staining was either negative or positive. Statistical analysis was performed in SPSS-13 software, p<0.05. RESULTS: no significant correlation between Ki-67 and classical prognostic factors (p>0.05) was reported; however, in relapsed CC, Ki-67 correlates with tumor grading (r=0.386, p<0.05). Significant correlation between E-cadherin and tumor size (r=-0.280, p=0.029), relapse (r=-0.386, p=0.002) and disease free survival (r=0.374, p=0.003) were demonstrated. Indirect statistically significant moderate correlation between Ki-67 and E-cadherin (r=-0.461, p<0.00001) was shown, mainly in invasive squamous CC (r=-0.549, p=0.0001), stage IB (r=-0.578, p=0.009), IIB (r=-0.585, p=0.003), relapsed CC (r=-0.525, p<0.01), HPV-infection (r=-0.504, p=0.033). CONCLUSIONS: CC aggressiveness, particularly in invasive squamous carcinoma, either 16 or 18 HPV-positive cases, FIGO stage IB and IIB, and cases with relapse, depends on two pivotal factors, tumor proliferation rate (Ki-67) and tumor invasiveness (E-cadherin).

[Expression of the Bax proapoptotic factor in asthmatic patients]. / Expresia factorului proapoptotic Bax la pacientii cu astm bronsic.

The alteration in the expression of the factors involved in the apoptotic mechanism of the respiratory cell epithelium of asthma patients remains an unsolved issue. The mitochondria holds a very important place in the apoptotic chain, making therefore the study of the proteins belonging to the Bcl-2 family that "controls" permeability of mitochondria's membrane extremely interesting, both for asthma as for other diseases. Our main concern in this study was to reveal the changes in expression of Bax factor at the level of the respiratory epithelium in asthma patients. We examined a number of 21 patients with different degrees of asthma. The tissue samples were accordingly processed for immunotyping with anti-Bax antibodies. Following microscopic examination we have found, even in mild cases of asthma, a decreased expression of Bax factor at the epithelial level, compared with the witness. The discovery of all these factors involved in cell apoptosis as well as their alteration in Asthma will be the future of therapeutic approach in these patients.

[Cutaneous squamocellular carcinoma--immunohistochemical aspects]. / Aspecte imunohistochimice in carcinoamele scuamocelulare cutanate.

UNLABELLED: p53 gene is frequently founded mutated in nonmelanoma skin cancer like basocellular carcinoma, squamocellular carcinoma and equally in actinic keratosis and Bowen disease. Our objective was to evaluate the role of p53 protein and proliferative factor Ki-67 on skin squamocellular carcinoma. MATERIAL AND METHODS: We investigated a number of 21 cases of cutaneous squamocellular carcinoma using immunohistochemical research for p53, proliferative factor Ki-67 (avidine-biotine-peroxidase method). RESULTS: The reaction for p53 was positive in 76.2% cases (16 of 21 cases), without semnificative statistical correlation with histologic differentions. Ki-67 values distribution was between 10% - 75%, with a mean value of 35%, with semnificative statistical correlation with histologic differentiations (p = 0.02). CONCLUSION: p53 immunoreactivity was high in the majority of skin squamous cell carcinoma examined, probable related to an inactivated protein resulting from mutations of the p53 gene or other unclear molecular mechanisms. Diagnostic and prognostic role of p53 and Ki-67 level is difficult to establish in the light of recent results. This study suggests the utility of this tests, but low number of cases of study limit the statistical validity of conclusions.

[Clinical and paraclinical diagnosis of pelvic endometriosis. Clinical study]. / Studiu privind diagnosticul clinic si paraclinic al endometriozei pelvine.

