ROMANIAN JOURNAL of MORPHOLOGY and EMBRYOLOGY

Vol. 55 No. 3, 2014

1. Imaging neuroinflammation after brain injuries by ultrasensitive MRI and two-photon laser-scanning microscopy

Anja Scheller, Denis Vivien, Frank Kirchhoff, Cyrille Orset, Raluca Elena Sandu, Aurel Popa-Wagner

Worldwide, stroke is the leading cause of disability in the aging population. Neuroinflammation is a common feature of acute stroke and is considered to be a major obstacle to endogenous neurogenesis and exogenously administered stem cells. Therefore, drug and cell therapies aimed at suppressing post-stroke inflammation have emerged as a promising approach to improve recovery after stroke. However, progress toward the development of efficient cell-based therapies for ischemic stroke has been disappointing mainly because the interplay between host neuroinflammation and stem cell-based therapies during the acute stroke and the recuperation phase is virtually unknown. The pathophysiological evolution of stroke events indeed seems driven by complex cellular interactions between several different cell types whose sequential recruitments have been insufficiently documented due to the lack of respective technologies, in particular, of non-invasive imaging modalities. The development of in vivo ultrasensitive magnetic resonance imaging (MRI) and two-photon laser-scanning microscopy has revolutionized our understanding of neuroinflammation. Therefore, the purpose of this review is to highlight the interplay between host neuroinflammation, which is considered to be a major obstacle to exogenous-mediated neuronal precursor cells, and exogenously administered stem cells.

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

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2. Microglia - performers of the 21st century

Nidal Atallah, Renata Vasiu, Adina Bianca Bosca, Denisa-Ioana Cretu, Carmen Georgiu, Anne-Marie Constantin, Alina Simona Sovrea

At the frontier between immunology and neuroscience, microglia, the enigmatic macrophages of the brain, have generated, in recent years, increasing interest. In response to even minor pathological changes in the brain, these extremely versatile glial cells occasionally enter in an over-activating state and produce pro-inflammatory cytokines and free radicals, thereby contributing directly to neuroinflammation and various brain disorders. This review provides an analysis of the latest developments in the microglia field, considering the important new research that illustrate their involvement in brain related diseases.

Corresponding author: Renata Vasiu, Associate Professor, MD, PhD; e-mail: renata.vasiu@yahoo.com

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3. Optical coherence tomography investigation of ischemic stroke inside a rodent model

Eugen Osiac, Tudor-Adrian Balseanu, Laurentiu Mogoanta, Dan Ionut Gheonea, Ionica Pirici, Maria Iancau, Smaranda Ioana Mitran, Carmen Valeria Albu, Bogdan Catalin, Veronica Sfredel

Although already in use in several medical domains, only recently optical coherence tomography (OCT) has been applied in the study of ischemic events. In this paper, we will focus on characterizing ischemic stroke, in a rat model, by OCT. Investigations were carried on a set of 25 rats, on which ischemic stroke was inflicted by a transient occlusion of the middle cerebral artery (tMCAO). Animals were sacrificed 1, 3, 7 and 28 days after occlusion. We tested the OCT s power of detection and discrimination of stroke area compared to both normal, contralateral hemisphere and non-affected brain tissue, together with the aid of histochemical and pathological examination. Our results show a great potential of OCT to be used as a detection tool in acute and chronic phases of stroke.

Corresponding author: Bogdan Catalin, MD, PhD; e-mail: bogdan.catalin@webmail.umfcv.ro

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4. Ultrasound and histopathological features of myocardial involvement in HIV infection in children

Anca Meda Georgescu, Cosmin Moldovan, Leonard Azamfirei, Dan Georgescu

Aim: HIV infection in children is an important clinical and pathologic entity, which embraces many forms of presentation and can involve multiple organs and systems. This study aimed at identifying the main forms of cardiovascular involvement in HIV-infected children with horizontally transmitted disease and describing them with the aid of ultrasound and histopathological examinations. Results: We recorded cardiovascular anomalies in 79 (67.52%) patients out of the 117 comprised in the study population, and noted the following prevalence distribution: systolic dysfunction in 49 (41.88%) patients, left ventricular hypertrophy (LVH) in 30 (25.6%) patients, right ventricular hypertrophy (RVH) in 15 (12.82%) patients, and dilated cardiomyopathy (DCM) in 22 (18.8%) patients. We also carried out post-mortem histopathological examinations in five patients, and observed the main modification incurred by the disease. Conclusions: Cardiac involvement during HIV infection differs significantly in different mechanisms of virus transmission, and the horizontal transmission of HIV yields a lower prevalence of this type of pathology. The general diagnostic picture can be significantly improved by adding histopathological examination to the ultrasonographic method of investigation.

Corresponding author: Cosmin Moldovan, Lecturer, MD, PhD; e-mail: cosmin.moldovan@umftgm.ro

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5. Rheumatoid myositis, myth or reality? A clinical, imaging and histological study

Codrina Ancuta, Daniela Cristina Pomirleanu, Carmen-Rodica Anton, Eovelina Moraru, Emil Anton, Rodica Marieta Chirieac, Eugen Ancuta

Rheumatoid myositis (RM) is still poorly characterized, albeit the concept of muscle involvement in rheumatoid arthritis (RA) is well-recognized as being driven by a wide range of causes including inflammation, drugs, impaired joint flexibility, sedentarism. Objective: To describe clinical, serological, imaging and histological pattern of RM. Materials and Methods: This is a retrospective study on eight RM selected from a cohort of one hundred and three RA systematically assessed for skeletal muscle involvement. Data collected included clinical, serum muscle enzymes, muscle imaging and biopsy (Hematoxylin-Eosin, modified Gomori trichrome staining). Results: Routine muscle histology indicated both non-specific muscle fiber damage (changes in fiber size and internal structure: pleomorphic mitochondria, dilated sarcotubular system, multiple internal or subsarcommal nuclei; abnormal fiber types distribution: trend towards type II; atrophy; degenerative/regenerative modifications) and the presence of inflammatory deposits in all patients (mild to moderate, patchy B- and T-cells infiltrates, mainly perivascular and endomysial, but also in the perimysial region classified as polymyositis-like deposits). High levels of serum muscle enzymes, abnormal EMG (short duration, small amplitude, polyphasic motor unit action potentials) without insertional activity and fibrillations, active inflammation on both Doppler ultrasound and MRI were commonly reported. Conclusions: Traditional analysis of muscle biopsy specimens (Hematoxylin-Eosin, modified Gomori trichrome staining) is faraway unsatisfactory, only documenting changes in muscle fibers size, architecture, internal structure, and, possibly, detecting perivascular, perimysial or endomysial inflammatory deposits. Upcoming research should address the value of muscle imaging for the diagnosis and evaluation of treatment response and muscle function in rheumatoid myositis.

