CHSJ

Archive issue

Current Health Sciences Journal

vol. 41 no. 2, 2015

1. Wireless Capsule Endoscopy in Correlation with Software Application in Gastrointestinal Diseases

ADRIANA FLORENTINA CONSTANTINESCU, MIHAELA IONESCU, I. ROGOVEANU, C.C. VERE, V.F. IOVANESCU, C.T. STREBA, M.E. CIUREA

Wireless capsule endoscopy is one of the most recent investigation techniques of gastrointestinal pathology. Unlike conventional upper and lower endoscopy, it has the advantages of being a noninvasive and painless procedure. One of the capsule endoscopy disadvantages is represented by the necessary time to analyze the video frames obtained. Software applications proposed in this purpose could offer support in the images evaluation. Different algorithms have been described in the literature, but further research is needed to establish the practical value of computer vision tools in gastroenterology.

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2. Airborne Pseudomonas species in Healthcare Facilities in a Tropical Setting

SUCHITHRA SUDHARSANAM, SHANTHI MATHIAS, MOHANBABU ETHIRAJ, GOPALSAMY SARANGAN, RAMYA BARANI, SANDHYA SWAMINATHAN, RAVI ANNAMALAI, PADMA SRIKANTH

Background and objectives: Pseudomonas species are nosocomial pathogens that are capable of colonising moist surfaces. Little is known whether they get airborne. The study was undertaken to 1) characterise Gram-negative bacteria in indoor air of different hospitals; 2) characterise Pseudomonas sp. by phenotypic and genotypic methods; 3) determine homology of study environmental Pseudomonas isolates and correlate with established pathogenic strains sequences. Methods: Samples were collected (duplicates) at the time of peak activity, by exposing media-containing plates (blood agar and MacConkey agar) for 30 minutes. Plates were incubated aerobically at 37°C for 24-48 h. Microorganisms were identified by standard microbiological procedures. Polymerase chain reaction targeting Pseudomonas specific 16S-rDNA was performed to obtain 618 bp amplicons. Representative strains were sequenced and compared with established sequences of pathogenic Pseudomonas strains from existing database for evolutionary details. Results: A total of six hospitals comprising 13 wards, 7 intensive care units (ICUs) and 8 operating rooms (ORs) were sampled over one-year period. A variety of Gram-negative bacilli were isolated, of which Pseudomonas sp. was predominant. Indoor air of 10 wards (77%), 5 ICUs (71%), 4 ORs (50%) harboured Pseudomonas. Similar strains of Pseudomonas stutzeri were isolated from indoor air of different hospitals. Phylogenetic analysis showed these environmental strains to be closely related to the pathogenic Pseudomonas stutzeri strain from the GenBank database. Conclusions: Isolation of airborne Pseudomonas stutzeri from different hospitals suggests a possible new reservoir in the hospital environment, indicating the need for appropriate engineering control measures to contain the spread of these nosocomial agents.

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3. Predictive Factors for the Evolution of Reactive Arthritis to Ankylosing Spondylitis

SIMONA BANICIOIU-COVEI, F.A. VREJU, PAULINA CIUREA

Purpose: The goal of this study is to identify the predictive factors involved in the evolution of the reactive arthritis to ankylosing spondylitis by following the presence and degree of sacroiliitis and also the appeareance of acute anterior uveitis.Material and Methods: The study was performed between 01.01.2011- 31.12.2014 on 112 patients, aged between 17 and 47 years old, in evidence of the Rheumatology Clinic of the Clinical Hospital Emergency Craiova. The patients were divided in 2 lots according to the radiological criteria: the first lot included 52 patients diagnosed with reactive arthritis, with the determination of the pathogen agent involved and the demonstration of sacroiliitis first degree by Nuclear Magnetic Resonance (IRM). The second lot included a number of 60 patients diagnosed with reactive arthritis, without presenting significant changes in the sacroiliac joints and at the cervico-dorsal- lumbar spine. Results: In the first batch, the performance of the IRM and monitoring the patients every 6 months over 3 years revealed the progression of sacroiliitis from first degree in which it was at the moment of diagnosing at second degree in a 6 month period. Three years after initiation of therapy for reactive arthritis, the patients from the second batch responded favorably to treatment, the performance of imaging not revealing signs of sacroiliitis.Conclusion: Association of sacroiliitis at baseline proved to be a negative prognostic factor in reactive arthritis, which can suggest the evolution to ankylosing spondylitis, monitoring these patients being necessary for at least 3 years from the point of diagnosing.

