Vol. 45 No. Cumulative Issue 1999-2004, 2004

1. From telepathology to virtual pathology institution: the new world of digital pathology

K. Kayser, G. Kayser, D. Radziszowski, A. Oehmann

Telepathology has left its childhood. Its technical development is mature, and its use for primary (frozen section) and secondary (expert consultation) diagnosis has been expanded to a great amount. This is in contrast to a virtual pathology laboratory, which is still under technical constraints. Similar to telepathology, which can also be used for e-learning and e-training in pathology, as exemplarily is demonstrated on Digital Lung Pathology ( at least two kinds of virtual pathology laboratories will be implemented in the near future: a) those with distributed pathologists and distributed (>=1) laboratories associated to individual biopsy stations/surgical theatres, and b) distributed pathologists (usually situated in one institution) and a centralized laboratory, which digitizes complete histological slides. Both scenarios are under intensive technical investigations. The features of virtual pathology comprise a virtual pathology institution (mode a) that accepts a complete case with the patient's history, clinical findings, and (pre-selected) images for first diagnosis. The diagnostic responsibility is that of a conventional institution. The Internet serves as platform for information transfer, and an open server such as the iPATH ( for coordination and performance of the diagnostic procedure. The size and number of transferred images have to be limited, and usual different magnifications have to be used. The sender needs to posses experiences in image sampling techniques, which present with the most significant information. A group of pathologists is "on duty", or selects one member for a predefined duty period. The diagnostic statement of the pathologist(s) on duty is retransmitted to the sender with full responsibility. The first experiences of a virtual pathology institution group working with the iPATH server working with a small hospital of the Salomon islands are promising. A centralized virtual pathology institution (mode b) depends upon the digitalization of a complete slide, and the transfer of large sized images to different pathologists working in one institution. The technical performance of complete slide digitalization is still under development. Virtual pathology can be combined with e-learning and e-training, that will serve for a powerful daily-work-integrated pathology system. At present, e-learning systems are "stand-alone" solutions distributed on CD or via Internet. A characteristic example is the Digital Lung Pathology CD, which includes about 60 different rare and common lung diseases with some features of electronic communication. These features include access to scientific library systems (PubMed), distant measurement servers (EuroQuant), automated immunohistochemistry measurements, or electronic journals (Elec J Pathol Histol, It combines e-learning and e-training with some acoustic support. A new and complete database based upon this CD will combine e-learning and e-teaching with the actual workflow in a virtual pathology institution (mode a). The technological problems are solved and do not depend upon technical constraints such as slide scanning systems. At present, telepathology serves as promoter for a complete new landscape in diagnostic pathology, the so-called virtual pathology institution. Industrial and scientific efforts will probably allow an implementation of this technique within the next two years with exciting diagnostic and scientific perspectives.

Corresponding author: Klaus Kayser, MD, PhD, e-mail:

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2. Thymus and thymoma: what's new?

M. Raica, Svetlana Encica, L. Mogoanta, C. Suciu, Anca Maria Cimpean, Cl. Margaritescu

The thymus is the prototype of lymphoid and epithelial organ that consists of lymphoid and epithelial cells. In spite of remarkable progresses made in the field of the immunohistochemical characterisation of the thymus parenchyma, the diagnosis of thymoma largely depends on the interpretation of conventional morphologic aspects. Histogenesis of this organ is a multi-step process, and many stages reproduce lesions and changes found in the adult thymus. The normal structure and its variants are extremely helpful to differentiate normal from pathologic aspects. Particular aspects of the thymus structures were shown in myasthenia gravis, despite the behaviour of thymoma in these patients is not clearly understood. Authors performed a detailed description of the conventional pathology of the thymoma, based on the new classifications, recently adopted. The immunohistochemical profile could be helpful in the diagnosis of many cases, and also seems to be useful in prediction of invasion that is the most important criterion in prognosis.

