Archive issue

1. Intestinal Microbiome, Small Intestinal Bacterial Overgrowth and Inflammatory Bowel Diseases - What are the Connections?

M. ANDREI(1), T. NICOLAIE(1), ADRIANA STOICESCU(1), ADRIANA TEIUSANU(1), S. GOLOGAN(1), M. DICULESCU(2)

IBD (inflammatory bowel diseases) represent chronic idiopathic inflammatory diseases, prone to relapse in the digestive tract; it is estimated that they result from the interaction of the intestinal microbiome with the intestinal immune system. The inflammatory microbiome exerts multiple beneficial roles. Perhaps the central element to developing IBD is dysbiosis; there is still an incompletely established association between intestinal microbiome changes in patients with IBD and SIBO (small intestinal bacterial overgrowth). Influencing the intestinal microbiome may play an adjuvant therapeutic role in the treatment of IBD. We present a synthesis of the connections between the entities mentioned above.

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2. Autophagy and Neovascularization in Colorectal Cancer

C.D. USCATU1,2 , T. DUMITRESCU3,4 , IONELA CRISTINA DELIU5 , ALEXANDRA DUMITRESCU6 , M. SCHENKER7 , F. MIXICH1

Purpose: This study aims to determinate the microvessel density at the base of the tumor, as well in tumor’s mass, in order to determinate the number of neovascularization vessels (marked with CD105) in comparison with presence or absence of autophagy puncta. Material/Methods: Standard immunohistochemistry was performed on 38 samples of colorectal adenocarcinoma, in order to determinate the presence of autophagy and neovascularization blood vessels with the help of LC3, CD34, CD31 and CD105 antibodies. Results: The autophagy process was observed in the cancerous cells and was noted as present in both regions of interest from the tumor. The mean number of blood vessels market with CD105 is higher in tumor mass then at its base, p value of the Student t test being highly significant (p<0.0001). Conclusions: The presence of autophagy puncta was notice in every case, both in the mass of the tumor and at its base. Microvascular density of new-grown blood vessels is higher in the mass of the tumor compared with the base of the tumor.

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3. The Estrogen-Induced Effects on Myometrium Are the Result of Activation of Two Different Receptor Types: GPER and Er?

OANA SORINA TICA(1), A. COMANESCU(1), MARIA BOGDAN(2), D.O. ALEXANDRU(3), A.A. TICA(4), T. CIUREA(5)

GPER (G protein coupled estrogen receptor 1), a particular estrogen binding site, is ubiquitously present in human tissues, but its precise physiological role is still very disputed. GPER is associated with normal and abnormal estrogen-dependent proliferations in female tissues and is involved in generation of rapid estrogenic answers. A very important fact is that GPER-induced genomic effects are additive to those mediated by “classic” estrogen receptors, but regarding the rapid effects, as we prove in this study, these can be significant different or even antagonistic.

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4. Musculoskeletal Ultrasound in Early Rheumatoid Arthritis - Correlations with Disease Activity Score

CARMEN MITRAN(1), ANDREEA BARBULESCU(2), F.A. VREJU(3), CRISTINA CRIVEANU(3), ANCA ROSU(3), PAULINA CIUREA(3)

Purpose: to assess synovial inflammation using musculoskeletal ultrasonography, in a cohort of patients with early rheumatoid arthritis, and establish the correlations with disease activity score. Material and methods: We enrolled 29 patients diagnosed with early RA, according to ACR-EULAR 2010 classification criteria, in Rheumatology Clinic, Emergency County Hospital Craiova, between September 2013-August 2014. We performed clinical evaluation, assessed laboratory tests and performed US for all the patients. Musculoskeletal ultrasonography was performed using an ESAOTE MyLab 25 ultrasound, by the same examiner, with multi-frequency linear array transducers (7-12Mh). The presence of synovitis was assessed both in grey scale (GSUS) and power-Doppler (PDUS), according to OMERACT-EULAR consent. Results: Assessment of synovial inflammatory activity by ultrasound examination, using both grey scale and PDUS, has shown active synovitis in all patients, with a mean number of joints with active synovitis of 5. Evaluating the grade, using PDUS, we found grade 1 in all patients, 2 in 14 and grade 3 in 3 patients. Inflammatory markers correlated significant with both GSUS and PDUS. Analysis of correlation between GSUS examination parameters and disease activity score, found a positive, moderate inter-relation, results found also for PD score. Conclusion: our results sustain the role of US for detecting inflammatory activity in early RA patients, and, in addition with clinical and biological assessment, represents a sensitive, reliable and reproducible method, easily approached, that should be included in our routine evaluation

