ROMANIAN JOURNAL of MORPHOLOGY and EMBRYOLOGY

Vol. 48 No. 2, April-June 2007

1. The correlation between the immunostains for p53 and Ki67 with bcl-2 expression and classical prognostic factors in colorectal carcinomas

Simona Gurzu, J. Jung, T. Mezei, Z. Pavai

The prognostic role of p53 and Ki67 in colorectal carcinomas (CRC) is very controversial in the literature. In our study, we tried to find if their immunostains are correlated with bcl-2 expression or other classical prognostic factors (sex, age, localization and size of tumor, the grade and staging of tumor). We studied 507 cases with CRC and chose 38 cases in which we realized these correlations. Fourteen cases were mucinous CRC, the other 24 cases being non-mucinous CRC (six well differentiated, 13 moderate and five poorly differentiated). For statistical analysis, we used the Statistical Program Graph Pad In Stat 3-Trial Version. We considered the significant association when p<0.05, with 95% confidence interval. Results. The median value was 75% for p53 expression, respectively 35% for Ki67 expression. Bcl-2 was positive in 47% of cases but not correlated with p53 or Ki67. We found a significantly statistical decrease p53 immunostain with grade of tumor (70% in well differentiated, respectively 40% in poorly differentiated CRC) and increase of Ki67 median expression (25% in well differentiated, respectively 60% in poorly differentiated CRC). Ki67 was correlated with age of patients, lymph node involvement, being more expressed in N2 (80%) than in N0 (22.5%) and with Dukes MAC staging (25% in B1, 60% in C2). P53 was correlated with age of patients and pT component, after pTNM staging (75% in pT2, 40% in pT4). P53 was not correlated with Ki67. Conclusion. The CCR prognostic is not determined only by proliferative capacity of tumoral cells.

Corresponding author: Gurzu Simona, MD, PhD, e-mail: simonagurzu@yahoo.com

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2. Involution of the thymus: a possible diagnostic pitfall

M. Raica, Anca Maria Cîmpean, Svetlana Encica, R. Cornea

There were investigated 22 cases from which the thymic tissue was removed either during surgery for cardiovascular malformations (n = 14), or for myasthenia gravis (n = 8). Histological sections were stained with routine morphologic methods, and immunohistochemistry was performed for cytokeratin, CD20, CD3, and S100 protein. Aspects characteristic for thymus involution were found in 11 cases without myasthenia gravis and in all cases with myasthenia gravis. Morphological changes of the thymus of involution are age-dependent. There were characterized stages of involution, with special reference to cortical-medulla inversion, lymphocyte depletion and sequestration. In advanced-stage of involution, epithelial cells are arranged in cords or compact islands, and could mimic a thymoma or a metastatic carcinoma. The immunohistochemical profile is similar but not identical to the active thymus. We noticed a decreased expression of cytokeratin, and a reduced number of CD3, CD20, and S100 positive cells. Morphologic and immunohistochemical peculiarities of the thymus of involution are discussed in relation with the specific pathology of the organ.

Corresponding author: Marius Raica, MD, PhD, e-mail: raica@umft.ro

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3. Quantification of apoptotic phenomenon on endometrial biopsies in postmenopausal patients under hormonal replacement therapy (HRT)

Dana Terzea, Cristina Iosif, Florina Vasilescu, F. Andrei, Camelia Dobrea, Alina Nicolae, Alina Georgescu, F. Mircea, M. Ceauşu, Ariana Vişan, Mihaela Mihai, Carmen Ardeleanu

Aim. To quantify the apoptotic phenomenon on endometrial biopsies in postmenopausal patients under hormonal replacement therapy (HRT). Material and methods. The study lot consisted of 30 endometrial biopsies on which we studied the apoptotic phenomenon through morphological and molecular biology techniques (TUNEL reaction). Examination of endometrial biopsies before and post-therapeutically has been made. Results and discussions. From morphological point of view, pre-therapeutically, endometrial biopsies presented apoptotic changes in about 1-3% of cells and under TSH, there have been observed apoptotic changes in about 1-2% of cells. In female reproductive system, we found out a raised rate of cellular proliferation and concurrently a raised rate of apoptosis. Apoptotic phenomenon can be observed in endometrium at every menstrual cycle. In proliferative endometrium apoptosis rate is low, but in endometrial carcinoma apoptosis rate grow up. Bcl2 and Bax are expressing in normal and hyperplastic endometrium, but in endometrial carcinoma Bcl2/Bax ratio decline. Conclusions. Quantification of apoptosis, using morphological and TUNEL reaction methods, on endometrial biopsies in postmenopausal patients before and after therapy indicate a low rate of apoptotic phenomenon.