Corresponding author: Eugen Ancuta, MD, PhD, MsC; e-mail: eugen01ro@yahoo.com

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6. Lectin purification from fruiting bodies of brown roll-rim fungus, Paxillus involutus (Fr.) Fr., and its application in histochemistry

Rostyslav Antonyuk, Alexander Lutsyk, Volodymyr Antonyuk

A lectin (agglutinin) from fresh fruit bodies of the brown roll-rim fungus - Paxillus involutus (Fr.) Fr. - has been purified with output approx. 60 mg/kg of raw material. Method of purification included the sedimentation of viscous polysaccharide by ethanol, removal of ethanol by dialysis, ion-exchange chromatography on DEAE-Toyopearl and affinity chromatography on Sepharose 6B column with immobilized mannose-specific Polygonatum multiflorum lectin. The obtained lectin preparation (abbreviated PIFA) is a glycoprotein with 6.5+/-1% carbohydrates, molecular mass of 64 kDa, consisting of four identical subunits. Lectin interacted only with N-acetyl-lactosamine and glycoproteins that contained Gal(beta1)-4GlcNAc disaccharide moieties; agglutinated erythrocytes of dog, sheep and horse, but not of humans. The specificity of PIFA binding to tissue samples of the rat has been investigated. Lectin selectively reacted with gastric parietal cells, submandibular salivary gland duct cells. In the kidney, PIFA labeled epithelial cells of renal tubules, collecting ducts, nuclei of podocytes and mesangiocytes. It was also revealed selective lectin binding to Purkinje cells of cerebellum. Brush border of absorptive cells in small intestine was also strongly reactive, while goblet cells both in small and large intestine were completely negative. Considering similarities in carbohydrate specificity of Paxillus involutus (PIFA) and Ricinus communis agglutinin (RCA-120), histochemical reactivity of these two lectins was compared. It was similar, yet not identical: differences included absence of PIFA binding to the brush border of renal tubules, higher interaction with absorptive cells of the small intestine, lower background staining of cerebellar cortex and renal corpuscles. A conclusion was made that due to the unique carbohydrate specificity PIFA lectin can cover prospective position in experimental histochemistry and diagnostic histopathology comparable to PNA (Peanut agglutinin) and SNA (Sambucus nigra agglutinin).

Corresponding author: Alexander Lutsyk, MD, PhD, Professor and Chair; e-mail: lutsyk@meduniv.lviv.ua, lutsykalexander@gmail.com

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7. Histopathological and imaging modifications in chronic ethanolic encephalopathy

Roxana Folescu, Carmen Lacramioara Zamfir, Alina Maria Sisu, Andrei Gheorghe Marius Motoc, Adrian Cosmin Ilie, Marius Moise

Chronic abuse of alcohol triggers different types of brain damage. The Wernicke-Korsakoff syndrome gets together Wernicke s encephalopathy and Korsakoff s syndrome. Another type of encephalopathy associated with chronic ethanol consumption is represented by the Marchiafava-Bignami malady or syndrome, an extremely rare neurological disorder, which is characterized by a demielinization of corpus callosum, extending as far as a necrosis. Because the frequency of ethanolic encephalopathy is increased and plays a major role in the sudden death of ethanolic patients, we have studied the chronic ethanolic encephalopathy both in deceased and in living patients, presenting different pathologies related to the chronic ethanol consumption. The present study investigated the effects of chronic ethanolic encephalopathy on the central nervous system based both on the histopathological exam of the tissular samples and the imaging investigation, such as MRI and CT.

Corresponding author: Roxana Folescu, University Assistant, MD, PhD; e-mail: roxanafolescu@yahoo.com

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8. Prevalence and histopathological types of skin carcinomas in Arges County, Romania

Marilena Monica Tantu, Eduard Belu, George Mihail Man, Dan Manu, Liliana Marcela Rogozea, Cristina Mariana Stocheci, Andrei Theodor Balasoiu, Casiana Stanescu, Carmen Daniela Domnariu

Non-melanoma skin cancers presented a significant incidence increase in the last decades, worldwidely. Even though the impact upon mortality is a relatively low one, through the incidence increase, their impact upon the public healthcare systems is a considerable one. In our study, we evaluated 109 cases of skin carcinomas hospitalized during 2012 in the Department of Plastic Surgery of the Emergency Hospital of Pitesti, Romania, for a surgical treatment. The gender distribution showed slight lesion predominance in women, being recorded 56 (51.38%) tumors in women and 53 (48.62%) in men. The highest incidence of skin carcinomas (75.23%) was recorded in the persons aged over 60-year-old. Of 109 cases of skin carcinomas, 80 (73.4%) carcinomas developed on indignant tegument areas, while 29 (26.6%) on premalignant skin lesions (fiberconjunctive papillomas, keratocantomas, keratosic verrucas). The histopathological study highlighted the fact that of 109 skin carcinomas, 87 (79.82%) were basal cell carcinomas and only 22 (20.18%) were squamous cell carcinomas. The immunohistochemical reaction to 34betaE12 cytokeratin was highly positive in the cells of the basal cell carcinomas and well-differentiated squamous cell carcinomas (except for the keratosic pearls) and moderately positive in the moderately differentiated squamous cell carcinoma.

Corresponding author: Andrei Theodor Balasoiu, MD, PhD candidate; e-mail: andrei_theo@yahoo.com

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9. The single versus combinatorial effects of MK-801, CNQX, Nifedipine and AP-3 on primary cultures of cerebellar granule cells in an oxygen-glucose deprivation model

Ana-Maria Zagrean, Ana Spataru, Mihai Ceanga, Leon Zagrean

The excitotoxicity cascade associated with energetic failure during and after cerebral ischemia involves the overactivation of glutamate receptors and intracellular calcium loading. We searched for synergistic neuroprotective effects of various drugs designed to prevent intracellular calcium influx in a model of oxygen-glucose deprivation (OGD) in cerebellar granule cells primary cultures. (5S,10R)-(-)-5-Methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine hydrogen maleate (MK-801), D,L-2-Amino-3-phosphonopropionic acid (AP-3), 6-Cyano-7-nitroquinoxaline-2,3-dione disodium salt (CNQX) and Nifedipine were tested alone or in combinations. Treatments were applied during a two-hour OGD exposure and cellular outcome was assessed throughout 20-hour reoxygenation by the measurement of Propidium Iodide (PI) fluorescence. All treatments were able to prevent neuronal damage. OGD resulted in a mortality of 36.3+/-2.3% and 61.3+/-3.1% after 10 and 20 hours of reoxygenation, respectively. The most effective single treatment was AP-3 (3.3+/-1.4%; 17.9+/-2.6% mortality after 10 and 20 hours), followed in order by Nifedipine (7.2+/-1.6%; 20.1+/-3.0%), CNQX (8.5+/-2.5%; 20.0+/-3.5%), and MK-801 (14.9+/-2.6%; 39.3+/-6.4%). The combination of AP-3 with MK-801 showed a moderate synergistic effect (11.8+/-2.0% mortality at 20 hours), while the combinations of CNQX with Nifedipine and CNQX with MK-801, as well as the triple mix CNQX, Nifedipine and MK-801 failed to show a further improvement in the reduction of cellular death. In conclusion, targeting two mechanisms of cellular demise (ionotropic receptors and metabotropic glutamate receptors) provided an advantage against several unimodal strategies (blocking calcium entry through ionotropic glutamate receptors and L-type calcium channels). Our results suggest that a multimodal combinatorial treatment strategy in cerebral ischemia may increase neuroprotective efficacy and call for further research.

