ROMANIAN JOURNAL of MORPHOLOGY and EMBRYOLOGY

Vol. 54 No. 3, 2013

1. Microglia: first responders in the central nervous system

B. Catalin, A. Cupido, Maria Iancau, Carmen Valeria Albu, F. Kirchhoff

Microglia has emerged not only as an essential inflammatory cell but also as a major player in the development of the adult brain. Microglia phagocytize extra-numerical synapses during postnatal development, maintain and strengthen the remaining subset of synapses, remodel synaptic circuits and clearing apoptotic newborn neurons. Thereby, microglia plays a crucial role for the establishment, plasticity and function of adult neural circuits. In addition to the key role in normal brain function, any imbalance in microglia activity has been associated with neurodegenerative diseases. Microglial cells respond rapidly to smallest pathological changes, this being a vital aspect in many tissue scaring and the local confinement of focal lesions. It is assumed that the high motility of microglial cells represents an important requirement to fulfill the numerous functions. In this review will highlight the role of microglial motility in the healthy and the injured brain, and discuss how impairment of microglia motility can affect normal brain function.

Corresponding author: Alexander Cupido, PhD; e-mail: alexander.cupido@uks.eu

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2. Dental pulp as a stem cell reservoir

Andreea Cristiana Didilescu, M. C. Rusu, G. Nini

Currently, much information is provided regarding the presence and the roles in tissue regeneration of stem cell niches residing in post-natal dental pulp. So far, three types of adult stem cells have been isolated from dental pulp. Correct evaluation of these cells is important in order to determine their potential use in clinical fields. The present study aims to review the origins and immunophenotype of these cells. The particularities of interstitial cells of the stem cell niches are also debated.

Corresponding author: Mugurel Constantin Rusu, Senior Lecturer, MD, PhD; e-mail: anatomon@gmail.com

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3. High variability in MMP2/TIMP2 and MMP9/TIMP1 expression in secondary liver tumors

Roxana Avadanei, Irina-Draga Caruntu, Cornelia Amalinei, Ludmila Lozneanu, Raluca Balan, Adriana Grigoras, Delia Ciobanu Apostol, Simona Eliza Giusca

The assessment of MMPs/TIMPs expression in various primary tumors has potential prognostic values. Considering the paucity of studies in secondary liver tumors, our aim was to study the expression of MMP2, MMP9, TIMP1, and TIMP2 in secondary hepatic cancer, focusing on their variability in the malignant cells. The study group included 25 cases of liver metastases of colorectal cancer, diagnosed and surgically treated at "Sf. Spiridon" University Hospital of Iassy, Romania. Immunohistochemistry for MMP2, MMP9, TIMP1, and TIMP2 has been performed, followed by the semi-quantitative assessment of the markers using a scoring system based on the positive tumoral cells percentage and the staining intensity. The expression of investigated markers revealed an increased staining variability. The scores showed that MMP2/TIMP2 and MMP9/TIMP1 immunoreactivity was extremely heterogeneous within the analyzed group, with a dominant weak expression for MMP2 and MMP9, in contrast to strong TIMP2 and TIMP1 expression. Ten different patterns of expression of the investigated markers have been identified. No major differences between the expression of MMP2 (14 positive cases) and MMP9 (16 positive cases) could be detected. Our results sustain the inverse correlation between MMP and the correspondent TIMP expression, supporting the hypothesis that MMPs/TIMPs balance has mainly an inhibitory effect on invasiveness. Our study demonstrated that tumoral cells are adapted to MMPs:TIMPs production in the liver microenvironment. The lack of significant differences between MMP2 and MMP9 expression shows that the activity of both MMPs is independent, without reciprocal influences.

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

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4. Study of the diagnosis and treatment of cancer located in the head and neck and correlation with expression of prognostic markers

Fl. Badulescu, Adriana Badulescu, Anda Crisan, Florina Carmen Popescu

A prospective study made by authors was initiated in 2000 in order to analyze differences in terms of response rate, overall survival and progression free survival for patients with advanced head and neck carcinoma treated with radiotherapy vs. radiochemotherapy, respectively and to analyze the role of modern molecular biomarkers in the prognosis of these patients (p53, EGFR and Ki67). It was detected one significant difference appeared between the two groups for response rate (p<0.0001) and median overall survival [18.8 months in Group A and 17.2 months in Group B, with a hazard ratio for survival of 0.88 (95% CI, 0.75-1.12, p<0.0001]. Progression free survival was not significant different between these two groups [6.9 months for Group A and 7.2 months for Group B, p=0.3179]. Multivariate analysis by sex, age, TNM stage, site of disease, revealed TNM stage and site of disease as clinical phenotypes with predictive value. Also, the molecular biomarkers p53, EGFR and Ki67 have a prognostic significance in head and neck cancer in addition to the established clinical prognostic factors such as the stage, site of the tumor and the type of treatment. Because of material reasons, we decided to carry out the immunohistochemical marking in the group of patients who were radiochemotherapy-treated and the results of multivariate analysis reached statistical significance threshold in terms of response rate to treatment, overall survival and disease-free interval. Furthermore, immunohistochemical examination was not performed for patients with rhinopharyngeal carcinoma with marked radiochemosensibility and reduced tumor aggressiveness reflected in significantly better therapeutic outcomes by treatment response rate, overall survival and disease free interval. In the present study of the 93 cases that underwent immunohistochemical staining for EGFR, the majority (66 cases) showed a positive reaction for this marker, in 36 cases are highly immunohistochemical staining, and in 30 cases were weakly positive. In addition, cell proliferation was intense in 26 cases in which Ki67 index was greater than 45%, p53 protein expression was positive in 18 cases, but the majority (22) of cases showed a positive reaction for this marker; index was greater than 55%, and in the most (22) cases p53 protein was not expressed. Radio/chemoradiotherapy regimen associated with molecular anti-EGFR target therapy is standard therapy in advanced squamous head and neck carcinoma.