Corresponding author: Marius Raica, MD, PhD, e-mail:

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3. Assessment of Gleason system use on different types of prostatic tissue samples

I. E. Plesea, B. Zaharia, S. D. Enache, G. Mitroi, P. Tomescu, O. T. Pop, P. Badea, A. Kozokic

One of the well-known and accepted methods of prostatic adenocarcinoma grading is Gleason system. The authors made a retrospective analysis of 221 prostatic adenocarcinomas divided into three groups (transvesical prostatectomies, transurethral resections and needle biopsies) following the type of surgical procedure used for drawing the tissue. Gleason scores and comparison between odd and even Gleason scores were assessed in the entire group and in each subgroup. High scores prevailed, meaning a tendency towards a low grade of differentiation. Even scores also prevailed meaning, on one hand, that, often, the examined specimen reveals only one pattern and, on the other hand, that surgical procedures as transurethral resections and unique needle biopsies cannot offer a sufficient material for examination, the multicentricity of prostatic carcinoma being well known.

Corresponding author: Iancu Emil Plesea, MD, PhD, e-mail:,

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4. HOPE - a novel tool for the pathologist

E. Vollmer, J. Galle, R. Sen Gupta, T. Goldmann

The growing demand for the detection of prognostic and diagnostic biomarkers by application of molecular-biological techniques has been slowed down to a large degree by the use of formalin and its influence onto antigenic structures and nucleic acids, although other reasons also exist. With regard to morphology a limitation in the possibilities, compared to frozen sections, takes place. With the introduction of the HOPE (Hepes glutamic acid buffer mediated Organic solvent Protection Effect) -fixation to date both have become possible; substantially enlarged possibilities concerning the detection of DNA, RNA and proteins in combination with excellent morphological results comparable to formalin-fixed tissues. We also highlight some aspects from our lung specific work during the last years. Immunohistochemical markers for differential diagnostics of malignancies in the lungs are discussed; their enhanced detectability using the HOPE-technique is pathbreaking for the future. The same holds true for the substantially enhanced detectability of germs (e.g. mycobacteria) in HOPE-fixed tissues.

Corresponding author: Ekkehard Vollmer, MD, PhD, e-mail:

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5. Hermaphroditism pathology

F. Jaubert, C. Nihoul-Fekete, S. Lortat-Jacob, N. Josso, M. Fellous

Hermaphroditism is a more general term for the discrepancy between the phenotype and the genotype of sex development than sexual ambiguity, which refers mainly to external genitalia anomalies. Hermaphroditism is studied on an historical and pathogenetical perspective. Short embryological summaries are integrated. The defects of sexual differentiation due to a hormonal deficiency are first studied: androgen insensitivity, steroid 5 alpha-reductase 2 deficiency, defects of testosterone synthesis, persistent mullerian ducts syndrome. Sexual determinism deficiencies come after: Turner syndrome, XX males, pure gonadal dysgenesis, and true hermaphroditism, mixed gonadal dysgenesis, Drash and Frasier syndrome. Tumors of dysgenetic gonads followed. Mixed tumors developed in dysgenetic gonads are gonadoblastoma and dysgerminoma. Sex cord tumors are androgen insensitivity associated tumors, Leydig cells tumors and adrenal cell inclusion tumors. New perspectives open by sex reversion genes are open.

Corresponding author: Francis Jaubert, MD, PhD, e-mail:

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6. The C-cells: current concepts on normal histology and hyperplasia

Angela Borda, Nicole Berger, M. Turcu, M. Al Jaradi, Sorana Veres

We describe the current concepts on the embryology, normal morphology and immunohistochemistry of a minor cell population of the thyroid, the C-cells. We also try to make delineation between the normal number of the C-cells and C-cell hyperplasia. The two types of C-cell hyperplasia, physiologic and neoplastic are defined and characterized from morphologic and genetic point of view. Their relation with thyroid pathology, especially with medullary thyroid carcinoma is discussed.

Corresponding author: Angela Borda, MD, PhD, e-mail:

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7. Cellular immune response in atypical tuberculosis diagnosed by PCR in paraffin embedded material

Carmen Ardeleanu, F. Andrei, M. Ceausu, D. Ene, M. Mihai, Georgeta Butur, Florica Staniceanu, Camelia Dobrea, P. Galbenu, F. Vasilescu

