Vol. 54 No. 1, January-March 2013

1. Cutaneous MALT-lymphoma: from cutaneous immunocytoma and pseudolymphoma to the current (and future) conceptions

A. Fernandez-Flores

The current report examines the evolution of the concepts of immunocytoma and pseudolymphoma in a historical perspective, paying special attention to their evolvement into the groups of marginal-zone lymphoma and cutaneous MALT-lymphoma. It also examines the current conception of the existence of at least two types of cutaneous MALT-lymphomas and their relation to the duality immunocytoma/pseudolymphoma from the old literature.

Corresponding author: Angel Fernandez-Flores, MD, PhD; e-mail:

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2. The anatomical relations of the sphenoid sinus and their implications in sphenoid endoscopic surgery

V. Budu, Carmen Aurelia Mogoanta, B. Fanuta, I. Bulescu

Alongside the past development of endoscopic sinus surgery, knowledge about the anatomy of the sinuses has become crucial for surgeons. The sphenoid sinus is one of the most variable of all sinuses. Its relations to vital vascular and nervous elements make its approach a challenge for endoscopic surgeons. These relations include the internal carotid artery, the optic nerve, the pituitary gland, the Vidian nerve, and other vascular and nervous elements depending on the size of the sinus. This paper will make a review of these anatomical relations and their importance in endoscopic sphenoid surgery.

Corresponding author: Vlad Budu, MD, PhD; e-mail:

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3. The value of PAX8 and WT1 molecules in ovarian cancer diagnosis

Ludmila Liliac, Maria Luisa Carcangiu, Silvana Canevari, Irina-Draga Caruntu, Delia Gabriela Ciobanu Apostol, M. Danciu, M. Onofriescu, Cornelia Amalinei

PAX8 and WT1 are transcription factors, each of them with distinct roles in organogenesis, morphogenesis, cell growth, and differentiation. Recently, their expression was also confirmed in a variety of malignancies, being included in the antibodies panel recommended for the female genital tract pathology. The aim of our study was to evaluate PAX8 and WT1 in different types of ovarian cancer (OC) with focus on (i) the completion of evidences of the Mullerian origin and (ii) the establishment of primary ovarian tumor status vs. metastasis. The study group consisted of 86 cases, with histopathological diagnosis covering the main subtypes of OC (low- and high-grade serous, low- and high-grade endometrioid, clear cell, mucinous, malignant Brenner tumor, malignant mixed Mullerian tumor, undifferentiated, and borderline). The investigation was based on immunohistochemical examination, performed by using specific antibodies applied on blocks obtained through Tissue MicroArray technique, and interpreted by scores assessing the nuclear positivity of tumoral cells. One case was not valuable due to technical difficulties. PAX8 expression was positive in 70 (81.39%) cases, the remaining 15 (17.44%) negative cases suggesting a non-Mullerian origin. WT1 expression was positive in 61 (71%) cases, mainly expressed in serous carcinoma, regardless of their differentiation degree, and negative in 24 (27%) cases. Our study provide supplementary evidences to support the association of PAX8 and WT1 immunostaining in the investigation of the complex biology of OC, PAX8 confirming the ovarian primary and WT1 allowing the refinement of the diagnosis in phenotype overlapping cases.

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

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4. In vitro expansion and characterization of corneal stem cells isolated from an eye with malignant melanoma

O. Samoila, Olga Soritau, M. Calugaru, Lacramioara Totu, S. Susman, Cristina Cristian, Carmen Mihaela Mihu

Purpose: The objective of this study was the identification, characterization and in vitro replication of the human corneal stem cells, taking into consideration the difficulties in obtaining sufficient corneal material from living donors. The study explored a variety of stem cell markers, usually found in embryonic or adult mesenchymal stem cells. Culture medium and replication substrates had to be identified, with no data available on this subject in our country (there are no other reports on corneal stem cells in Romania, to our knowledge). Materials and Methods: Corneal epithelial limbus was harvested from an enucleated eye, containing also a choroid malignant melanoma. Stem cells from the limbus were isolated and cultivated in vitro. Expression of specific stem cell markers was evaluated with immunocytochemistry. Results: Corneal stem cell expansion in primary culture was slow, achieving 70-80% confluence after 28 days. Stem cells were easily isolated in standard medium, showed fibroblastoid morphology and were positive for certain stem cell specific markers in immunocytochemical staining: Oct3/4, SOX2, Nanog, SSEA4, CD44, CD90, CD133, and CD34. They also expressed pan-cytokeratin. Donor age (72 years) and the presence of a malignant tumor close to limbal stem niche could have had an impact on the proliferation rate and the characteristics of the corneal stem cells. Conclusions: Isolated limbal cells were adult type stem cells with an epithelial orientation. The characterization of these cells with immunocytochemistry allowed us to observe surface markers that other stem cells also express.

Corresponding author: Ovidiu Samoila, MD; e-mail:

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5. Monitoring M-BCR-ABL expression level in CML patients by RQ-PCR: experience of a single Center

Annamaria Szanto, Zsuzsanna Pap, I. Benedek, Erzsebet Benedek-Lazar, Judit Beata Kopeczi, Aliz-Beata Tunyogi, Kinga Vasile, Emoke Horvath, Z. Pavai

Chronic myelogenous leukemia (CML) is characterized by the Philadelphia chromosome and the BCR-ABL fusion gene that encodes an abnormal tyrosine kinase. Development of specific tyrosine kinase inhibitors completely changed the management of these patients. Materials and Methods: Between April 2008 and July 2012, at the Molecular Biology Laboratory, University of Medicine and Pharmacy of Targu Mures, Romania, we monitored the M-BCR-ABL transcript level by real time quantitative PCR in case of 15 CML patients diagnosed at the Hematology and Transplant Center of Targu Mures. Results: Modification of M-BCR-ABL expression level shows statistically significant correlation (p=0.013) with the clinical course of these patients. Conclusions: Molecular biology techniques have an important role in monitoring CML patients and regular analysis is recommended.