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4. Osteoporosis in Systemic Lupus Erythematosus - Correlations with Disease Activity and Organ Damage

ANDREEA BARBULESCU, F.A. VREJU, CRISTINA CRIVEANU, ANCA ROSU, PAULINA CIUREA

Purpose: to assess the incidence of osteoporosis in a cohort of SLE patients and to determine the possible correlations with disease activity, organ damage and glucocorticoid therapy. Material and methods: We analyzed a cohort that included 25 consecutive SLE patients, diagnosed according to ACR revised criteria, and 21 controls, with the same demographic characteristics. We assessed demographic, life style related, clinical, biologic and immunologic data; also, we registered information about the treatment and calculated disease activity and damage scores. Bone mineral density was measured both in lumbar spine and total hip, using dual-energy X-ray absorptiometry. Results: Evaluation of T score profile, both in lumbar spine and total hip, established a percentage of 36 (9) patients with osteoporosis, 40 (10) patients with osteopenia and 24 (6) cases with normal values. Mean T score in lumbar spine was -1.28+1.31 SD and for the total hip -1.21+1.34 SD. Analysis of correlation between T score, both in lumbar spine and total hip, and SLEDAI, established a moderate, negative correlation; for SDI we obtained a moderate correlation, both for lumbar spine and total hip, statistically significant. Conclusions: our results show an increased percentage of osteoporosis among SLE patients and a relation to disease and treatment variables, imposing a periodic evaluation, in order to establish an early diagnosis, the proper therapeutic measures, and to prevent the major consequence, vertebral and non-vertebral fractures.

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5. Liver Fatty Acid Binding Protein And Hemojuvelin

MARIA BOGDAN, ISABELA SILOSI, PETRA SURLIN, A.A. TICA, OANA SORINA TICA, T.A. BALSEANU, ANNE-MARIE RAUTEN, D. CIOLOCA, A. CAMEN

Purpose. The purpose of the present study was to investigate whether the co-administration of aripiprazole and fluoxetine could produce impaired liver function in Wistar rats by means of liver fatty acid binding protein (L-FABP) and hemojuvelin (HJV) serum levels. Furthermore, the experiment intended to assess the salivary levels of L-FABP and HJV and to determine whether they correlate with the serum levels of the two markers. Materials and Methods. Adult male Wistar rats were randomly assigned to four groups: control (saline 10ml/kg), aripiprazole (4.05 mg/kg), fluoxetine (10 mg/kg) and aripiprazole + fluoxetine (4.05 mg/kg + 10 mg/kg). The drugs were administered by gavage, daily at the same hour, along a 6 week period. L-FABP and HJV levels were determined in serum, from intraventricular blood, and in saliva. Also from intraventricular blood, serum levels for aspartate aminotransferase (ASAT) and alanine amino transferase (ALAT) were assessed. Results. Positive and statistically significant correlations between serum and salivary levels of L-FABP and HJV were found. Aripiprazole + fluoxetine group experienced increased serum L-FABP levels than aripiprazole and fluoxetine groups, and salivary L-FABP as compared to aripiprazole group; but it registered decreased levels for serum and salivary HJV, for ASAT and ALAT than aripiprazole and fluoxetine groups, and for salivary L-FABP compared to fluoxetine group. Conclusions. The data indicate that: aripiprazole coprescribed with fluoxetine do not cause additional alterations in liver function; L-FABP and HJV levels can be helpful as biomarkers for impaired function of hepatocytes; and that their salivary determination can replace serum determination.

