ROMANIAN JOURNAL of MORPHOLOGY and EMBRYOLOGY

Vol. 57 No. 1, 2016

1. Telocyte - a particular cell phenotype. Infrastructure, relationships and putative functions

Nicolae Mirancea

Here we review on the infrastructure, relationships and putative role of a new cell phenotype termed telocyte. Ultrastructural and immunochemical characteristics of telocytes, their identification in different tissues, homo- and heterocellular telocyte s contacts and considerations concerning their putative role in normal as well as pathological conditions are largely presented. A special section of the paper is devoted to our opinion/hypothesis concerning the possibility that, to some extent, endoneurial fibroblast dendritic phenotype (existed in the peripheral nerve endoneurium) can be assimilated to the telocyte or, at least to the telocyte-like. In this respect, we report about a body of evidence that endoneurial fibroblasts dendritic cells share some infrastructural characteristics with telocyte phenotype. Telocytes involvement in pathology, tissue engineering and regenerative medicine is also debated.

Corresponding author: Nicolae Mirancea, Professor, PhD; e-mail: nick_mirancea@yahoo.com

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2. pH sensitive core-shell magnetic nanoparticles for targeted drug delivery in cancer therapy

Iulia Ioana Lungu, Marius Radulescu, George Dan Mogosanu, Alexandru Mihai Grumezescu

In the last decade, nanobiotechnology has evolved rapidly with an extensive impact on biomedical area. In order to improve bioavailability and minimize adverse effects, drug delivery systems based on magnetic nanocomposites are under development mainly for cancer imaging and antitumor therapy. In this regard, pH sensitive core-shell magnetic nanoparticles (NPs) with accurate controlled size and shape are synthesized by various modern methods, such as homogeneous precipitation, coprecipitation, microemulsion or polyol approaches, high temperature and hydrothermal reactions, sol-gel reactions, aerosol/vapor processes and sonolysis. Due to their unique combined physico-chemical and biological properties (such as higher dispensability, chemical and thermal stability, biocompatibility), pH responsive core-shell magnetic NPs are widely investigated for controlled release of cytostatic drugs into the tumor site by means of pH change: magnetite@silicon dioxide (Fe3O4@SiO2), Fe3O4@titanium dioxide (TiO2), beta-thiopropionate-polyethylene glycol (PEG)-modified Fe3O4@mSiO2, Fe3O4 NPs core coated with SiO2 with an imidazole group modified PEG-polypeptide (mPEG-poly-L-Asparagine), polyacrylic acid (PAA) and folic acid (FA) coating of the iron oxide NP core, methoxy polyethylene glycol-block-polymethacrylic acid-block-polyglycerol monomethacrylate (MPEG-b-PMAA-b-PGMA) attached by a PGMA block to a Fe3O4 core, PEG-modified polyamidoamine (PAMAM) dendrimer shell with Fe3O4 core and mesoporous silica coated on Fe3O4, mostly coated with an anticancer drug. This review paper highlights the modern research directions currently employed to demonstrate the utility of the pH responsive core-shell magnetic NPs in diagnosis and treatment of oncological diseases.

Corresponding author: Marius Radulescu, Chem Eng, PhD; e-mail: radulescu_marius@yahoo.com

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3. Age-related hypoxia in CNS pathology

George Mihai Badescu, Madalina Filfan, Ovidiu Ciobanu, Danut-Adrian Dumbrava, Aurel Popa-Wagner

Although neuropathological conditions differ in the etiology of the inflammatory response, cellular and molecular mechanisms of neuroinflammation are probably similar in aging, hypertension, depression and cognitive impairment. Moreover, a number of common risk factors such as obesity, hypertension, diabetes and atherosclerosis are increasingly understood to act as silent contributors to neuroinflammation and can underlie the development of disorders such as cerebral small vessel disease (cSVD) and subsequent dementia. On the other hand, acute neuroinflammation, such as in response to traumatic or cerebral ischemia, aggravates the acute damage and can lead to a number of pathological such as depression, post-stroke dementia and potentially neurodegeneration. All of those sequelae impair recovery and most of them provide the ground for further cerebrovascular events and a vicious cycle develops. Therefore, understanding the mechanisms associated with vascular dementia, stroke and related complications is of paramount importance in improving current preventive and therapeutic interventions. Likewise, understanding of molecular factors and pathways associated with neuroinflammation will eventually enable the discovery and implementation of new diagnostic and therapeutic strategies indicated in a wide range of neurological conditions.

Corresponding author: Aurel Popa-Wagner, Professor, PhD; e-mail: aurel.popa-wagner@med.uni-rostock.de

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4. Particularities of associating viral hepatitis with pregnancy and mental disorders

Victor Gheorman, Anca Livia Chirita, Elena-Madalina Dumitrescu, Ion Rogoveanu, Octavian Istratoaie, Valeriu Gheorman, Razvan-Cosmin Pana

Pregnancy generates particular circumstances for all co-existent conditions. Associating pregnancy with liver diseases has distinct particularities. The authors will perform a presentation of the etiopathogenic, diagnosis and therapeutic conduct particularities regarding the association between pregnancy, chronic liver diseases and mental disorders. The three pathological entities are analyzed separately, followed by a study of a triple association. Associating pregnancy and mental disorders has been better studied due to a higher frequency of mental disorders, especially postpartum, but the triple association pregnancy, chronic hepatitis with viral etiology mainly, and mental disorders has been less analyzed. There is concluded that pregnancy, through the physiological changes it undergoes, as well as its pathology, represents a clearly influencing factor of the association with a chronic liver disease or with a mental disorder.

Corresponding author: Valeriu Gheorman, Associate Professor, MD, PhD; e-mail: valeriu_gheorman@yahoo.com

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5. Messy granuloma: an additional pattern of granulomatous reaction

Angel Fernandez-Flores, Jose Luis Rodriguez-Peralto

Aim: There are six main types of granulomas in dermatopathology. The aim is to present and describe a seventh pattern of granuloma that we came across while studying biopsies of cutaneous leishmaniasis. Materials and Methods: We investigated 17 cases of cutaneous leishmaniasis. A granulomatous response was identified in 14 of them. The features that were common to all of them were noted. These common features were compared with the morphological characteristics of the already known six types of granuloma in 155 cutaneous biopsies from our archives. We also looked for figures of granulomas in cases of cutaneous leishmaniasis from the literature. Results: The granulomas identified in all of our cases of leishmaniasis were composed of epithelioid elongated glassy histiocytes, arranged in a randomly disorganized pattern. We dubbed these messy granulomas. Additionally, the granulomas presented with necrotic eosinophilic material that did not collect in the center, but rather intermingled with the elongated histiocytes. We found many examples in the literature in which the granulomas could be reclassified as messy. Conclusions: Messy granuloma - although not pathognomonic of leishmaniasis - seems to be quite characteristic of leishmaniasis, at least of cutaneous leishmaniasis.

Corresponding author: Angel Fernandez-Flores, MD, PhD; e-mail: dermatopathonline@gmail.com

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6. IL-8, IL-8RA (CXCR1) and IL-8RB (CXCR2) expression in pilomatricoma

Corina Maria Cianga, Petru Cianga, Gabriela Florenta Dumitrescu, Anca Sava

Pilomatricoma is a rare benign tumor of the hair follicle matrix cells, which associates during its evolution a foreign body-like inflammatory process. We have investigated three such tumors, two of them displaying a rather poor stroma, while the third was distinctive due to its stroma and large numbers of inflammatory cells infiltrating the tumor. The analysis of IL-8 (interleukin-8), CXCR1 (IL-8RA - IL-8 receptor alpha) and CXCR2 (IL-8RB - IL-8 receptor beta) expression showed that these molecules are present not only in many different types of inflammatory and endothelial cells, but also in several tumor basaloid, transitional and even few ghost cells. Taking into consideration the roles played by IL-8 and its two receptors, CXCR1 and CXCR2, this pattern of expression offers some insights on the potential roles of these molecules in tumor survival, cell proliferation and angiogenesis. Furthermore, given the expression of IL-1 (interleukin-1) and TNF (tumor necrosis factor) receptors by the tumor cells, IL-8 production by such cells might be under the control of these pro-inflammatory cytokines.

Corresponding author: Petru Cianga, MD, PhD; e-mails: petrucianga@hotmail.com, petru.cianga@umfiasi.ro

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7. Histological and immunohistochemical study on sentinel lymph node in colorectal cancer - values and limitations

Stelian Stefanita Mogoanta, Firmilian Calota, Ion Vasile, Antoanela Valentina Crafciuc, Mircea Constantin Gherghinescu, Konstantinos Sapalidis, Daniel Sorin Ilie, Daniela Adriana Ion

Colorectal cancer is currently one of the most common malignancies in both men and women. Surgical resection remains the essential element in the local control of the disease but the development of novel diagnostic and therapeutic tools can enhance the results of radical surgery. The indication for adjuvant treatment majorly depends on a correct pathological assessment of the surgical specimen - a correct pTNM staging. For patients diagnosed with stage III disease (characterized by the presence of lymph node metastases), adjuvant chemotherapy increases the survival rate, while in stage II disease, in most cases, the chemotherapy is contraindicated, due to increase morbidity without real benefit. This is why an accurate pN stage becomes essential. It is proven that classic pathological exam sometimes fails to identify lymph node micrometastases or isolate tumor cells, which might explain local or distant relapses in stage II patients. In our study, we evaluated a total of 39 surgical specimens of cTNM stage II patients operated for colon or rectal cancer. In the attempt to enhance the accuracy of pTNM staging we used ex vivo lymph node mapping combined with sentinel node analysis on serial sections in both classical histological and immunohistochemical (IHC) staining. We have demonstrated that the IHC staining on sentinel lymph node can improve the accuracy of pTNM staging, when used as a complementary diagnostic test, by identifying micrometastases and isolated tumor cells.