Corresponding author: Ana-Maria Zagrean, Associate Professor, MD, PhD; e-mail: azagrean@umf.ro, azagrean@gmail.com

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10. Potential value of in situ cellular immune response in HPV subtype 16 and 18 positive cervical cancer

Eugen Ancuta, Stefan Butureanu, Florin Zugun-Eloae, Carmen-Rodica Anton, Codrina Ancuta, Damian Ditescu, Emil Anton

There is no doubt that the association between infection of the cervical epithelium by carcinogenic Human Papilloma Virus (HPV), particularly types 16 and 18, and cervical cancer (CC) is responsible for the activation of the immune response (IR). Research on tumor infiltrating lymphocytes at the primary tumor site could give us important information on how the immune cells are fighting against cancer. Aim: The aims of our study were to assess HPV status and to evaluate the significance of in situ cellular IR in CC. Materials and Methods: We performed a two-step retrospective analysis of IR in 18 CC: evaluation of HPV 16 and 18 infections by in situ hybridization and immune biomarkers (CD20, CD3, CD45) by immunohistochemistry. Immune cell profile, densities (assigned scores 0 if no inflammatory infiltrate, 1+ low, 2+ intense), tissue distribution and classical negative prognosis factors in relationship with survival and relapse were further assessed. Results: We successfully demonstrated HPV 16 and/or 18 in all cases. We reported statistical significant correlations (p<0.005) between CD3, CD20, CD45 and survival (r=0.800), relapse (r=-0.892), clinical stage (r=-0.914), tumor size (r=-1) as well as the association between survival and CC subtype (r=0.548), FIGO stage (r=-0.914), tumor size (r=-0.800) and grading (r=0.61). Conclusions: The density of different immune cells is significantly involved in guiding prognosis of the CC in high-risk 16 and 18 HPV positive women; low cellular densities for CD3, CD20 and CD45 meaning limited immune response reflect negative disease outcomes promoting local relapse and decreased survival in such settings._x000D_

Corresponding author: Codrina Ancuta, Senior Lecturer, MD, PhD; e-mail: codrina_ancuta@yahoo.com

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11. Prenatal diagnosis and perinatal outcome in congenital diaphragmatic hernia. Single tertiary center report

Stefania Tudorache, Luminita Cristina Chiutu, Dominic Gabriel Iliescu, Raluca Georgescu, George Alin Stoica, Cristiana Eugenia Simionescu, Eugen Florin Georgescu, Raducu Nicolae Nemes

Purpose: To evaluate the perinatal results for fetuses and neonates with left-sided congenital diaphragmatic hernia (CDH) and the role of the prenatal diagnosis in the pregnancy outcome. Materials and Methods: We reviewed data from fetuses and neonates with left-sided CDH, managed from January 2009 and December 2013 in the University Clinic Hospital, Craiova, Romania. The following data were analyzed: the gestational age at the time of diagnosis, fetal karyotyping, presence of associated structural malformations, ultrasound (US) data (circumference and area of right lung, lung-to-head ratio - LHR, observed/expected LHR, hepatic herniation), the type of antenatal care, the pregnancy outcome, the place of birth and the conventional autopsy data, if performed. Perinatal outcomes were obtained by reviewing hospital documents. Results: Twenty-one cases were identified. No fetal surgery was performed in our series. Mean gestational age at time of diagnosis was 29 weeks of amenorrhea (WA) (range, 16-37 WA). Associated structural malformations were noticed in nine (42.8%) cases, in which three fetuses had a normal karyotype and two had chromosomal abnormalities, and four fetuses were not investigated. Isolated congenital diaphragmatic hernia was confirmed in 12 (57.1%) cases. All early second trimester diagnosed cases were terminated. The overall mortality rate was 61.9%. Rates of fetal deaths, early neonatal deaths, late neonatal deaths, and survival were 28.5%, 19%, 14.2%, and 38%, respectively. The perinatal mortality rate was 19% in cases with isolated congenital diaphragmatic hernia. Conclusions: The overall and perinatal mortality rate in congenital diaphragmatic hernia was still high in our series. Early perinatal deaths are associated with early diagnosis and with the presence of other structural defects. The prevalence of chromosomal abnormalities in perinatal death could not be determined from these data. In isolated congenital diaphragmatic hernia, mortality is related to the presence of herniated liver and severe pulmonary hypoplasia, this being well correlated with antenatal ultrasound parameters used for the estimation of fetal lung volumes. The antenatal diagnosis allowed better counseling of the parents, description of associations and improving the neonatal care.

Corresponding author: Luminita Cristina Chiutu, Associated Professor, MD, PhD; e-mail: luminita.chiutu@gmail.com

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12. Effect of apitherapy products against carbon tetrachloride-induced toxicity in Wistar rats

Calin Vasile Andritoiu, Vasile Andritoiu, Magdalena Cuciureanu, Delia Nica-Badea, Nela Bibire, Marcel Popa

The present paper aimed to evaluate the influence of apitherapy diet in Wistar rats with carbon tetrachloride induced hepatotoxicity, by the means of biochemical determinations and histopathological changes of liver, spleen, pancreas and testicular tissue. The experiment was carried out on six groups of male Wistar rats. Hepatic lesions were induced by intraperitoneal injection of carbon tetrachloride (dissolved in paraffin oil, 10% solution), 2 mL per 100 g, every two days, for two weeks. Hepatoprotection was achieved with two-apitherapy diet formulations (containing honey, pollen, propolis, Apilarnil, with/without royal jelly), that have been administered for six up to nine weeks. The biochemical results revealed that the two-apitherapy diet formulations had a positive effect improving the enzymatic, lipid, and protein profiles, coagulation, mineral parameters and also the bilirubin levels, after six weeks of treatment. The histopathological results demonstrated the benefit of the two-apitherapy diet formulations on reducing the toxicity of liver, spleen and pancreas in laboratory animals, after six and nine weeks, respectively. In conclusion, apitherapy products have a hepatoprotective effect in carbon tetrachloride-induced hepatopathy.

Corresponding author: Nela Bibire, Associate Professor, PhD; e-mail: nelabibire@yahoo.com

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13. Iron oxide nanoparticles modulate the interaction of different antibiotics with cellular membranes

Claudia Mihaela Istrate, Alina Maria Holban, Alexandru Mihai Grumezescu, Laurentiu Mogoanta, George Dan Mogosanu, Tudor Savopol, Mihaela Moisescu, Minodora Iordache, Bogdan Stefan Vasile, Eugenia Kovacs

