ROMANIAN JOURNAL of MORPHOLOGY and EMBRYOLOGY

Vol. 46 No. 2, 2005

1. Primary malignant lymphoma of the testis

Amelia Petrescu, Camelia Dobrea, Mariana Vasilica, Fl. Andrei, L. Niculescu

Primary testicular lymphomas are rare entities representing 1-2% of non-Hodgkin lymphoma (NHML) and 1-7% of malignant testicular tumors and they are the most common testicular tumors in men older than 50 years of age. This study included eight cases of inpatients diagnosed by echography and NMR with testicular tumors. The age of patients was between 46 and 81 (with a mean of 52). The tumors were unilateral, with disease limited to testicle and accompanied by pain except one case with bone involvement. Orchectomy was performed as first therapeutic and diagnostic purpose. All patients were clinically staged according to the Ann Arbor criteria in IE and IIID stage and received a doxorubicin based chemotherapy regimen (CHOP, MTX, CVP, and Leukeran). A standard chemotherapy protocol has not been used because of reduced number of patients. Tumor fragments were fixed in 10% formalin, paraffin embedded, sectioned and standard H.E. stained. Immunohistochemistry for L26, Alphafetoprotein, NK1, CD30, and CLA was performed. Microscopy revealed in all cases a stromal proliferation with medium size cells, monomorphic shape and prominent nucleoli. Alphafetoprotein was positive in seminal tubes and negative in tumor, NK1 in small lymphocyte and negative in tumor and L26 diffuse positive in tumor. CLA diffuse positive in tumor. We were able to follow up only four patients. Conclusions: The diagnosis was of NHML in six cases and for two secondary involvement of hematopoietic malignancy (myeloid sarcoma and leukemia). Lymphoma cases were typed using REAL classification as small and large B-cell lymphoma. Unfavorable evolution with 6 months relapse and one death prove a more aggressive evolution of primitive testicular lymphoma.

Corresponding author: Amelia Petrescu, MD, PhD, e-mail: etamy58@yahoo.com

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2. Cytokinic panel in rheumatoid arthritis and correlation with histological patterns of synovitis - active type of disease

Carmen Avramescu, C. C. Vere, Cl. Margaritescu, Adriana Turculeanu, Maria Balasoiu, Suzana Rogoz

Rheumatoid arthritis (RA) is a chronically inflammatory disease of the articular synovial, with severe, progressive and irreversible articular destruction. RA pathogeny implies an autoimmune mechanism, the role of cytokines resulting from the exaggerated production of some cytokines that act as pro-inflammatory agents, being directly responsible of certain symptoms and articular destructions, and/or inadequate inhibition of certain cytokines that act as anti-inflammatory agents. Objective. We want to determine certain pro-inflammatory cytokines (tumoral-alpha - TNF-alpha necrosis factor, interleukine-6 - IL-6, interleukine-8 - IL-8), anti-inflammatory cytokines (interleukine-10 - IL-10) and immunomodulators (interleukin-2 - IL-2) in some RA patients serum at the active stage of the disease and correlation with histological patterns of synovitis - active type of disease. Material and methods. The 37 patients have been grouped in stage I (9 patients), II (15 patients) and III (13 patients) according to the anatomical staging of the ARA (American Rheumatism Association), the serum levels of the cytokines being determined by ELISA technique. From the 37 patients clinical investigated only 12 were histopathological examined. Results. The serum levels of IL-2 have been found low in patients with RA in the active stage of the disease, the lowest values having been determined at the patients in the 3rd stage of the disease, who also have the highest values of pro-inflammatory cytokines. In the case of IL-10 the lowest values have been found at patients in the advanced stages of the disease. In the serum of RA patients with follicular synovitis TNF-alpha was a dominant cytokine compared to patients with diffuse disease, but the greatest serum level was found in patient with granulomatous synovitis. Concentrations of IL-6 and IL-2 were highest in the serum of RA patients with follicular synovitis in comparison to patients with diffuse synovitis and could distinguish RA patients with these two histological variants of the disease. Conclusion. In the active stage of the disease, the symptoms are a consequence of the interleukin pro and anti-inflammatory game: high serum levels of pro-inflammatory cytokines are accompanied by low serum levels of anti-inflammatory cytokines. Cytokines can be used as monitorization markers of the acutization period in RA, increase in serum levels of pro-inflammatory cytokines showing the progression from the inactive phase to a new period of activity of the disease. The association between distinct histological appearance of rheumatoid synovitis and serum cytokine profile and diverse clinical activity of disease seems to confirm its heterogeneity.