Corresponding author: Anda Crisan, Assistant, MD, PhD; e-mail: anda_crisan2005@yahoo.com

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5. Immunohistochemical expression of RBP2 and LSD1 in papillary thyroid carcinoma

Lingling Kong, Guoan Zhang, Xu Wang, Jian Zhou, Sen Hou, Wen Cui

Purpose: To investigate the prognostic significance of LSD1 and RBP2 expression in patients with papillary thyroid carcinoma. Materials and Methods: LSD1 and RBP2 expressions were detected by immunohistochemistry in surgically resected samples from thyroid adenoma, papillary thyroid carcinoma and paracancerous tissues. Results: To be members of histone demethylases, LSD1 and RBP2 were both localized mainly to the thyroid cell nucleus. Despite the fact that both RBP2 and LSD1 expressions were higher in papillary thyroid carcinoma than in paracancerous tissues (U=-3.855, p=0.000; U=-5.575, p=0.000) and thyroid adenoma (U=-1.972, p=0.049; U=-3.190, p=0.001), they did not show us statistical correlation (r=-0.149, p=0.270). Like LSD1 (U=-2.286, p=0.022), RBP2 expression was less frequently in paracancerous tissues than in thyroid adenoma (U=-1.985, p=0.047). Neither LSD1 nor RBP2 expression was significantly associated with age, gender, stage status, tumor size, and lymph node metastases (p>0.05). Conclusions: Both LSD1 and RBP2 are well related with the occurrence and malignant transformation of papillary thyroid carcinoma. Though the positive expression of both LSD1 and RBP2 can be used to estimate the potentiality of thyroid carcinoma and help for the adjuvant treatment, LSD1 is a more sensitive molecular marker than RBP2 on thyroid cancer diagnosis.

Corresponding author: Lingling Kong, Associate Professor; e-mail: Kong001199@yahoo.com; Wen Cui, Professor; e-mail: cuiwenmd@163.com

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6. Comparative study of placenta acute fetal distress and diabetes associated with pregnancy

V. Gheorman, Lavinia Gheorman, C. Ivanus, R. C. Pana, A. M. Goganau, Anca Patrascu

The authors analyze the main histopathological changes of placentas from pregnancies ended with fetal distress at birth and the tasks associated with diabetes. The parallel between the two types of placentas not trying to prove the existence of pathognomonic lesions. Are set out both the similarities between the two titles of placentas lesions (such as changes in microcirculation and so on) as well as particular aspects. The authors analyze a group of 19 pregnant women hospitalized in Obstetrics and Gynecology Clinics of Emergency County Hospital of Craiova, Romania, in September 2010-September 2011, who were born and who were diagnosed with diabetes. In the same period, were studied 21 pregnant women whose pregnancy ended with the birth of a child with fetal distress. Such were identified as placental lesions suggestive of fetal distress as diverse etiology of placental vascular changes and the placenta in pregnancy associated diabetes as immaturity and vascular edema and fibrinoid changes and glycogen stores. The authors have proposed to highlight some lesions suggestive of two groups of diseases but independent groups were analyzed and conclusions were drawn after discussing results. This study is justified by insufficient knowledge of the causes that lead to fetal distress regardless of its etiology. In conclusion, the authors mention both placenta's common changes as specifically changes of the placenta for each type of disorder.

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

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7. Morphological expression of the anterior spinal artery and the intracranial segment of the vertebral artery: a direct anatomic study

L. Ballesteros, P. Forero, I. Quintero

Given its importance for clinical diagnosis and management, the purpose of this study was to determine the morphological expression of the intracranial segment of both vertebral and spinal arteries. Ninety-five fresh cadaveric specimens autopsied at the Instituto de Medicina Legal of Bucaramanga (Colombia) were perfused with polyester resins at the vertebrobasilar vascular bed, by canalizing the proximal segment of the internal carotids and vertebral arteries. The intracranial segment of the vertebral arteries (VA) had a length of 33.2+/-5.2 mm and a caliber of 3.03+/-0.81 mm. The left vertebral artery had a caliber of 3.12+/-0.85 mm, this being greater than the caliber of the right artery in 59.6% of the cases. The VA junction was seen at the level of the ponto-medullary sulcus in 44.2%, above this level in 30.5%, and below it in 25.3%. The distance from the origin of the anterior spinal branches (ASBs) to the VA junction was 7.34+/-2.71 mm, with a greater distance corresponding to the right ASBs, a difference that did not reach statistical significance (p=0.32). The anterior spinal arteries (ASA) had a bilateral origin in 65.3% of the cases, with the caliber being balanced in 37 (39%) and dominant at the right side in 15.8%. A right unilateral origin of the ASA was observed in 13.7% and a left unilateral origin in 9.5% of the specimens. There is disagreement in the literature as to the level of the junction of the two vertebral arteries. The unilateral origin of the spinal arteries is more frequent than most prior studies have indicated.