Corresponding author: Stelian Stefanita Mogoanta, Lecturer, MD, PhD; e-mail: ssmogo@yahoo.com

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8. Testosterone stimulates proliferation and preserves stemness of human adult mesenchymal stem cells and endothelial progenitor cells

Maria Cristina Corotchi, Mirel Adrian Popa, Maya Simionescu

Human adult stem and progenitor cells are promising cell types widely studied for their clinical benefits. A reduced number of stem cells present in the human body are associated with numerous dysfunctions. Since androgens have a profound effect on different cell types, we questioned whether testosterone (T), one of the main androgens, influence and are involved in the proliferation of stem cells and/or affect their stemness potential. Isolated mesenchymal stem cells (MSCs) and endothelial progenitor cells (EPCs) were cultured and then stimulated with different concentration of testosterone (10-100 nM). The cellular proliferation rate, adhesion, and viability were measured in real-time using xCELLigence system and DNA-cell proliferation assay. The immunophenotype of the stimulated cells versus non-stimulated cells was determined by flow cytometry. The maximal effect on MSCs and EPCs proliferation was obtained at 40 nM testosterone; this concentration was used in further experiments. The cellular index measured in real-time by impedance-based dynamic measurements revealed that 40 nM testosterone had a proliferative effect on both MSCs and EPCs, having a proliferative index of cca. 50% above the control (non-stimulated) cells. Furthermore, flow cytometry assay indicated that testosterone stimulation did not alter the phenotype of MSCs and EPCs, both cell types preserving the expression of the characteristic surface markers. Testosterone stimulation increases the proliferation and preserves stemness of MSCs and EPCs suggesting that, besides other factors, the hormone may engineer these cells and increase their therapeutic potential.

Corresponding author: Maria Cristina Corotchi, Research Assistant, PhD Student; e-mail: cristina.corotchi@icbp.ro

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9. The prognostic significance of p53, Bax, Bcl-2 and cyclin E protein overexpression in colon cancer - an immunohistochemical study using the tissue microarray technique

Carmen Stanca Melincovici, Carmen Mihaela Mihu, Mariana Marginean, Adina Bianca Bosca, Andrei Coneac, Ioana Moldovan, Maria Crisan

In colon cancer, biological markers continue to have a limited prognostic value, the results being controversial. Studies of cell-cycle regulatory proteins and anti-apoptotic proteins aim to identify groups of patients that develop more aggressive tumors and might benefit from an individualized therapy management. The present study evaluates the prognostic role of the p53, Bax, Bcl-2 and cyclin E immunoexpression in colon cancer, using the tissue microarray (TMA) method. Tissue samples were obtained from 31 patients operated for colon cancer, embedded in TMA paraffin blocks and immunohistochemically stained for p53, Bax, Bcl-2 and cyclin E. We evaluated the relationship between the overexpression of these proteins and the clinico-pathological parameters, as well as the effect of these molecular markers on the survival rate. 65.22% of the patients were p53 positive, 39.13% Bcl-2 positive, 78.26% Bax positive and 34.78% cyclin E positive. Bcl-2(+) patients had significantly better differentiated tumors (p=0.043). Significantly poorly differentiated tumors were: Bax(+) patients (p=0.031), Bcl-2(-)/p53(-) patients (p=0.042), Bcl-2(-)/Bax(+) patients (p=0.029), and Bcl-2(-)/p53(-)/Bax(+) patients (p=0.016). The individual expression of the studied proteins did not influence the survival rate. A significantly lower survival rate was found in the following groups of patients: Bcl-2(-)/p53(-) (40% vs. 83.3%, p=0.027), p53(-)/Bax(+) (40% vs. 83.3%, p=0.027), Bcl-2(-)/p53(-)/Bax(+) (25% vs. 84.2%, p=0.003). The current study identified groups of patients with a significantly lower survival rate, which consequently are at an increased risk to develop tumors with a more aggressive biological behavior.

Corresponding author: Carmen Mihaela Mihu, Professor, MD, PhD; e-mail: carmenmihu2004@yahoo.com

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10. Histopathological changes induced by selective inactivation of menin on the thyroid gland in RET/PTC3 and E7 transgenic mice. A study of 77 cases

Oana-Maria Capraru, Nicole Berger, Nicolas Gadot, Myriam Decaussin-Petrucci, Chang Zhang, Angela Borda, Tibor Szilagyi, Francoise Borson-Chazot, Samia Selmi-Ruby

Multiple Endocrine Neoplasia Type 1 (MEN1) does not involve the thyroid gland, but animal studies have shown that mice with inactivation of menin could develop thyroid pathologies. The objective was to evaluate if the selective inactivation of menin in murine thyroid glands expressing RET/PTC3 and E7 oncogenes, might induce an increased index of proliferation and a more rapid development of thyroid hyperplasia and/or tumors. The thyroid glands of 77 mice aged 4-18 months (31 expressing the E7 oncogene and 25 the RET/PTC3 oncogene) were analyzed for histological changes and Ki67 proliferation index. Fifty-two mice had selective inactivation of menin in the thyroid gland (16 mice with RET/PTC3 oncogene and 19 mice with E7 oncogene). As compared to wild type, mice with inactivation of menin presented an increased Ki67 proliferation index. Mice presenting the E7 oncogene showed larger thyroid glands with a pattern of diffuse hyperplasia. Mice expressing the RET/PTC3 oncogene presented larger thyroid glands compared to the wild type mice but smaller compared to E7 mice. The lesions in the RET/PTC3 group were proliferative papillary cystic changes (60%), cribriform (16%), solid (8%) and a combination of these patterns in the rest of the thyroid glands. The inactivation of menin in the thyroid gland of young mice does not seem to change the histological pattern, but it influences the proliferation of follicular cells. Further molecular studies especially in aged mice are needed to better understand the correlation between certain oncogenes and the inactive status of menin.

Corresponding author: Angela Borda, Professor, MD, PhD; e-mail: prorector_int@umftgm.ro

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11. Basal cell carcinoma develops in contact with the epidermal basal cell layer - a three-dimensional morphological study

Ionica Pirici, Marius Eugen Ciurea, Ion Mindrila, Ioan Avramoiu, Alexandru Pirici, Monica Georgiana Nicola, Otilia-Constantina Rogoveanu

Basal cell carcinoma is the most common malignant tumor of the skin, and it develops most frequently on the areas of the body that make its treatment and care extremely difficult, especially in cases of neglecting or aggressive growth and invasion. Both typical mild cases as well as locally aggressive tumor types do not tend to metastasize, and it has been postulated that they should share some common biological and morphological features that might explain this behavior. In this study, we have utilized a high-resolution three-dimensional reconstruction technique on pathological samples from 15 cases of common aggressive (fibrosing and adenoid types) and mild (superficial type) basal cell carcinomas, and showed that all these types shared contact points and bridges with the underlying basal cell layer of the epidermis or with the outmost layer of the hair follicle. The connections found had in fact the highest number for fibrosing type (100%), compared to the superficial (85.71%) and adenoid (55%) types. The morphology of the connection bridges was also different, adjacent moderate to abundant inflammatory infiltrate seeming to lead to a loss of basaloid features in these areas. For the adenoid type, tumor islands seemed to be connected also to each other more strongly, forming a common tumor lace, and while it has been showed that superficial and fibrosing types have higher recurrence risks, all together these data might iterate a connection between the number of bridging points and the biological and clinical manifestation of this skin tumor.

Corresponding author: Marius Eugen Ciurea, Associate Professor, MD, PhD; e-mail: meciurea@gmail.com

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12. Antimicrobial coatings based on zinc oxide and orange oil for improved bioactive wound dressings and other applications

Marius Radulescu, Ecaterina Andronescu, Andreea Cirja, Alina Maria Holban, Laurentiu Mogoanta, Tudor-Adrian Balseanu, Bogdan Catalin, Tiberiu Paul Neagu, Ioan Lascar, Denisa Alexandra Florea, Alexandru Mihai Grumezescu, Bianca Ciubuca, Veronica Lazar, Mariana Carmen Chifiriuc, Alexandra Bolocan

This work presents a novel nano-modified coating for wound dressings and other medical devices with anti-infective properties, based on functionalized zinc oxide nanostructures and orange oil (ZnO@OO). The obtained nanosurfaces were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), selected area electron diffraction (SAED), differential thermal analysis-thermogravimetry (DTA-TG), X-ray diffraction (XRD), and Fourier transform infrared (FT-IR) spectroscopy. The obtained nanocomposite coatings exhibited an antimicrobial activity superior to bare ZnO nanoparticles (NPs) and to the control antibiotic against Staphylococcus aureus and Escherichia coli, as revealed by the lower minimal inhibitory concentration values. For the quantitative measurement of biofilm-embedded microbial cells, a culture-based, viable cell count method was used. The coated wound dressings proved to be more resistant to S. aureus microbial colonization and biofilm formation compared to the uncoated controls. These results, correlated with the good in vivo biodistribution open new directions for the design of nanostructured bioactive coating and surfaces, which can find applications in the medical field, for obtaining improved bioactive wound dressings and prosthetic devices, but also in food packaging and cosmetic industry.