Corresponding author: Carmen Avramescu, MD, PhD, e-mail: c.avramescu@yahoo.com

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3. Morphometric investigation of carotid body in Sudden Infant Death Syndrome

Z. Pavai, Klara Toro, Eva Keller, J. Jung

Background. The pathomechanism of sudden infant death syndrome (SIDS) has not been clarified yet. The high rate of early progenitor cells in carotid body has been reported as a pathognomic feature for SIDS. Aim and Study design. The morphometric analysis was done by NIKON Eclipse microscope with a morphometric program Lucia G. Subjects. This study was designed to investigate the structure and developmental state of carotid body in SIDS and non-SIDS cases. A comparison was made between the rates of dark and early progenitor cells. Outcome measures. The Kruskal-Wallis test showed a significantly higher number of progenitor cells in the SIDS group than in controls (p = 0.0003). Results and conclusion. In this study on Hungarian SIDS cases, we confirmed the observation that infants who died suddenly have an underdeveloped carotid body.

Corresponding author: Zoltan Pavai, MD, PhD, e-mail: zpavai@umftgm.ro

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4. C-erbB2 oncoprotein: prognostic marker in breast cancer

Anca Rosian, Elena Lazar, Alis Dema

Aim. The invasive mammary cancer is the most frequent malign tumor of a woman. Among the potential prognostic factors are included the biomarkers which measure or are associated with biological processes involved in the tumoral progression. We study the prognostic importance of the c-erbB2 oncoprotein in the invasive mammary cancer. Material and methods. We included in the study 56 female patients suffering of invasive mammary cancer. The tissue fragments were fitted in formalin, included to paraffin, commonly colored Hematoxylin-Eosin (HE); for the determination of the c-erbB2 we used the immunohistochemical method of Avidin-Biotin complex (ABC). Results. From the total of 56 tumors, 38 (67.9%) presented different degrees of positivity; 18 (32.1%) did not express the oncoprotein c-erbB2. Conclusions. The positivity of c-erbB2 oncoprotein was correlated with the size of the tumor; the carcinomas which are not infiltrative ductal, knows as having a better prognostic, were c-erbB2 negative.

Corresponding author: Anca Rosian, MD, PhD, e-mail: morfopat@umft.ro

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5. The place and the role of histological examination in diagnostic algorithm of urinary system tuberculosis

Em. Trasca, E. T. Trasca, R. Buzulica, Gh. Dragoi, Iuliana Nicolescu

The diagnosis of tuberculosis of renal and urinary tract is made by identifying Koch bacillus in special cultures and using histological examination of surgical removed pieces. Not in all cases, the usual pathological techniques are very specific. Using special stain for acid-fast bacilli it can be certified the etiological diagnostic. Histological changes of renal parenchyma and/or upper urinary tract (renal pelvis and urether) structures in 57 patients clinical and paraclinical pyonephrosis diagnosed where studied. All the surgical removed pieces were studied using usual pathology methods. In order to find renal tuberculosis we performed on surgical pieces special staining (Ziehl-Nielsen), we noted the pathological finding in each case and we found seven cases with certainly tuberculosis etiology.