Corresponding author: Luis Ballesteros, Professor, MD, MSc; e-mail: lballest56@yahoo.es

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8. CD105 microvessels density, VEGF, EGFR-1 and c-erbB-2 and their prognostic correlation in different subtypes of cervical adenocarcinoma

Ileana Barbu, Stefania Craitoiu, Cristiana Eugenia Simionescu, Ana Maria Dragnei, Cl. Margaritescu

In this study, we have investigated the immunohistochemical expression of endoglin (CD105), vascular endothelial growth factor (VEGF), epidermal growth factor receptor (EGFR-1) and epidermal growth factor receptor 2 (c-erbB-2) and their prognostic correlation in 13 cases of cervical adenocarcinomas with mucinous, endometrioid and serous type differentiation. Our study revealed that for uterine cervix adenocarcinoma the most intense angiogenic activity occurs at the invasion front of these tumors. In addition, we noticed a trend towards increased CD105 MVD values in those cases in which were recorded the highest VEGF and c-erbB-2 reactivity. Thus, we concluded that in cervical adenocarcinomas occurs an intense process of angiogenesis, mainly at the invasion front, controlled by interrelations between VEGF and EGFR family members, especially the c-erbB-2 receptor. Further studies are needed to elucidate whether specific angiogenic molecular profiles exist in different histopathological subtypes of uterine adenocarcinomas and which is their impact on prognosis and therapeutic outcomes for these patients.

Corresponding author: Claudiu Margaritescu, Associate Professor, MD, PhD; e-mail: c_margaritescu2000@yahoo.com

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9. Childhood rhabdomyosarcoma. Anatomo-clinical and therapeutic study on 25 cases. Surgical implications

Smaranda Diaconescu, M. Burlea, Ingrid Miron, S. G. Aprodu, Doina Mihaila, Claudia Olaru, L. Miron

Rhabdomyosarcomas (RMS) are the most frequent soft tissue sarcomas of childhood. Despite advances in knowledge about biological pathways of tumorigenesis, risk stratification and multimodal treatment, the immediate and long-term prognosis of these lesions in many countries with limited resources is still poor. Patients and Methods: Twenty-five histologically confirmed pediatric RMS were recorded during the period of study. Demography, clinical presentation, diagnostic means, pretreatment staging and post-surgical grouping, histological type, therapy and outcome were evaluated. Results: The mean age was 6.7 years; the group included 12 boys and 13 girls. Twelve lesions were localized in the genitourinary tract, eight in the trunk and extremities, two cases each in head and neck and retroperitoneum and one case in biliary tract. Primary surgical attempt was performed in 15 patients but only in nine of them underwent complete resection (three with free margins) other six cases achieving removal with residual disease. In 10 cases, solely biopsy was possible. Twenty-four patients received chemotherapy but only four cases performed radiation therapy. Overall survival rate was only 36% (nine cases). Conclusions: As mean feature children from our series had late presentation with locally extended (bulky and node positive) lesions and unfavorable sites. Improved multimodal management of RMS in recent years will probably lead to better survival curves in an increasing number of cases and an outstanding outcome in children with locally advanced disease.

Corresponding author: Sergiu Gabriel Aprodu, Associate Professor, MD, PhD; e-mail: sgaprodu@yahoo.com

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10. Oral treatment of metabolic acidosis in hemodialyzed patients and the implications on the hemodynamic status

I. A. Checherita, Cristiana David, A. Ciocalteu, I. Lascar, Laura Budala

Metabolic acidosis slowly develops during renal impairment natural evolution towards ESRD and represents an important contributing factor of CKD progression. Although, several clinical and experimental trials reported the major impact of metabolic acidosis on CKD evolution, the pathophysiology mechanism remains a matter of debate. Furthermore, international guidelines do not impose a specific treatment scheme for metabolic acidosis in CKD patients, and metabolic acidosis is not fully compensated once hemodialysis starts. Therefore, the aim of our study was to determine an adequate follow-up of metabolic acidosis therapy benefits and risks in HD patients. Patients and Methods: 164 HD patients were evaluated according to the following protocol: bioumoral laboratory tests, the measure of different important parameters (residual diuresis, UF, BP, LVMI, volemia status). The assessed data were statistic analyzed using non-paired Student's t-test for continuous variables and chi-square (chi-square) test for qualitative parameters (p-value <0.05 was considered statistically significant). Results: HD individuals were followed-up depending on their predialysis-alkaline reserve value. After therapy started, predialysis-alkaline reserve mean level increased from 19.4 mEq/L to 22.6 mEq/L (p<0.001). Furthermore, we observed a significant decrease of nitrogenous waste products values (T=10.87<1.66) and intradialytic hypotension events (p<0.001). Conclusions: Our findings emphasize the beneficial effects of correcting metabolic acidosis using the proposed treatment scheme with direct impact on hemodynamic status improvement.

Corresponding author: Ionel Alexandru Checherita, MD, PhD; e-mail: fizij@yahoo.com

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11. Immunohistochemical expression of EGF, c-erbB-2 and EGFR in intestinal variant of gastric adenocarcinomas

Anca Oana Docea, P. Mitrut, Daniela Cernea, C. C. Georgescu, D. Olimid, Cl. Margaritescu, Daniela Dumitrescu

In this study, we aimed to determine EGF, c-erbB-2 and EGFR expression in 25 specimens of intestinal gastric adenocarcinomas by standardized immunohistochemistry and to establish correlations with the major clinico-morphological parameters of these patients. We observed EGF reactivity in 22 (88%) cases, a c-erbB-2 protein expression in eight (32%) cases and an EGFR reactivity in 13 (46.42%) cases. The EGF expression was significantly correlated with the tumor degree of differentiation, but not with other investigated clinico-morphological parameters and nor with c-erbB-2 and EGFR1 expression. However, we noticed the existence of a dependence between c-erbB-2 and EGFR1 expression in the main tumor mass. Such immunoprofile suggests the possible intervention of autocrine and paracrine loops in the developing of intestinal variant of gastric adenocarcinomas.