The interaction of nanomaterials with cells and lipid bilayers is critical in many applications such as phototherapy, imaging and drug/gene delivery. These applications require a firm control over nanoparticle-cell interactions, which are mainly dictated by surface properties of the nanoparticles. The aim of this study was to investigate the interaction of Fe3O4 nanoparticles functionalized with several wide use antibiotics with opossum kidney (OK) cellular membranes in order to reveal changes in the membrane organization at different temperatures. We also investigated the in vivo biodistribution of the tested nanoparticles in a mouse model. Our results showed that, at low temperatures (31-35 degrees C), plain Fe3O4 nanoparticles induced a drop of the membrane fluidity, while at physiological or higher temperatures (37-39 degrees C) the membrane fluidity was increased. On the other hand, when nanoparticles functionalized with the tested antibiotics were used, we observed that the effect was opposite as compared to control Fe3O4 nanoparticles. Although most of antibiotics, used as plain solutions or linked on magnetite nanoparticles, proved heterogeneous effect on in vitro OK cells membrane fluidity, the aminoglycosides streptomycin and neomycin, used both as plain solutions and also combined with nanoparticles kept the same effect in all experimental conditions, increasing the membrane fluidity of OK cells plasma membrane. In vivo results showed that the antibiotic functionalized nanoparticles have a similar biodistribution pattern within the mouse body, being transported through the blood flow and entering the macrophages through endocytosis. Functionalized magnetite nanoparticles manifested a preferential biodistribution pattern, clustering within the lungs and spleen of treated mice. These results demonstrate that antibiotics manifest a different effect on plasma membrane fluidity depending on their type and temperature. Magnetite nanoparticles may interfere with antibiotic-cellular interactions by changing the plasma membrane fluidity. The fact that the antibiotic functionalized magnetite nanoparticles have a similar biodistribution pattern, are transported through the blood flow, and they increase the cellular uptake of the drug, suggest that they may be used for further studies aiming to develop personalized targeted delivery and controlled release nanoshuttles for treating localized and systemic infections.

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

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14. Correlation between corneal thickness and optic disc morphology in normal tension glaucoma using modern technical analysis

Laurentiu Coman, Monica Costescu, Mihail Alecu, Oana Andreia Coman

The purpose of this study was to evaluate the relationship between central corneal thickness (CCT) and optic disc morphology in normal tension glaucoma (NTG). Patients with NTG underwent eye examination, optic disc imaging with Heildelberg Retina Tomograph II (HRT II) and ultrasound corneal pachymetry. The morphological parameters of the optic discs were used to classify the eyes into four groups: generalized enlargement (GE) type, myopic glaucomatous (MY) type, focal ischemic (FI) type and senile sclerotic (SS) type. A correlation between CCT and optic disc morphology obtained by HRT II was calculated. Multiple comparison and post hoc tests were performed in order to determine the significance of the differences between the four groups. The strongest correlation was between CCT and the parameters of optic disc imaging obtained at HRT II in the GE type of optic disc.

Corresponding author: Laurentiu Coman, MD, PhD; e-mail: laurentiucmn@yahoo.com

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15. Clinical dental adhesive application: the influence on composite-enamel interface morphology

Ruxandra Matei, Mihai Raul Popescu, Mircea Suciu, Anne-Marie Rauten

Although the adhesion phenomenon is crucial in achieving and maintaining a composite building on dental structure, this phenomenon is not completely understood. On the other hand, adhesion is dependent on the interface quality (the interface between enamel and adhesive). In this study, the authors approached the subject of the influence of adhesive clinical application on the composite-enamel interface, which was less investigated by the scientists. On intact extracted human teeth were prepared enamel areas, and then filled with light-curing composite. The teeth were sectioned and prepared for microscopic investigation, at 10x, 100x and 200x magnifications.

Corresponding author: Mihai Raul Popescu, Associate Professor, DMD, PhD; e-mail: popescumihairaul@yahoo.com

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16. Intraocular biodistribution of intravitreal injected chitosan/gelatin nanoparticles

Andreea Dana Moraru, Marcel Costuleanu, Anca Sava, Danut Costin, Catalina Peptu, Marcel Popa, Dorin Chiselita

The aim of this study was to evaluate intraocular biodistribution of a fluorescent polymeric nanosystem composed of chitosan and gelatin after intravitreal administration in rat eyes. The nanoparticles based on chitosan and gelatin were synthesized using a reverse emulsion-double cross-linking technique (ionic and covalent) and their structural characteristics are presented. Two units of 1% suspension of fluorescein-labeled nanoparticles in saline solution were injected intravitreal in rat eyes. The histological cross-sections obtained at 24 and 72 hours were analyzed by confocal microscopy and compared to a similar number of control cross-sections. The scanning electron microscopy of the nanoparticles obtained by double cross-linking in reverse emulsion technique revealed spherical, smooth, highly porous particles with no tendency to form aggregates. The chitosan-fluorescein conjugate was present in all the ocular tissues both at 24 and at 72 hours. The nanoparticles were present in the retina in a larger quantity and persisted longer than in the other ocular tissues. They were mainly fixed paravascular. The double cross-linking in reverse emulsion technique was efficient in synthesizing a biocompatible polymeric nanosystem. The in vivo study of intraocular biodistribution of fluorescein-labeled nanoparticles revealed their affinity for the retina after intravitreal administration.

Corresponding author: Anca Sava, MD; e-mail: dr_anca_sava@yahoo.com

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17. Molecular study of weight gain related to atypical antipsychotics: clinical implications of the CYP2D6 genotype

Laura Alexandra Nussbaum, Victor Dumitrascu, Anca Tudor, Raluca Gradinaru, Nicoleta Andreescu, Maria Puiu

Atypical antipsychotics, especially some of them, influence cellular lipogenesis, being associated with metabolic side effects including weight gain. Due to the increasing use of atypical antipsychotics in children and adolescents, their metabolic and endocrine adverse effects are of particular concern especially within this pediatric population that appears to be at greater risk. Genetic factors with a possible influence on atypical antipsychotics adverse effects include CYP2D6 polymorphisms. Our study, performed in 2009-2014, with a two-year enrolment period during which we recruited children and adolescents with a diagnosis of schizophrenia or bipolar disorder on treatment with the antipsychotics (Risperidone, Aripiprazole or Olanzapine), included 81 patients, aged between 9 and 20 years, median age being 15.74 years. The gender percentage was 54% girls/46% boys. The CYP2D6 genotyping was performed after enrolment of the last patient. Based on the CYP2D6 genotype, three activity groups were identified and compared and we found that the patients with wt/4 genotype, intermediary metabolizer (carrier of one functional and one non-functional allele) have significantly higher weight gain values than the patients who did not exhibit allele 4. The CYP2D6 genotype in children and adolescents with schizophrenia and bipolar disorder, proved to be a good predictor for the response to atypical antipsychotics and the side effects registered. The significant correlations between the CYP2D6 polymorphisms and the weight gain/BMI (body mass index) increase, as major side effects induced by antipsychotics proved the fact that the pharmacogenetic screening is needed in the future clinical practice, allowing for individualized, tailored treatment, especially for at-risk individuals.