The aim was to evaluate the cellular immune response in atypical tuberculosis and granulomatous inflammation consistent with tuberculosis (TBC), negative histochemically for acid-fast bacilli and analysed by PCR for Mycobacterium tuberculosis (MT) detection in paraffin-embedded tissue. Thirty six samples of differently localized atypical tuberculous lesions and granulomatous tuberculoid lesions negative for acid fast bacilli and 4 positive cases on Ziehl-Nielsen stain were analysed by PCR for MT detection and were tested immunohistochemically (IHC) for the cellular immune response in the granulomas and perigranulomatous tissue. The samples selected were: 7 pulmonary and 33 extrapulmonary specimens, especially lymph nodes. Histologically, the atypical tuberculous lesions contained supurative necrosis, defective granulomas and cellular polymorphism. The epithelioid cells showed frequent mitoses. The immunoprofile of cells was polymorphous. L26 positive small lymphocytes were found in nodular lymphoid aggregates surrounding granulomas. A significantly increased number of positive UCHL1 cells were found in 33 out of the 40 analysed cases, with a larger percentage of CD 4 positive T cells (81.8% of cases). CD44 was positive in multinucleated giant cells (17.5% of cases), epithelioid cells (60% of cases) and lymphocytes (30% of cases). CD68 was localized in multinucleated giant cells and epithelioid cells, in a 4%, respectively 62.5% of cases. The PCR was performed in all 40 cases; the tissue samples were heterogeneous (lung, lymph nodes, lever, nasopharynx, etc.) and needed a good quality extraction of DNA. Performing a control PCR for Beta Globin tested the extraction; a good result was obtained in 31 cases (77.5%); from these, 19 cases had amplification for IS 6110. The cellular immune response in the atypical tuberculous lesions was similar in cases with and without acid-fast bacilli, but positive for PCR. In the most cases with negative PCR reaction, it was due to a deficient fixation of the material. The T-lymphocytes were numerous in all types of tuberculous granulomas, with the prominence of CD4-positive subtype. The immunoprofile of the epithelioid cells, positive for CD44 and CD68, presenting frequently mitoses suggests an activate state in a possible relationship to the T-cell-mediated immune response in tuberculosis.

Corresponding author: Carmen Ardeleanu, MD, PhD, e-mail:

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8. Variation of cephalometric parameters in peri-puberty cephalometric aspects of sexual dymorphism

A. Motoc, Marilena Motoc, S. Bolintineanu, Corina Musuroi, Andreea Popescu, M. Niculescu

The data regarding the global and segmental growth and development of the child are useful in pluri-disciplinary studies which view the human body as an entity where the biological aspects (both the normal and the pathological ones) interlace with the socio-cultural ones, thus offering the possibility to any specialist in the field of humanities and morphological studies to cover a wide range in contemporary anthropological research. The development of the skull takes place in two active stages separated by a stage of relative quiet during the age of seven and puberty when the skull has a dolicocephalic aspect. During the second stage of active growth (which starts with puberty) there is a transversal growth of the basis of the cranium followed by an anteroposterior growth. The degree of gender-related differences as far as the dimensions and proportions of the skull are concerned vary greatly according to race, therefore we may say that general ethnic differences are more pronounced than gender-related ones.

Corresponding author: Andrei Motoc, MD, PhD, e-mail:

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9. Morphologic evaluation of radiotherapy in cervical carcinoma - a computerized morphometric approach

Irina-Draga Caruntu, D. Visan, C. Cotutiu

Our work analyses the effects of radiotherapy in cervical carcinoma by computerized morphometry techniques that evaluate the morphologic changes in the specimens taken from pre and post-irradiation biopsies of eight cases. These cases were diagnosed as squamous cell carcinoma, undifferentiated (1 case), moderately (5 cases) and well differentiated (2 cases). For each case, 40 relevant microscopic fields were chosen, reflecting the morphologic aspects before (20 fields) and after irradiation (20 fields). The digitised images of these fields were used for measuring the tumoral and stromal areas by a program we implemented in the Zeiss KS400 environment. The program provides a mean tumoral/stromal area for each case, computed by averaging the results of the 20 measurements performed on the pre and post-irradiation images. In terms of a stereological interpretation, each mean tumoral area expresses the percentage of the tumoral volume and can be used as an index for quantifying the efficiency of the radiotherapy. Such an index allows developing comparative discussions on cases that belong either to the same diagnosis entity, or to different diagnosis entities. These discussions yield the idea that the morphologic classification of a case is not sufficient to accurately predict the efficiency of irradiation, and a systematic approach to the morphometric features, before and after irradiation, may contribute to the refinement of the prediction.