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5. Immunoexpression of N-cadherin, Twist and Vimentin in Bladder Urothelial Carcinomas

A.E. STEPAN(1), MIRELA CIUCA(2), CRISTIANA SIMIONESCU(1), DESDEMONA STEPAN(3), C. MARGARITESCU(1)

Purpose: Epithelial mesenchymal transition consists in the acquisition of neoplastic epithelial cells of a mesenchymal phenotype the process being involved in cancers invasion and metastasis. In this study were analyzed the expression of N-cadherin, Twist and Vimentin in bladder urothelial carcinomas according to the main prognostic parameters. Material/Methods: The study included 20 bladder urothelial carcinomas which were analyzed histopathological, immunohistochemical and statistical. Results: N-cadherin was identified in 45% of cases, which belonged to high-grade carcinomas with deep invasion and lymph node metastases. Twist immunoreaction was identified in all cases and was significantly increased in advanced stages carcinomas. Vimentin was present only in the advancing edge in 25% of cases, which belonged to highly invasive carcinomas. Urothelial carcinoma metastases were N-cadherin and Twist and Vimentin negative. We found a linear positive distribution of N-cadherin and Twist values. Conclusion: the used markers are useful for identifying aggressive urothelial carcinomas in the context of reciprocal stimulation mechanisms inside of urothelial epithelial-mesenchymal transition process.

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6. Particularities of Renal Manifestations in Chronic Viral Hepatitis

V.F. IOVANESCU(1), C.T. STREBA(2), D.O. ALEXANDRU(3), A.F. CONSTANTINESCU(1), C.C. VERE(2), I. ROGOVEANU(2), E. MOTA(4)

Introduction: Chronic viral hepatitis represent major health problems worldwide, with an evolution that is sometimes marked by a series of extrahepatic manifestations. Among these, kidney disease may occur, either as glomerulonephritis, and/or renal dysfunction. Material and methods: The purpose of this study was to achieve a complete assessment of the liver and kidney function in a series of patients diagnosed with chronic viral hepatitis, in order to identify a possible kidney disease in this context. 104 patients with chronic viral hepatitis B and C, aged between 25 and 80, were included in the study. These patients were assessed by a series of tests that allowed us to perform a complete evaluation of both liver and kidney function: liver cytolysis enzymes, serum bilirubin, markers of cholestasis, markers of viral infection, urinalysis, albuminuria, electrophoresis of urinary proteins, urea, creatinine, and abdominal ultrasound. Results: Glomerular injury was a rare finding in patients with chronic hepatitis. Patients with chronic hepatitis B had glomerular disease in 5,88 % of cases, while 10 % of the patients with hepatitis C presented this type of renal disease. Acute kidney injury was not identified in our patient group. None of the patients in the hepatitis B study group presented chronic kidney disease. CKD was present in two patients with chronic hepatitis C that had no other associated conditions.

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7. Contribution of Nail Fold Videocapillaroscopy in Patients with Early Inflammatory Arthritis

M. FLOREA(1), ANCA ROSU(1), F.A. VREJU(1), ANCA EMANUELA MUSETESCU(1), CRISTINA CRIVEANU(1), PAULINA CIUREA(1)

Early inflammatory arthritis (EIA) a condition defined by joint swelling, with or without morning stiffness, and/or swelling of metacarpophalangeal and/or metatarsophalangeal joints. Nail fold video-capillaroscopy (NVC) is important for the evaluation of microcirculation in vivo. There are limited data about the role of NVC in EIA. Objectives: To study the capillaroscopic pattern in patients with EIA. Patients and methods: 27 patients with EIA - 21 women, 6 men, mean age of 41.6±4.2 yrs, mean disease duration of 6.9±3.1 months. Anamnesis and clinical examination were perform to identify clinical signs associated with connective tissue diseases. All the patients were nonsmokers and had no personal medical history of diabetes or non-immune mediated occlusive vascular disease. Blood and urine samples were collected for bio-chemical and immunological evaluation. All the patients were interrogated by NVC. Results: Raynaud phenomenon was found in 9 patients, puffy fingers in 2 pts, telangiectasia in 2 pts, Sicca symptoms in 6 pts, malar rash in 2 pts, photosensitivity and psoriasis plaque in one patient. Combined information from clinical exam, NVC and immunological assessment allowed a specific diagnosis in 5 patients. Conclusions: Nail fold capillaroscopy assessment can provide further information and has diagnostic value in some cases of early arthritis. Nail fold capillaroscopy assessment contribution to the differential diagnosis in patients with early arthritis is sometimes significant, especially in poorly clinical and immunological defined cases.