Corresponding author: Emoke Horvath, Lecturer, MD, PhD; e-mail:

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6. Foxp3 and IL17 expression in tumor infiltrating lymphocytes (TIL) and tumor cells - correlated or independent factors?

Florina Vasilescu, D. Arsene, Florina Cionca, Maria Comanescu, V. Enache, Cristina Iosif, D. O. Alexandru, D. Georgescu, Camelia Dobrea, Adina Balan, Carmen Ardeleanu

Tumor infiltrating lymphocytes (TIL), as a microenvironment component were studied in various epithelial tumors, with contradictory results. Recent data about regulatory T-cells (Treg) revealed new explanations for pro- and anti-tumor implications of TIL. Tregs immunoprofile was recently completed with Foxp3 expression. A T-cell fraction (Th) is producing cytokine IL17 and is now considered acting in tumor progression. Our study aimed to analyze immunohistochemically (IHC) Foxp3+ and IL17 expression in resected lung adenocarcinomas, since they could become possible targets in the antitumor immunotherapy. The studied material was represented by paraffin-embedded tumor fragments from 59 patients with TIL identified on HE staining. The antibodies used were Foxp3 and IL17. The statistical analysis used logistical regression on SPSS19 software (Chicago, IL, USA). TIL was usually mild or scarce. A positive statistic correlation resulted between the amounts of TIL in peritumoral and intratumoral location but without correlation to histopathological grading. Foxp3 and IL17 were present in TIL lymphocytes, tumor cells and fibroblasts; IL17 was expressed also in periendothelial cells (PEC). Foxp3 positivity was significantly correlated for lymphocytes/tumor cells, lymphocytes/fibroblasts and tumor cells/fibroblasts, suggesting their concerted action. Tumor cells and lymphocytes Foxp3 expression was inversely correlated with the amount of TIL. Between lymphocytic Foxp3 and PEC IL17, we found a weak negative correlation. The TIL had a quite positive correlation with PEC IL17. In these conditions, Foxp3 could be a mediator of the tumor cells inhibitory aggression upon the immune system and could be used as a molecular target for biological antitumor therapy.

Corresponding author: Carmen Ardeleanu, Senior Researcher, MD, PhD; e-mail:

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7. Histopathologic features of Spitzoid lesions in different age groups

Adriana Diaconeasa, D. Boda, C. Solovan, D. M. Enescu, Alina Maria Vilcea, Sabina Zurac

Spitz nevus is one of the most difficult melanocytic lesions to diagnose in regard of malignancy, even for experienced dermatopathologists. We analyzed 28 tumors with Spitzoid morphology from 15 children (three little children 2-4-year-old, 12 peripubertary children 9-17-year-old) and 13 adults; there were 21 Spitz nevi, five atypical Spitz tumors and two Spitzoid melanomas in order to establish the diagnostic value of several morphologic parameters in different age groups. No significant differences in respect of age and/or tumor type occurred for gender, location, dimension, symmetry, sharp lateral demarcation, junctional nests orientation, adipose tissue extension, side-to-side cytologic symmetry, uniform melanin deposits, nuclear pleomorphism, presence of mitoses, inflammation and epidermal alterations. Pagetoid growth and/or melanin deposits in the keratin layer were significantly higher in little children. In adults, presence of isolated cells within the lateral margins allows differentiating Spitz nevus from atypical Spitz tumor and Spitzoid melanoma. Deep located mitoses were statistically associated with Spitzoid melanoma in adults. Ulceration was statistically more frequent in peripubertary patients than in adults, probably due to trauma. In conclusion, presence of worrisome morphologic features (pagetoid growth, isolated cells within the lateral margins or ulceration) is correlated to patient's age and less to tumor type; there is no unique morphologic feature to relay on when evaluating a Spitzoid tumor, the final diagnosis being the results of interpretation of multiple clinical, morphologic, immunohistochemical and molecular data and not least dermatopathologist's personal experience.

Corresponding author: Sabina Zurac, Associate Professor, MD, PhD; e-mail:

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8. From basic lesions to a pathological staging of pulmonary fibrosis

Irina Stoia-Djeska, Raluca Amalia Ceausu, Anca Maria Cimpean, A. Nicodin, V. Tudorache, M. Raica

Idiopathic pulmonary fibrosis is a severe disease, with unpredictable evolution that frequently leads to respiratory failure and death, despite some progresses made in the field of therapy. Basically, the bad prognosis and failure of therapy are the consequence of the lack of data about the molecular events that have as result the extensive fibrosis. Although the basic lesions were defined many years ago, the pathological classification of pulmonary fibrosis is controversial. In the present work, we analyzed the prognostic impact of basic microscopic lesions on a possible new classification that could be related to the patient outcome. For this purpose, we have investigated 20 cases with idiopathic pulmonary fibrosis and samples of lung parenchyma were obtained by video assisted thoracoscopy. The specimens were processed by usual histological technique and sections were stained with Hematoxylin-Eosin, Masson's trichrome and Gordon-Sweet silver staining. There were evaluated the lung architecture, the chronic inflammatory infiltrate, macrophages and fibrosis. The distribution and severity of each parameter was converted into points and finally graded from I to IV, with corresponding score from 1 to 12. We found four cases with degree II, 12 with degree III, and four with degree IV. Our results support the hypothesis that the evaluation of basic lesions could be the basis for a more objective classification and staging of lung fibrosis and, possibly, a better prognostic method and, eventually, a predictor for the response to targeted therapy.