Corresponding author: Raluca Gradinaru; e-mail: ralucagradinarutm@yahoo.com

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18. An endoscopic and pathological survey of digestive tract disorders in patients infected with human immunodeficiency virus monitored in the Clinic of Infectious Diseases from Tirgu Mures, Romania

Nina-Ioana Sincu, Simona Mocan, Lucia Carmen Chiriac, Simona Bataga

Gastrointestinal symptoms are among the most frequent complaints of patients infected with human immunodeficiency virus (HIV). Purpose: An endoscopic and histopathological survey of digestive tract diseases among HIV-infected patients monitored in the Clinic of Infectious Diseases I from Tirgu Mures, Romania. Materials and Methods: Retrospective, observational study, on a group of 38 HIV-positive patients admitted to the Clinic of Infectious Diseases I from Tirgu Mures, Romania, during 2006-2013, undergoing upper/lower endoscopy. We collected data regarding the results of endoscopy and histopathological examination, CD4+ T-lymphocytes levels, microbiological examinations and outcome. Statistical analysis, performed by using Microsoft Office Excel 2007 and GraphPad Prism 5 programs, included contingency tables analysis and comparing means. Results: Our study depicted a variety of digestive disorders among HIV-infected patients, ranging from opportunistic infections to HIV enteropathy and non-HIV-associated conditions. The presence of Candida esophagitis implied significantly lower levels of CD4+ T-cells (p=0.0043). We found a statistically significant negative association between antiretroviral therapy and the presence of opportunistic infections (p=0.0375, OR=0.2030, 95% CI 0.0423-0.9741). Thirteen (34.21%) patients died, mostly due to tuberculosis and central nervous system infections. All were diagnosed with acquired immunodeficiency syndrome (AIDS). Conclusions: HIV-infected patients experience a wide variety of digestive tract disorders, both AIDS-defining illnesses and non-HIV-associated conditions. Gastrointestinal opportunistic infections occur more often among patients with low CD4+ T-cells levels and in those not receiving antiretroviral therapy. Although digestive conditions did not represent direct causes of death in our study, they may predict an unfavorable outcome in AIDS-stage patients.

Corresponding author: Lucia Carmen Chiriac, Lecturer, MD, PhD; e-mail: carmen.l.chiriac@gmail.com

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19. An animal model of peripheral nerve regeneration after the application of a collagen-polyvinyl alcohol scaffold and mesenchymal stem cells

Silviu-Adrian Marinescu, Otilia Zarnescu, Ioana-Ruxandra Mihai, Carmen Giuglea, Ruxandra Diana Sinescu

Extensive nerve injuries often leading to nerve gaps can benefit, besides the gold standard represented by autologous nerve grafts, by the inciting field of tissue engineering. To enhance the role of biomaterials in nerve regeneration, the nerve conduits are associated with Schwann or Schwann-like cells. In this study, we evaluated rat sciatic nerve regeneration, by using a biodegradable nerve guide composed of Collagen (COL) and Polyvinyl Alcohol (PVA), associated with mesenchymal stem cells (MSC). After the exposure of the rat sciatic nerve, a nerve gap was created by excising 1 cm of the nerve. Three experimental groups were used for nerve gap bridging: autografts, nerve conduits filled with medium culture and nerve conduits filled with MSC. The methods of sensory and motor assessment consisted of the functional evaluation of sciatic nerve recovery - toe-spread, pinprick tests and gastrocnemius muscle index (GMI). The histological and immunocytochemical analysis of the probes that were harvested from the repair site was performed at 12 weeks. Successful nerve regeneration was noted in all three groups at the end of the 12th week. The functional and immunocytochemical results suggested that COL-PVA tubes supported with mesenchymal stem cells could be considered similar to autologous nerve grafts in peripheral nerve regeneration, without the drawbacks of the last ones. The functional results were better for the autografts and the ultrastructural data were better for the nerve conduits, but there were not noticed any statistical differences.

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

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20. Inhibin alpha-subunit, Melan A and MNF116 in pheochromocytomas

Diana Paun, Marius-Cristian Neamtu, Elena Taina Avramescu, Madalina Gabriela Georgescu, Rodica Petris, Dana Terzea, Catalina Poiana, Sorin Paun

Aim: The aim of this study was to make immunohistochemical analyses with Inhibin alpha-subunit, Melan A and MNF116 (pan-Cytokeratin antibody) in pheochromocytomas, because immunohistochemistry is useful for the distinction between adrenal tumors. Patients and Methods: We used 20 patients with pheochromocytomas submitted to laparoscopic (n=19) or classical (n=1) surgery and we have explored immuno-staining with Inhibin alpha-subunit, Melan A and MNF116 in these tumors. This can be helpful when we cannot make the distinction between adrenal tumors. Results: Pheochromocytomas did not stain with Inhibin alpha-subunit, Melan A and MNF116. Conclusions: In our study, Inhibin alpha-subunit, Melan A and MNF116 were not sensitive for pheochromocytomas._x000D_

Corresponding author: Madalina Gabriela Georgescu MD, PhD; e-mail: madalina.georgescu@gecad.com

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21. Craniofacial morphology in patients with Angle Class II division 2 malocclusion

Dana Cristina Bratu, Raluca-Adriana Balan, Camelia-Alexandrina Szuhanek, Silvia-Izabella Pop, Emanuel Adrian Bratu, George Popa

Cephalometric analysis is frequently used in orthodontics for diagnostic purposes, in order to evaluate the positional relationship of the upper and lower jaw to the cranial base, as well as to objectively asses the way dental arches relate to one another and to the skeletal base. As with other dento-skeletal anomalies, the normal growth process can induce changes in these parameters. The purpose of this study is to evaluate the skeletal and dental changes that occur in growing patients with Angle Class II division 2 malocclusion. The study also focuses on analyzing and comparing several parameters in three groups of young patients of different ages, diagnosed with Class II division 2 malocclusion, in order to determine whether the anomaly worsens or improves during the patients growth period. A total of 25 lateral skull teleradiographs were analyzed using cephX. The patients were divided into three groups (Group 1: 6-8 years, Group 2: 9-14 years, Group 3: 15-18 years). We used the cephalometric parameters described in Bjork-Jarabak and Tweed analyses, as well as the relationship of the upper and lower central incisors to the skeletal landmarks. The statistical methods used in this study were the analysis of variance (ANOVA) and the unpaired Student s t-test (p<0.05). We concluded that, during the physiological growth process, the Angle Class II division 2 malocclusion has the following cephalometric characteristics: the maxillary central incisors were in accentuated retroclination, the interincisal angle was very obtuse, the gonial angle showed lower than normal values towards the end of the growth period, the lower anterior face height was definitely decreased, the mandibular body length was shorter than normal in the early growth period and the tendency towards a hypodivergent skeletal pattern remained stable during growth.

Corresponding author: Raluca-Adriana Balan, University Assistant, DMD, PhD; e-mail: raluca_balan22@yahoo.com; George Popa, DMD, PhD Student; e-mail: georgepopa86@yahoo.com

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22. Pathological assessment of tumor biopsy specimen and surgical sentinel lymph node dissection in patients with melanoma

Gheorghe Noditi, Cristian C. Nica, Horatiu Pompiliu Petrescu, Codrut Ivan, Zorin Petrisor Crainiceanu, Tiberiu Bratu, Alis Dema

Introduction: Actual trends of cutaneous malignant melanoma show a faster increase then other forms of cancer. Early detection and diagnosis, and accurate pathologic interpretation of the biopsy specimen is extremely important for the treatment and prognosis of clinically localized melanoma. The surgical approach to cutaneous melanoma patients with clinically uninvolved regional lymph nodes remains controversial. Materials and Methods: A retrospective study of melanoma cases was conducted in the Casa Austria Department of Plastic and Reconstructive Surgery, Emergency County Hospital, Timisoara, Romania. We have analyzed the medical records of 21 patients that underwent surgical treatment for different stages of melanoma in the period 2008-2012. For histopathological diagnosis of melanoma and the sentinel lymph node(s) status, tissular fragments were routinely processed. For the difficult cases, additional immunohistochemical investigation was done. Results: A positive family history was noted in two cases. The presence of different sizes and localization of pigmented nevi was found in 38% of the cases. Different types of melanoma like superficial spreading melanoma, nodular melanoma or lentigo malignant melanoma and acral lentiginous melanoma was described. The surgical treatment consisted in all cases in wide excision of the primary tumor and prophylactic dissection of sentinel lymph node after lymphoscintigraphy examination. A positive biopsy of the sentinel lymph node was noted in 4.9% of the cases. Conclusions: The surgical treatment combining the wide excision of the primary tumor with respect to safe oncological limits with the prophylactic dissection of sentinel lymph node after lymphoscintigraphy examination had the confirmation done by the pathologic interpretation of the biopsy specimen showing that all the patients had a Breslow index more than 1.5 mm.