Corresponding author: Claudiu Margaritescu, Associate Professor, MD, PhD; e-mail: c_margaritescu2000@yahoo.com

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12. Histological diagnosis and risk of renal vein thrombosis, and other thrombotic complications in primitive nephrotic syndrome

M. Harza, G. Ismail, G. Mitroi, Mihaela Gherghiceanu, A. Preda, G. Mircescu, I. Sinescu

Background: The risk of thromboembolic events is increased in patients with nephrotic syndrome (NS) as compared with other medical conditions and is a severe complication associated with significant morbidity and mortality. We aimed to assess the risk of renal vein thrombosis, and other venous thromboembolic events (VTE) in a large cohort of patients with NS and to identify the disease-specific risk for VTE. Patients and Methods: We performed a prospective observational study including consecutive adult patients with primitive NS admitted to our department. Clinical and biological data were obtained every six months during follow-up. Occurrence of VTE confirmed by imaging techniques was the primary study outcome. Results: We enrolled 191 patients (47+/-15 years, 53% men) with a median follow-up of 24 [IQR:12,36] months. During follow-up, 23 VTE occurred, of which 65.2% in the first six months. The disease-specific risk of VTE during the follow-up period was different across the histological groups, with the lowest risk in minimal change disease and IgA nephropathy and the highest in membranous nephropathy and membranoproliferative glomerulonephritis patients. In the subgroup of membranous, the severity of the subepithelial electron dense deposits did not correlate with the risk for VTE (p=0.5). Conclusions: In this prospective study, the risk of VTE was higher in the first six months of follow-up in NS patients. The histological pattern seems to influence the risk of VTE in this setting.

Corresponding author: Gener Ismail, MD, PhD; e-mail: gener732000@yahoo.com

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13. Microanatomic aspects of arterial blood supply in rectal carcinomas - predictive models

M. V. Hinganu, Delia Hinganu, L. L. Francu

Rectum is divided into three distinctive regions (pelvic peritoneal, pelvic subperitoneal and perineal) regarding the regions where it is crossing through. Those three parts are individually not only due to their relation but due to their blood supply, also. The differences occur among them when one of them is involved into a neoplastic process. Both types of pelvic rectal tumors behave quite in the same way but those involving perineal rectum are much different. This is because they purchase a smaller number of anastomosis; when a tumor monopolizes a wide vessel the possibility to grow and become a metastasis is much more likely. These two processes (growth and metastasis) are directly related to the size of its supplying artery. On the other hand, a pelvic rectal tumor is more likely to metastasis by blood flow then a perineal rectal one. The last one will rather send clone cells by lymphatic drainage or will disseminate into the soft tissues around it. In this study, we want to propose an anatomical mathematical model for each of the rectal tumors, depending on their stages also. We used specimens from 24 patients and analyzed them using arteriography; we connected the results of mathematical counting of micro vessels density in a specific area with already known medical aspects regarding their diagnosis, treatment and evolution. The goal of the study concerns the prognosis of the patients (with or without surgical treatment) and the example is useful in rectal tumors staging.

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

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14. K-ras gene mutation status in colorectal cancer: comparative analysis of pyrosequencing and PCR-RFLP

Maria Dobre, Maria Comanescu, D. Arsene, Cristina Iosif, G. Bussolati

Background: In patients with high-stage colorectal carcinomas (CRC), anti-EGFR therapy is known to be effective only in cases with a wild-type K-ras gene status. Different procedures have been proposed for such evaluation. Materials and Methods: The mutation status of K-ras gene, codons 12, 13 and 61 was determined in 250 CRC cases using the pyrosequencing assay. In addition, we compared the performance of the pyrosequencing procedure with that of PCR-RFLP in a subset (n=100) of the CRC samples the latter only in codons 12 and 13. Results: Using pyrosequencing, 46.4% of the 250 CRC cases were found mutated. Most mutations were located in codon 12 (36.4% from all cases) and several were located in codon 61 (3.2%). All mutation identified by PCR-RFLP were confirmed by pyrosequencing and, in addition, one more mutated sample was identified in the subset of 100 samples. Conclusions: Both methods are highly specific and can profitably be used in the molecular diagnosis of colorectal cancer in order to establish the adequate therapy.

Corresponding author: Maria Comanescu, MD, PhD; e-mail: mariacomanescu@yahoo.com

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15. Hairy cell leukemia - a rare type of leukemia. A retrospective study on 39 patients

Amelia Maria Gaman

Hairy cell leukemia (HCL) is a chronic B-cell lymphoid leukemia characterized by pancytopenia, splenomegaly, myelofibrosis and the presence in peripheral blood, bone marrow and spleen of atypical lymphoid cells with a hairy aspect. This is a retrospective analysis of 39 patients hospitalized in the Clinic of Hematology, "Filantropia" Municipal Hospital, Craiova, Romania, between 1997-2012, devised by age, sex, and HCL type. Characteristic features of diagnosis (including clinical features, laboratory data: complete blood cell count, differential count, peripheral blood and bone marrow infiltration with atypical lymphoid cells with cytoplasm fine prolongations, immunophenotyping of peripheral blood, bone marrow, spleen or lymph node biopsies with histopathological exams and immunohistochemistry), types of therapy (focused on IFN-alpha), complications (infections, hemorrhage, autoimmune, second malignancies) and survival rate were monitored. Conclusions of the study revealed the importance of histopathology and immunohistochemistry for diagnosis, of the therapeutic options in the absence of purine nucleoside analogues, the most frequent complications and the decrease of their incidence correlated with therapy and increased count of neutrophils.