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6. ATG5 rs2245214 C/G Polymorphism Frequency in a Romanian Population

FLORIN BURADA, D.N. FLORESCU, M.G. CUCU, ALINA LILIANA CIMPOERU, D.I. GHEONEA

Purpose. The aim of this study was to assess the frequency of a key autophagy gene ATG5 rs2245214 C/G polymorphism in a Romanian volunteer cohort, as there are no data regarding an Eastern European population. Material/Methods. DNA was extracted from peripheral blood of 105 Romanian unrelated volunteers. The ATG5 rs2245214 C/G polymorphism was genotyped by Real-Time PCR using allelic discrimination TaqMan assay. RealTime PCR was performed on a ViiA™ 7 Real Time PCR System. Hardy–Weinberg equilibrium of allele frequencies at individual loci was assessed using the Chi-squared test. Results. The genotype frequencies in controls were distributed in accordance with Hardy–Weinberg equilibrium (?² = 1.07; p = 0.3). We found CC genotype in 53 subjects (50.48 %), CG genotype in 40 (38.10 %) and GG genotype in 12 subjects (11.42 %).The G risk allele was found in 52 individuals, and the frequency of the minor G allele was 0.3. Conclusion. This is the first report on a Romanian population regarding the frequency of the ATG5 gene rs2245214 polymorphism. Our results are slightly different to the distribution pattern from other Caucasian populations and larger studies including various ethnic groups are required.

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7. The Role of Podoplanin in the Lymphangiogenesis of Oral Squamous Carcinomas

RALUCA NICULINA CIUREA, A.E. STEPAN, CRISTIANA SIMIONESCU, C. MARGARITESCU, V. PATRASCU, M.E. CIUREA

Oral squamous cells carcinoma (OSCC) mortality rate ranges between 29-40/100,000 inhabitants. Regional lymph node metastases occur in 30-40% cases and are associated with unfavorable prognosis and decreased survival. Recentely it was suggested that the tumor-associated lymphatic vessels formation plays an active role in metastasis proces of several human malignancies, including OSCC. In the present study we investigated comparatively podoplanin immunoexpression in OSCC in both the tumor cells and lymphatic vessels reported to clinicopathological prognostic parameters. Material and methods The study included a total of 25 cases of OSCC. We investigated clinico-pathological parameters as age, gender, tumor site, and degree of differentiation, size and depth of invasion (pT), lymph node status (pN). Histologic classification was done according to the WHO criteria. For immunohistochemical (IHC) analysis we used podoplanin (Dako, clone D2-40). As visualization system it was used LSAB2 (Dako, Redox, Romania, code K0675) and chromogen DAB (Dako, Redox, Romania, code K3468). Negative controls were obtained by omitting the primary antibodies. IHC quantification was done intensity score and by lymphatic microdensity (LVD), intratumoral and on the advancing edge. For the statistical analysis we used Student t-tests, ANOVA, chi square and Pearson, using SPSS 10 software. Results Podoplanin immunoexpression in tumor parenchyma presented with an average of 43%, varying intensity. We found a higher intensity in weak and moderately differentiated SCC then in well differentiated ones and no difference intratumoral aand advancing edge. In relation to the degree of tumor differentiation the mean LVD D2-40 was higher in the advancing edge. SCC who presented lymph node metastasis mean values for LVD D2-4 was higher then at advancing edge and higher than those of non-metastatic carcinomas. Conclusions Podoplanin immunoexpresion suggests the involvement both in tumor growth and the acquisition of an lymphangiogenic phenotype invasive by autocrine mechanisms.

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8. Prosthetic Treatment Possibilities for Extended Partially Edentulous and Completely Edentulous Patients, Depending on the Clinico-Morphological Characteristics of the Oral Mucosal Substrate

MONICA MIHAELA CRAITOIU, C. MARGARITESCU, MONICA SCRIECIU, ROXANA PASCU, ALMA MARIA FLORESCU, VERONICA MERCUT

The number of partially and completely edentulous patients requiring removable prosthetic treatment has increased constantly and denture wearers often have an associated oral status pathology. The purpose of this article is to present removable prosthetic treatment options that we chose for the extended partially edentulous and completely edentulous patients who required prosthetic rehabilitation treatment. We conducted this study on 52 extended partially edentulous and completely edentulous patients. We recorded in the examination chart the characteristics of the mucosal membrane covering the denture supporting structure and the peripheral structures and we performed a statistical analysis using Microsoft Excel. The prosthetic treatment phases were individualized according to morphological characteristics of the oral mucosal substrate.