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

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23. The expression of CK19, vimentin and E-cadherin in differentiated thyroid carcinomas

Cristina Mariana Calangiu, Cristiana Eugenia Simionescu, Alex Emilian Stepan, Daniela Cernea, Roxana Eugenia Zavoi, Claudiu Margaritescu

Thyroid carcinomas constitute lesions with an ascending incidence, for which many prognosis assessment systems were developed. This study focused on analyzing the immunoexpression of CK19, vimentin and E-cadherin in a number of 43 differentiated thyroid carcinomas, of which 39 papillary carcinomas and four follicular carcinomas, and assessed the relationship of these markers with clinico-pathological parameters of interest, such as age and gender of patients, the histological type and subtype, tumor size and extension, metastases in regional lymph nodes and tumor stage. CK19 immunostaining indicated higher scores in conventional and follicular papillary carcinomas compared with tall cell variant. In relation to the size and extension of the tumor, we found significantly higher values of vimentin and E-cadherin scores in T1-T2 carcinomas compared with T3-T4 category and a positive linear distribution of these markers, which sustain their involvement in common mechanisms of tumor progression.

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

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24. Morphological changes of the peritoneal membrane in patients with long-term dialysis

Traian Taranu, Laura Florea, Dumitru Paduraru, Stefan Octavian Georgescu, Laurian Lucian Francu, Cristinel Ionel Stan

Morphological alterations of peritoneum in chronically dialyzed patients involve fibrosis and angiogenesis as pathogenic mechanisms. The aim of this retrospective study was to evaluate morphological changes of peritoneum in chronic peritoneal dialysis (PD) at 4, 8, 12, and 14 years. Peritoneal changes were investigated in 110 patients with end stage renal failure, which were included in a PD program. Intraoperative biopsies were grouped in four study Groups (A: 1-48 months, B: 49-96 months, C: 97-144 months, and D: 145-168 months), and were processed histologically and stereologically. Mesothelial denudation was found in percentage volumes of 5.49% - Group A, 16.10% - Group B, 16.68% - Group C and 19.88% - Group D. Reduplication of the basement membrane was observed in patients with over five years of PD. Interstitial stromal fibrosis recorded percentage volumes of 25.49% (Group A), 26.10% (Group B), 35.85% (Group C) and 56.63% for the patient with 14 years of PD. Subendothelial hyalinizing vasculopathy was recorded in percentage volumes of 2.22%, 6.63%, 9.16% up to 9.20%. Vascular permeability reduction was recorded as decreasing percentage volumes from 22.59% to 12.81%, 7.77% and 7.37%. Perivascular inflammation was marked in the serosa of the patients in Group A (4.55%). Calcifications recorded percentage volumes of 1.63% at eight years, 3.74% at 12 years and 4.03% at 14 years of PD. Peritoneal morphological changes appear at 3-4 years of PD and progressively aggravate with long-term PD.

Corresponding author: Laura Florea, MD; e-mail: lflorea68@yahoo.com

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25. Anatomoclinical aspects of conjunctival malignant metastatic melanoma

Claudia Florida Costea, Kreolla Anghel, Gabriela Dimitriu, Gabriela Florenta Dumitrescu, Ziyad Faiyad, Ana-Maria Dumitrescu, Anca Sava

Conjunctival malignant melanoma is a rare tumor with a high risk of local recurrence, lymph node and systemic metastases. The aim of this study was to correlate tumor thickness, tumor ulceration, high mitotic rate, epithelioid cells with the presence of metastases and death from conjunctival malignant melanoma. We report the case of a 33-year-old patient who presented with a left eyelid ptosis associated with an eyelid prominence, foreign body sensation in the eye, and bloody discharge, symptoms occurring about one month earlier. Ophthalmologic examination revealed in eyelid conjunctiva two vegetant and ulcerative tumors of 8/6 mm and 3/3 mm. The two tumors were surgically removed with safety margins. The diagnosis of amelanotic malignant melanoma of the conjunctiva with brain metastasis was made by routine morphological methods and immunohistochemical reactions (HMB45, vimentin, S100 protein). Systemic metastases (skin, brain, lung, liver, kidney, peritoneal) and peripancreatic lymph node metastases were detected at 1.9 years after the diagnosis of conjunctival malignant melanoma. The patient died three months after the surgical excision of brain metastasis. Early diagnosis is essential to prevent tumor recurrence, ocular invasion, systemic and lymph node metastases, and preserving visual function.

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

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26. Micropapillary urothelial carcinoma: an aggressive variant of urothelial carcinoma

Andrada Loghin, Calin Chibelean, Martha Orsolya, Adela Nechifor-Boila, Alin Nechifor-Boila, Angela Borda

Micropapillary urothelial carcinoma (MPC) is a rare variant of urothelial carcinoma (UC) with an aggressive clinical course, an advanced stage at first presentation and a high metastatic potential. The aim or our study is to present five illustrative cases of MPC, diagnosed among the 21 patients with UC treated by radical cystectomy in the Department of Urology, County Hospital of Tirgu Mures, Romania, between January 1, 2011 and December 31, 2013. The morphological and immunohistochemical features of this rare and aggressive variant of UC, as well as a brief review of the literature are all presented. All five cases were associated with lymph node metastases with micropapillary features, regardless of the microscopic aspect of the tumor on the surgical specimens - transurethral resection (TUR) or cystectomy. Three of them had a micropapillary component in the TUR, on the cystectomy specimen, or in both, along with lymph nodes metastases. In two cases, the MPC features were present only in the lymph node metastasis, with a conventional UC on the TUR and on the cystectomy. Immunohistochemical staining demonstrated that both micropapillary and associated conventional UC were positive for CK7 and CK20. Ki67 was expressed in 40% of tumor cells and CD34 was positive in the endothelial cells and negative in the flattened spindled cells lining the retraction spaces around tumor cell nests. MPC is a highly aggressive variant of UC with specific morphological characteristics. Any amount of micropapillary component found in UC is significant, and should be reported because it encompasses an aggressive clinical behavior and a poor prognosis.