Corresponding author: Amelia Maria Gaman, Associate Professor, MD, PhD; e-mail: gamanamelia@yahoo.com

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16. Anatomical description of the deep venous channel from the anterior vestibular wall of the right atrium

F. M. Filipoiu, Mihaela Balgradean, I. Brezean, V. Jinga

The deep venous channels in the walls of the right atrium are not routinely described in medical treatises although their existence has been remarked from some time. Our study demonstrates that the right atrium is opened for some of the anterior veins of the heart through a venous channel located deep in the myocardium of the vestibule of the right atrium. We suggest that no Thebesian veins drain into this channel. We achieved anatomical dissection in 14 adult human cadaveric hearts, which had been fixed in formalin solution, and we found the channel in 75% of cases. These channels were measured, photographed and presented in detail. We highlighted the importance of the proximity with the tricuspid annulus in order to avoid incidents during tricuspid annuloplasty. The authors intend that through a qualitative study to draw attention to such a structure often ignored. This vascular structure and its role in cardiac physiology and pathology have not been investigated yet. Although not constant, specialists in cardiology and cardiac surgery should be informed about this basic detail on the endocardium morphology.

Corresponding author: Mihaela Balgradean; e-mail: mbalgradean@gmail.com

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17. Masticatory tensile developed in upper anterior teeth with chronic apical periodontitis. A finite-element analysis study

C. M. Petcu, D. Nitoi, Veronica Mercut, Mihaela Jana Tuculina, A. A. Iliescu, Cristiana Ileana Croitoru, Oana Andreea Diaconu, Mihaela Georgiana Iliescu, Lelia Mihaela Gheorghita, A. Iliescu

Commonly is accepted that a non-vital tooth has a higher risk of root fracture but there is a relatively little knowledge about the specific biomechanical behavior of non-vital frontal teeth with chronic apical periodontitis. The aim of our study was to evaluate the deformation and tensile generated in these teeth while vertically loading them because it is crucial to assess the moment when the absorbed occlusal forces exceed the elasticity of root dentine. Using the method of finite-element analysis, we highlighted the distribution patterns of the compressive and tension tensile, as well as their concentration areas. The vertical forces of 100 N generate deformations of no clinical risk in teeth with chronic apical periodontitis. The tensile developed in these teeth are higher than those in the vital teeth are but do not exceed the value of the elastic modulus of the radicular dentin. By increasing the force to 300 N occur elastic deformations, which cannot be neglected anymore. Even so, the 300 N forces do not generate deformations of the alveolar bone. The highest tensile at loading with 300 N was generated in vertical direction but in the cervical area of the tooth also developed tensile in lingual and mesiodistal direction that must be taken into consideration because they are near the risk limit of the elasticity modulus. The crack lines or fractures can appear both in case of excessive or even usual but accumulative occlusal forces that gradually alter the mechanical resistance of the tooth.

Corresponding author: Cristian Marian Petcu, Junior Assistant, DDS; e-mail: petrus_dorel.petcu@yahoo.com

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18. A complex assessment of patients with knee osteoarthritis and Baker's cyst: observational study

Rodica Traistaru, Roxana Popescu, Corina Gruia, Otilia Rogoveanu

In our observational study, we performed the clinical and functional examination, analyzed imagistic and histological findings and evaluated the correlation between previous aspects in patients with primary knee osteoarthritis (PKOA) and Baker's cyst (BC). The correlations were made to better understanding of BC in patients with PKOA and optimal choosing for treatment. Seventy patients with painful PKOA (ACR criteria) and BC were assessed. We evaluated knee pain using a 100 mm VAS and functional status using the pain, stiffness and functional subscales of WOMAC index. All patients were imagistic examined (ultrasonography and MRI). Thirty-eight patients with PKOA and simple BC respond to conservative treatments. Thirty-two patients with PKOA and complex BC need surgical removal (arthroscopic decompression +/- open excision in larger cysts). For these patients, it is performed histological assessment. Any medical team that manages a PKOA patient with BC may develop the treatment plan based upon not only the size of BC, symptoms and other associated conditions but also on the WOMAC scoring and complex anatomic and histological data about BC.

Corresponding author: Otilia Rogoveanu, Lecturer, MD, PhD; e-mail: rodicatraistru@hotmail.com

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19. Histopathological examination in evaluation of long-term results after osteocartilaginous transplantation

R. Radulescu, A. Badila, O. Nutiu, R. Manolescu, Silvia Terinte, Maria Sajin, I. M. Japie

Objective: To assess long-term outcomes of osteocartilaginous transplantation for non-degenerative lesions of hyaline articular cartilage in the knee, by performing minibiopsies from the transplanted area and examining them histopathologically. Patients and Methods: Forty-four patients with post-traumatic cartilage injuries of the bearing surfaces of the knee were enrolled in a prospective study, that included treatment with autologous osteocartilaginous grafts at the level of the lesion, "second look" arthroscopy and targeted minibiopsies at one year and five years postoperatively (six minibiopsies per patient). The collected tissue fragments were examined by optic microscopy. In order to integrate the histopathological findings in the clinical context, the function of the knee was also quantified by calculation of the International Cartilage Repair Society Score preoperatively, at one year and at five years postoperatively. Results: Five years post-transplant the outcomes for 36 patients were available. One year post-transplant, the histopathological examination revealed the presence of hyaline cartilage in 165 of the 216 (76.39%) tissue samples collected and fibrocartilage in 51 (23.61%) respectively. Five years after surgery, the proportions of these findings were 159/216 (73.61%) for hyaline cartilage and 57/216 (26.39%) for fibrocartilage. The difference was not statistically significant (p>0.1).The evolution of the ICRS clinical score was from 38.57+/-3.42 preoperatively to 80.31+/-3.85 (p<0.0001) after one year and to 81.35+/-4.57, respectively at five years after surgery. Conclusions: Autologous osteocartilaginous transplantation brings hyaline articular cartilage at the level of the injured area. Approximately three quarters of the surface lesion remains covered by high quality hyaline cartilage that maintains its macroscopic structure and architecture for a long period of time.