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9. Evaluation of Angiogenesis in Colorectal Cancer

IONELA CRISTINA DELIU, PAULINA CIUREA, DANIELA NEAGOE, MARIA CRISTINA BEZNA, IOANA GHEONEA, C.D. USCATU, T. DUMITRESCU, T. CIUREA

Purpose: Angiogenesis is an important step in the process of cancer growth. The purpose of this study was to determine the neoangiogenesis with CD31, CD34 and CD105, and tried to observe the differences between these three antibodies. Material/Methods: The blood vessels stained with CD31, CD 34 and CD105 were counted, and we reported their number per square millimeter to obtain microvascular density (MVD). For angiogenesis quantification we determined the neoformation blood vessels with CD105. The CD31 and CD34 were used as control markers, in order to observe the difference between neoformation blood vessels and mature vessels. Results: Comparing the average effective vessels marked with the 3 markers, Student t test showed that the mean number of blood vessels market with CD 34 is higher than blood vessels market with CD31 and CD 105. The value of the Student t test was highly significant in all three cases (p<0.001). By calculating the Pearson correlation coefficient for the relationship CD31-CD105 we obtained a value r = 0.440, which corresponds to p = 0.0013 <0.05, indicating a statistically significant direct correlation between the two factors. Conclusions: An important number of vessels (around 40%) that can be found in tumor area are neoformation vessels, this concept being an important assessment for the choice of the correct and effective treatment in colorectal adenocarcinoma.

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10. Correlations Between Intratumoral Microvessel Density and Histopathological Type or Neoadjuvant Radiotherapy for Rectal Carcinoma

T. DUMITRESCU, C.D. USCATU, D.O. ALEXANDRU, C. MESINA, S.S. MOGOANTA, ILEANA DINCA, E. MORARU, ALEXANDRA DUMITRESCU, D.I. GHEONEA, RALUCA ELENA NICOLI, I. VASILE

Purpose: This study aims to evaluate intratumoral microvessel density in rectal carcinoma cases with different histopathological type (adenocarcinoma and mucinous carcinoma) and different preoperatory neoadjuvant radiotherapy status (irradiated / non-irradiated) ,thus analyzing any possible statistical correlation between these parameters. Material and methods: Our prospective study consists in standard immunohistochemistry procedures using CD34, CD31 and CD105 antibodies, which were performed on 25 samples of rectal carcinoma, in order to determine intratumoral microvessel density. Results: The 25 case study group was divided either by histopathological type or by prior radiotherapeutical treatment as follows: 9 cases of mucinous carcinoma versus 16 cases of adenocarcinoma and 13 cases of rectal cancer that have not received neoadjuvant radiotherapy versus 12 cases of rectal cancer with preoperatory radiotherapy. Conclusions: The number of intratumoral microvessels is higher in non-irradiated rectal tumors and in adenocarcinomas, this remark being statistical significant (with only one exception – CD34 staining in non-irradiated versus irradiated tumors) for all types of vessels (new-grown and mature). This result is due to the benefic effect of neoadjuvant radiotherapy on decreasing angiogenic activity, thus having an important prognostic value for rectal cancer.