Corresponding author: Alexandru Mihai Grumezescu, Chem Eng, PhD; e-mail: grumezescu@yahoo.com

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13. Remodeling of basement membrane in patients with asthma

Adriana Grigoras, Constantin Cristian Grigoras, Simona Eliza Giusca, Irina-Draga Caruntu, Cornelia Amalinei

The bronchial remodeling specific for the asthmatic disease consists in irreversible changes of the bronchial wall, including glandular and smooth muscle fibers hyperplasia and/or hypertrophy, goblet cells hyperplasia, and thickening of basement membrane (BM). We aimed to analyze the BM thickness in asthma patients, in order to validate the relationship between its changes and the disease severity defined in agreement with the Global Initiative for Asthma (GINA) criteria. The study group has been formed of 38 patients with different degrees of severity of asthma established by spirometry using Forced Expiratory Volume in one second (FEV1), and two patients without asthma symptoms as controls. The specimens harvested by fibrobronchoscopy have been processed by paraffin embedding followed by Hematoxylin-Eosin (HE) and Periodic Acid-Schiff (PAS) staining. For each case, the BM measurement has been realized by a point-by-point method. Statistical analysis has been performed using SPSS 17 software, by applying non-parametric correlation tests. The quantitative assessment revealed a progressive increase in BM thickness during the course of the disease, from a mean value of 11.2 micro-m in stage 1 to that of 15.6 micro-m in stage 4. Even if this process has been noticed starting with the first stage of asthma, the differences in the BM size were statistically significant only for stages 1 and 3 (p=0.047), stages 1 and 4 (p=0.000), stages 2 and 3 (p=0.000), and stages 3 and 4 (p=0.000). Spearman s test has shown an opposite correlation between the BM thickness and asthma severity defined by FEV1 values (r=-0.86, p<<0.01, 95% CI). Our study demonstrates that the collagen deposition at the epithelium-connective interface is initiated in early stages of asthma and continues in a progressive modality, the BM thickening being correlated with the disease severity. Thus, we support the concept of biological consequences of BM thickening in asthma pathogenesis, a mechanism still incompletely deciphered.

Corresponding author: Irina-Draga Caruntu, Professor, MD, PhD; e-mail: irinadragacaruntu@gmail.com

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14. Postmenopausal osteoporosis - clinical, biological and histopathological aspects

Oana-Roxana Pavel, Mihaela Popescu, Liliana Novac, Laurentiu Mogoanta, Laurentiu Petrisor Pavel, Razvan-Marius Vicas, Magdalena-Rodica Traistaru

Osteoporosis is one of the most common disorders in postmenopausal women, affecting the quality of life and increasing the risk for fractures in minor traumas. Changes in the bone microarchitecture causes static changes in the body and affects motility. In this study, we analyzed two groups of women, one with physiological menopause and one with surgically induced menopause. The diagnosis of osteoporosis was suspected based on the clinical symptoms and confirmed by assessing bone mineral density by the dual-energy X-ray absorptiometry (DEXA). Comparing some clinical and biological aspects there was noted that a much higher percentage of women with surgically induced menopause exhibited increases in body mass index, changes in serum lipids, cholesterol, triglycerides, blood glucose, serum calcium, magnesemia and osteocalcin. In contrast, no significant differences were observed in the histopathological aspects of bone tissue examined from these two groups. In all patients, there was identified a significant reduction in the number of osteocytes and osteoblasts, the expansion of haversian channels, reducing the number of trabecular bone in the cancellous bone with wide areola cavities often full of adipose tissue, non-homogenous demineralization of both the compact bone and the cancellous bone, atrophy and even absence of the endosteal, and the presence of multiple microfractures. Our study showed that early surgically induced menopause more intensely alters the lipid, carbohydrate and mineral metabolism, thus favoring the onset of osteoporosis.

Corresponding author: Mihaela Popescu, Lecturer, MD, PhD; e-mail: mihaela.n.popescu99@gmail.com

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15. Experimental study regarding the biocompatibility test of the Prolene (polypropylene abdominal mesh) product

Elena-Violeta Radu, Ionut-Simion Coman, Oana-Ilona David, Stefan-Iulian Bedereag, Ruxandra-Diana Sinescu, Valentin-Titus Grigorean, Mihai Popescu, Cristian-Dumitru Lupascu, Nicolae-Dan Straja, Ioan-Petre Florescu

The polypropylene mesh, although is one of the most used prosthetic biomaterials for abdominal wall defects, proved not to be completely inert, generating from precocious foreign body inflammatory reactions (varying by individual reactivity, the amount of used material and its structure), to late complications such as chronic infections, stercoral fistulae or mesh migration. The present paper was aimed at studying the behavior of implants of this material in three different areas of the body of experimental animals, as follows: intramuscular, intraperitoneal and extraperitoneal. The observation time was 21 days and 90 days. We observed foreign body reactions induced locally by the mesh that remains temporary, generating a moderate number of macrophages and foreign body giant cells. The material did not systemically affect the healing and the scaring of the surgical wounds, but in all three implant areas, the polypropylene mesh generated locally a fibrous proliferation reaction of neoformation tissue, which wrapped and secured the implanted product on all surfaces.

Corresponding author: Ruxandra-Diana Sinescu, MD, PhD; e-mail: ruxandrasinescu@gmail.com

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16. Morphological and clinical characteristics of the torus palatinus and torus mandibularis in a sample of young and adults Romanian people

Monica Scrieciu, Veronica Mercut, Razvan Mercut, Carrol Birjovanu, Mihaela Cristina Stan, Iulia Roxana Marinescu, Mihaela Niculescu, Daniel Iorgulescu, Marilena Bataiosu

The oral exostoses are protuberance located on the alveolar surfaces of the jawbones with nodular, flat or pedunculated shape. The purpose of this study was to highlight the variability of the morphological and clinical characteristics of torus palatinus (TP) and torus mandibularis (TM) in a sample of young and adults Romanian people. The study was conducted on 74 participants examined in Dental Prosthetics Clinic of the Faculty of Dentistry, University of Medicine and Pharmacy of Craiova, Romania, during October-December 2014. The morphological characteristics of the tori were non-metrical evaluated by the standard procedures of the clinical examination. Descriptive statistics only including means, averages and percentage incidence have been used to describe the results. Of the 74 study participants, 31 (41.89%) were males and 43 (55.40%) were females. Six had only TP, seven had only TM and three participants had both TP and TM. The most of the palatal tori had spindle shaped, located in all area of the hard palate The round mandibular tori with big size were located in the area of both premolars, and those with elongate shape were located in the canine-premolars area. The palatal tori were more frequently in women and the frequency of mandibular tori was equally in men and women. Most of the palatal tori had spindle shape and most of the mandibular tori were solitary bilateral.

Corresponding author: Monica Scrieciu, Associate Professor, MD, PhD; e-mail: scrieciu_monica@yahoo.com

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17. Experimental measurement of flexion-extension movement in normal and corpse prosthetic elbow joint

Daniela Tarnita, Danut-Nicolae Tarnita

This paper presents a comparative experimental study of flexion-extension movement in healthy elbow and in the prosthetic elbow joint fixed on an original experimental bench. Measurements were carried out in order to validate the functional morphology and a new elbow prosthesis type ball head. The three-dimensional (3D) model and the physical prototype of our experimental bench used to test elbow endoprosthesis at flexion-extension and pronation-supination movements is presented. The measurements were carried out on a group of nine healthy subjects and on the prosthetic corpse elbow, the experimental data being obtained for flexion-extension movement cycles. Experimental data for the two different flexion-extension tests for the nine subjects and for the corpse prosthetic elbow were acquired using SimiMotion video system. Experimental data were processed statistically. The corresponding graphs were obtained for all subjects in the experimental group, and for corpse prosthetic elbow for both flexion-extension tests. The statistical analysis has proved that the flexion angles of healthy elbows were significantly close to the values measured at the prosthetic elbow fixed on the experimental bench. The studied elbow prosthesis manages to re-establish the mobility for the elbow joint as close to the normal one.

Corresponding author: Danut-Nicolae Tarnita, Professor, MD, PhD; e-mail: dan_tarnita@yahoo.com

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18. An optical investigation of dentinal discoloration due to commonly endodontic sealers, using the transmitted light polarizing microscopy and spectrophotometry

Ioana Suciu, Ecaterina Ionescu, Bogdan Alexandru Dimitriu, Ruxandra-Ioana Bartok, Georgiana-Florentina Moldoveanu, Irina-Maria Gheorghiu, Ileana Suciu, Mihai Ciocirdel

The aim of this study was to establish the degree of tooth crown staining by commonly used endodontic sealers. Crown discolorations by tooth canal sealers [AH Plus (Dentsply DeTrey Gmbh, Konstanz, Germany); Endofill (Produits Dentaires SA, Vevey, Switzerland); Apexit (Dentsply DeTrey Gmbh, Konstanz, Germany); and MTA Fillapex (Angelus, Londrina, Brazil)] were tested on extracted human premolars. The samples were divided into five groups of five samples each, after root canal sealing. Five teeth were used as control groups. The spectrophotometric method was performed in order to quantify in terms of color change of the coronal part (it was also recorded a track on how the color changes over time). For the microscopic study of the extracted dental specimens subjected to this study, polarized transmitted light microscopy was used. This method involves the development of special microscopic preparations, called thin sections. In our case, the thin section was performed on 20 prepared and obturated recently extracted teeth. The degree of discoloration was determined after one week and three months using spectrophotometry and polarized light microscopy. All sealers usually cause some degree of discoloration on the cervical aspect of the crowns that increases in time. AH Plus and Endofill caused the greatest discoloration, followed by Apexit and MTA Fillapex.