Corresponding author: Dana Terzea, MD, e-mail: danaterzea@gmail.com

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4. The value of the immunohistochemical exam in the diagnosis of the secondary malignant tumors to the thyroid gland

Delia Ciobanu, Carmen Vulpoi, B. Găluşcă, Niculina Florea, Simona-Eliza Giuşcă, Irina-Draga Căruntu

The aim of the study was to analyze several cases of secondary tumors to the thyroid, by focusing on the role of the immunohistochemical (IHC) exam in specifying the origin of the tumoral process. The studied group included 16 patients, investigated by fine-needle aspiration biopsy, frozen sections at the surgical moment, routine histopathological exam and immunohistochemical staining, using different antibodies, in accordance with the histological aspects. The final diagnosis was established as follows: metastases of squamocellular carcinoma with different degree of differentiation (seven cases), metastases of adenocarcinoma (four cases), metastases of renal cell carcinoma (two cases), metastases of Hodgkin (one case) and non-Hodgkin lymphoma (two cases). In four cases, the primary tumors were identified after the diagnosis of their metastases in thyroid. The immunohistochemical staining was useful in the diagnosis of squamocellular carcinoma metastases, poorly differentiated (CK19 positive), of renal cell carcinoma with clear cells (CK18, CK19 and CD10 positive) and in the establishing of the tumoral origin for adenocarcinomas (CK7 positive - respiratory tract, CK20 positive - digestive tract). Secondary tumors to the thyroid are rare tumors, with miscellaneous histological aspects, reason for which the diagnostic may be difficult. In these cases, IHC is a useful method, allowing to the identification of the primary tumor.

Corresponding author: Delia Ciobanu, MD, PhD, e-mail: deliaku@yahoo.com

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5. Angiogenesis and c-erbB-2 (HER2/neu) overexpression status in primary breast cancer patients: an analysis of 158 needle core biopsies

S. Vameşu

Angiogenesis, formation of new blood vessels from a preexisting vascular bed is a complex multistep process, which may also permit metastasis. The subset of patients with breast cancer demonstrating a c-erbB-2 (HER2/neu)-positive status has aggressive tumors and a poor prognosis. To investigate how tumor angiogenesis correlates with c-erbB-2 (HER2/neu) overexpression in breast carcinoma diagnosed on core biopsy, microvessels were counted (and graded the density of microvessels) within the initial invasive carcinomas of 158 patients. Using light microscopy, the number of microvessels was counted manually in a subjectively selected hot spot (in the most active areas of neovascularization per 400x field), and their values were separated as above or below median (low and high), without knowledge of the outcome in the patient or any other pertinent variable. When the mean values of MVD of the various groups defined by c-erbB-2 (HER2/neu) overexpression were compared, significant difference was noted (P = 0.014818). When tumors were classified as high or low MVD, based on a cut-off value (30.70175 microvessels/square-mm), cases with high MVD were significantly more numerous. MVD did show a relationship with groups defined by c-erbB-2 (HER2/neu) status (P = 0.002699) or c-erbB-2 (HER2/neu) score (P = 0.027589). The correlation of angiogenesis with c-erbB-2 (HER2/neu) overexpression may be a potential therapeutic target for the treatment and prevention of breast cancer, using antiangiogenic molecules.

Corresponding author: Sorin Vameşu, MD, PhD, e-mail: sorinvamesu@yahoo.com

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6. Molecular changes in superficial bladder cancer