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

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9. Follicular non-Hodgkin's lymphoma: correlation between histology, pathophysiology, cytogenetic, prognostic factors, treatment, survival

Amelia Maria Gaman

Background: Folicullar lymphoma (FL) is one of the most common types of all non-Hodgkin's lymphomas (25-40%), characterized by a slowly progressive enlargement of lymph nodes, impairment of hematopoiesis, increased risk to infections, a relatively good prognosis, but usually incurable. Histopathologically, FL has been graded according to the proportion of centroblasts and stratified into grades 1-3: FL grade 1-2 (low-grade), which include cases with few centroblasts, and FL grade 3, divided into grades 3a and 3b, based on the absence of centrocytes in the latter category. Several studies have identified some differences between grade 3a and grade 3b of FL, with most cases of FL grade 3b being more closely related to diffuse large B-cell lymphoma (DLBCL) at molecular level. Several multicenter prospective randomized trials demonstrated an improved outcome when Rituximab (R) was added to chemotherapy for the treatment of follicular non-Hodgkin's lymphomas and a beneficial effect in the quality of life after Rituximab maintenance therapy at these patients. Aim of study: To establish some correlation between histology, prognostic factors, treatment and evaluate whether maintenance therapy with anti-CD20+ monoclonal antibodies prolonged progression free survival compared to observation only at the patients with follicular lymphomas treated with R-chemotherapy regimens. Patients and Methods: We studied nineteen patients with follicular non-Hodgkin's lymphomas (grades 1-3) treated with R-CHOP/R-miniCHOP regimens hospitalized in the Clinic of Hematology from Craiova (Romania), between 2008-2011. After these treatments, nine patients with stage III/IV follicular lymphomas were treated with Rituximab maintenance therapy (eight cycles Rituximab 375 mg/sqm, i.v., once every three months for two years) vs. observation only at 10 patients. Results: In our study, low-grade FL was correlated with a good prognosis at patients with FLIPI score 0-2; the statistical analysis revealed that the progression free survival (PFS) was prolonged at the patients with stage III/IV follicular lymphomas who received Rituximab maintenance therapy compared to the ones with observation only with 1.9 years. Conclusions: Low-grade (1-2) FL was correlated with a good prognosis in patients with FLIPI score 0-2; Rituximab maintenance therapy compared with observation only is safe and prolonged progression free survival at patients with follicular lymphomas treated with R-chemotherapy as first line therapy.

Corresponding author: Amelia Maria Gaman, Associate Professor, MD, PhD; e-mail:

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10. Epidemiological and morphological data of ocular melanocytic lesions

Flavia Baderca, C. Solovan, Loredana Boghian

Ocular melanocytic lesions comprise a spectrum of lesions ranging from benign nevi to invasive melanoma. Clinical and histopathological appearance of conjunctival lesions ranges from freckle to lentigo and to nevi. Between these types, conjunctival nevi and conjunctival melanosis are the most frequent. Conjunctival and uveal melanocytes are derived from the neural crest, as their cutaneous counterparts, whereas the pigment epithelial melanocytes are derived from the neuroepithelium or the layers of the optic cup. Melanomas can develop in one of several places within the eye, and can be divided in uveal melanomas and conjunctival melanomas. The purpose of the study was to investigate the epidemiological and morphological data of ocular melanocytic lesions, especially intraocular melanoma, through analysis of the ocular biopsies received in the Department of Pathology, Emergency City Hospital, Timisoara, Romania, for the period of five years. We did not observed any gender predilection neither in benign nor in malignant tumors. In our study, whatever the tumor location was, the most common type of melanomas was mixed with both, epithelioid and spindle cells. In some cases, immunohistochemical investigations are useful to appreciate the benign or malignant character of the tumor.

Corresponding author: Loredana Boghian, MD, PhD student; e-mail:

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11. Anatomy and variations of palmaris longus in fetuses

S. Albay, Yadigar Kastamoni, Busra Sakalli, S. Tunali

The aim of this study was to assess the absence of the palmaris longus, the proportion of the lengths of tendon and muscle belly, the development of the tendon and the belly during the fetal period, look for any difference between sides and gender. Fifty-eight spontaneously aborted human fetuses (26 female, 32 male, 116 upper extremities) were studied. The presence or absence of the palmaris longus was determined. The lengths of the belly and tendon were measured, and belly/tendon length ratio was calculated. Correlation with gestational age, body side and gender were studied. The muscle was absent in 44 forearms (37.93%; 20 right side, 34.48%; 24 left side, 41.38%); being bilateral in 19 of 58 fetuses (32.76%) and unilateral in six (10.34%). The unilateral absence rate was higher on the left side with a statistically significant difference. The absence of palmaris longus was more common in females, and the difference was statistically significant. The belly/tendon length ratio was 1.04+/-0.35 on the right side and 1.09+/-0.3 on the left. It did not show any difference according the fetal age. A sound knowledge on the anatomy and variations of palmaris longus is of great importance during surgical interventions; because it is the first choice for tendon grafts, by the virtue of its structure and function. Thus, this study is of academic interest for anatomists and hand surgeons alike.