Corresponding author: Ioana Suciu, Associate Professor, DMD, PhD; e-mail: joa_suciu@yahoo.com

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19. Morphological aspects of the rectal neovascularization in colorectal cancer - anatomical-surgical and imaging implications

Delia Hinganu, Iuliana Eva, Cristinel Ionel Stan, Marius Valeriu Hinganu

Colorectal neoplasia has an increasing incidence among the population, and this fact compels in achieving an early diagnosis and treatment protocols. The extramural vascular invasion (EMVI) score is a method used for staging cancer. It defines the presence of malignant cells in the blood vessels, outside its own vascular tunic. The purpose of the study is to evaluate the extramural vascular invasion, and thus the impact of this determination in diagnosis, treatment and prognosis of colorectal neoplasia. This was done using magnetic resonance images (MRI) of colonoscopic diagnosed patients with colorectal cancer and subsequently comparing these results with the control group of patients without malignancy. The EMVI criteria taken into account were correlated with the images obtained in patients in the study group. In each of the evaluated patients, we found both, arterial and venous blood vessels in 0 stage - EMVI correlated with T2 or T3 stage tumor. In T3 B stages, we could objectify suggestive images for stage 4 EMVI near and distant from the tumor. Blood and lymph vessels have a high density to their origin area and it decreases anteriorly. This distribution is consistent with a predisposition to the formation of a vascular rectal cancer, especially on the posterior wall. In conclusion, anatomical-imaging staging of colorectal cancers using EMVI score is very close to the colonoscopic diagnosis and to the accuracy of the classic cancer staging. Corroborating EMVI score with a vascular radiology reference pattern improves the accuracy for determining an appropriate treatment and assessment of prognosis.

Corresponding author: Delia Hinganu, University Assistant, MD, PhD; e-mail: delia_f24@yahoo.com

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20. Study of CK18 and GDF5 immunoexpression in oral squamous cell carcinoma and their prognostic value

Mihai-Catalin Afrem, Stefania Craitoiu, Mihaela-Cezarina Hincu, Horia Octavian Manolea, Vasile Nicolae, Monica Mihaela Craitoiu

Oral cavity cancer remains one of the most common cancers worldwide, with an increased incidence in young adults, although there has been lately a decrease tendency in the incidence of this form of cancer. Lingual localization has a very high mortality and tends to be more aggressive becoming frequently metastatic at the regional lymph nodes. The purpose of this study is to investigate the immunohistochemical expression of cytokeratin 18 (CK18) and the reactivity to GDF5 (CDMP-1), called the morphogenetic protein-1, cartilage-derived, in lingual squamous cell carcinoma and the correlation between the immunoreactivity of this panel of antibodies, and the clinical stage, the degree of differentiation and the invasion pattern. In this regard, we studied the immunohistochemical behavior of these markers in 15 cases of lingual squamous cell carcinoma. In our study, we observed the correlation of CK18 and GDF5 expression with the clinical stage, differentiation degree and invasion pattern, the highest levels of immunoreactivity being recorded in poorly differentiated forms, in high-level invasion patterns and in the most advanced stages. The markers used can become therapeutic targets, which could help increase the quality of life and life expectancy for these patients.

Corresponding author: Stefania Craitoiu, Professor, MD, PhD; e-mail: scraitoiu@yahoo.com

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21. Assessment of chromosomal aneuploidies in sperm of infertile males by using FISH technique

Nicoleta Ioana Andreescu, Mirela Cosma, Simona Sorina Farcas, Monica Stoian, Daniela-Georgiana Amzar, Maria Puiu

Reproductive failure is one of the most important issues for the population at age of procreation and approximately 15% of the couples who try to conceive a baby encounter reproductive difficulties. In this study, we used multicolor fluorescent in situ hybridization (FISH) probes for chromosomes 13, 18, 21, X and Y to evaluate the aneuploidy incidence in sperm cells. The study group included 35 males with infertility and oligoasthenoteratozoospermia (OAT) and 20 males with normal fertility and normal semen characteristics for which the conventional cytogenetic investigation using peripheral blood revealed a normal karyotype. The overall chromosome disomy and nulisomy in OAT group was higher than the one identified in the control group. By comparing the incidence of the disomy in the OAT group, the highest incidence was the sex chromosome disomy, followed by the disomy of chromosomes 13, 21 (equal values) and then 18. The nulisomy incidence in the OAT group was higher for sex chromosomes, followed by the nulisomy of autosomes 13, then 21 and 18. As in these days, for patients with OAT, intra-cytoplasmic sperm injection (ICSI) is frequently used, it is important to inform the patients if they might have an increased risk of aneuploidies in embryos.

Corresponding author: Nicoleta Ioana Andreescu; e-mail: nicollandreescu@yahoo.com

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22. Immunohistochemical study of the epithelial and stromal components of Warthin s tumor

Luminita Daguci, Alex Emilian Stepan, Veronica Mercut, Constantin Daguci, Marilena Bataiosu, Oana Cella Andrei, Ionela Teodora Dascalu, Alma Maria Florescu, Cristiana Eugenia Simionescu

Warthin s tumor is a benign monomorphic adenoma with unclear origin with the highest incidence in the sixth and seventh decades. The analysis of tumor markers involved in the pathogenesis of Warthin s tumor can improve the patients prognosis. This study included 29 cases of Warthin s tumor, which were histopathologically and immunohistochemically examined for different compartments of tumors. For immunohistochemical study, we used as specific markers for epithelial compartment CD117, CEA and AMA, respectively S100 and D2-40 for the stromal compartment. The evaluation of immunoreactions was performed by semiquantitative analysis. The analysis of the CEA, CD117 and AAM immunoexpression allowed observing various patterns of immunostaining for tumor double-layered epithelia, which has the tendency of being similar to that in the normal ductal epithelia. S100 protein positivity similar to Langerhans cells suggests that delayed hypersensitivity can be involved in tumor development. The presence of D2-40 expression in majority of tumor subcapsular vessels, similar to lymph nodes structure, confirms the hypothesis that Warthin s tumor has its origin in regional lymph nodes.

Corresponding author: Alex Emilian Stepan, Lecturer, MD, PhD; e-mail: astepan76@yahoo.com

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23. Etiological factors associated with temporomandibular joint disorder - study on animal model

Andrea Maria Chisnoiu, Radu Chisnoiu, Marioara Moldovan, Liana Maria Lascu, Alina Monica Picos

The etiology of temporomandibular disorder (TMD) is multidimensional: biomechanical, neuromuscular, bio-psychosocial and biological factors may contribute to the disorder. The main objectives of our study were investigation and analysis of the degree of involvement for several presumptive etiological factors of TMD: biomechanical stress (BS), estrogen hormones (EH) and emotional stress (ES). Six groups (n=10) of mature female Wistar rats were included in the study. Single presumptive etiological factor was applied in three groups (BS, EH and ES groups) and also association of presumptive etiological factors were applied in two groups (BSEH and BSES groups). No etiological factor was applied for the control group. Animals were sacrificed after a 60 days period and histological analysis of the temporomandibular joint (TMJ) tissues was performed. The changes in the mandibular cartilage, articular disc, temporal bone and synovial tissue were observed under optical magnification and quantified. All samples developed changes in the thickness of the condylar cartilage comparing to control group. The reduction was highly statistical significant for the EH, ES and BSES groups (p<0.001) and statistical significant for the BS and BSEH groups (p<0.05). The most important modifications with severe cartilage thickness reduction have been obtained in case of BSES group. In conclusion, biomechanical, emotional stress and estrogens can be considered as possible etiological factors in TMD.

Corresponding author: Alina Monica Picos, DMD, MD, PhD; e-mail: alinapicos@yahoo.com

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24. Clinical study on thermography, as modern investigation method for Candida-associated denture stomatitis

Laura Iosif, Cristina Teodora Preoteasa, Catalina Murariu-Magureanu, Elena Preoteasa

Candida-associated denture stomatitis is an infectious inflammatory condition of the oral mucosa, with frequent recurrences. The aim of this study was to assess the use of infrared thermography as investigation method for Candida-associated denture stomatitis (as inflammatory disorder of the maxillary denture bearing area), by comparing disease and non-disease groups. An observational study was conducted on maxillary edentulous patients treated by acrylic dentures, with and without Candida-associated denture stomatitis. Diagnostic test methods used were clinical examination for denture stomatitis and conventional microbiological culture method for oral candidiasis. Thermography analysis of the maxillary denture bearing area was made using the ThermaCAM PM350 infrared camera (Inframetrics, Flir Systems) and ThermaGram Pro 95 software, data being acquired by usage of standard protocol of thermographic registrations. The sample included 52 patients, 21 with and 31 without Candida-associated denture stomatitis. The temperature of the maxillary mucosa corresponding to the denture bearing area was found to be statistically significantly higher in Candida-associated denture stomatitis (mean 36.20 degrees C), compared to healthy oral mucosa (mean 34.85 degrees C). The thermal threshold value of 35.44 degrees C was identified as best differentiating a pathological from normal state of the maxillary mucosa corresponding to the denture bearing area. In conclusion, infrared thermography, a rapid non-invasive investigation method, has the premises to bring valuable data in inflammatory disorders of the maxillary denture bearing area, as Candida-associated denture stomatitis that may be used for screening, diagnostic or monitoring purposes.