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8. Transthoracic Needle Biopsy (TNB) under Different Guiding Methods - the Experience of the Thoracic Surgery Clinic of Craiova after the First 235 Cases

A. DEMETRIAN(1), A. DOBRINESCU(1), SERENADA BALA(2), CAMELIA DEMETRIAN(3), IOANA ANDREEA GHEONEA(4), DANA MARIA ALBULESCU(4)

Transthoracic needle biopsy (TNB) is a fast and safe method used to establish definitive diagnosis for most thoracic lesions, whether the lesion is located in the pleura, the lung parenchyma, or the mediastinum. Diffuse disease and solitary lesions are equally approachable.TNB can avoid (when technically possible) more complex diagnostic interventions such asmediastinoscopy, thoracoscopy and exploratory thoracotomy. This article focuses on the advantages of TNB which is a safe, affordable and quick method to obtain histopathological confirmation of intrathoracic tumors. Material and MethodsThe study included a total of 235 cases over a period of 4 and a half years (01.01.2011-30.04.2015). We investigated the demographic and clinical parameters, the guiding methods, the histological results and the complications of the procedure. ResultsThe median age of the patients was 62 years and the predominent sex was male. We could obtain a tissue biopsy in 99% with a histopathological confirmation of 88%. The most frequently used guiding method was the previous CT scan of the patient and the anathomical landmarks (53%). The main histopathological result was squamous cell carcinoma.ConclusionsTNB is generally a safe procedure with limited morbidity and extremely rare mortality. It is an affordable and quick method to obtain histopathological confirmation of intrathoracic tumors.Most TNBs can be performed by using local anesthesia without conscious sedation and virtually any location in the chest can be safely addressed.

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9. Formulation and In Vitro Evaluation of Alendronate Sodium/PLGA Microspheres for Applications in Bone Related Disorders

ANDREEA-GABRIELA DECA(1), IONELA BELU(2), O. CROITORU(3), MARIA-VIORICA BUBULICA(3), C.V. MANDA(3), J. NEAMTU(3)

Purpose - Bisphosphonates are a group of drugs that can be used in the development of new therapies for bone disorders. Microencapsulation by solvent evaporation is a method that results in microspheres with controlled release of the drug. The purpose of the present study was to include sodium alendronate (AL) in poly (lactic-co-glycolic acid) (PLGA) microspheres with a good encapsulation rate. Furthermore it was intended to obtain a prolonged release of the drug over a period of time. Material and methods - Microspheres were prepared by water/oil/water solvent evaporation method. The microspheres were characterized by FTIR analysis using an Avatar Nicolet spectrophotometer. Both encapsulation ratio and in vitro drug release were assessed by chromatographic analysis. Results - The method chosen for sodium alendronate incorporation resulted in microspheres with a good entrapment efficiency. The in vitro drug release from PLGA/AL microspheres was maintained up to 21 days. This suggests that the microspheres had a sustained releasing tendency. Conclusion - Given the prolonged release, PLGA/AL microspheres may be used in a controlled release system with applications in bone disease treatment.

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10. Clinical Study Regarding Arrhythmogenic Risk Factors and Oxidative Stress Inductibility in Young People

MARIA CRISTINA BEZNA(1), DOINA CARSTEA(1), MARINELA BEZNA(2), IONELA CRISTINA DELIU(2), D.O. ALEXANDRU(3), PAULINA CIUREA(4)

Rhythm disorders in young people are often reported and when they are persistent, repetitive or with a severity degree, an ethiopathgenical assessment of arrhythmogenic risk factors and their implications is required. Purpose: Arrhythmogenic risk factors evaluation and the possibility of determining oxidative stress in the arrhythmic pathology in young people. Material and Methods: The study was conducted on 184 young subjects, aged 16-26 years old; the points of interest were: the presence or absence of cardiac dysrhythmias, the existence of proarrhythmogenic risk factors and determination of oxidative stress status modifications. Results: Of the studied young subjects, 39% presented heart rhythm disturbances, repetitive or persistent (atrial extrasystolic arrhythmia, sinus tachycardia, ventricular extrasystolic arrhythmia, paroxysmal atrial fibrillation, paroxysmal supraventricular tachycardia, associated dysrhythmias, atrial flutter, sinus bradycardia), which have been associated with the following risk factors: coffee consumption 82%, stress 80%, physical effort 72%, energy drinks consumption 72%, hyperlipidic diet 69%, familial predisposition 69%, alcohol intake 53%, frequent sleep deprivation 50%, smoking 31%, overweight 31%. The observed risk factors may be involved in the increasing of oxidative stress level, and, for this reason, the determination of oxidative stress biomarkers is required. The association of arrhythmogenic risk factors, with the expression of oxidative stress markers and the existence of enzymatic genetic polymorphism of redox systems, requires proper monitoring for the further risk of endothelial lesions induction, leading to aterosclerosis. Conclusions: Arrhythmogenic risk factors and biomarkers of oxidative stress are important, especially in young people cases, for monitoring the cardiovascular risk, for primary prevention and early treatment.