Corresponding author: Soner Albay, Assistant Professor, MD; e-mail:

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12. Endometriosis - clinical approach based on histological findings

C. Cristescu, Andreea Veliscu, B. Marinescu, Anca Patrascu, E. T. Trasca, O. T. Pop

Endometriosis is a benign disease defined by the presence of endometrial glands and stroma outside of the uterus and is associated with both pelvic pain and infertility. The most common sites of endometriosis, in decreasing order of frequency, are the ovaries, anterior and posterior cul-de-sac, posterior broad ligaments, uterosacral ligaments, uterus, fallopian tubes, sigmoid colon, appendix, and round ligaments. The main treatment is surgical, but often-microscopic islands of endometrial tissue remain, which proliferate and are responsible for relapses. We tested the efficacy of two drugs (Medroxyprogesterone and Triptorelinum), administered for six months to prevent recurrence after surgery. Treatment with Medroxyprogesterone was 100% effective in terms of relapse, while Triptorelinum could not prevent recurrence of endometriosis.

Corresponding author: Anca Patrascu, Associate Professor, MD, PhD; e-mail:

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13. CLDN3 expression and significance - breast carcinoma versus ovarian carcinoma

Carmen Ionescu Popescu, Ludmila Liliac, Raluca Amalia Ceausu, Raluca Balan, Adriana Grigoras, Irina-Draga Caruntu, Cornelia Amalinei

Claudins (CLDNs) are transmembrane proteins, as normal constituents of the architecture of tight junctions. Recent studies support their involvement in carcinogenesis, as changes in CLDNs structure result in alterations in tight junctions' structure and function, facilitating malignant transformation. We aimed CLDN3 investigation in both breast and ovarian carcinoma, targeting the identification of its expression differences. The immunohistochemical assessment was performed on 20 cases of breast carcinomas (Group 1) and 19 cases of epithelial ovarian carcinomas (Group 2). Firstly, the specific panel for the molecular classification was applied for specimens of the first group. Then, all the specimens were immunostained for CLDN3 and a semi-quantitative evaluation was made, based on the percentage of positive cells and the intensity of staining. In Group 1, in the ER positive category, CLDN3 was overexpressed in five cases (four cases of luminal A and one case of luminal B subtype, respectively), negative in three cases (luminal A subtype) and weakly expressed in a single case (luminal A subtype); in ER negative category, CLDN3 expression was strong in four cases (one case of Her2/neu subtype and three cases of basal-like subtype), negative in two cases (normal breast-like subtype) and weak in five cases (one case of Her2/neu subtype, one triple-negative subtype, and three basal-like subtype). In Group 2, CLDN3 was overexpressed in 15 cases, histopathologically diagnosed as serous (10 cases), mucinous (two cases), endometrioid (two cases), and mixed carcinomas (one case); a weak expression was noticed in a single case, of the serous subtype; CLDN3 was undetectable in three cases (one serous, one clear cell, and one endometrioid type). Our comparative analysis of CLDN3 profile in breast and ovarian cancer clearly indicates organ specificity.

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

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14. Comparative study of microvascular density in experimental third-degree skin burns treated with topical preparations containing herbal extracts

G. D. Mogosanu, Florina Carmen Popescu, Cristina Jana Busuioc, I. Lascar, L. Mogoanta

During the healing process of third-degree skin burns, a very complex response involves different cells and tissues linked together by intra- and extra-cellular mechanisms. For the restoration of damaged tissues, angiogenesis is the key point in the formation of new blood vessels. By their emollient, astringent, antiseptic, anti-inflammatory, biostimulator, epithelizing and cicatrizing effect, active principles from natural products contribute to the acceleration of the wound-healing process. In our study, we investigated the angiogenesis process in experimental model of third-degree skin burns treated with three topical preparations (cold-creams) containing 10% herbal extracts, comparing with 1% sulfadiazine cream and cold-cream base respectively. By their biostimulator, epithelizing and cicatrizing effect, cold-creams with herbal extracts are locally modulators of the cellular response and support the wound healing. The phytocomplex stimulates the favorable evolution of the burnt skin wounds and the development of neoangiogenesis capillaries.

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

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15. New ways of bronchial stump closure after lung resection: experimental study

V. Bud, B. A. Suciu, V. Butiurca, Klara Brinzaniuc, Ruxandra Copotoiu, C. Copotoiu, Anca Sin

Introduction: Lung surgery has long been associated with a high rate of mortality and morbidity. Bronchial stump fistula is one of the leading causes of high morbidity and high mortality rate. The objectives of our study are to asses the effect of new ways of closure the bronchial stump after lung resections. Materials and Methods: For our study, we used eighteen adult rabbits. We performed left trans-thoracic inferior lobectomy to each animal because left lung functional capacities are smaller than right lung functional capacities. The bronchial stump was closed in three different ways: simple running suture of 5.0 monofilament polypropylene (Sweet procedure), running suture of 5.0 monofilament polypropylene (horizontal "U" shape model) and Vycril patch, running suture of 5.0 monofilament polypropylene (horizontal "U" shape model) and bovine pericardium patch. All surviving rabbits were sacrificed in 30 day postoperatively. Results: In our study, we did not notice any deaths among our experimental animals and we did not observe any bronchial stump fistula. In the group of rabbits in which the bronchial stump was closed using a patch of heterologous bovine pericardium, the histological exams revealed a cvasinormal bronchial wall with a normal bronchial epithelium and basal membrane integrity and no inflammatory lympho-plasmocytary infiltrate cell, absence of macrophages at this level. Conclusions: Using the heterologous bovine pericardium patch to reinforce the bronchial stump could be beneficial because of the faster healing and scarring process. To our knowledge, no studies have previously evaluated this procedure of bronchial stump closure. Our findings could therefore, serve as an impetus for further research in this area.