Corresponding author: Cristina Teodora Preoteasa, Assistant Professor, DMD, PhD; e-mail: cristina_5013@yahoo.com

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25. Impression conjunctival cytology in sicca syndrome - correlations between clinical and histological findings related to dry eye severity

Carmen-Luminita Mocanu, Sanda Jurja, Andreea-Gabriela Deca, Florica Birjovanu, Andrei Olaru, Denissa-Greta Popa, Alin-Stefan Stefanescu-Dima, Maria-Rodica Manescu

Keratoconjunctivitis sicca represents a progressive deterioration of ocular surface produced by a deficient secretion of lachrymal film (quantitative disorder) or excessive tear evaporation (qualitative disorder). The cytological analysis of conjunctival impression in 42 patients with dry eye syndrome established a strong correlation between the clinical grade of severity of disease and the grade of squamous metaplasia, including goblet cell loss. The cellular anomalies were represented by modifications of keratinization, epithelial cells anisocytosis, anisochromia, the nuclear condensation and the cytoplasmic vacuolization. Pyknotic nuclei and anucleated cells were only seen in the most severe dry eye. The modifications in epithelial cells and conjunctival goblet cells reveal cellular sufferance, with an evident parallelism between these anomalies and clinico-functional signs in dry eye. Conjunctival impression provides an easy and quick identification of the lachrymal film alterations with high specificity and sensitivity, giving valuable information about the qualitative disorder.

Corresponding author: Andreea-Gabriela Deca, University Assistant, Pharm, PhD; e-mail: gabriela_deca@yahoo.com

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26. E-cadherin expression in primary colorectal cancer and metastatic lymph nodes

Madalina Palaghia, Catalina Mihai, Ludmila Lozneanu, Delia Ciobanu, Ana-Maria Trofin, Alexandra Rotariu, Filip Tarcoveanu, Cristina Cijevschi Prelipcean

Colorectal cancer (CRC) is one of the most prevalent malignancies and fourth cause of cancer death worldwide but the current TNM staging system together with clinicopathological characteristics are not sufficient to identify cases that have a poor prognosis. The aim of the current study was to compare E-cadherin expression in primary CRC and lymph node metastases with the one exhibited by normal colic mucosa and normal lymph nodes, and to evaluate its association with disease severity. The authors retrospectively analyzed 65 patients that underwent colectomy for CRC at the First Surgical Clinic, Sf. Spiridon Emergency Hospital from Iassy, Romania, over a 10 years period, from January 2004 to December 2013. In all cases, immunohistochemical staining against E-cadherin was performed on primary CRC and associated lymph nodes slices. Primary CRC presented a higher rate (64.62%) of abnormal E-cadherin expression (no staining, cytoplasmic or mixed staining) compared to normal colic mucosa (16.67%) (p<0.001). Both primary CRC and corresponding metastatic lymph nodes displayed a predominantly membranous expression (pure or mixed) of E-cadherin (67.69% and 89.23%, respectively). Well and moderately differentiated tumors displayed an increased E-cadherin expression (44 of 56 cases) compared to poorly differentiated tumors that lacked E-cadherin expression in six out of nine cases. In conclusion, E-cadherin expression abnormalities in CRC are rather qualitative than quantitative and E-cadherin is an important marker of tumor aggressiveness and spreading potential.

Corresponding author: Catalina Mihai, MD, PhD; e-mail: catalinamihai@yahoo.com

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27. New technical procedure involving Achilles tendon rupture treatment through transcutaneous suture

Danut-Nicolae Tarnita, Daniela Tarnita, Dan Cristian Grecu, Dan Marian Calafeteanu, Bogdan Capitanescu

The Achilles tendon is the widest tendon of the human body. Achilles tendon belongs to the extrasynovial tendons group and this allows it a faster recovery, thanks to local hematoma from the peritenon, necessary for the scarification. We concluded that in Achilles tendon rupture treatment it is essential to maintain the tendon covering skin integrity, the peritendinous integrity, to maintain the local hematoma formed during and after tendon rupture, reattaching the ruptured tendon heads and maintain them in this position by suturing them and by relaxing the sural triceps muscle. The percutaneous suture requires five pairs of mirror micro-incisions (5 mm) on one side and the other of the tendon. It is necessary for one of the pairs to be placed to the rupture level. With a surgical needle, we arm the proximal and distal heads of the tendon by different threads. By traction and muscular relaxation, we bring in contact the two ruptured heads and then we knot together the arming threads. The inferior member was cast immobilized in relaxing position for the sural triceps muscle for a 45 days period. Using this technique, we have operated 15 cases in our Clinic. In all the cases, we obtained a healing by first intention of the tegument micro-incisions. After the cast immobilization suppression, during 30 days the patients were in a recovery program. At the end of this program, they have recovered completely the dorsal and plantar flexion and the walking. In four months after the surgery, the esthetic of the area is completely restored, this technique being the only surgical technique that realizes this recovery.

Corresponding author: Dan Cristian Grecu, Associate Professor, MD, PhD; e-mail: grecudan@ortopezi.ro

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28. Rheumatoid nodules and quality of life in rheumatoid arthritis females - complex assessment

Magdalena-Rodica Traistaru, Diana Kamal, Diana-Maria Trasca, Maria-Camelia Foarfa, Corina-Lavinia Gruia, Otilia-Constantina Rogoveanu

Rheumatoid arthritis (RA) represents the most commonly diagnosed arthropathy that affect many tissue types and organs, characterized by a clinical, functional and therapeutic pathogenic complexity and it affects especially diarthroidal joints. Rheumatoid nodules (RNs) are one of the most frequent extra-articular manifestations of RA, and usually reflect an advanced stage of the disease and a poor prognosis. The complexity of histological, clinical and functional aspects in RA has a real impact on the quality of life in all patients diagnosed with this disorder. Our prospective study presents the RNs involvement in the rehabilitation program performed in order to enhance the quality of life in the 25 RA female patients. We made a complex assessment and realized a correlation between pain, disability and histological aspect of RN, before and after the rehabilitation program. Also, we evaluated the clinical and functional effectiveness of a complex rehabilitation program and changes in impairment and activity limitation in women with RA and RNs. The immunohistological complexity of RNs reflects the intensity of the inflammatory-immune process and completes the assessment of RA patients with RNs. It allows for medical assistance quantification, even for patients that have a poor evolution prognosis.

Corresponding author: Magdalena-Rodica Traistaru, Associate Professor, MD, PhD; e-mail: rodicatraistru@hotmail.com

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29. The influence of irradiation on autophagy process in normal and malignant colorectal epithelia

Constantin Daniel Uscatu, Francisc Mixich, Theodor Viorel Dumitrescu, Elena-Raluca Nicoli, Ionela Cristina Deliu, George Mihail Man, Iancu Emil Plesea

Aim: The authors assessed the influence of preoperative radiotherapy on autophagy process using a quantitative assessment of LC3 expression on both normal and tumoral colorectal tissues. Materials and Methods: Normal and malignant tissue samples were taken from 50 patients that underwent surgery for colorectal adenocarcinoma of which 11 received preoperative radiotherapy. Tissue samples were included in paraffin and sections were immunomarked for LC3 expression. LC3 percentage was assessed with dedicated software on 10 randomly selected fields with 40x objective from both normal and malignant tissue samples of each patient. The resulting data were assessed and compared with a statistical apparatus. Results: LC3 was overexpressed in tumoral tissue as compared with normal one. The LC3 percentage is different from person to person and the higher it is in normal epithelium, the higher will be in tumoral epithelium of the same person, regardless the irradiation. The LC3 expression levels are decreasing from tumoral non-irradiated epithelia to normal irradiated epithelia. LC3 expression in tumoral cells is granular, with particular perinuclear disposal and often annular pattern. Conclusions: The autophagy process has a basal level in the normal tissue, with interindividual variability. The autophagy process proved to be upregulated in the tumoral cells, with a particular morphological expression, namely the presence of cytoplasmic coarse granules disposed in an annular pattern. Preoperative radiotherapy is downregulating the autophagy process both in normal and tumoral tissue but to a lesser extent in the latter.

Corresponding author: Iancu Emil Plesea, Professor, MD, PhD; e-mail: pie1956@yahoo.com

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30. Diagnostic and therapeutic aspects in malignant sinonasal lymphoma

Mihail Tusaliu, Viorel Zainea, Carmen Aurelia Mogoanta, Ana Alexandra Dragu, Cristina Maria Goanta, Maria Nitescu, Marina-Ruxandra Otelea, Vlad Andrei Budu

Among head and neck malignancies, lymphoma is the second most frequent cause. They can develop in the head and neck lymph nodes or as extranodal determinations. The onset of malignant lymphoma outside of lymph node registers increasingly frequent, currently over 30% of malignant lymphoma are diagnosed as taking place outside the lymph nodes. The aim of this paper is bringing in discussion and the presentation of current elements on the diagnosis and treatment in case of malignant sinonasal lymphoma, a pathological entity with a growing incidence. This study is a retrospective one, from January 2008 through December 2013, and included 31 patients admitted to Prof. Dr. Dorin Hociota Institute of Phonoaudiology and Functional ENT (Ear, Nose and Throat) Surgery, Bucharest, Romania, and diagnosed with malignant sinonasal lymphoma. Despite progress in terms of immunological techniques, cytogenetic and molecular histological outlining of malignant lymphomas, correct diagnosis and appropriate therapy of malignant sinonasal lymphoma is still a problem for the clinician and pathologist, as multidisciplinary collaboration of ENT-Hematology-Pathology-Radiotherapy being essential in this regard.