Corresponding author: Emil Trasca, MD, PhD, e-mail: lucianbuzulica@yahoo.com

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6. A rare case of glioblastoma multiforme

I. Radu, A. Nicola, Daniela Micu, D. Minea

A male, 39 years, enters rapidly in coma, is admitted in the Neurological Hospital Brasov and deceases after 4 hrs. in hospital. At necropsy, macroscopic appearances of the cut surface showing unique tumor mass in all brain from frontal lobe to cerebellum, with extensive cortical invasion. The tumor has a variable coloration, a yellowish central necrosis and cystis containing turbid fluid. Histopathological aspect is extremely variable with regional heterogeneity: striking cellular pleomorphism with a lack of cell processes, anaplastic glial cells, and some multinucleated giant cells, large necrotic areas with pseudopalisading of surrounding tumor cells and microvascular proliferation, as glomeruloid tufts.

Corresponding author: Ioan Radu, MD, PhD, e-mail: raduic@k.ro

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7. Neurofibroma, schwannoma or a hybrid tumor of the peripheral nerve sheath?

Elena Doina Murarescu, Luminita Ivan, Maria Sultana Mihailovici

Classically, the different subtypes of the peripheral nerve sheets tumors display typical morphological features, which differentiate them from the other nervous tumors. Either schwannoma or neurofibromma have characteristic histological and immunohistochemical features. We present a case of peripheral nerve sheets hybrid tumor, which have had histological and immunohistochemical features of neurofibroma and schwannoma.

Corresponding author: Elena Doina Murarescu, MD, PhD, e-mail: doinamurarescu@yahoo.com

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8. Cell cycle regulatory factors in juxtatumoral renal parenchyma

Daniela Nicoleta Petrusca, Amelia Petrescu, Camelia Vrabie, L. Niculescu, V. Jinga, Carmen Diaconu, Lorelei Brasoveanu

The aim of this study was to evaluate regulatory cell cycle factors in juxta-tumoral renal parenchyma in order to obtain information regarding early primary changes occurred in normal renal cells. Specimens of juxta-tumoral renal parenchyma were harvested from the tumoral kidney in 10 patients with no history of treatment before surgery. The expression of p53, Bcl-2, Rb and PCNA was studied by immunohistochemical methods in paraffin-embedded tissues. The apoptotic status was evaluated by flow-cytometry analysis following propidium iodide incorporation. The p53 protein expression was recognized in most of the cases (80%) with different intensities. High intensity apoptotic process detected in juxta-tumoral parenchyma seemed to be p53 dependent and well correlated with the low Bcl-2 expression. 70% of cases were Rb positive. In this type of tissue, Rb has only an anti-proliferative and anti-tumoral role. PCNA was present in half of the cases being low expressed due to the tissue regenerating mechanism. Our data suggest that the high intensity of programmed cell death in this type of tissue is supported by the status of cell regulatory factors that control this process. Previous studies have demonstrated that healthy renal tissue has neither apoptosis nor mitotic activity. Juxta-tumoral renal tissue is also displaying normal morphology and DNA content (diploidy) but the microenvironmental status induced by the tumor presence prompts cells to choose death rather than malignant transformation. Further studies are necessary to emphasize if these results have a clinical relevance for the outcome of therapeutical approaches in renal carcinomas.

Corresponding author: Daniela Nicoleta Petrusca, MD, PhD, e-mail: dpetruscaro@yahoo.com

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9. Diagnostic significance of the immunoexpression of CD34 and smooth muscle cell actin in benign and malignant tumors of the breast