Corresponding author: Bogdan Andrei Suciu, MD, PhD; e-mail:

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16. Sinus lift and dental implant treatment as an option for enhancing the quality of life of our patients

S. Bran, Fl. Onisor-Gligor, D. Lucaciu, R. S. Campian, Ondine Patricia Lucaciu, Diana Dudea

The loss of teeth in the posterior maxillary area is a real handicap for patients, which significantly affects their quality of life. This study aims to perform an exhaustive multidisciplinary analysis of bone reconstruction at this level using alloplastic osteoinductive material, followed by dental implant treatment. The results of the study show a 92.7% rate of implant integration and a 35% rate of replacement of the augmentation material with new bone after six months. The recovery of the lost functions is complete, and after a maximum length of time of 18 days, with a mean of 4.36 days, patients no longer perceive the reconstruction as a foreign body, which means a real improvement in their quality of life.

Corresponding author: Florin Onisor-Gligor, University Assistant, MD, PhD; e-mail:

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17. Early healing after ridge preservation with a new collagen matrix in dog extraction sockets: preliminary observations

C. Cioban, Raluca Zaganescu, Alexandra Roman, A. Muste, F. Beteg, R. S. Campian, Bianca Bosca

The aim of the present animal study was to investigate the early healing processes developing in the post-extraction sockets preserved with a new-marketed collagen matrix as, to our knowledge, such investigations have not been reported so far. In both quadrants of the mandible of a mongrel dog, the distal sockets of the second premolars served as experimental sites for ridge preservation. The experimental site 1 was protected with a resorbable membrane and then with the collagen matrix. The experimental site 2 was filled with a xenograft and then covered with the collagen matrix. The samples were harvested after one month of healing. In both experimental sites, the bundle bone lining the inner surface of the alveolus was replaced with trabecular bone containing areas of woven bone. A continuous layer of osteoblasts could be observed on the surface of woven bone areas. Osteoclasts encased within resorptive lacunae lined the outer portions of bone walls for the experimental site 1. The trabecular bone occupied only the apical third of the socket in experimental site 1, but it was obviously more abundant in the experimental site 2, occupying also the central compartment of the socket. Moreover, the trabeculae of the bone occupying the inner area of the alveolus were thicker for the experiment site 2 than for experiment site 1, suggesting an increased osseous deposition in the latter situation. Our preliminary results suggest that the association collagen matrix plus xenograft may be a valuable method for ridge preservation.

Corresponding author: Alexandra Roman, Professor, PhD; e-mail:

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18. Dendritic cells: friends or foes of laryngeal cancer?

C. Doros, N. Balica, Anca Maria Cimpean

Background: Dendritic cells are important keyplayers of various malignant tumors but less studied in laryngeal malignancies. We assessed the immunohistochemical expression and distribution of dendritic cells in different types of laryngeal carcinomas and tried to find if they could influence evolution and prognosis of such malignancies. Materials and Methods: Immunohistochemistry was applied on 49 laryngeal tumors. Dendritic cells were identified by using S100 protein staining. The specimens were then evaluated for dendritic cells presence, number and distribution. Results: S100 positive cells were identified in all cases of squamous cell carcinoma, being absent in the case of adenoid cell carcinoma. Squamous cells carcinomas had the highest numbers of S100 positive cells. For them, we encountered an inverse correlation between peri- and intra-tumor S100 positive cells density. Intraepithelial dendritic cells density was lower for undifferentiated squamous cell carcinoma, also as for stroma of well-differentiated squamous cell carcinoma. Poorly differentiated carcinoma had a higher density of stromal S100 positive cells. S100 positive cells were identified in tumor area with squamous differentiation in all cases, and in peritumor area in 41 cases (83.67%). S100 positive cells density was correlated with tumor grade but not with invasion. Conclusions: Taking together, our results suggest that migration of stromal dendritic cells inside tumor areas could be an important component of the antitumor immune response induction and thus, S100 positive dendritic cells may be considered as a favorable prognostic factor in laryngeal carcinomas.

Corresponding author: Anca Maria Cimpean, Associate Professor, MD, PhD; e-mail:

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19. EGFR and Her2/neu immunoexpression in papillary urothelial bladder carcinomas

M. Enache, Cristiana Eugenia Simionescu, A. Stepan

The study analyzes the immunoexpression of EGFR and Her2/neu in 45 cases of papillary urothelial bladder carcinoma. The cases have been investigated through histopathological and immunohistochemistry techniques, the quantification of results considering the number of marked cells and intensity of the reactions. Immunoexpression of EGFR has been observed in 53.3% of cases, the reaction score having high values in moderate carcinomas and poorly differentiated carcinomas with invasion in the muscularis propria or in the entire wall. Immunostain of Her2/neu has been positive in 42.2% of cases, the poorly differentiated carcinomas presenting high scores regardless of tumor stage. The analyzed markers proved useful for identifying urothelial papillary carcinomas with a poorly differentiated grade, which are in an advanced stage.

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

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20. COX-2 and Ki-67 immunohistochemical markers in the assessment of long-standing ulcerative colitis associated dysplasia

O. C. Fratila, Tiberia I. Ilias

Introduction: Malignancy in LUC (long-standing ulcerative colitis) presumably evolves through a chronic inflammation-dysplasia-adeno-carcinoma sequence in which a multitude of yet not fully understood factors takes part. Aim: To assess ulcerative colitis (UC) associated dysplasia and to distinguish regenerative changes from premalignant ones using immunohistochemical (IHC) markers. Materials and Methods: We studied 80 LUC biopsy specimens: 20 high-grade dysplasia (HGD), 20 low-grade dysplasia (LGD), 20 indefinite for dysplasia, 20 regenerative atypia. We used anti-COX-2 and Ki-67 antisera (Dako, Carpinteria, USA) to perform immunohistochemical staining by the labeled Streptavidin-Biotin method, and then assessed and graded staining intensity and distribution using previously described scoring systems. Statistical analysis was made using chi-square test and SPSS application. A p-value <0.05 was considered significant. Results: In LGD, most of the cases had middle and top Ki-67 localization of the staining. For HGD, we found to be characteristic the top and surface staining of the crypts and no case of basal immunostaining. COX-2 immunostaining was positive (total score >=3) in 72.5% of all the UC cases studied. In non-dysplastic lesions (regenerative atypia), COX-2 expression was negative and as the pathologic process progressed towards dysplasia/malignant transformation, COX-2 expression became positive with a progressive increase of the total score. Conclusions: A combination of enhanced colonoscopic surveillance and IHC markers those are more sensitive for dysplasia might be the optimal way to manage the increased colorectal cancer (CRC) risk in LUC patients. Further studies to find additional prognostic parameters will provide valuable insights into the behavior of LUC.