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11. Current Role of Endoscopic Ultrasonography in Rectal Cancer Evaluation During Multidisciplinary Therapy

ELENA TATIANA CARTANA(1), CLAUDIA GEORGESCU(2), DIANA TUDORASCU(3), C.C. VERE(1), A. SAFTOIU(4)

We report the case of a patient presenting in the Gastroenterology Department with intermittent rectal bleeding during the past year. A diagnosis of a low rectal adenocarcinoma was based on colonoscopy examination with biopsies, and staging procedures included transrectal endoscopic ultrasonography and magnetic resonance imaging of the abdomen and pelvis (cT2N0M0). Consequently the patient was referred for pre-operative chemoradiotherapy, achieving a complete clinical response as documented by repeated EUS and MRI examinations. Transanal endoscopic microsurgery with pathological assessment of the resected specimen revealed residual adenocarcinoma, highlighting the limitations of current imaging methods, and the constant need of technological improvements.

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12. The Role of Real Time Elastography in the Diagnosis of a Patient with Liver Cirrhosis and Carcinoma of the Uncinate Process of the Pancreas - Case Study

DANIELA LARISA SANDULESCU(1), S.C. DINESCU(2), IOANA ANDREEA GHEONEA(3), SINETA CRISTINA FIRULESCU(2), S. SANDULESCU(4)

Hepatic cirrhosis represents the most advanced stage of any chronic liver disease characterized by progressive fibrosis. We report the case of a patient with carcinoma of the uncinate process of the pancreas and an occult form of liver cirrhosis. We concluded, based on the bioclinical profile, that the jaundice syndrome had an underlying mixed mechanism, obstructive and hepatic. Although, the clinical suspicion of pancreatic cancer was backed up by ultrasound, computed tomography and magnetic resonance imaging, we did not obtain, through these imaging investigations, any indicative features of liver cirrhosis. In order to further evaluate the presence of liver cirrhosis, we assessed liver stiffness using two non-invasive methods: Transient Elastography and Real Time Tissue Elastography (RTE). We observed highly suggestive features of liver cirrhosis only through RTE, although its diagnostic accuracy still needs large validation studies. Intraoperative assessment confirmed the diagnosis of liver cirrhosis, changing also the type of surgical approach and patient prognosis.

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13. Overlap Syndrome-Systemic Sclerosis, Systemic Lupus Erythematosus and Dermatomyositis - Case Report

CLAUDIA CIRSTEA(1), ANDREEA BARBULESCU(2), LOREDANA ELENA STOICA(3), F.A. VREJU(4), PAULINA CIUREA(4)

Overlap syndrome is often defined as an entity that satisfies the classification criteria for at least two distinct connective tissue disease. We present the case of a female patient, 39 years old, hospitalized in the Dermatology clinic of Emergency County Hospital in september 2014, for ulcerative lesions on the left hallux and the second and third left toes, associated with pain and difficulties in walking. After performing the clinical exam and paraclinical tests, we decided for both intravenous synthetic analogue of prostacyclin PGI2-iloprost and local therapeutic measures, with a favourable outcome of the ulcerations. Due to the fact that overlap syndrome is an entity with several visceral involvement and unpredictable evolution, we must not disregard the skin manifestation, so that we can prevent a severe evolution and improve the outcome. The case represents a permanent challenge for the multidisciplinary team that examine, survey and periodically adjust the treatment, based on the biological status and predominant symptoms.