Corresponding author: Calin Chibelean, Lecturer, MD, PhD; e-mail: calinchibelean@yahoo.com

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27. Desmoplastic melanoma - challenges in the diagnosis and management of a rare cutaneous tumor

Irina Margaritescu, Aurel Doru Chirita

Desmoplastic melanoma (DM) represents a distinctive rare variant of spindle cell melanoma with a predilection for chronically sun-exposed skin of the elderly. This neoplasm is notoriously difficult to diagnose, both clinically and histopathologically. Therefore, DM is deeply infiltrative at the time of presentation. Histologically, the tumor presents as a proliferation consisting of non-pigmented spindle cells arranged in poorly formed fascicles. The neoplastic cells have a deceptively bland appearance with slightly pleomorphic and hyperchromatic nuclei, inconspicuous nucleoli and low mitotic activity. DM can mimic a whole range of benign and malignant neoplasms with spindle cell and fibrous appearance. Even though S100 remains the first-choice marker for DM, currently, there is no reliable marker with both high sensitivity and specificity for its detection. However, emerging melanoma markers, such as SOX10, have shown promising results in the diagnosis of DM. An accurate diagnosis of DM should always be based on the integration of all the clinical, histological and immunohistochemical features. Once diagnosed, DM should be aggressively excised with at least 2 cm lateral margins and down to the fascia. We present a case of DM that appeared on a non sun-exposed site. The tumor recurred multiple times in spite of repeated surgery involving wide local excisions and histologically reported negative margins. Recurrences are almost always associated with the presence of neurotropism. In our case, the neurotropism was obvious only in the second recurrence. We highlight the difficulties encountered in the diagnosis and management of both the initial tumor and its recurrence.

Corresponding author: Irina Margaritescu, MD, DipRCPath; e-mail: irina.margaritescu@gmail.com

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28. Role of optic microscopy for early diagnosis of Menkes disease

Dana Craiu, Stephen Kaler, Mihai Craiu

We report the case of a male patient with a normal development in the first three months of life, presenting for global regression, central axial hypotonic syndrome, pyramidal syndrome, focal epileptic seizures, and a particular aspect of the hair - almost absent, short, sparse, lightly colored, at age of five months, becoming coarse, twisted (kinky hair) by the age of 21 months. Different diseases associate similar neurological and macroscopic aspect of the hair (biotinidase deficiency, argininosuccinic aciduria, aminoaciduria, giant axonal neuropathy, trichothiodistrophy and Menkes syndrome). The microscopic aspect of the patient s hair showing normal hair, silver colored hair, hair shafts twisting 1800, trichoclasis, and trichoptilosis, was highly characteristic for Menkes disease. Diagnosis was further supported by the low concentration of serum copper and ceruloplasmin and exclusion of other metabolic disorders with similar macroscopic aspect of the hair. Molecular genetic testing by multiplex PCR indicated deletion of exon 22 in the ATP7A gene situated in Xq21.1 region, consistent with the clinical and biochemical phenotype. Physicians should use microscopic evaluation of the hair more often when suspicion of Menkes disease is raised, aiming a narrow further diagnostic workup and early positive diagnosis and genetic advice for the affected families.

Corresponding author: Dana Craiu, Associate Professor, MD, PhD; e-mail: dcraiu@yahoo.com

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29. Changing the root morphology in a case of periodontal impairment at a maxillary molar used as a sustaining tooth

Constantin Daguci, Luminita Daguci, Marilena Bataiosu, Mihai Raul Popescu, Adina Bunget, Ruxandra Margarit, Oana Cella Andrei, Adina Dumitrache, Nicolae Vasile, Ruxandra Sfeatcu

Research evolution regarding dental research determines the increase of patient s desire to preserve their dentition for a long time. The loss of a molar tooth as the last teeth present on the dental arch, raises issues regarding how the prosthesis should be placed, still patients are usually reluctant to use as last resort the prosthetic solution. This article describes the technique of root amputation to save a maxillary molar used as bridge abutments.

Corresponding author: Constantin Daguci, Lecturer, DMD, PhD; e-mail: dagucicristi@yahoo.com

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30. Clinical and histopathological studies using fibrin-rich plasma in the treatment of bisphosphonate-related osteonecrosis of the jaw

Octavian Dinca, Sabina Zurac, Florica Staniceanu, Mihai Bogdan Bucur, Dana Cristina Bodnar, Cristian Vladan, Alexandru Bucur

The authors report their experience using platelet-rich fibrin (PRF) therapy for the treatment of ten patients presenting bisphosphonate-related osteonecrosis of the jaw (BRONJ). The aim of our study was to evaluate the effect of this therapy on recurrent BRONJ and to describe the clinical and histopathological/immunohistochemical staining features of PRF treatment. As such, we describe the method we used and report the results observed in the areas treated as well as side effects. The reported results recommend the safety and efficacy of PRF in treatment of BRONJ.

Corresponding author: Cristian Vladan, Teaching assistant, MD, DDS, PhD; e-mail: crystalvladan@gmail.com

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31. Cutaneous mastocytosis, problems of clinical diagnosis of four cases

Virgil Patrascu, Andreea-Oana Enache, Raluca Niculina Ciurea, Corneliu Cristian Georgescu, Alina Maria Vilcea, Loredana Elena Stoica, Marius Eugen Ciurea

Mastocytosis is a rare disease characterized by a pathological increased of mast cells in one or more tissues, particularly in the skin, bone marrow, liver, spleen, lymph nodes and gastrointestinal tract. Cutaneous mastocytosis represents over 90% of cases found with predilection in children. The aim of the paper was to summarize the authors clinical, histopathological and immunohistochemical observations on patients with cutaneous mastocytosis. We present four cases of cutaneous mastocytosis, sporadic form, customized by clinical presentation and age of onset: two installed in the neonatal period, a case with onset in infancy and another in adulthood. For the assessment of the severity and the effectiveness of the treatment, we used SCORMA Index. We performed in each patient histopathological examination of the skin (Hematoxylin-Eosin and Giemsa stains), the dosage of mediators (serum tryptase level, serum histamine levels, urinary histamine metabolites) and the balance of expansion (complete blood cell count, liver biological investigations, abdominal ultrasound, skeletal radiography, chest radiography). For the adult with mastocytosis, we performed abdominal scanner and cytological study of the bone marrow. Following investigations carried out in each case, we mentioned the diagnosis of cutaneous mastocytosis, and also excluded several diseases confounded by clinically and histologically aspect. Considering the fact that the balance expansion was negative, we excluded the diagnosis of systemic mastocytosis. The presence of anemia and protein energetic malnutrition in children with mastocytosis involves carrying out balance extension for the exclusion of a systemic form of the disease. Histopathological examination of the skin using special stains, the dosage of mediators (serum tryptase level, serum histamine levels, urinary histamine metabolites) and balance expansion establish the diagnosis of cutaneous mastocytosis and also exclude many confusions because of the clinical presentation.

Corresponding author: Virgil Patrascu, Associate Professor, MD, PhD; e-mail: vm.patrascu@gmail.com

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32. A rare tumor revealed by abdominal trauma: case presentation

Camelia Cristina Diaconu, Dorel Arsene, Alice Lavinia Balaceanu, Daniela Bartos

Pseudomyxoma peritonei is a rare and poorly understood form of disease characterized by mucin deposits in the peritoneum. The term includes a broad range of neoplasms with different patterns of evolution, from benign to borderline or even to malignant lesions. The disease may be asymptomatic until advanced stages. We present the case of a 65-year-old patient who presented for pain in the right hemiabdomen, after a trauma by falling from small height. Abdominal imaging studies oriented the diagnosis to a traumatic disease. At laparotomy, a mucinous tumor attached to the right colon was discovered. The main particularity of the case is that the origin of the pseudomyxoma could not be identified.