UNLABELLED: Pelvic endometriosis is a difficult diagnosis in gynecological practice, due to different symptoms and advanced stages in which the patients arrive to us. MATERIAL AND METHOD: This study retrospectively analyses 73 cases of confirmed endometriosis admitted in our service during 5 years. We gathered information about their clinical data and diagnosis, and intraoperative laparoscopic or laparotomic findings. RESULTS: The majority of our cases were in the 3rd age decade (52%). The clinical symptoms were: infertility (34%), pelvic or abdominal pain (71%), vaginal bleeding (14%), or an accidental finding after laparotomy or laparoscopy for other reasons (7%). The intraoperative evaluation of the cases showed: endometrial genital foci in 59%, with half of them located at the ovary; extragenital sites in 30 cases (bladder--4 cases, Douglas pouch 19 cases, abdominal wall 5 cases, para-cervix 2 cases). Associated pathology included: adhesions in 46%, retroverted uterus in 15%, ovarian non endometriotic cysts (18%), polycystic ovaries (12%) and tubal pathology (6%). CONCLUSION: Our study confirm the difficulty of a clinical diagnostic of endometriosis, but the main symptoms remain infertility and pelvic-abdominal pain. The laparoscopy or laparotomy were the main tools for the diagnostic and management of our cases.

[Lymphangioleiomyomatosis (LAM)--an uncommon cause of bilateral spontaneous pneumothorax]. / Limfangioleiomiomatoza cauza rara de pneumotorax spontan bilateral.

LAM, a rare lung disease typically affecting women of reproductive age, is characterized by abnormal proliferation of smooth--muscle cells and progressive loss of pulmonary function due to destruction of lung parenchyma. Two cases of bilateral successive recurrent spontaneous pneumothorax and haemoptysis are presented. Repeated conventional and video-assisted surgery was required in both cases, for drainage of the recurrent pneumothorax and resection of subpleural bulla, with good immediate postoperative evolution. Immunohistochemical studies of resected specimens revealed LAM cells in the lung parenchyma with receptors for oestrogen and progesterone. HMB45 monoclonal antibodies in the LAM cells were identified in one case. The follow-up of the patients revealed no signs of recurrence at 84 and 18 months respectively, although pulmonary transplantation should be considered in case of further deterioration of respiratory function.

T-cell primary cutaneous lymphomas. A clinicopathological and immunohistochemical study.

Within the large framework of the lymphoproliferative diseases, the primary cutaneous lymphomas are distinct pathologic conditions, defined by particular morphologic, immunologic, genetic, and clinic criteria. The study aimed to create the first clinicopathological and immunohistochemical profile of primary cutaneous lymphoma for a Romanian region. We investigated a series of 16 cases (diagnosed during a 5-year period) in accordance with the general principles of primary cutaneous lymphoma management. The methods included the clinic and morphologic exams, the latter relying on standard and immunohistochemical staining. The results revealed that all studied cases were T-type lymphomas, in terms of the WHO-EORTC classification. Most of these cases were diagnosed as mycosis fungoides; the group also included cases of Sezary syndrome, as well as rare entities such as: mycosis fungoides associated with follicular mucinosis and subcutaneous panniculitis-like T-cell lymphoma. Our discussions focused on the role of the clinicopathological assessment for the primary cutaneous lymphoma diagnosis and emphasized the importance of the immunohistochemical investigation. Compared with the previous Romanian researches on this topic, presenting only isolated cases, the current study develops a new level of analysis, based on the rigorous monitoring of a relatively large geographical area, for a long time horizon.

[The detection of dysplastic lesions of the gallbladder mucosa in polypoid cholelithiasis]. / Evidentierea leziunilor displazice din mucoasa veziculei biliare în colecistitele calculoase polipoide.