Anca Maria Cimpean, M. Raica, Diana Narita

Purpose. Our aim was to investigate the distribution of CD34 and smooth muscle cell actin positive myofibroblasts in the stroma of the normal mammary gland, benign and malignant tumors. The observations were especially focused on the diagnostic value of the cumulative results obtained with these immunoreactions. Methods. Our study included 112 female patients with suspect breast masses obtained by surgery or biopsy. We performed morphological study and immunohistochemistry for CD34 and SMA. Results. We have found normal breast tissue, sclerosing adenosis, fibroadenomas, fibrocystic diseases, phyllodes tumor, DCIS, ductal invasive, lobular, squamous, medullary, mucinous, and papillary carcinomas. We also found apocrine metaplasia, florid ductal hyperplasia, atypical hyperplasia, papilloma and LCIS associated with the malignant tumors. All the normal breast tissues and most of benign lesions were positive for CD34 and negative for SMA. The exceptions were represented by a case of fibroadenoma and the phyllodes tumor, with CD34 positivity and a focal acquisition of SMA; fibrocystic disease with associated apocrine metaplasia adjacent to a squamous carcinoma with loss of CD34 expression and focal acquisition of SMA. All our DCIS and invasive carcinomas have lost CD34 expression and gained SMA positivity. Some particular behavior had the mucinous and squamous carcinomas. Conclusions. Although there were some exceptions especially when one of the two markers was interpreted separately and in some cases associated with sclerotic stroma, we conclude that the combined expression of CD34 and alpha-SMA is of potential diagnostic value in the distinction between benign and malignant tumors in some difficult cases.

Corresponding author: Anca-Maria Cimpean, MD, PhD, e-mail: ancacimpean1972@yahoo.com

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10. Morphological changes in dental pulp after the teeth preparation procedure

Anca Vitalariu, Irina-Draga Caruntu

The aim of this study was to evaluate the immediate changes in the pulp-dentin complex that result from crown preparation, and their correlation with the thickness of remaining dentin and the preparation technique (with or without water spray cooling). Thirty upper intact premolars scheduled for extraction for orthodontic reasons were high speed prepared, extracted immediately after preparation and divided in five groups. The analysis of the pulp morphology demonstrated that there are several differences according with the preparation technique. The most severe changes appear after the profound preparation without water-cooling, the odontoblastic layer being extremely affected. Also, vascular reactions and inflammatory infiltrate (in the absence of bacteria) were present. Our study revealed that the histologic changes in the pulp and dentin following complete crown preparation occur anyway and they are considered difficult to avoid, even if an adequate technique of preparation is used.

Corresponding author: Irina-Draga CÄ?runtu, MD, PhD, e-mail: dicarunt@mail.dntis.ro

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11. Immunological and histopatological correlations in exocrine pancreatic cancer

Adriana Turculeanu, Carmen Avramescu, Maria Balasoiu, Em. Plesea, Cristiana Simionescu, Carmen Florina Popescu

The CA19-9 antigen is a tumoral marker that can be found in high concentration within a maligned digestive pathology. Objective. Study of CA19-9 antigen from the immunological point of view, at the patients with pancreatic cancer, and his relation with the histopathological aspects. Material and method. The determinations have been done at the time of the diagnosis and after the treatment of the patients with pancreatic cancer (25). Method: indirect ELISA. Results. The values of the CA19-9 marker have been increased at the time of the diagnosis either in pancreatic cancer of the head and mid. At the moment of the diagnosis the CA19-9 marker has higher values (150-400 U/mL) in cancer of the pancreatic body than in cancer of the head of the pancreas (40-200 U/mL). Correlating the size of the tumor with the value of the CA19-9 marker in the case of pancreatic cancer, we have been shown the highest serum values (300-400 U/mL) at patients whose tumor was 3 cm bigger. Four weeks post-surgery the CA19-9 values decreased (37-100 U/mL) or reached normal levels (<37 U/mL). At patients with local recurrence or metastasis, especially hepatical, CA19-9 values have increased (100-400 U/mL in case of recurrence and 800 U/mL in case of metastasis). The highest values of the CA 19-9 marker were found in cases of mid pancreas adenocarcinoma. Conclusions. CA19-9 marker values higher than normal cause problems to digestive cancer (especially pancreatic). CA19-9 is a good marker for monitoring of these cancers after treatment; the favorable development is associated with lower values in comparison to the determinations before treatment, or with normal values, unfavorable evolution (local recurrence or metastasis) being associated with very high values. The high values of CA 19-9 suggest an adenocarcinoma.