Corresponding author: Camelia Cristina Diaconu, Associate Professor, MD, PhD; e-mail: camiluciemi@yahoo.com

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33. Aneurysmal bone cyst - clinical and morphological aspects

Radu Radulescu, Adrian Badila, Robert Manolescu, Maria Sajin, Ion Japie

Aim: To assess the results of surgical treatment in aneurysmal bone cysts. Materials and Methods: 31 patients with aneurysmal bone cysts underwent surgical treatment in our department. In almost half of cases, the lesion was located in the femur. In 12 cases, a pathological bone fracture was the first clinical sign. The treatment consisted in curettage, abrasion of the cavity inner walls using a motorized burr and filling with morsellized bone grafts (autografts +/- allografts) or bone substitutes (four cases). Microscopically, the diagnosis relies on cystic spaces filled with blood, divided by fibrous septae consisting in immature bone trabeculae, hemosiderin filled macrophages and fibroblasts. We performed multiple bioptic probes from different levels of the lesion. Results: Macroscopically, the osseous lesion appeared as a multi-loculated blood-filled cavity (cavities separated by septa) in 30 (96.77%) cases and as a solid tumor in one (3.23%) case. At 12 months after surgery, grafts osteointegration was present in 24 cases. At an average follow-up time of six years and four months, refilling with bone grafts was necessary in two cases and no local recurrence was observed. Conclusions: Aneurysmal bone cyst is most frequent in the second decade of age. Its prevalence in female gender is double compared to the male gender. The positive diagnosis relies on the histopathological examination. Because of the strong relationship with a number of precursor lesions (giant cell tumor, fibrous dysplasia, non-ossifying fibroma, chondroblastoma, osteoblastoma) multiple bioptic probings are mandatory, in order to diagnose, if possible, a primary lesion, which may modify the therapeutic attitude. Treatment by curettage, abrasion of the cavity inner walls and filling with morsellized grafts has very good results. The risk of recurrence is very low.

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

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34. Antenatally diagnosed congenital pyloric duplication associated with intraluminal pyloric cyst - rare entity case report and review of the literature

Cristina Oana Marginean, Claudiu Marginean, Emoke Horvath, Liliana Gozar, Horea Gheorghe Gozar

Duplication of the digestive tract is a very rare malformation in children whereas the congenital pyloric duplication is extremely rare, few cases being described in the literature. We present the case of a male infant, aged seven days, who was diagnosed at 17 weeks of gestation with a cystic mass in the abdomen and who presented vomiting in the sixth day of life. Ultrasound and CT diagnosed a pyloric duplication cyst. The malformation was visualized intraoperatively and removed successfully without incidents. The infant s postoperative evolution was excellent, with an upward weight curve without a dumping syndrome. Finally, we present a review of the published cases of congenital pyloric duplications in the world, this one being the fifth one reported of the prenatally diagnosed cases.

Corresponding author: Claudiu Marginean, Associate Professor, MD, PhD; e-mail: marginean.claudiu@gmail.com

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35. Medico-historical overview and histopathological comments about a hyalinizing trabecular tumor case of thyroid gland

Cristian Barsu, Marina Barsu

Introduction: Hyalinizing Trabecular Tumor (HTT) of the thyroid gland is a rare and controversial tumor, whose rigorous identification is extremely important due to the difficulty of establishing a correct diagnosis using only the routine histological stains (Hematoxylin-Eosin, Periodic Acid-Schiff, van Gieson, etc.). Using only usual stains, difficult problems of differential diagnosis arise. In this paper, we report a clinically unexpected case of HTT, the first in Romania, whose diagnosis has been established only by immunohistochemistry reactions. Materials, Methods and Results: In assessing a case with nodular goiter for diagnose purpose, we have found a nodule difficult to diagnose. We have been determined to appeal to some additional immunohistochemical techniques. Conclusions: According to the immunohistochemistry reactions made, we found that particularly the Ki-67 immunohistochemical stain is obvious indispensable in establishing the diagnosis of HTT. We present also a medico-historical overview concerning some first descriptions of this rare entity. There are also mentioned some data regarding the behavior of this rare thyroid tumor.

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

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36. Unusual morphological pattern and distribution of the ansa cervicalis: a case report

George K. Paraskevas, Konstantinos Natsis, Zoe Nitsa, Alexandra Mavrodi, Panagiotis Kitsoulis

Ansa cervicalis presents great anatomic variability regarding its origin and formation, the number of its roots and its distribution, as well. In the current case, we report an aberrancy in the form and distribution of ansa cervicalis branches to the infrahyoid muscles and the sternocleidomastoid muscle, which is unique, since, to the best of our knowledge, a similar case has not been recorded in the literature. During regular dissection, we detected that the ansa cervicalis loop, which was formed underneath the superior belly of the omohyoid muscle, provided a branch for the sternothyroid muscle, from which two recurrent rami were arisen. These two rami joined together forming an unusual triangular nerve formation. The neural trunk formed by the union of the aforementioned two rami perforated the inferior belly of the omohyoid muscle and afterwards was directed towards the ipsilateral sternocleidomastoid muscle. The awareness of such an unusual variability to the surgeons of the head and neck region would be of great importance, since it is crucial not to damage the ansa cervicalis or its branches in order to prevent any possible phonation disorders. Additionally, ansa cervicalis is proved to be extremely useful in the re-innervation of the larynx following paralysis of the recurrent laryngeal nerve._x000D_

Corresponding author: George K. Paraskevas, Assistant Professor, MD, PhD; e-mail: g_paraskevas@yahoo.gr

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37. Internal jugular vein cannulation complications and elimination of the muscular triangle of the neck due to aberrant infrahyoid muscles

Athanasios Raikos, Ashwin Agnihotri, Saif Yousif, Panagiota Kordali, Minu Saberi, Beate Brand-Saberi

We report on a rare case of anatomical variations of the infrahyoid muscles with prominent clinical significance. The aberrant anatomy was on the right side of the neck and involved the omohyoid and sternohyoid muscles. The superior belly of the omohyoid was duplicated in width due to an aberrant belly anteriorly and merged with fibers of the inferior belly inferiorly and the sternohyoid muscle medially. An additional aberrant muscle slip extended between the inferior third of the sternohyoid muscle and united with the inferior belly of the omohyoid. The intermediate tendon between the two bellies of the omohyoid was absent, whereas the so-called muscular triangle of the neck was diminished. Due to the arrangement and fusion of myofibers the muscle could be termed as omo-sternohyoid muscle. A profound hematoma was noted in the aberrant muscle at the area overlying the internal jugular vein indicating difficulty in obtaining jugular venous access for catheter placement. Clinicians and surgeons should be aware of muscular anatomic variations when intervening in the lateral neck area as the classical anatomical landmarks might be misinterpreted and confuse._x000D_

Corresponding author: Athanasios Raikos, Assistant Professor, MD, PhD; e-mail: a.raikos@yahoo.com

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38. Book Review: Jacques Poirier: Petit dictionnaire du charlatanisme medical (Small dictionary of medical fraud)

Catalin Ilie

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Corresponding author: -

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