Corresponding author: Mihail Tusaliu, MD, PhD; e-mail: mtusaliu@yahoo.com

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31. Tuberculous constrictive pericarditis complicated with tuberculous mediastinitis - case report

Milena Adina Man, Mimi Floarea Nitu, Lelia Strambu, Cristina Florescu, Costin Teodor Streba, Antigona-Carmen Trofor

Constrictive pericarditis is a rare and severe disease. A 37-year-old patient was admitted in the hospital for dyspnea, precordial pain, right-sided cardiac failure. Chest X-ray showed cardiac enlargement and an opacity suggestive for pleural effusion. Echocardiography revealed an adhesive-effusive-constrictive pericarditis, a very thickened pericardium and bilateral pleural effusion. After a pericardiectomy done to restore cardiac compensation and to identify etiological factors, a tuberculous pericarditis (TBP) was diagnosed. After surgery and starting anti-TB treatment, the patient presented altered clinical status, dyspnea, dry cough, fever and delayed callus formation at sternum level. Thoracic scan revealed mediastinal air collections, pericarditis and pleurisy. Thus, the TBP diagnosis was extended to mediastinal TB and anti-TB therapy was continued. After four months of treatment, another thoracic scan showed disappearance of the mediastinal air-leakage bubbles, multiple new micronodules in both lungs and lymph nodes of up to 15 mm; also increasing pericardial and pleural effusions. This case was interpreted as a TB treatment failure situation. A retreatment regimen was started, resulting in a slow favorable outcome. Pericardial TB is a rare condition, usually with delayed diagnosis and poor treatment benefits. Whenever possible, earlier diagnostic can contribute to better management of these cases.

Corresponding author: Mimi Floarea Nitu, Professor, MD, PhD; e-mail: dr_nitumimi@yahoo.com

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32. Mastoiditis and facial paralysis as initial manifestations of temporal bone systemic diseases - the significance of the histopathological examination

Alma Aurelia Maniu, Oana Harabagiu, Laura Otilia Damian, Eugen Horatiu Stefanescu, Bogdan Marius Fanuta, Andreea Catana, Carmen Aurelia Mogoanta

Several systemic diseases, including granulomatous and infectious processes, tumors, bone disorders, collagen-vascular and other autoimmune diseases may involve the middle ear and temporal bone. These diseases are difficult to diagnose when symptoms mimic acute otomastoiditis. Case reports: The present report describes our experience with three such cases initially misdiagnosed. Their predominating symptoms were otological with mastoiditis, hearing loss, and subsequently facial nerve palsy. The cases were considered an emergency and the patients underwent tympanomastoidectomy, under the suspicion of otitis media with cholesteatoma, in order to remove a possible abscess and to decompress the facial nerve. The common features were the presence of severe granulation tissue filling the mastoid cavity and middle ear during surgery, without cholesteatoma. The definitive diagnoses was made by means of biopsy of the granulation tissue from the middle ear, revealing granulomatosis with polyangiitis (formerly known as Wegener s granulomatosis) in one case, middle ear tuberculosis and diffuse large B-cell lymphoma respectively. After specific associated therapy facial nerve functions improved, and atypical inflammatory states of the ear resolved. Conclusions: As a group, systemic diseases of the middle ear and temporal bone are uncommon, but aggressive lesions. After analyzing these cases and reviewing the literature, we would like to stress upon the importance of microscopic examination of the affected tissue, required for an accurate diagnosis and effective treatment.

Corresponding author: Alma Aurelia Maniu, Lecturer, MD, PhD; e-mail: almacjro@yahoo.com

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33. Morphological study of cephalothoracopagus deradelphus type conjoined twins. A case report

Monica Mihaela Cirstoiu, Florin Mihail Filipoiu, Elvira Bratila, Costin Berceanu, Florin Catalin Cirstoiu, Vlad Andrei Budu, Ioan Alexandru Bulescu, Octavian Munteanu

Cephalopagus is a rare variety of conjoined twins. They are fused with their heads, thoracic and upper abdominal cavities. The exact mechanism for development of conjoined twins cannot be clearly explained. It appears that there is an alteration in the normal developmental process of monozygotic twins, which fail to separate from each other. We present the morphology of a cephalothoracopagus, revealed through anatomical dissection, emphasizing the arrangement of the viscera in the thoracic and abdominal cavities. They are fused with their heads, thoracic and upper abdominal cavities. The lower abdomen and pelvic cavities are free. Each twin has two upper and lower limbs, normally shaped. Each twin has a heart and two lungs. There is a single pharynx, esophagus and stomach, but normal lower abdominal systems. The genital and urinary systems are apparently normal. Due to the fusion of the heads and abnormal arrangement of the superior central nervous system, surgery is not attempted in these cases, the prognosis being very poor.

Corresponding author: Ioan Alexandru Bulescu, MD, PhD; e-mail: ibulescu@gmail.com

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34. EGIST of the greater omentum - case study and review of literature

Mihail-Gabriel Dimofte, Vlad Porumb, Dan Ferariu, Nicoleta-Camelia Bar, Sorinel Lunca

The development of immunohistochemical methods has outlined a particular group of tumors, with very specific features and treatment, originating in the Cajal cells of the muscularis propria or related stem cell-like precursors present in the wall of the digestive tract called gastrointestinal stromal tumors (GISTs). A sub-segment with similar features may develop outside the digestive tract, namely extra-gastro-intestinal stromal tumors (EGISTs). From the small category of EGISTs, we report on a case of a primary epithelioid EGIST of the greater omentum, which is seldom reported in literature. The tumor was diagnosed in a man with non-specific symptoms who presented for abdominal enlargement. The tumor was characterized and there was a preoperative suspicion of a non-digestive tumor located in the greater omentum. The tumor was surgically removed showing no contact with adjacent organs. Immunohistochemical examination was consistent with a primary EGIST of the greater omentum. Treatment with Imatinib mesylate was started and at two-year follow-up, the patient is disease free. The case raises problems regarding pathogenesis, immunohistochemical features, behavior, evolution and prognosis of omental EGIST, for which no significant or conflicting data are available in the literature.

Corresponding author: Vlad Porumb, Assistant Professor, MD, PhD student; e-mail: porumbvlad@ymail.com

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35. The TRAP (twin reversed arterial perfusion) sequence - case presentation

Claudiu Marginean, Maria Oana Marginean, Daniel Muresan, Luminita Zahiu, Emoke Horvath

We present a particular case of TRAP (twin reversed arterial perfusion) syndrome, which has a very rarely association of the simultaneous existence of a rudimentary malformed heart and brain, and also other malformations like abdominal wall abnormality, absent bladder with present kidneys, and absence of the lungs, which appear only in a few cases on the receptor twin from this sequence, malformations incompatible with life. A Caucasian 26-year-old pregnant woman, at the first pregnancy, with a monochorionic-diamniotic pregnancy, 26 weeks of gestation was referred to our hospital, for polyhydramnios. The patient delivered a living female newborn, weighing 950 g, with an Apgar score of 2 at one minute - the donor fetus and a second female newborn with multiple malformations, no signs of life and who weighed 2300 g - the receptor fetus. The anatomopathological examination confirmed the TRAP sequence associated with severe facial dysmorphism, bilateral phocomelia and cardiac malformations (rudimentary hypoplastic, univentricular) and a vascular anastomosis between the two umbilical cords. Anemia and cardiac complications which can lead to cardiac failure, appear early during pregnancy and caused the death of the pumping twin. We emphasize that in our case of TRAP sequence, the ultrasound examination established the diagnosis of the syndrome with high accuracy. Therefore, we can conclude that the existence of a rudimentary heart and a vascular anastomosis between the two umbilical cords supports the apparition of TRAP sequence. The early diagnosis of this pathology, the observation of the pregnancy with the help of weekly ultrasounds and the intrauterine interventions can increase the survival chances of the donor fetus from the TRAP sequence.

Corresponding author: Maria Oana Marginean, MD; e-mail: oanamarg93@gmail.com

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36. Squamous esophageal carcinoma and mucinous adenocarcinoma of the colon - an unusual association

Cecil Sorin Mirea, Manuela Ioana Vasile, Ionica Daniel Vilcea, Ion Vasile, Emil Moraru, Mihai Calin Ciorbagiu, Maria Cristina Sfeclan, Catalina Marin, Vasile-Cosmin Obleaga, Ioana-Andreea Gheonea, Alina Maria Vilcea

The existence of a simultaneous cancer of the esophagus and colon is a rare situation that recognizes an increased incidence in recent years in the world, probably as a result of the improved measures of diagnosis and treatment, as well as the development of screening programs. The aim of this work is to present a case of synchronous esophageal squamous carcinoma with mucinous adenocarcinoma of the hepatic angle of the colon. The patient was hospitalized to our Surgical Clinic with the thoracic squamous esophageal carcinoma diagnosis. On admission, symptoms were dominated by overall dysphagia, patient showing a weight loss of 10 kg for the last 30 days. Preoperative imaging tests did not revealed regional or distant metastatic disease. Preoperative colonoscopy was incomplete (only until the splenic angle of the left colon) due to the insufficient mechanical preparation. On laparotomy, a carcinoma of the hepatic angle of the colon, partially stenosing was discovered. An upper pole esogastrectomy with intrathoracic esogastrostomy and a right colectomy with ileotransversostomy were practiced, at the same operative session. Postoperative evolution was poor and the patient died on the ninth day from the surgery during an alcohol withdrawal crisis.