Camelia Doina Vrabie, Angela Petrescu, Maria Waller

Urinary bladder cancers represent a spectrum of diseases that can be grouped into three general categories: superficial, invasive and metastasis. Each differs in clinical behavior, prognosis and primary management. For superficial tumors, the aim is to prevent recurrences and progression to an incurable stage, recognizing that surgical removal of the bladder (over treatment for most tumors) is curative up to a point. For more invasive disease, the issue becomes how to determine which tumors can be cured with a single therapy such as surgery, and which, by virtue of a high metastatic potential, requires an integrated systemic approach to achieve cure. For metastatic disease, combination chemotherapy is the standard yet, despite responses in more than 50% of cases, overall cure rates remain low, and progression has been minimal over the past few years. We analyzed histopathological and immunohistochemical 70 patients with bladder carcinomas searching the stage, the grade and other associated lesions. The results showed that 70% were papillary transitional carcinomas infiltrated in lamina propria (T1), and almost 22.85% represent non-invasive papillary carcinomas (Ta); we found only five cases in Tis stage (7.15%). The immunohistochemistry investigated three antibodies: p53 oncoprotein, bcl-2 oncoprotein and retinoblastoma protein (pRb). We noticed the antibodies distribution related to stage: carcinoma in situ (Cis or Tis) high percent of p53 (69) and bcl-2 (37.5%). Concerning the superficial tumors we found low values of p53 in T1 (45%) versus invasive tumors (51%); oncoprotein bcl-2 is higher in T1 (35%) versus non-invasive one (6%).

Corresponding author: Camelia Doina Vrabie, MD, e-mail: angelap@vbabes.ro

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7. Lymphatic vessels identified with podoplanin. Comparison of immunostaining with three different detection systems

Anca Maria Cîmpean, M. Raica, D. A. Izvernariu, Diana Tătucu

The aim of the study was to find the optimal immunostaining protocol for monoclonal mouse anti human podoplanin antibody clone 18H5, which is less studied. We tested three detection systems for different antibody dilutions and antigen retrieval methods applied on normal and tumor breast tissues, lip squamous cell carcinoma, kidney tumors and testis carcinoma. The interpretation was linked to the background staining, specificity for the lymphatic endothelium and cross-reactivity. The ABC method gave a high background even the antibody dilution was increased. No immunostaining was obtained with Envision detection system for any dilution or type of antigen retrieval. We obtained the best results using LSAB+ working system for an antibody dilution of 1:500 with no background and podoplanin expression was restricted to the lymphatic endothelium. We consider that the standardization of immunostaining protocol is the first step for an optimal interpretation of the podoplanin expression using less studied antibody clones.

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

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8. Clinical, histological and immunohistochemical characteristics in patients with tumors of urinary bladder and renal failure

R. Buzulică, E. Traşcă, Iuliana Nicolescu, Polixenia Stancu, E. T. Traşcă

Chronic renal failure in patients with tumors of urinary bladder appears because of either familial tubulo-interstitial pielonephritis evolution (Balkan Endemic Nephropathy) or invasion and obstructive uropathy. A group of 27 clinical cases with tumors of urinary bladder and renal failure was available for our study between the years 2004-2005. Simultaneity of the two serious diseases, neoplasia and terminal renal failure made the prognostic more difficult and determined some other complications to appear. Identification of some clinical and histopathological features in due time led to a therapeutic algorithm favorable to a prolonged survival.

Corresponding author: Radu Buzulică, MD, e-mail: lucianbuzulica@yahoo.com

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9. Comparative microscopic study of the ovarian blood vessels

Emma Gheorghe, V. Tomuţa, T. Mehedinţi, Mihaela Hîncu, V. Broască

The present study puts forth the morphometric analysis of modifications that occur in the ovarian structures (stroma, parenchyma, blood vessels) before and after menopause onset. Our study was performed on 30 human ovary specimens from patients with no history of genital pathology that were split in two groups (15 ovaries harvested from fertile women and 15 ovaries harvested from women at menopause). The specimens were processed following the classic histological technique of paraffin embedding and stained using Hematoxylin-Eosin technique and trichromic Goldner-Szeckelly technique. The samples were examined and assessed using Nikon E-600 light microscope. The images captured by computer were processed using LUCIA G 4.10 software. To evaluate the histological modifications, we chose the following parameters: Area Fraction of blood vessels and Circularity of blood vessels. The study shows that modifications of the ovary components are synchronized. The vascular changes that occur after menopause onset lead to a poor vascular supply of all ovarian components: germinal epithelium, ovarian follicles and ovarian stroma. The comparative morphometric analysis on premenopause and postmenopause ovarian specimens sustains the idea that the ovarian involution starts from the ovarian vessels, which are the first structures affected by hyaline degeneration.