Corresponding author: Irina Draga Caruntu, MD, PhD, e-mail:,

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10. The histopathological and immunohistochemical diagnosis in small cell lung carcinoma

Ana-Maria Dragan, G. Simu, T. Vaida

The small cells lung carcinoma (SCLC) represents 10-25% of the bronchogenical carcinomas, prevails at middle ages and is closely associated with tobacco smoke. It belongs to the group of carcinomas with neuroendocrine differentiation, the patients presenting various paraneoplasic syndromes. The study was performed on 491 patients with a clinical and radiological suspicion of bronchopulmonary cancer. In this study, the histopathological examination gives results similar to those mentioned in the medical literature for the small cells lung carcinoma, prevailing in the case of young patients. The immunohistochemical examinations are particularly precious in confirming the diagnosis, especially in the diagnosis of the lymphonodular micrometastases.

Corresponding author: Ana-Maria Dragan, MD, PhD, e-mail:

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11. The immunohistochemical profile of luminal epithelial neoplastic component from pleomorphic adenomas of salivary glands

Cl. Margaritescu, Maria Florescu, M. Raica, Cristiana Simionescu, L. Mogoanta, E. Preda

The immunohistochemistry study made on 30 cases of salivary glands pleomorphic adenomas aims to establish the antigen profile of luminal epithelial neoplastic component proliferates into these tumors. We ascertain that luminal epithelial neoplastic cell were almost exclusively positive to antibody peculiar to epithelial differentiation (AE1/AE3, MNF 116, CK 8, CK 19, CK20, EMA) and occasional to S-100 protein and negative to vimentin, alpha-smooth muscle actin, calponin, Glial Fibrillary Acidic Protein (GFAP). This kind of immunoreactivity was similar to ductal cells from striated channels of salivary glands, suggesting the origin of such pleomorphic adenomas from this place.

Corresponding author: Claudiu Margaritescu, MD, PhD, e-mail:,

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12. Evaluation of T-lymphocyte subtypes in the diagnosis of celiac disease

Maria Sajin, M. Craiu, M. Iordachescu, Alina Stanescu, Ana Maria Sajin

The objectives of this study were tot verify comparatively clinical age reported antecedents and the morphological aspect of the intestinal mucosa in patients sensitive to gluten, hospitalised at IOMC ("Alfred Russescu" Hospital) in the last 7 years. Celiac disease is an inflammation most frequently affecting the proximal small intestine, depending on the presence of gluten in the diet, whose pathogenesis seems to be immunological in nature. 107 cases were divided in three groups following clinical manifestations types at hospitalization time: typical digestive, untypical digestive and extra digestive manifestations. Intestinal biopsies, made with Crosby probe, in children aged between 1.3 and 8 years (one single case was diagnosed as late as at the age of 15), regardless of gender. Then we analyzed morphologically (HE usual and PAS histochemical staining) and immunohistochemically (lymphocytes B, T with possible subtypes). The lesions were counted at the first biopsy according to the Marsh score. The immunohistochemical tests have indicated the prevalence of T-lymphocytes (UCHL1, CD3, CD4, CD8, gamma-delta) both in the luminal epithelium with various degrees of aggression in lamina propria and also spread in stroma. B-lymphocytes (L26) are distributed prevalently nodular in stroma. In conclusion, it is CD4 T-cells that are present in particular in the control of the gluten immune response in patients with Marsh I and Marsh III lesions.

Corresponding author: Maria Sajin, MD, PhD, e-mail:

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13. Histological study of the femural head and neck microscopic architecture in persons with senile osteoporosis

Nina Ionovici, L. Mogoanta, D. Grecu, Adriana Bold, D. N. Tarnita, S. D. Enache

For our study were available fragments of bony tissue from 24 patients (15 females and 9 males) aged between 55-82 which needed hip arthroplasty after they had undergone femural neck fractures. Biologic pieces have been fixed, then decalcified and processed by wax embedding. We noted that the spongious osseous tissue appeared as rarefied after the remoulding processes; osseous traveas grew unplainly thinner, with their large areolar cavities filled of bony yellow marrow. The osseous cortex presented deformed osteomas with large irregular Havers ducts. The osteocytes appeared rarefied, of small size with a picnotic and hypercrome nucleus.