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14. Clinical and Pathological Analysis of Two Cases of Cutaneous Malignant Melanoma

ANTONIA BLENDEA(1), DIANA MONICA GEORGESCU(2), STEFANA OANA PURCARU(3), DANIELA ELISE TACHE(3), I. FLOREA(4), LOREDANA ELENA STOICA(5), I. TOLEA(6), CLAUDIA VALENTINA GEORGESCU(7)

The purpose of this study was the clinical and histo-immunohistochemical analysis of two cases: a cutaneous pigmented facial malignant melanoma and a lumbar congenital nevus with malignant transformation. A series of clinical elements raised the suspicion of some malignant melanocytic lesions and the histopathological analysis through the paraffin embedding technique confirmed the clinical suspicion. The immunohistochemical analysis using the streptavidin-biotin-peroxydase method of the facial malignant melanoma showed: S100 protein intense and diffuse positive, Tyrosinase diffuse positive, HMB45 strong and focal positive, Cyclin D1 positive in approximately 40% and Ki-67 positive in almost 70% of the tumor cells. The malignant melanoma developed on the nevocellular nevus displayed: S100 protein intense and diffuse positive, both in the nevus cells and in the malignant melanocytes as well, Tyrosinase intense and diffuse positive in the malignant melanocytes, poor and focal positive in the nevus cells and HMB45 intense and focal positive in the malignant cells and positive in the isolated nevus cells. Cyclin D1 was positive in about 70% of the malignant cells, but negative in the nevus area and Ki-67 was found positive in relatively 30% of the malignant melanocytes, also in less than 1% of the nevus cells. The pattern and the intensity of the Tyrosinase and HMB45 immunoexpression are important in the differentiation of the nevus cells from the malignant melanocytic cells. The immunoexpression of Cyclin D1 does not correlate directly with the proliferating activity of the malignant melanocytic cells in all types of malignant melanomas.

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15. A Case Presentation Emphasizing the Value of Full-field ERG in Retinal Dystrophies

ANDREEA C. CORICI(1), D.O. ALEXANDRU(2), OANA M. CORICI(1), MARIA IANCAU(1)

Purpose: A case presentation is used to discuss the value of full-field ERG in the diagnosis of cone-rod dystrophy. Case report: A 49 year old female patient known with amblyopia and myopic astigmatism since childhood, visited us complaining of decreased visual acuity and photophobia. Her BCVA was 0.6 in the RE and 0.4 in the LE. The fundus examination revealed normal colored and sharply limited papilla, normal macula and narrower vascularization. The Ishihara test was 10/15 in both eyes. The fundus autofluorescence and optical coherence tomography were normal. The DA 3 ERG was decreased and slightly delayed as well as the DA 0.01 ERG. Regarding the cone response, the LA 3.0 and 30 Hz flicker ERG were markedly reduced and delayed. The ERG findings indicated cone-rod dystrophy. At the 8 years follow-up examination, the BCVA decreased to 0.4 and 0.2 in the right and left eye, respectively, the Ishihara test was 1/15 in both eyes and the fundus examination showed a macula with no reflex, otherwise no changes compared to the first examination. The fundus autofluorescence and OCT were still normal. The ERG showed a much decreased and delayed rod response compared to the first examination, while the cone response was undetectable. Conclusion: Our case demonstrates the importance of standard full-field ERG in diagnosing CRD. The Full-field ERG responses were decreased and delayed, or even undetectable, while morphological examinations were still unchanged.

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16. Imaging Evaluation of the Orbit in Graves Ophthalmopathy

T.N. SAS(1), GEORGIANA-CRISTIANA CAMEN(1), A. STEFANESCU(2), IOANA ANDREEA GHEONEA(1)

Diagnosis of Graves ophthalmopathy is based on the typical eye signs and symptoms, the exclusion of other orbital pathologies and evidence of thyroid autoimmunity. We report the case of a non-smoker 61 years old woman, known with Graves disease, which performed a 3T MRI examination in order to evaluate the orbits. The MRI exam revealed eye protrusion, increased orbital fat with compression on the optic nerve and increased volume of the extraocular muscles. We concluded that MRI, using high-field 3T imaging techniques, is very important for diagnosis of these anomalies, making the differential diagnosis with other pathology and also providing additional information in the decision for therapy.

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17. Endomicroscopy with Fluorescent CD105 Antibodies for ”In Vivo” Imaging of Colorectal Cancer Angiogenesis

ADRIANA CIOCALTEU(1), D. PIRICI(2), CLAUDIA VALENTINA GEORGESCU(3), V. SURLIN(4), A. SAFTOIU(5)

The aim of this case report was to evaluate the feasibility of in vivo acquisition of microscopic images using fluorescent CD105 antibodies for molecular imaging in human colorectal cancer. After excluding the presence of tissue autofluorescence, the antibody solution was topically administered through a spray-catheter. The targeted area was analyzed by eCLE and images were recorded. The fractal dimension of tumor vessels and the vessel density were determined using ImageJ software. Immunohistochemistry was used as a gold standard. In vivo CLE analysis of CD105 expression enabled the study of tumor vascular network, revealing a chaotic structure.

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