Corresponding author: Adrian Badila, MD, PhD; e-mail: adrian_emilian_badila@yahoo.com

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20. Anatomo-clinical aspects of the basal cell carcinoma at the level of the cephalic end

Anca Bordianu, I. P. Florescu, Anca Muresan, Elena Bernad, M. Craina, Izabella Sargan

More than one third of all diagnosed cancer cases from all over the world are skin cancers, and the majorities are basal cell carcinomas. The incidence of skin tumors around the world is growing, the main concern being the increased frequency of skin tumors among young persons. Therefore, an early detection is required, starting with the general practitioner, dermatologist, oncologist, plastic surgeon and not least, the pathologist. We studied a sample consisting of 149 patients diagnosed with basal cell carcinoma at the level of the cephalic end and neck, all of them undergoing surgery - surgery individualized for each case. Some cellular and molecular modifications also resulted from infecting the cells with high carcinogenic risk HPV, considered by some scientists as responsible for the developing and progression of some neoplasias, including melanocytes and non-melanocytes skin tumors.

Corresponding author: Anca Bordianu, MD; e-mail: anca.bordianu@gmail.com

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21. Morphometric analysis of bone vascular channels during the biointegration of autologous bone grafts

H. P. Petrescu, G. Dinu, G. Noditi, M. Craina, Delia Berceanu-Vaduva, M. Berceanu-Vaduva, D. Vermesan

Vascularization of the transplanted bone tissue is a key factor for success and precedes the formation of bone tissue. Vascularized bone grafts have been widely used in bone transplantation for their efficiency. Maturation of the bone tissue at the place of the transplant involves the change in the vascular patterning, from plexiform irregular vascular networks, to regular, polygonal networks following the structure of osteons. Seven dogs were performed bone transplantation and intravenous injection with China ink in order to highlight the network of blood vessels during bone formation at the place of the transplant. The area occupied by blood vessels increased during osteogenesis, and their length decreased in evolution, suggesting a maturation of the vascular pattern.

Corresponding author: Horatiu Pompiliu Petrescu, Assistant Professor, MD, PhD; e-mail: lalusha87@yahoo.com

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22. The effect of chronic toxicity of pethidine on the spinal cord: an experimental model in rabbits

C. Pestean, M. Taulescu, C. Ober, C. Catoi, V. Miclaus, L. Oana, C. Bodolea

The aim of this study was to evaluate the toxicity of chronic spinal analgesia with pethidine in a rabbit model. We introduced epidural catheters in twenty New Zealand white rabbits, divided into two groups, and we administered 0.5 mg/kg pethidine or the same volume of normal saline through the catheters, for three consecutive days. Throughout the experiment, the animals were evaluated in terms of neurological status using the Tarlov score. After the rabbit's euthanasia, 4 micro-m sections of spinal cord stained with Hematoxylin-Eosin were analyzed by a pathologist blinded to the study for neurohistopathological changes. The results were statistically analyzed with Prism 5 software for Windows. No significant differences were noticed between the two groups in as far as body temperature (p=0.295) and weight (p=0.139) were concerned. In the group of animals, which received epidural pethidine, nine rabbits showed histological changes suggestive for neurotoxicity at the lumbar level of the spinal cord. These findings were significantly different compared with the control group which received only saline (no microscopic lesions revealed; p=0.0006). When combining the data from both groups or using the pethidine group alone, there was a significant correlation between the presence of neurological injury (Tarlov score) and the presence of the histopathological lesions in the spinal cord (r=-0.709, p=0.0002 and r=-0.635, p=0.013, respectively). Based on our findings, the chronic epidural administration of pethidine in rabbits induces moderate to severe histological changes on the spinal cord, but further investigations are needed to make a definitive statement about the histological effect of pethidine on the neurological tissue.

Corresponding author: Marian Taulescu, Teaching Assistant, DVM, PhD; e-mail: taulescumarian@yahoo.com

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23. Microscopic aspects of angiogenesis and lymphangiogenesis in oral squamous cell carcinoma

Adina Bunget, A. Fronie, Emilia Afrem, Dorina Corlan Puscu, H. Manolea, Anca Rodica Dan, Malina Coman, Vanda Roxana Nimigean

Despite various great scientific and financial efforts, head and neck carcinomas represent a public health problem, being the eighth cause of cancer death worldwidely. The rate of tumor growth, its local expansion, as well as the metastasis of cancerous cells depend on the tumor vascularization, on the ability of blood vessels to provide a constant supply of nourishing substances and oxygen and to eliminate the residual products resulted from tumor growth. That is why angiogenesis and lymphogenesis are considered to be essential processes within the neoplastic process. The assessment of tumoral neoformed blood vessels in oral squamous carcinomas, using the CD34 antibody, showed a significant growth of the microvascular density, the average number being 504.66+/-177.65 vessels/mm(2). The diameter of angiogenesis vessels varied between 3.42 and 121.27 micro-m. The density of lymphogenesis vessels was 508.78+/-235.93 vessels/mm(2), while the diameter varied from 2.82 to 165.28 micro-m. Both angiogenesis and lymphogenesis vessels were more numerous in the areas where the inflammatory infiltrate was more abundant, which suggests that chronic inflammation plays the part of a promoter factor of neoplastic lesions.