Corresponding author: Adriana Turculeanu, MD, PhD, e-mail: turculeanu_balasoiu@yahoo.com

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12. Postmenopausal choriocarcinoma: a case report

Madalina Marcu, Alina Chefani, Maria Sajin

Postmenopausal uterine choriocarcinoma is very rare and benefits of curative chemotherapy. We present here the case of 62-year-old women with uterine bleeding. Emergency surgery revealed a uterine tumor and histopathology findings were consistent with choriocarcinoma. Immunohistochemistry tests confirmed betahCG and cytokeratin expression by malignant cells, thus establishing the positive diagnosis.

Corresponding author: Madalina Marcu, MD, PhD, e-mail: marcu.madalina@gmail.com

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13. Quantitative versus qualitative in the analysis of cervical squamous cell carcinoma

Irina-Draga Caruntu, Raluca Balan, C. Visan

Our work aimed to reveal and explore connections between the cellular morphometric features and the differentiation degree of cervical squamous cell carcinoma. Six cases were studied, for which biopsy was performed before the radiation treatment. The microscopic exam diagnosed two cases of well-differentiated squamous cell carcinoma and four cases of moderately differentiated squamous cell carcinoma. The morphometric technique was developed in the Zeiss KS400 software environment, by implementing a script, which allowed measuring the cytoplasmic and nuclear areas and calculating the nucleo-cytoplasmic ratios. For each studied case, characterized by a number of 500-measured cells (randomly selected), the paper presents the mean, minimum and maximum values of the two types of areas, as well as the histograms of the nucleo-cytoplasmic ratios. These numerical results showed that the quantitative approach could play an important role in enlarging the perspectives on the evaluation of the differentiation degree in invasive squamous cell carcinoma. Special attention was paid to the nucleo-cytoplasmic ratios, since we supposed they are able to provide enough information for quantifying several levels in the differentiation of the cells. Although we have obtained encouraging results, our current work cannot be regarded as fully confirming this assumption, because of the small number of cases. By involving larger groups of patients in our further researches, we expect to get a solid confirmation, and, thus, to support the refinements of the radiotherapy schemes.

Corresponding author: Irina-Draga Caruntu, MD, PhD, e-mail: dicarunt@mail.dntis.ro

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14. HPV and p53 expression in dysplastic lesions and squamous carcinomas of the oral mucosa

Cristiana Simionescu, Cl. Margaritescu, Claudia Valentina Georgescu, M. Surpateanu

We studied 18 premalignant and malignant lesions of the oral mucosa, three dysplasias, and 15 oral squamous carcinomas with varied localizations: lips, palate, and tongue. Immunoassays for HPV were positive in 42.8% of the squamous carcinomas, without correlation with the pattern or the degree of differentiation of the lesions. P53 immunoexpression was positive in two of the three studied cases of dysplasia (66.6%) and in 13 from the 15 cases of squamous carcinomas (86.6%). The p53 immunopositivity in both lesional categories, with a higher rate in carcinomas, indicates the fact that the p53 gene mutations can appear precocious in the oral carcinogenesis, but cannot be used as malign potential prediction factor.

Corresponding author: Cristiana Simionescu, MD, PhD, e-mail: csimionescu2004@yahoo.com

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15. Signet-ring prostatic carcinoma. Case presentation and review of the literature

Renata Vasiu, C. D. Olinici, I. Coman

The authors present the case of a 71-year-old man with the histopathological diagnosis of a signet-ring prostatic carcinoma, examined in usual and special stains as PAS and mucicarmin. The literature, the clinical aspect, the microscopic features, and the pathogenesis concerning this subject are reviewed.

Corresponding author: Renata Vasiu, MD, PhD, e-mail: anapatol@yahoo.com

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16. In memoriam of Professor Maria Florescu (1944-2005)

Committee of The Romanian Society of Morphology, Craiova

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Corresponding author: -

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