Corresponding author: Emma Gheorghe, MD, PhD, e-mail: gheorghe_emma@yahoo.com

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10. Multiple anomalies involving testicular and suprarenal arteries: embryological basis and clinical significance

S. R. Nayak, Jiji P. J, Sujatha D'Costa, Latha V. Prabhu, A. Krishnamurthy, Mangala M. Pai, Prakash

Variations in the origin of arteries in the abdomen are very common but with the invention of new operative techniques within the abdominal cavity, the anatomy of abdominal vessels has assumed much more clinical importance. During routine dissection of the abdominal cavity, we came across multiple arterial anomalies involving testicular and suprarenal arteries. On the right side, there was double testicular artery (medial and lateral) and the right inferior suprarenal artery aroused from the medial testicular artery. The right inferior phrenic artery (IPA) and middle suprarenal artery took origin from a common trunk just above the origin of right renal artery (RRA). On the left side, the left testicular artery was arching over the lower tributary of the left renal vein proper (LRVP). Apart from the developmental and morphological interest in arching gonadal arteries, they are of practical importance from a clinical and surgical viewpoint. The embryological and clinical significance of above variations has been described.

Corresponding author: Soubhagya Ranjan Nayak, MSc, e-mail: ranjanbhatana@gmail.com

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11. Ilio-femoral aneurysm masquerading as an inguinal abscess

Latha V. Prabhu, Hema Kini, S. R. Nayak, Mangala M. Pai, Narayana K

True isolated atherosclerotic aneurysms of the superficial femoral artery are rare. In this paper, we report a case of ilio-femoral aneurysmatic dilatation masquerading as an inguinal abscess in 40-year-old male. This interesting case was accidentally found during dissection of pelvic vessels, in the Department of Anatomy, Kasturba Medical College, Mangalore. After going through the history of the patient. We found that he had the history of swelling in both groins being drained 20 years back, and also the multiple heterosexual exposures. The swelling was pulsatile, tender, and soft to firm in consistency and measured 7x3 cm. All physical signs were suggestive of an abscess and he was treated conservatively by ampicillin, gentamycin, metronidazole, and anti-inflammatory drugs. Fine needle aspiration yielded only frank blood. Subsequently, general condition of the patient deteriorated and died due to hypovolemic shock. Autopsy revealed a ruptured fusiform aneurysm measuring 10x5 cm at the junction of right external iliac with the femoral arteries measuring 3 cm in diameter. This case is an example of difficulties in diagnosing the rare arterial aneurysm, thus leading to misdiagnosis as an inguinal abscess. When only the blood is drawn during fine needle aspiration, one should suspect the swelling as an aneurysmatic dilatation.

Corresponding author: Soubhagya Ranjan Nayak, MSc, e-mail: ranjanbhatana@gmail.com

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12. Vaginal superficial myofibroblastoma. Case report and review of the literature

C. D. Olinici, Doiniţa Crişan, Adriana Zolog, Maria Puşcaş

Superficial myofibroblastoma of the lower female genital tract is a rare, recently described tumor, which has a distinctive clinico-pathological profile. We report a 63-year-old patient who presented with a polypoid vaginal mass. Histopathological examination revealed a moderately cellular tumor composed of bland spindled and stellate cells. Immunohistochemical investigations showed reactivity for vimentin, desmin and CD34. This tumor should be differentiated from other mesenchymal lesions, which may arise in this area.

Corresponding author: Corneliu Dorin Olinici, MD, PhD, e-mail: anapatol@yahoo.com

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13. Portal cavernomatous transformation leading to variceal hemorrhage in Sturge-Webber syndrome. A rare, but possible association