AIM: To investigate the hyperplastic and/or dysplastic lesions in the gallbladder mucosa. METHODS: We used the immunohistochemical Ardeleanu-Hsu-Bussolati and Gugliotta methods, with avidin and peroxide biotin, adapted in the laboratory of University Hospital "St. Spiridon" Iasi. There were studied ten cases of polypoid cholelithiasis, with the following markers: Ki67, PCNA, and p53. RESULTS: The immunohistochemical methods using Ki67, PCNA and p53 protein allowed the evaluation of the abnormal proliferation abilities characteristic for some hyperplastic or dysplastic lesions, found in our cases of cholelithiasis. CONCLUSIONS: The utilization of these markers permitted us to establish a good correlation between the high activity of Ki67 and the morphological aggression indicators in hyperplastic and dysplastic gallbladder epithelium. There is also a correlation between the percentage of the cells marked with anti-PCNA antibody and the existence of p53 protein in these lesions.

[Different phenotypes generated by changes in bcl-2 and p53 expression in breast cancer]. / Fenotipuri diferite generate prin modificari in expresia bcl-2/p53 in cancerul mamar.

We have identified by immunohistochemistry/ immunocytochemistry the expression of bcl-2 molecule in 55 primary breast carcinomas and in 30 corresponding axillary lymph nodes metastases, together with a set of molecules known as prognostic factors: estrogen receptors, progesterone receptors, and p53 protein. Our results demonstrated a significant correlation (p < 0.05) between bcl-2 and hormonal receptors expression in tumors, but not in axillary metastases (p < 0.1), a significant inverse correlation between bcl-2 and p53 expression in primary tumors (p < 0.02), but a significant direct correlation in axillary metastases (p < 0.02). The bcl-2+/p53- phenotype, associated with normal breast epithelium, is present in 79.17% primary tumors, but only in 15.38% axillary lymph nodes metastases. A larger number of lymph nodes metastases expressed a bcl-2+/ p53+ more aggressive phenotype compared with primary tumors (58.82% versus 48.39%). This shows that changes in the expression of bcl-2, p53, estrogen and progesterone receptors can lead to an increased cellular aggressiveness and thus to an increased tumoral invasive and metastasizing potential.

[Evaluation of the angiogenic process in breast carcinoma--correlations with cancer aggressiveness]. / Evaluarea angiogenezei în cancerul mamar. Corelatii cu agresivitatea tumorala.

Angiogenesis represents an essential event required by tumors to support their growth. The aim of our study was to investigate the presence of neovascularization in 44 primary breast carcinomas and in 24 axillary lymph nodes metastases and to establish a possible correlation between the presence of tumor angiogenesis, some clinical and pathological features of the cases and the expression of p53, an important cell cycle regulator. To identify the new blood vessels, we used immunohistochemistry for von Willebrand factor, a marker of the endothelial cells. The results showed that 77.27% of the primary breast carcinomas and 75% of the lymph nodes metastases are positive for von Willebrand factor and this positivity is significantly correlated (p < 0.05) with the expression of p53, supporting the idea that angiogenesis is a marker for tumor aggressiveness and p53 could be involved in this process.

[Early diagnosis in hypertrophic cardiomyopathy--case report]. / Importanta diagnosticului precoce în cardiomiopatia hipertrofica.

Disproportionate heart muscle enlargement compared with little or no chamber enlargement are characteristic for hypertrophic cardiomyopathy (HCM). The clinical course of HCM is highly variable. Many patients are asymptomatic or mildly symptomatic and may be relatives of patients with known disease. Unfortunately, the first clinical manifestation of the disease may be sudden death, frequently occurring in children and young adults, often during or after physical exertion. We present 2 cases of HCM who emphasize the variability of clinical and ECG modifications and underline the importance of routine echocardiography in patients with atypical cardiac symptoms in order to identify the disease and prevent high risk for sudden death.

[Transcriptional and electron microscopic analysis of extracellular matrix proteoglycans in acute acetaminophen intoxication]. / Analiza la nivel transcriptional si electronomicroscopic a expresiei proteoglicanilor de matrice extracelulara în intoxicatia acuta cu Acetaminofen.