Corresponding author: Ovidiu Cristian Fratila, Associate Professor, MD, PhD; e-mail:

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21. Cerebral microvascular changes induced by rich cholesterol and saturated fatty acid diet in Wistar rats

R. Stanescu, M. R. Stanescu, Adriana Bold, Garofita Olivia Mateescu

The impact of an excess of fatty acids in the diet on cardiovascular diseases has been studied and discussed both in human and animal studies. Generally, excessive saturated fats increase the risk, while unsaturated fats are considered less harmful. Our aim was to perform an experimental study in order to analyze how fatty diet quality (unsaturated vs. saturated fatty acids) influences atherogenesis. Materials and Methods: In our experimental study, 18 adult Wistar rats were randomly divided into two equal groups. One group was subjected to a rich unsaturated fatty acid diet (untar) and the other group to a rich saturated one (palm oil). Three animals from each group were sacrificed after 12, 18, and 48 weeks. The brain was removed and microscopically examined after Hematoxylin-Eosin, Orcein and Masson's trichrome classical staining, and after immunohistochemical marking using the anti-alpha smooth muscle actin antibody. Results: Rats sacrificed after 12 weeks revealed modicum lesions, as intimal vacuoles or minute intraluminal thrombosis, and cerebral parenchymal edema. After 18 weeks, some of rats subjected to a rich saturated fatty acid diet presented vacuoles found in all arteriolar wall layers, and a tendency towards parietal thrombosis. In rats subjected to a rich unsaturated fatty acid diet, the subintimal arteriolar vacuolization was associated with an intramural and adventitial fibrosis. In rats sacrificed after 48 weeks, lesional polymorphism was pronounced, but in rats subjected to a rich unsaturated fatty acid diet complete luminal thrombosis was followed by a an organized thrombus with multiple capillary channels. Although in Wistar rats atherosclerosis appeared only after intensive changes in diet, different experimental studies showed that, in transgenic rats, rich saturated fatty acid diet induced progressive atherosclerotic lesions, resembling those observed by us, but also some aspects described in human pathology. Conclusions: Our experimental study reveals differences in atherogenesis under saturated vs. unsaturated fatty diet.

Corresponding author: Radu Stanescu, MD, PhD student; e-mail:

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22. Hematogenous placental infection in acute respiratory infections

Claudia Sandu, Roxana Folescu, Elena Pop, A. G. M. Motoc

The study focuses on the macroscopic and microscopic aspects of the placentae resulting from abortions or febrile births and their correlation with acute disorders of the upper or lower respiratory apparatus in pregnant women in various stages of pregnancy. The viral, bacterial or mycotic disorders were considered responsible for triggering septic abortion, premature or full-term deliveries, followed by septic complications of the child/fetus or of the mother. When the mother's acute respiratory infection is induced by highly virulent pathogens, in patients with low immunity or lacking adequate medical treatment, the infection may spread through the mother's bloodstream to the placenta. The study was conducted on 90 placentae. Microscopic analysis of the tissue samples revealed acute inflammatory infiltration. Two of the study cases should be mentioned here: a four-month pregnant woman suffering from septic abortion and a nine-month pregnant woman whose fetus died in the womb because of acute pneumopathy on a non-breathing lung. Both pregnant women had the same type of disorder and neither followed any medical treatment prescribed by a physician. The prevention of placental infection is closely connected to the prevention of acute respiratory diseases or their proper treatment after their onset.

Corresponding author: Claudia-Mariana Sandu, University Assistant, MD, PhD; e-mail:

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23. Phases of the cutaneous angiogenesis process in experimental third-degree skin burns: histological and immunohistochemical study

Cristina Jana Busuioc, G. D. Mogosanu, Florina Carmen Popescu, I. Lascar, H. Parvanescu, L. Mogoanta

Skin burns represent a major problem of public health because of their frequency and because of their seriousness, too. The healing process of the burnt wound is extremely complex, as it requires a well-coordinated collaboration among different tissues and cellular strings. From the morphological point of view, the stages of the repairing process of the skin wounds include processes of inflammation, proliferation and tissular remodeling. Angiogenesis has a role of extreme importance within the healing process of third-degree skin burns. That is because the vascularization remake is necessary for feeding the tissue of granulation with nutritive substances and oxygen. The angiogenesis started relatively fast. Three days after the producing of the burn, there could be identified strings of CD34+ endothelial precursor cells at the edges and deep into the wound, all these having contact with the normal blood vessels or with those lees affected by the thermal aggression. After the lumenization of the newly-formed capillary vessels, there appeared the pericytes within their membrane. The CD34+ endothelial precursor cells (EPc), as well as the pericytes, participate at the synthesis of the base membrane of the angiogenesis vessels. The density of the angiogenesis vessels on the surface unit within the tissue of granulation grew from three to 12 days. After that, they reduced progressively while the tissue of granulation was becoming mature. The angiogenesis vessels go through a process of reshuffling and maturation at the same time with the maturation of the tissue of granulation, but these processes did not appear to be finished when the skin was completely healed, and the epidermis was totally recovered.