Corresponding author: Laurentiu Mogoanta, MD, PhD, e-mail:,

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14. Value of immunohistochemistry in confirming undifferentiated ovarian carcinomas

Claudia Valentina Georgescu, Maria Florescu, Cristiana Simionescu

The present immunohistochemical study was carried out on ten cases of ovarian tumors diagnosed with usual staining as undifferentiated ovarian carcinomas. A panel of antibodies was chosen in order to confirm the epithelial origin of these tumors and to eliminate some possible ovarian metastasis. The cytokeratin AE1/AE3, EMA and BerEP4 positivity supported their epithelial origin even if thirty per cent of the cases co-expressed cytokeratin and vimentin. Even if there is not a specific marker to confirm the ovarian origin of the tumors, the CA125 and CEA positivity suggested that these carcinomas might represent a serous or a mucinous dedifferentiation. Associated with these antibodies, the calretinin, CK7 and CK20 stainings allowed the separation of these tumors from peritoneal mesotheliomas with ovarian extension and from ovarian metastasis originating in the gastrointestinal tract. The evaluation of the PCNA labelling index confirms the high degree of cell proliferation and the aggressiveness of these tumors.

Corresponding author: Claudia Valentina Georgescu, MD, PhD, e-mail:

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15. Histological and histochemical alteration in certain Black Sea bivalves submitted to accidental pollution

Rodica Mehedinti, Mihaela Hancu, Malina Coman, T. Mehedinti

Due to their microphage capacity, two-valve shells have great biologic importance: they clean the environment of harmful microorganisms and decaying organic compounds which result in water purification, water filtering by means of adaptive structures of digestive system and, indirectly, by those of respiratory, circulatory and excretive system is of major significance. Waste materials derived from animal metabolism coalesce in larger aggregates which sink and deposit onto the bottom, thus enabling the bivalvular shells to contribute to developing and securing adequate conditions for the productive cycle of marine environment and even twining themselves into food for animals. Our study makes a histological, histochemical and histometric survey of branchia (ctenidia) of Mytilus galloprovinciallis so as to enlarge the scant body of knowledge on the topic in reference literature.

Corresponding author: Mihaela Hancu, MD, PhD, e-mail:

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16. Overexpression of cytokeratin 34beta E12 in thymoma: could it be a poor prognosis factor?

Anca Maria Cimpean, M. Raica, Svetlana Encica

We included in our study nine normal human thymuses and sixteen thymomas (1 type A, 2 type AB, 3 type B1, 3 type B2, 5 type B3 and 2 type C). The 25 patients were between 7 days and 75 years old, and were submitted to open surgery for correction of congenital heart defects or for mediastinal tumor mass. Biopsies were formalin fixed for 24 hours and then embedded in paraffin using routine procedure. Five micrometers step sections were mounted on silanized slides. Slides from each case were stained with hematoxylin and eosin method for morphological diagnosis. Additional sections were immunostained using monoclonal antibodies against high molecular weight cytokeratin clone 34beta E12. We found a very strong positive reaction for this type of cytokeratin in thymomas type B2, B3 and inconstant in type C comparative with normal thymus. Also, the number and distribution of positive epithelial cells in normal thymus versus thymomas were different. We found positive cells into capsular vessels of thymomas. This could be an invasion marker for apparent encapsulated thymomas. Strong positive reaction in almost all epithelial cells of thymomas types mentioned above was correlated in part with invasion. We concluded that expression of 34beta E12 cytokeratin is a useful marker for thymomas with high grade of malignacy, correlated with vascular and capsular invasion.

Corresponding author: Marius Raica, MD, PhD, e-mail:

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17. Histopathologic and immunohistochemical study in one case of cystadenoma of parotid gland becoming malignant

Cristiana Simionescu, Maria Florescu, Cl. Margaritescu, Magdalena Marinescu

Cystadenoma and cystadenocarcinoma of the salivary glands are rare tumors, approximately half of cases being diagnosed at the level of parotid. The studied tumor associated both benign and malign zones, with deep infiltrating character in the adjacent structures. In both areas the growing pattern was predominantly cystic and papillary, and the proliferated neoplastic cells were majority cubical or columnar, with low grade of nuclear pleomorphism in the becoming malign areas, discordant with the infiltrative character of the neoplasm. Immunohistochemically, we investigated the tumor with AE1-AE3, CEA, SMA, S-100 and PCNA.