UNLABELLED: Acetaminophen (APAP) OTC regimen is not compatible with kidney and liver toxic effects. In order to further understand the determinism of the repair process, we have studied the expression of some target extracellular matrix proteoglycan components in an animal model. MATERIALS AND METHODS: An acute APAP toxic dose (800 mg/bw) was administered to 45 mice aged 1 month. At 24 hours, tissue samples were processed for light microscopy, electromicroscopy and RT-PCR analysis. RESULTS: Extensive necrotic lesions involving hepatocytes and tubular epithelial cells associate strong positivity for proteoglycans, detected as convoluted filaments with lateral projections, frequently associated with collagen fibers, cell surface and other matrix components. Overexpression for perlecan, syndecan 1 and 4 and low expression for biglycan was assessed by RT-PCR. CONCLUSIONS: Extracellular matrix and cell surface components are early involved in both lesional mechanism and repair reaction post acetaminophen injury.

[The significance of Fas/FasL molecular system in breast cancer]. / Semnificatia expresiei sistemului molecular Fas/FasL în cancerul mamar.

Fas (CD95/APO-1) and its natural ligand, FasL, are molecules expressed on cellular membranes, being involved in the induction of programmed cells death or apoptosis. Recently, it has been shown that malignant cells originating from solid tumors tend to inhibit the expression of Fas, as an escape mechanism from the immune cells' attack and to express FasL, as a counterstrike mechanism against the immune effector cells. The purpose of this study was to investigate, by immunohistochemistry, the presence of Fas and FasL in 15 breast carcinomas and to establish possible associations between the expression of these molecules and the histological type and grading of the tumors. Our results showed that 7 breast tumors have lost the expression of Fas and 11 tumors were positive for the Fas-ligand expression, important arguments for the mechanisms of immune escape and tolerance induction. Furthermore, 7 of the 11 FasL+ tumors were poorly differentiated invasive ductal carcinomas, suggesting a possible association between FasL expression and tumor aggressivity.

The MHC class I related Fc receptor, FcRn, is expressed in the epithelial cells of the human mammary gland.

The major histocompatibility complex (MHC) class I related neonatal Fc receptor (FcRn) plays multiple roles, being involved in transporting immunoglobulin G (IgG) and protecting this antibody class from catabolism. The presence of this receptor was previously demonstrated in the lactating murine mammary gland. In the current study we have investigated FcRn expression in various histologic types of human breast carcinoma and lymph node metastases. We used immunohistochemical methods to demonstrate the presence of FcRn in epithelial cells, whereas this Fc receptor could not be detected in mammary gland endothelial cells. The presence of the receptor was also found in the metastasizing epithelial cells within the lymph nodes, and this provides a useful marker for their identification.

Combined modality treatment for locally advanced breast cancer.

PURPOSE: To assess the value of aggressive loco-regional surgery in desperate situations with locally advanced breast cancer. METHODS: In this study there were considered 31 patients with locally advanced breast cancer who underwent surgery in a 5-year period. 10 of them received 3 cycles of chemotherapy and radiotherapy before surgical intervention; the rest of the 21 patients had systemic or local contra-indications for neo-adjuvant therapy. We describe clinical aspects and technical difficulties. Surgical intervention focused on tumour removal and lymph node dissection. Skin defect was covered with flaps according to the Mortimer-Show technique. Postoperatively, the outcome was influenced in a favourable way by the use of Detralex, a micronized flavonoid; all but 2 patients received chemotherapy and locoregional radiotherapy. RESULTS: 25 patients survived free of disease; from 6 patients who suffered recurrence, 2 are still living and 4 have died. CONCLUSIONS: In some forms of locally advanced breast cancer, aggressive surgery offers improvement in the quality of life and increases survival.

[Overexpression of c-erbB-2 gene product is associated with poor prognosis factors in breast carcinoma]. / Supraexpresia produsului genei c-erbB-2 se asociaza cu factori de prognostic rezervat în carcinomul mamar.