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

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24. Correlations between anthropometric and serologic elements of metabolic syndrome and histopathologic features of nonalcoholic fatty liver disease

Krisztina Szalman, Ligia Bancu, Anca Sin

Aim of the study: Studying the correlation between elements of metabolic syndrome and histological changes of the liver in nonalcoholic fatty liver disease. Patients and Methods: Thirty-nine patients with nonalcoholic fatty liver disease were included in our study. Inclusion criteria were: presence of liver steatosis on ultrasound in patients with waist circumference over 94 cm in men and over 80 cm in women and with serologic elements of metabolic syndrome. Exclusion criteria were: chronic viral hepatitis, autoimmune hepatitis, Wilson disease, hemochromatosis, regular alcohol consumption. Body mass index, waist circumference, fasting plasma glucose, serum triglyceride and cholesterol levels and serum ALT were determined. On liver biopsy specimens, performed in each patient, the NASH score, representing the sum of fibrosis, steatosis, lobular inflammation and ballooning, was calculated. Results: Necroinflammation was mild in 15 patients, medium in 19 patients and severe in five patients. Mild fibrosis was present in four cases, medium in 14 cases, severe in six, and two patients were diagnosed with cirrhosis. We found statistically significant correlation between waist circumference and the grade of histological activity, the presence of diabetes and both fibrosis grade and histological activity, and the serum ALT and histological activity. Conclusions: Noninvasive assessment of the severity of hepatic histological changes in nonalcoholic fatty liver disease could be made by anthropometric parameters or by serologic components of metabolic syndrome, but it is not an accurate method to identify patients with high-risk for disease progression. These noninvasive parameters cannot replace liver biopsy.

Corresponding author: Ligia Bancu, Associate Professor, MD, PhD; e-mail:

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25. Evaluation of microvascular density in inflammatory lesions and carcinoma of palatine tonsil

Carmen Aurelia Mogoanta, Daniela Adriana Ion, V. Budu, Gabriela Mutiu, D. Salplahta, Emilia Afrem

The tonsil carcinoma with squamous cells seems to be one of the neoplastic lesions with a growing incidence worldwide, even in those geographical areas where the smoking incidence has been reduced. In the disease etiopathogenesis, more factors are incriminated. Among these, the most frequently mentioned are smoking, alcohol consumption and the infection with the Human Papilloma Virus. Among the morphological modifications incriminated for the tumoral appearance and development, there is also included the angiogenesis process that involves the apparition of new blood vessels out from the pre-existent ones, vessels that bring a plus of oxygen and nutritive substances for the tumoral cells. Taking into consideration the fact that the tumoral process is most often accompanied by an inflammatory reaction, in our study we also determined the microvascular density in the carcinoma with squamous cells in the palatine tonsil and in chronic tonsillitis, compared to the vascular density in normal tonsil stroma. We quantified the reaction of the mast cells in the stroma of the two types of lesions, too. The microvascular density in the carcinoma with squamous cells in the palatine tonsil was a lot greater than the microvascular density in chronic tonsillitis. The maximum number of blood vessels in tumoral lesions as well as their area, quantified through the "hot spot" technique on the surface unit, was around two times greater than in chronic tonsillitis. The number of mast cells was significantly larger in chronic tonsillitis and in tonsil carcinoma, too, but the reaction of these cells in the inflammatory affections was more intense than in the neoplastic lesions.

Corresponding author: Carmen Aurelia Mogoanta, MD, PhD; e-mail:

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26. Micromolecular multiple myeloma with chronic kidney failure in a young female patient on continuous hemodialysis

Ionela Rotaru, M. Badea, Alina Daniela Tanase, Camelia Foarfa

The maximum incidence of multiple myeloma appears in the 6th-7th decade of life and although the number of patients aged les than 60 years is increasing in recent years, the diagnosis of a monoclonal gammopathy in a young patient, under the age of 40 years remains a rarity. Literature data cite an incidence of approximately 2.2% in patients less than 40-year-old and an incidence of 0.3% in patients less than 30-year-old of all cases diagnosed with multiple myeloma. We present the case of a 32-year-old patient, being on continuous hemodialysis for chronic kidney failure for about a year, at the Hematology Clinic of Craiova, Romania. We investigate the origin of a serum monoclonal component revealed when performing serum protein electrophoresis. Bone marrow examination revealed the presence of a plasma cell infiltrate of 18%, which associated with the presence of a serum monoclonal component and in the conditions of renal failure as a complication of the disease, has allowed the diagnosis of multiple myeloma.

Corresponding author: Ionela Rotaru, Assistant Professor, MD; e-mail:

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27. Morphological characteristics of a mucinous adenocarcinoma of the prostate: differential diagnosis considerations

Manuela Enciu, Mariana Aschie, Mariana Deacu, I. Poinareanu

Mucoid adenocarcinoma of the prostate is a very rare variant, account less than 1% of prostatic adenocarcinomas. In this respect, the most common histopathological type of prostate cancer is acinar adenocarcinoma. Diagnosis of this variant is very important due to peculiarities: aggressive biologic behavior, poor response to radiotherapy. Although these tumors are not as hormonally responsive as acinar adenocarcinomas, some of them respond to androgen withdrawal. Before making a diagnosis of primary mucinous adenocarcinoma, is necessary to exclude an extraprostatic malignant neoplasia, especially from the bladder or bowel. We present the case of a male patient who suffered a transurethral prostatic resection surgery. Histopathological examination revealed malignant tumor cells floating in a pool of mucus or even acini mixed with signet ring cells. A particular aspect is that floating mucus cells have a non-papillary pattern similar to colloid carcinoma of the breast. The use of monoclonal antibody revealed positive immunoreaction of malignant cells for prostatic specific markers and excluded neoplastic invasion of the bladder or bowel carcinoma.