Corresponding author: Claudiu Margaritescu, MD, PhD, e-mail:,

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18. Histologic and immunohystochemical study of the vascular and perivascular structures in patients with chronic obstructive bronchopneumopathy (COBP)

L. Mogoanta, Claudia Valentina Georgescu, Carmen Florina Popescu, Carmen Aurelia Mogoanta, S. Frasie

Histologic and immunohistochemical changes of the lung vascular and perivascular structures in 37 clinical and paraclinical COBP diagnosed were studied. We noted the deterioration of an important part of the pulmonary capillary network because of the destruction of interalveolar septae, with an intense vascular congestion within restant capillaries. The venules and arterioles have had a fibrosis wall by the replacing of the elastic and muscular fibers with inextensible fibers of collagen. The perivascular stroma was infiltrated by fibroblasts, macrophages, lymphocytes and plasma cells.

Corresponding author: Laurentiu Mogoanta, MD, PhD, e-mail:,

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19. Aspects of the cell population in the skin of the guinea pig exposed to hyperbaric-hyperoxic environment

Mihaela Hancu, Rodica Mehedinti, Malina Coman, T. Mehedinti

Knowing that in hyperbaric-hyperoxic environment, the production of free oxygen radicals is increased, with the direct implications on cellular metabolism, our study follows the influence of the hyperbaric environment, a very complex organ that is exposed to directly to this environment. We used a group of 10 guinea pigs split into to lots (test and control lots). The test lot was exposed for consecutive days to an ambient pressure of 6 atm. for a period of 30 min. After 50 days both groups was given lethal doses of pentobarbital, afterwards skin was harvested and fixed into Bouin's fluid and formalin, then paraffin embedded. The slides stained with Hematoxylin-Eosin and trichromic Van Gieson. The epidermal fields examined by us revealed no significant variations of nuclear number, though the mean saturation of nuclei was significantly greater in test animals than in the control group, while the nuclear area fraction was higher in the control group. The melanin load of the epidermal cells was increased in the control animals. The presence of intraepidermal vacuoles was identified, their amount being almost double in test animals than in the control group.

Corresponding author: Mihaela Hancu, MD, PhD, e-mail:

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20. Histopathologic aspects in microglandular hyperplasia of endocervix

Maria Florescu, Cristiana Simionescu, Claudia Valentina Georgescu, Magdalena Marinescu

In this study, we analyzed 373 cases of microglandular hyperplasia of endocervix belonging to patients from the III-VII decades of life. Most of the lesions had the typical aspect of proliferation of small glandular lumen lined by endocervix type epithelium, with or without areas of basal proliferation or immature squamous metaplasia. In eight cases, the growing patterns were peculiar, with mucinous or florid aspects, three of the cases rising differential diagnosis problems with cervical carcinoma.

Corresponding author: Cristiana Simionescu, MD, PhD, e-mail:

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21. Comparative study between prostatic carcinoma G1-G3 and Gleason grading systems

B. Zaharia, I. E. Plesea, S. D. Enache, P. Tomescu, A. Panus, O. T. Pop, P. Badea, A. Kozokic

The need of an accurate and prognostic valuable method for grading the prostatic carcinoma resulted in the last decades, a large number of systems none of the succeeding in being unanimously accepted by the pathologist's family. The authors selected and reassessed using Gleason system a group of 221 cases with prostatic adenocarcinoma previously diagnosed using a combined G1-G3 system. The results were converted from Gleason system to G1-G3 system, and then compared with the initial diagnostics. The group was divided, following the type of surgical procedure used for drawing the tissue, into three groups: transvesical prostatectomies (TVP), transurethral resections (TUR) and needle biopsies (B). The concordance between the two systems was good, even the number of TUR and B specimens was significant (85% of the studied group) showing that any of them can be successfully used in current practice, depending only on pathologist's experience.

Corresponding author: Iancu Emil Plesea, MD, PhD, e-mail:,

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