Oncogenes, the abnormal forms of proto-oncogenes, were shown to be involved in malignant transformation and in tumor progression. c-erbB2/HER2/neu is member of EGFR family and encodes the p185 protein, which functions as a tyrosine-kinase. Gene amplification and/or p185 overexpression were reported to be associated with poor prognostic in cancer. Our purpose was to investigate p185 immunohistochemical expression in breast carcinomas and in the corresponding axillary lymph nodes metastases and to identify possible correlation between p185 and other factors of poor prognostic, such as loss of hormonal receptors expression. In our study, 40.91% of cases were erbB-2 positive, p185 expression being maintained from the primary tumors to axillary metastases and associated with positive nodal status and with the absence of hormonal receptors expression (p < 0.05). These findings support the hypothesis the c-erbB2 is an advantageous acquisition for the aggressive behavior of the tumor cell and for its ability to invade and metastasize.

[Loss of hormonal receptors expression from primary breast carcinoma to axillary lymph node metastases]. / Pierderea expresiei receptorilor hormonali de la tumora primara la metastaza ganglionara în carcinomul mamar.

Breast tumors growth is regulated by female sex steroid hormones. The level of the estrogen and progesterone receptors (ER and PR) expression by the malignant cells is important for the evaluation of the tumor prognostic and the benefit of a hormonal therapy. The aim of our study was to identify the expression of estrogen and progesterone receptors in primary breast tumors and in the corresponding axillary lymph nodes metastases, in 24 cases. The results showed that more than 30% of poorly differentiated breast carcinomas lost their expression of hormone receptors from the primary tumors to axillary metastases, an event which can be associated with an aggressive tumoral behaviour and resistance to hormonal therapy.

[Could biological aggressivity markers of breast cancer alter the therapeutic course? Preliminary results]. / Aprecierea agresivitatii biologice a tumorilor maligne mamare ar putea schimba agresivitatea actului terapeutic? Rezultate preliminarii.

The aim of this study is to justify an individual therapeutical attitude in breast cancer, related to diversity of breast tumors, aggressiveness grade and metastatic potential. Between January 2000--December 2001, 150 patients were admitted with breast cancer (stage II and III) and underwent surgery in our department. We selected 75 cases in our study. In 51 (68%) cases the first therapeutical method was surgery, in 15 (20%) cases surgery was performed after chemotherapy, in 2 (2.66%) cases after radiotherapy and after chemotherapy and radiotherapy in 7 (9.33%) cases. We evaluated several classical factors and new immunohistochemical markers with an important value for diagnosis, prognosis and therapy: oestrogen and progesterone receptors, c-erb B2, pS2 and p53 proteins, von Willebrand factor. Several factors had a predictive role regarding the response to chemotherapy. These predictive factors will improve the histopathological diagnosis. The oncoproteins and hormonal receptors also will evaluate with more accuracy the metastatic risk and will assure a better therapy decision.

[Detection of lymph nodes micrometastases in breast carcinoma using immunohistochemistry for cytokeratin 8]. / Depistarea micrometastazelor ganglionare în carcinomul mamar prin imunohistochimie pentru citokeratina 8.

The lymph nodes status is one of the most important prognostic factors in breast cancer. The routine hematoxilin-eosin staining is efficient for the metastases detection only when there is a large number of tumor cells, while a small number of metastatic cells can easily remain undetectable. For those situations, the immunohistochemistry for cytokeratins, markers of the epithelial cells, is a very sensitive method. We have investigated the cytokeratin 8 expression in 10 primary breast carcinomas and in the corresponding axillary lymph nodes, comparing with hematoxilin-eosin. The routine examination has detected axillary lymph nodes metastases in six cases, confirmed by the immunohistochemistry for cytokeratin 8. Four cases were diagnosed as negative for the axillary lymph nodes metastases by the hematoxilin-eosin staining. In all those four cases, immunohistochemistry for cytokeratin 8 has detected a small number of tumor cells, either spread in the lymph nodes tissues, either confluent as small islets.