Corresponding author: Mariana Aschie, Professor, MD, PhD; e-mail:

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28. The Fahr syndrome and the chronic lymphocytic thyroiditis

Anca Sava, Gabriela Dumitrescu, Danisia Haba, Diana Hodorog, Claudia Mihailov, Elena Sapte

Fahr syndrome (FS) refers to basal ganglia calcification that is associated with many neurological and psychiatric abnormalities and appears as secondary to other diseases. We described a case of FS patient who was admitted in the Department of Neurology of "Prof. Dr. Nicolae Oblu" Clinical Emergency Hospital, Iassy, Romania, with seizure and mood disorders. On CT, the cause of seizure was found to be the bilateral calcifications of cerebellum, basal ganglia, thalamus and internal capsule. As the patient died after 15 days of hospitalization due to new seizures and gastrointestinal infection, an autopsy was made. Grossly, there were bilateral symmetrically gritty yellow areas in basal ganglia, thalami, internal capsule, cerebral cortex, cerebellar folia, dentate nucleus, and brain stem. A detailed histopathological examination revealed five types of calcium deposits within the walls of capillaries, small and medium-sized arteries from the intracerebral affected areas, chronic lymphocytic thyroiditis and fibro-adipose tissue instead of parathyroids. We consider that intracerebral symmetrical calcifications were the results of the hypoparathyroidism determined by an ancient autoimmune parathyroiditis that evolved to fibrosis as at microscopy we found an autoimmune thyroiditis.

Corresponding author: Anca Sava, MD, PhD; e-mail:

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29. The dedifferentiation of neuroendocrine tumor metastases: myth or reality?

Catalina Poiana, M. C. Neamtu, Elena Taina Avramescu, Mara Carsote, Raluca Trifanescu, Dana Terzea, Oana Maria Neamtu, Rucsandra Danciulescu Miulescu

Dedifferentiation is a process that may be found in metastasis from the neuroendocrine tumors. We present the case of a female within the seventh decade of life incidentally diagnosed with a mature teratoma with element of cartilages, bronchia, mucinous glands, and a poorly differentiated neuroendocrine carcinoma (Ki67 of 30%). After six months of chemotherapy and another six months of disease free interval, a metastasis of the great omentum was removed. The dedifferentiation was diagnosed based on much higher Ki67 (of 70%). The loss of estrogen receptor of 40% from the initial site to 3% into metastasis indicates an exclusive neuroendocrine aggressive pattern. The ovarian carcinoid is a rare situation, and metastasis to the great omentum with dedifferentiation is even seldom.

Corresponding author: Raluca Trifanescu, MD; e-mail:

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30. The importance of full tests of incidentally detected premalignant lesions

A. P. Carstea, Doina Carstea, E. F. Georgescu, Corina Gruia, V. Surlin

Early detection of asymptomatic phase or paraneoplastic manifestations in precancerous lesions and an early, correct and accurate diagnosis in terms of pathology of the lesion in question, makes important chances of healing and prolonged patient's life expectations. We present the case of a young patient who came to the emergency room and then admitted in the cardiology department with a heart rhythm disorder. The medical investigations that followed (gastric endoscopy, biopsy, histological and IHC exams), finds gastric polyps, which proved to be gastrointestinal stromal tumors (GIST).

Corresponding author: Andrei Carstea, MD, PhD student; e-mail:

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31. Talon cusp affecting primary dentition in two siblings: a case report

S. K. Mallineni, N. M. Manan, C. K. Lee, N. M. King

The term talon cusp refers to a rare developmental dental anomaly characterized by a cusp-like structure projecting from the cingulum area or cement-enamel junction. This condition can occur in the maxillary and mandibular arches of the primary and permanent dentitions. The purpose of this paper is to report on the presence of talon cusps in the primary dentition of two southern Chinese siblings. The 4 years and 2 months old girl had a talon cusp on her maxillary right primary central incisor, while her 2 years and 9 months old brother had bilateral talon cusps on the maxillary primary central incisors. The presence of this rare dental anomaly in two siblings has scarcely been reported in the literature and this may provide further evidence of a hereditary etiology.

Corresponding author: Nigel M. King, Winthrop Professor; e-mail:

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32. Lobodontia: the unravelling of the wolf teeth

A. Ather, Hunaiza Ather, Shashi Rashmi Acharya, R. A. Radhakrishnan

Abnormalities of tooth shape and size generally coexist, as both are determined during the morphodifferentiation stage of odontogenesis. This report describes a case of lobodontia exhibiting multiple dental anomalies of which, "fang like" cuspids, multituberculism of molars, hypodontia along with severe generalized microdontia are distinctive. Radiographic features, diagnostic convolutions, differential diagnosis and clinical significance have also been highlighted.

Corresponding author: Amber Ather, Assistant Professor, BDS, MDS; e-mail:

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33. Letter to the Editor. Parotid Warthin tumor with mucous differentiation

Adriana Handra-Luca, Mouna Bendib


Corresponding author: Adriana Handra-Luca, MD, PhD; e-mail:

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34. Conference Announcement: 2nd Dermatopathology Course in El Bierzo, July 24-26, 2013, Ponferrada, Spain

A. Fernandez-Flores


Corresponding author: Angel Fernandez-Flores, MD, PhD; e-mail:

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35. Conference Announcement: The XIth National Symposium on Microscopic Morphology with international participation, May 22-24, 2013, Craiova, Romania

L. Mogoanta


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

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