Corresponding author: Ionica Daniel Vilcea, Associate Professor, MD, PhD; e-mail: id.vilcea@yahoo.com

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37. AIDS-related lymphoma in a young HIV late presenter patient

Manuela Arbune, Costinela Valerica Georgescu, Doina Carina Voinescu

Non-Hodgkin s lymphoma (NHL) is an acquired immunodeficiency syndrome (AIDS)-indicative disease. Nowadays, NHL is rarely reported in Europe as indicative disease for human immunodeficiency virus (HIV) testing. We present the case of a 22-year-old Romanian male patient without past medical history, except the swelling of a submental lymph node 11 months ago. The excised node was histologically examined but the patient neglected to take his result. He was admitted for fever, asthenia, and weight loss over 10% of his weight, and night sweats in the last four months. The immunohistochemical analysis of the preserved lymph node samples suggested reactive hyperplasic lymphadenitis with suppuration and necrosis (lymphoid follicles CD20+, CD10+, BCL6+; germinal centers CD23+, CD68+, Ki67+; and interfollicular CD3+). Clinical, biological and imaging evaluations were performed. The diagnostic of lymphoma stage IV Ann Arbor was sustained. Severe immunosuppression and a positive HIV test were found. The patient received antiretroviral treatment, but he developed paraplegia consecutive to a vertebral metastasis, liver and kidney failure and died sooner than two months from the diagnostic time. Pathological examination confirmed NHL with diffuse lymphocyte infiltrate of multiple organs. Advanced lymphoma is a rare indicator condition of HIV diagnostic. Delayed diagnostic of lymphoma implies ethical issues on communication deficiencies between the heath providers and patients, concerning the significance of biopsy. Infectious co-morbidities with necrosis and suppurative lesions are confounder conditions in NHL histological and immunohistochemical diagnosis.

Corresponding author: Costinela Valerica Georgescu, Associate Professor, MD, PhD; e-mail: costinelag@gmail.ro

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38. Heterotopic mesenteric ossification of ileostomy - intraabdominal myositis ossificans

Konstantinos Sapalidis, Titika-Marina Strati, Lazaros Liavas, Efstathios Kotidis, Triantafilia Koletsa, Fani Tsiompanou, Ioannis Kanellos, Dinu Florin Albu

Heterotopic mesenteric ossification is a rare clinical entity with less than 40 reported cases. Moreover, heterotopic mesenteric ossification around a stoma is described in fewer cases. Case presentation: An 81-year-old male patient, who underwent a sigmoidectomy two years ago for a sigmoid colon cancer, was admitted in our Department in order to be investigated for a local recurrence of his disease. Because of unresectable pelvic lesions, the patient was taken in the operating room for a palliative double barrel ileostomy. The patient presented two weeks after the ileostomy clinical signs suggesting its obstruction. A revision of the ileostomy is performed. Macroscopically, the end part of the ileostomy was inflamed and resected. Histological findings revealed heterotopic ossification of the mesentery. Conclusions: The surgeon should be alert to the possibility of heterotopic ossification, especially within a previously operated wound and be prepared for the difficulties it may cause during reopening of an incision, during the operation itself, at closure of the wound and, if not removed, also in the postoperative setting.

Corresponding author: Konstantinos Sapalidis, Assistant Professor, MD, PhD; e-mail: sapalidis@med.auth.gr

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39. Multiple asymptomatic cutaneous pilar leiomyoma versus spontaneous eruptive keloids - a case report

Silviu Horia Morariu, Mircea Suciu, Mihai Alexandru Badea, Mihai Dorin Vartolomei, Corneliu Florin Buicu, Ovidiu Simion Cotoi

Cutaneous piloleiomyoma, angioleiomyoma and genital leiomyoma are variants of superficial cutaneous leiomyoma. The main purpose of this paper was to present clinical, histopathological and immunohistochemical diagnosis criteria for an unusual case of pilar leiomyoma in an 18-year-old male patient. The initial clinical aspect was very similar to spontaneous eruptive keloids: red-violet color, painless, aspect of crab pincers of some of the lesions, epidermis atrophy, telangiectasia, located on acneiform zones and compliance with cephalic extremity. The patient had no history of trauma, surgery or acne. Local treatment of one lesion was performed with cryotherapy using liquid nitrogen (-172 degrees C) together with intra-lesion steroid injections, occlusive dressings and silicone gel. Local therapy did not showed notable results, moreover the lesion become painful. Skin biopsy with histological and immunohistochemical analysis revealed the diagnosis of multiple cutaneous pilar leiomyoma without atypia. The particularity of the case stands in the atypical onset followed by explosive increasing of lesions number and the appearance of pathognomonic pain after local therapy.

Corresponding author: Mircea Suciu, Associate Professor, MD, PhD; e-mail: suciu.mircea@umftgm.ro

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40. Clinical and histological aspects of malignant inverted sinonasal papilloma. Two case reports

Vasile Nicolae, Mariana Sabau, Mihai Iulian Fagetan, Vlad Petrescu Seceleanu, Daniel Iorgulescu, Dana-Elena Dumitra, Alexandru-Dan Sabau, Sorin Ibric-Cioranu, Anca Sava, Viorel Ibric-Cioranu

The inverted sino-nasal papilloma is a benign epithelial lesion, ranging from 1% to 5% of the tumors that reside in this area. It is characterized by a tendency towards local bone destruction, recurrence and malignant transformation. This is a presentation of two cases diagnosed and surgically treated for malignant inverted papiloma, which started in the epithelium of the maxillary sinuses. The cases had in common the late presentation of the patient to the surgeon, thus the local destruction was significant with extension towards the eye socket and the base of the skull. The treatment of choice was the surgical resection of the maxillary bone with a prosthetic reconstruction of the defect followed by radiotherapy. After a careful analysis of the two cases, a series of statements regarding the diagnosis and treatment of this rare pathology were recommended.

Corresponding author: Sorin Ibric-Cioranu, PhD Student; e-mail: maxfaxsurg@yahoo.com

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41. Orbital lymphoma with isolated occult bilateral adrenal involvement: report of an extremely rare case

Tatjana Terzic, Dejan Rasic, Ruzica Nedeljkov-Jancic, Dragana Pesic, Snezana Lesovic, Vladimir Jurisic

Orbital lymphoma is very rare malignant neoplasm, usually diagnosed in early stage of disease as primary lymphoma, but dissemination occurs in approximately 33% of cases. Isolated bilateral adrenal lymphomatous involvement is extremely rare, described in 0.83% of cases. We present autopsy case of a 63-year-old man with bilateral orbital diffuse large cell lymphoma, clinical stage IEA, successfully treated by one cycle of chemo- and radiotherapy, but after administration of the second cycle, the patient developed signs of gastrointestinal hemorrhage and died two months after the diagnosis. Autopsy findings exclude lymphoma involvement of any organ except histopathological infiltration of both adrenal glands without evidence of a mass lesion.

Corresponding author: Tatjana Terzic, PhD; e-mail: tatjana.terzic@sbb.rs

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42. Ultrasonography role in the evaluation of a giant cell tumor of the flexor pollicis longus tendon

Marius Eugen Ciurea, Ananu-Florentin Vreju, Dragos-George Popa, Raluca Niculina Ciurea, Cristina Dorina Parvanescu, Andreea-Beatrice Chisalau, Anca Rosu, Paulina-Lucia Ciurea

Hand tendons lesions represent a challenge for an accurate diagnosis, an optimal treatment strategy, the description of the lesion and its location being an important step. The non-invasive ultrasound evaluation was demonstrated to be an important diagnostic method in these types of lesions, especially in those situations where clinical evaluation failed to reveal the pathological changes and therefore has an important role in the adequate management.

Corresponding author: Ananu-Florentin Vreju, Teaching Assistant, MD, PhD; e-mail: florin_vreju@yahoo.com

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43. A case of extrapleural solitary fibrous tumor of the thigh with eight years follow-up

Cristian Sorin Hariga, Sanda Claudia Achim, Adina Carmen Savu, Valentin Enache, Cristian Radu Jecan

Solitary fibrous tumor (SFT) is a rare neoplasia. Now classified as SFT, it was first described in 1942 as hemangiopericytoma (HPC), and its origin was supposed to be the pericytes. The location outside the pleura is considered uncommon and the tumor situation on the thigh is much more unusual. In this article, we present a case of a SFT located in the hip. Even if the limb situation of the SFT is considered rare, some tumors in lower limb were reported in the past few years. Our initial clinical diagnosis, in this case, was a variant of lipoma (fibrolipoma), so no further investigations were performed before surgery at that time, and the surgery was planned as a simple procedure. The mimics of SFT with a large variety of benign tumors, especially in long-term evolution cases, make the preoperative diagnosis much more difficult. After the excision of the tumor and histological and immunohistochemical (IHC) tests, the diagnosis was finally stated. Even the findings in the exam conducted us to a most likely benign evolution, the eight years follow-up allowed us to strongly correlate the evolution with laboratory findings in this case. Further follow-up in this case can and will be performed.