Corresponding author: Adrian Fronie, Assistant Professor, MD, PhD; e-mail: adrianfronie@yahoo.com

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24. Synchronous diagnosis of signet ring cell colon carcinoma and transitional carcinoma of the left distal ureter

Liliana Streba, Corina Gruia, S. S. Mogoanta, P. Mitrut, Letitia Adela-Maria Streba, Adriana Bold

The presence of synchronous multiple primary carcinomas, although recognized, remains very uncommon. Their etiopathogeny is unclear. The wide spread of imaging techniques currently facilitates diagnosis of simultaneous tumors. We present the rare case of a signet ring cell colon carcinoma coexisting with a transitional cell carcinoma (TCC) of the ureter, which debuted with symptoms of intestinal obstruction in a 75-year-old male patient, with no prior relevant history. Computed tomography showed signs of a tumor mass on the transverse colon, as well as left hydronephrosis with a tumor in the lower third of the ureter, which prompted for immediate emergency surgery. After the appropriate surgical maneuvers, tissue collected from both tumors underwent usual histological preparation and Hematoxylin-Eosin staining, as well as multiple immunostaining with a complex panel of markers. The patient had a favorable postoperative course and during a six-month follow-up, we did not detect any signs of illness.

Corresponding author: Letitia Adela-Maria Streba, Associate Professor, MD, PhD, MSc; e-mail: letitiastreba@yahoo.com

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25. Abnormalities in embryological development in total anomalous pulmonary venous connection. A case report

Mihaela Balgradean, Eliza Cinteza, C. Cirstoveanu, Augustina Enculescu, Doina Plesca

Pulmonary venous system development starts early in embryonic life. Abnormalities in the development of pulmonary venous system occur either by the absence of common pulmonary vein communication to the splanchnic plexus or by the absence of its incorporation into the dorsal wall of the left atrium. We present the case of a 10-day-old male newborn, diagnosed with TAPVC, operated, with long recovery and, who died by pneumonia, heart failure, and obstructive pulmonary disease (one pulmonary vein obstructed and another one with severe stenosis). Total anomalous pulmonary venous connection (TAPVC) reflects one of the most severe forms of congenital heart disease, with important clinical consequences.

Corresponding author: Eliza Cinteza, MD, PhD; e-mail: elizacinteza@yahoo.com

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26. Bilateral extensor medii proprius with split tendon of extensor indicis proprius, a rare anatomical variant

Jing Li, Zhen Feng Ren

The extensor medii proprius (EMP) is anomalous extensor muscle of the hand. During the routine dissection of a 78-year-old Chinese male cadaver, bilateral EMP and extensor indicis proprius (EIP) were observed in the upper limbs. The EMP originated from the distal third of the ulna and its tendon was inserted into the dorsal aponeurosis of the middle finger on both hands. The tendon of EIP was split into two slips on the dorsum of hand and inserted to the radial and ulnar side of the extensor digitorium communis (EDC)-index, respectively. Awareness of such anatomical variations in the extensor compartment of the forearm could help in the identification and repair of these structures.

Corresponding author: Jing Li; e-mail: lijing800912@yahoo.com.cn

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27. MRI diagnosis of perivascular epithelioid cell tumor (PEComa) of the liver

Hua-qiang Sheng, Dong-feng Chen, Xin-hai Sun, Xian Li, Jie Xu, Xi-bin Hu, Mei-Qin Li, Tao Wu, Ren-ya Zhang, Kai-zhi Li

Introduction: Perivascular epithelioid cell tumor (PEComa) has been rarely reported in the liver. Patient, Methods and Results: We present a liver PEComa case diagnosed by magnetic resonance imaging (MRI) findings. The patient was incidentally found to have an abnormal mass in the liver. MRI revealed early and strikingly homogeneous enhancement of the lesion. Partial hepatectomy was performed, and a pathological examination revealed signs of typical of PEComa. The patient was closely monitored for 12 months after the surgery, with no clinical or radiographic evidence of recurrence or metastatic disease. Conclusion: MRI diagnosis is applicable for PEComa.

Corresponding author: Hua-qiang Sheng, Professor; e-mail: huaqiang_sheng@163.com

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28. Malignant pleural mesothelioma presenting with symptomatic brain metastases: report of a case

A. Marzullo, Anna Scattone, Roberta Rossi, Antonietta Cimmino, Alessandra Punzi, F. Corsi, Domenica Cavone, Teresa Lettini, Gabriella Serio

We describe a unique case of brain metastases presenting as first symptom of a malignant mesothelioma (MM). MM is a highly aggressive tumor of the serous membrane that is generally believed to be rarely metastasizing. Recently, the reports of long surviving cases and larger literature reviews have suggested that cerebral metastases are not so uncommon. An extensive histochemical and immunohistochemical panel is needed to achieve a correct differential diagnosis, especially in the epithelioid type. Pathologists should be aware that brain metastases could have a mesothelial origin.