E. F. Georgescu, Ligia Stănescu, Daniela Dumitrescu, Reanina Ionescu, Iuliana Georgescu

Sturge-Weber syndrome is a rare disorder consisting of a port-wine nevus in the distribution of the ophthalmic branch of the trigeminal nerve and central nervous system malformations. Facial cutaneous vascular malformation, seizures, and glaucoma are among the most common symptoms and signs. The syndrome results from malformation of the cerebral vasculature located within the pia mater, most commonly over the occipital region. These malformations led to venous hypertension and subsequent hypoperfusion on the underlying cortex, causing chronic cerebral ischemia, atrophy, calcification and neurological deterioration. We describe 18-years-old young girl hospitalized for upper digestive hemorrhage that revealed a cavernomatous transformation of portal vein. At the same time, she presents extensive congenital, bilateral port wine stains on the face, epilepsy and glaucoma of the right eye. Computer tomography showed intracranial vascular abnormalities with calcifications, particularly in the right occipital lobe. The clinical presentation and imagistic assessment confirmed the diagnosis of Sturge-Weber syndrome associated with upper non-cirrhotic portal hypertension generated by a malformation of portal vein. Conclusions. Upper digestive hemorrhage is a quite rare eventuality in the Sturge-Webber syndrome. Moreover, portal tract malformations with cavernomatous transformation are exceptionally cited in the literature. Despite this rare association, abdominal investigation, as well as computed cranial tomography should be performed in all cases of children that present a facial cutaneous vascular malformation.

Corresponding author: Eugen Florin Georgescu, MD, PhD, e-mail: efg@usa.net

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14. Cutaneous metastases carcinoma. Case report and pathological considerations

Mariana Costache, Olga Simionescu, Maria Sajin, Alina Chefani

Diagnostic of cutaneous metastases is relatively rare; this is important because sometimes may be the first manifestation of an undiscovered internal malignancy. Usually dissemination may take place through the lymphatics or the blood stream. The case of a patient, female, which has on the inferior 1/3 part of the abdominal wall skin a multinodular tumor mass, which evolves from about three months ago and accompanying by lymphedema of the legs, is presented.

Corresponding author: Mariana Costache, MD, e-mail: m_costache_dermatopat@yahoo.com, angelap@vbabes.ro

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15. Kaposi's sarcoma associated with AIDS

Ligia Stănescu, Camelia Foarfă, Ana Claudia Georgescu, Iuliana Georgescu

In 1872, Moritz Kaposi, first described "Idiopathisches multiples Pigmentsarkom der Haut", which has become known as Kaposi sarcoma (KS). In the present KS is considerate an opportunistic neoplasm rather than a genuine cancer. It is a disease with clinical aspects extremely different, associate with some immunological deficits. The discovering in 1994 of a new type of human herpes virus called human herpes virus type 8 (HHV8) in the KS lesions sustains also a viral etiology. Four forms of Kaposi's sarcoma are recognized: classical, endemic (associated with AIDS), epidemic and iatrogenic (usually after transplant). All these forms have the same histopatologic aspects and are associated with HHV. However, these differ in prognosis and treatment. The authors present a KS case associated with AIDS occurring at a patient in the childhood. The particularities of the case are the presence of only two cutaneous lesions, from which one giant tumor, and the other nodular in aspect and the appearance of an infection HIV in the childhood with involvement of others risk factors except homosexuality. It is important, on one side the importance of the histopathologic exam of an angiomatous tumor for the establishing the diagnosis of KS even when is solitaire and appear in the child, and the other side the absolute necessity to search an eventual concomitant infection with HIV in the presence of a KS.

Corresponding author: Ligia Georgeta Stănescu, MD, PhD, e-mail: ligstanescu@yahoo.com

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16. An intrajugular paraganglioma. Unusual presentation of a classical tumor

D. Arsene, Carmen Ardeleanu, L. Dănăilă

Paragangliomas arise from the extraadrenal neuroendocrine system. They are locally aggressive tumors, causing adjacent invasion, bone destruction and compression related symptoms. We present a 35-years-old woman with a peculiar paraganglioma lacking all these features, and strictly located within the jugular vein. Differential diagnosis is detailed since other entities could have dissimilar clinical behavior. To the best of our knowledge, this is a very unusual site of occurrence for paragangliomas, and only two other comparable cases have been described.

Corresponding author: Dorel Arsene, MD, PhD, e-mail: dorelarsene@yahoo.com, dorelarsene890@hotmail.com

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17. Endometriosis of the abdominal wall

Maria Comănescu, Anca Potecă, Anca Lăzăroiu, Maria Sajin

Endometriosis is characterized by the presence and proliferation of ectopic endometrial tissue. It occurs most often in the pelvic area but sometimes it presents as a lump in the abdomen. We report three cases of endometriosis of the abdominal wall, presented at the Emergency University Hospital of Bucharest during the last year. The diagnosis was made by the histopathological analysis of the surgical specimen.

Corresponding author: Maria Comănescu, MD, e-mail: mariacomanescu@yahoo.com

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