Corresponding author: Cristian Radu Jecan, Lecturer, MD, PhD; e-mail: radujecan@yahoo.com

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44. Atypical case of Sjogren s syndrome with psychiatric and peripheral neurological disorder

Nicoleta Anamaria Pascalau, Felicia Liana Cioara, Elena Rosca, Gabriela Mutiu, Liana Oana Pobirci, Cristian Marius Jinca, Laura Monica Georgescu, Razvan-Marius Vicas

Sjogren s syndrome is a rare disorder of the immune system characterized by the chronic lymphocytic infiltration of the organs with exocrine secretion (lachrymal, salivary glands), but also of other tissues of the body, that can be primary or secondary and can appear alone or in association with other systemic diseases: rheumatic arthritis, systemic erythematous lupus, scleroderma or polymyositis/dermatomyositis. The case that we are presenting is that of a 40-year-old man, who came to the Department of Rheumatology with articular, muscular, ocular, psychological and neurological symptoms. After multiple biological, immunological, histological, neurological, psychiatric, ophthalmological, digestive investigations, it was reached the conclusion that the patient presents a rare autoimmune disease (primary Sjogren s syndrome) involving mainly peripheral neuromuscular and psychological (small frequency) and the patient was given specific immunomodulatory, anti-inflammatory and anti-depressive treatment, to which he responded well. Thus, after 18 months of investigation, severe depressive episodes and difficult collaboration of the patient with the medical team, it was possible to reach the definitive diagnosis and to perform the appropriate treatment.

Corresponding author: Nicoleta Anamaria Pascalau, MD, PhD; e-mail: n.pascalau@yahoo.com

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45. Diffuse large B-cell lymphoma and polycythemia vera discovered at the onset - a rare association and its possible importance in lymphoma prognosis

Viola Maria Popov, Camelia-Marioara Dobrea, Mihai Popescu, Iancu Emil Plesea, Valentin-Titus Grigorean, Ruxandra-Diana Sinescu, Victor Dan Eugen Strambu

Diffuse large B-cell lymphoma (DLBCL) represent the most frequently non-Hodgkin s lymphoma (NHL) (over 30%), especially in developing countries. Many associations of NHL with another neoplasia were described following chemotherapy or radiotherapy regimens. The coexistence of DLBCL with myeloproliferative neoplasms (MPNs) JAK2V617F positive at the onset was very rare reported in the literature. We describe a clinical case of a 52-year-old man who presented both diagnoses at the onset - DLBCL and MPN - polycythemia vera (PV) type. The patient was treated with two CHOP cycles (Cyclophosphamide, Hydroxydaunorubicin, Oncovin, Prednisolone) followed by six R-CHOP (Rituximab-CHOP) cycles, together with a platelet-reducing agent, achieving remission for 20 months, followed by a relapse which is under treatment. The clonally expansion of an abnormal pluripotent hematopoetic stem cell could be responsible for both, PV and DLBCL. However, recent reports suggested the possible involvement of two different clones. The clinical significance and the role of JAK2 mutation in the evolution of patients with NHLs, including DLBCL are still unknown. Further genetic and clinical studies have to point out common gene mutations for the two diseases and their connection with the diseases behavior under the treatment. Conclusions: The coexistence of NHLs and especially DLBCLs and MPNs JAK2 positive is very rare. Although DLBCL alone has good prognosis, other prognostic factors should be checked when it is associated with PV. The presence of JAK2V617F seems to be a candidate but whose role in DLBCL evolution, natural or under treatment has to be cleared up.

Corresponding author: Mihai Popescu, Professor, MD, PhD; e-mail: mihaipopescu2000@yahoo.com

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46. Pulsatile tinnitus caused by a dilated left petrosquamosal sinus

Dragos Catalin Jianu, Silviana Nina Jianu, Traian Flavius Dan, Andrei Gheorghe Marius Motoc, Marioara Poenaru

The emissary veins, like the petrosquamosal sinus (PSS), are residual valveless veins, which connect the intracranial dural venous sinuses and the extracranial venous system. Rarely, they may cause pulsatile tinnitus (PT). A 22-year-old woman developed in the first week of puerperium worsening headaches, vomiting, and diplopia, and the accentuation of a PT in the left ear that she presented for eight years. The clinical examination findings nine days after delivery were unremarkable, with the exception of a left sixth nerve palsy, and a peculiar sensibility of the left temporo-mandibular joint. High-resolution computed tomography (HRCT) revealed an osseous canal in the air cells of the left temporal bone compatible with a PSS. CT and magnetic resonance (MR) imaging/MR-venogram detected signs of thrombosis of the superior sagital sinus, and of the left lateral sinus. Laboratory tests revealed severe inherited thrombophilia. We used antithrombotic therapy (body weight-adjusted subcutaneous low-molecular weight heparin for three weeks, followed by indefinite therapy with warfarin), and the headaches, vomiting, and diplopia resolved within four days of treatment. A follow-up MR-venogram performed two weeks later indicated complete recanalization of the intracranial dural venous sinuses. The PT was improved after two weeks of medical therapy, so she could adapt to it without intervention on the PSS. The early initialization of an efficient medical therapy had a great impact on her favorable evolution. PSS could be identified in her case on HRCT.

Corresponding author: Andrei Gheorghe Marius Motoc, MD, PhD; e-mail: amotoc@umft.ro

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47. Correlations between clinics, dermoscopy and histopathology in a female with two dermatofibromas - a case report

Silviu-Adrian Marinescu, Alin-Laurentiu Tatu, Ioana-Ruxandra Mihai, Carmen Giuglea

Dermatofibromas are benign fibrohistiocytic tumors that involve the dermis. They have often a polymorphous clinical aspect, being frequently confused with other lesions varying from vascular tumors to melanoma. An important tool in diagnosis is represented by dermoscopy, which facilitates the recognition of dermatofibromas characteristic structures such as central white patch and peripheral pigment network. Although dermatofibromas are often solitary lesions, we report the case of a woman who presented two fibrohistiocytary masses, involving the calf and the thigh, the last one with an atypical aspect leading to the confusion with a malignant tumor. Furthermore, the lesions were different at both dermoscopic and histopathological examinations, needing a second histopathological opinion. The aim of our study is to emphasize the role of histopathology, which represents the diagnostic golden standard in suspicious cases and the possible connections between dermatoscopy and histology. Sometimes, even histopathology may be very difficult, needing repeated sections and even special stains.

Corresponding author: Ioana-Ruxandra Mihai, MD, PhD; e-mail: andamihai07@yahoo.com

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48. Skin spiradenocarcinoma - case presentation

Laura Florentina Rebegea, Dorel Firescu, Mihaela Dumitru, Anca Patrascu

Malignant eccrine spiradenoma is an extremely rare skin tumor of sweat gland origin. The available literature data indicates that spiradenocarcinoma nearly uniformly arises from a benign preexisting spiradenoma. Etiology is unknown but previous trauma is believed being an implicated factor. The article presents the clinical observation of a 34-year-old female patient, diagnosed with cutaneous spiradenocarcinoma of the left thigh, treated multidisciplinary: surgery and chemoradiotherapy. The presentation includes clinical, histopathological and therapeutic diagnosis aspects, arisen difficulties of histological diagnosis and literature data regarding treatment options for this type of tumor. In this case, surgery followed by second surgery intervention was performed due to the continuing development of local disease. After surgery, the patient performed external beam radiotherapy (EBRT) to tumoral bed (total dose of 50 Gy) and inguinal lymph nodes (total dose of 50 Gy), followed by six cycles of chemotherapy (CMT) (Carboplatin + Paclitaxel) with complete response. At the moment, the patient shows no evidence of recurrence or metastatic disease during the follow-up.

Corresponding author: Dorel Firescu, Professor, MD, PhD; e-mail: dorelfirescu@yahoo.com

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49. Fighting for Anatomy. Overview regarding two prestigious Romanian anatomists of the 20th century: Victor Papilian and Grigore T. Popa

Cristian Barsu

During many centuries, the progress of Anatomy was based on the perseverant and laborious activities done by anatomists. Their work can be considered as a fight for Anatomy. A particular problem arouse when this fight is excessively done and it limits the ability to correctly analyze the contribution made by other scientists in the same domain. This situation was identified at different personalities who lived in the same time and were involved in the same fundamental field of research. If, theoretically, the similarities between scientists should get them closer, in order to have a better communication, the antagonisms can lead them to rivalry. Our paper exemplifies a historical case in which the personages are Victor Papilian (1888-1956) and Grigore T. Popa (1892-1948). The resemblances between these two famous Romanian anatomists from the first half of the 20th century induced an evident scientific rivalry. Papilian and Popa brought a significant contribution in anatomy. Each of them was very much appreciated by his students. It is interesting that both of them had achievements in literature. We present the reasons of their disagreement and its consequences. Paradoxically, not the contrasts, but the resemblances between their strong characters produced a sort of animosity between them. This attitude diminished in time and their successors - Ioan Albu from the Cluj Faculty of Medicine and Ion Iancu from the Jassy Faculty of Medicine - had a long lasting and successful cooperation.

Corresponding author: Cristian Barsu, Associate Professor, MD, PhD; e-mail: cristianbarsu@yahoo.com

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50. Conference Announcement: The XIVth National Symposium on Microscopic Morphology with international attendance, 4-6 May, 2016, Tirgu Mures, Romania

Laurentiu Mogoanta

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Corresponding author: -

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