Corresponding author: Gabriella Serio, MD, PhD; e-mail: gabriella.serio1@uniba.it

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29. Myxoid/round cell conjunctival liposarcoma. A case report

Stela Giuri, M. Raica, M. Munteanu

Purpose: To present a rare case of conjunctival myxoid liposarcoma, subtype round cells, that had a seven years follow up. Clinical observation: A 61-year-old female patient presents with a palpable, non-painful tumor, on the superior temporal bulbar conjunctiva of the right eye. The initial examination detects a fleshy tumor, orange in color, under the superior temporal bulbar conjunctiva, as well as two oval-shaped hyperpigmented conjunctival lesions, near the limbus at 10 o'clock, causing moderate blepharoptosis. Vision was normal, there was no diplopia, proptosis, afferent pupillary defect or lymphadenopathy; there was no orbital involvement in MRI. An isolated 15/15 mm tumor, with no connections with the eye socket, was excised. Histopathology revealed a poorly differentiated myxoid liposarcoma. Five recurrences occurred, of which four were treated by local excision and the last required exenteration. Repeat histopathology detects lipoblasts, small round cells, with immunohistochemistry positive for CD34, S100 and vimentin. The last two rapidly evolving and large recurrences, as well as pulmonary metastasis and finally death of the patient, underlined the aggressive character of round cell conjunctival liposarcoma. Conclusions: Conjunctival myxoid liposarcoma is characterized by numerous local recurrences, but the speed of the succession and volume of the recurrences may suggest a change in the underlying histopathological aspect, that is definitory for the therapeutical and prognostic approach of the case.

Corresponding author: Mihnea Munteanu, Associate Professor, MD, PhD; e-mail: mihneam1@gmail.com

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30. Progressive intraparenchymal lung nodules dissemination in a heavy smoker and seropositive rheumatoid arthritis suspected of tuberculosis relapse

Oana Cristina Arghir, Mimi Nitu, Mihaela Trenchea, Camelia Ciobotaru

Anthony Caplan first described rheumatoid lung nodules associated with pneumoconiosis in coal-miners (Caplan, 1953). Intraparenchymal lung nodules were later described in rheumatoid arthritis (RA) patients who were never exposed to coal dust and/or without pneumoconiosis. Rheumatoid lung nodules are usually detected in unselected patients: 0.2% in chest radiography and 4% in high-resolution computed tomography (Nannini et al., 2008). Patients could be reluctant to perform surgical lung biopsy for an accurate histopathological diagnosis. We present a peculiar association between a seropositive RA and a presumptive active tuberculosis (TB) disease in a 59-year-old male patient, ex-smoker with a previously healed pulmonary TB disease. The purpose of this report is to describe an unusual case of a presumptive relapse of the nodular TB disease, which progressed to an extensive nodular bilateral dissemination under anti-tuberculosis therapy, mimicking a metastatic carcinoma. The diagnosis of rheumatoid necrobiotic lung nodules was confirmed after open biopsy left pulmonary was performed. Formalin-fixed paraffin-embedded pulmonary rheumatoid nodules were processed for histology and stained with Masson's trichrome. Central structure of the removed pulmonary nodules is typical of a rheumatoid nodule with central necrosis surrounded by a palisade of macrophages. The accumulation of anthracotic pigment was noticed inside the pulmonary nodules in a RA patient without professional exposure to coal or mineral dust. This rare entity is an appearance of the rheumatoid nodules lung syndrome and anthracosis in a heavy tobacco former smoker.

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

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31. Sebaceous carcinoma of the eyelid: anatomoclinical data

Claudia Florida Costea, D. Petraru, Gabriela Dumitrescu, Anca Sava

Sebaceous gland carcinoma of the eyelid is a rare slow-growing tumor and is one of the most aggressive malignancies of the eyelid. Diagnosis is often delayed because it can be confused with other periocular lesions. We report the case of a 78-year-old female who presented for the anesthetic aspect of a nodular tumor on the right upper eyelid occurring one year earlier. The patient was treated for three months for recurrent chalazion. Ophthalmologic examination revealed a nodular ulcerated tumor of 1 cm in size adherent to adjacent tissues. Surgical excision was performed with a safety margin of 4 mm. The diagnosis of moderately differentiated sebaceous carcinoma was made by routine morphological methods and immunohistochemical reactions (EMA and Ki-67). Being a rare tumor with considerable morbidity and mortality, early diagnosis and proper treatment are essential for a favorable prognosis and preservation of visual function.

Corresponding author: Claudia Florida Costea, MD, PhD; e-mail: costea10@yahoo.com

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32. Atypical debut manifestations in juvenile idiopathic arthritis

Dalia Dop, Carmen-Elena Niculescu, Ligia Stanescu, D. Niculescu, A. Stepan

Considering that destructive articular lesions may occur in the first stages of the illness, it is difficult but necessary to establish a diagnosis of Juvenile Idiopathic Arthritis (JIA) in due time. The authors present the case of a 9-year-old girl admitted to the Pediatrics Clinic of the "Filantropia" Municipal Hospital in Craiova, Romania, on November 26, 2002, for bilateral pain in the tarsometatarsal and carpometatarsal joints that had begun approximately four weeks before. After the clinical examination and paraclinical investigations, a diagnosis of unspecified arthritis is established and the adequate treatment is begun. Two months later, the patient returns to the clinic with bilateral knee pain and swelling. The results of laboratory tests indicate the persistence of anemia and of the inflammatory syndrome. The diagnosis of JIA is established. The evolution of the patient is unfavorable, both from a clinical point of view (a large number of articulations affected, a persistent rash, hepatomegaly) and a paraclinical one (increased acute phase reactants and radiological changes occurring two years after the onset of the illness).

Corresponding author: Dalia Dop, MD; e-mail: dalia_tastea@yahoo.com

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33. Letter to the Editor. Does "signet-ring stromal tumor" exist?

Teresa Pusiol, Doriana Morichetti, Maria Grazia Zorzi

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Corresponding author: Teresa Pusiol, Director; e-mail: teresa.pusiol@apss.tn.it

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