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11. Statistical Correlations of the Spontaneous Abortion with Trombophilia and Other Associated Pathologies

MAGDALENA MARIA MANOLEA, ANDA LORENA DIJMARESCU, SIMONA NEAMTU, D.O. ALEXANDRU, R. RICA, A. GOGANAU, LILIANA NOVAC

15-20% of all known pregnancies progress with a miscarriage or an ectopic pregnancy. The recurrent miscarriage is a delicate clinical situation because with all the advances in genetic and immunologic research the incriminating factors have not been discovered yet. The treatment is also subject to controversies, being perceived as either ineffective or aggressive. A number of 376 pregnant women with a gestational age smaller than 12 weeks have been selected, 226 of them with spontaneous abortion. In the study group of patients with previous spontaneous abortions we have found a series of associated pathology. From the cases with secondary non idiopathic recurrent miscarriage we have identified 4 cases of thrombophilia with antiphospholipid syndrome, 2 cases of thrombocytosis, one case of autoimmune thyroiditis, one case of uterine tumor, one case of MTHFR C homozygous thrombophilia and one case of FVL heterozygous thrombophilia. Knowing the correlations between the recurrent miscarriage and the hereditary thrombophilia as well as the options of treatment for increasing the chances of having a pregnancy with a normal evolution direct the doctors in testing the patients with recurrent miscarriage for hereditary thrombophilia.

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12. Sodium Ion Effect on Separation Of Butyrylcholinesterase from Plasma by Ion-Exchange Chromatography

ANGELA GABRIELA POSTOARCA, MIHAELA IONESCU, ALICE PIPEREA-SIANU, I. SARBU, LAVINIA GEORGETA HINESCU

Purpose of the paper was the optimization of mobile phase in natrium chloride gradient in the chromatographic separation of butyrylcholinesterase from human plasma. Materials/Methods. Butyrylcholinesterase (BuChE) was isolated from human plasma using a diethylamminoethyl-cellulose column, by elution with 0.02M acetate buffer pH=4.0, gradually increasing NaCl percentage from 10% to 80%. The procedure lasted approximately 28 days. Results. Absorbance of the successive collected fractions at 280 nm presented a maximum at 60 % NaCl concentration. Activity of obtained BuChE was maximum at the same concentration. Another observed effect of NaCl was the decrease of resistance of the column to flow of the elution fluid. In the absence of NaCl the flow rate was 7 mL/h. Increasing of NaCl concentration induced a continuous increase of the flow to a value of 21 mL/h at 60% NaCl solution. After this concentration the flow remained practically constant. The effect on the ionic exchange is essentially an effect on chromatographic partition coefficient, leading, as a rule, to a peak having a Gaussian form. Fitting separately of ascending and descending parts of the apparent peak, led to practically the same exponential coefficient. Conclusions. Separation of proteins and particularly of BuChE on a chromatographic DEAE Cellulose column can be considered as a method for separation and purification of BuChE from human plasma. Optimum concentration of NaCl is 60 %. Exponential fittings in the neighbourhood of maximum indicated a prevalence of effects of NaCl on the chromatographic partition face to effects on gel-sol equilibrium of stationary phase.

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13. Simultaneous Analysis of Clopidogrel Bisulfate, Acetylsalicylic Acid and Atorvastatin calcium in Tablets by HPLC Method

ADELA-MARIA SPIRIDON, J. NEAMTU, IONELA BELU, ADINA TURCU-STIOLICA, O. CROITORU

The concept of multidrug therapy with clopidogrel, acetylsalicylic acid and atorvastatin provides major advances for patients with acute coronary syndromes (ACS) treated invasively and conservatively to reduce the risk of ischemic events and improve survival. Dual antiplatelet therapy (DAPT) with acetylsalicylic acid and clopidogrel is currently the treatment of choice in acute coronary syndromes and prevention of thrombosis after coronary stent implementation. Present paper presents a simple, rapid and precise reversed phase high performance liquid chromatographic method for simultaneous estimation of clopidogrel hydrogen sulfate, acetylsalicylic acid and atorvastatin calcium from tablet dosage form. The cromatographic analysis was carried out using a ThermoFinnigan Chromatograph with UV detection and separation on an HDS Hypersil C18 column. The elution was isocratic with mobile phase consisting in mixture of 0.01M KH?PO? buffer: acetonitrile: methanol. Chemicals were represented by clopidogrel bisulfat, atorvastatin calcium and acetylsalicylic acid standards with high purity. The quality control samples used in the accuracy and precision evaluation were spiked at low, medium and high concentration levels. Preliminary tests were performed to select optimum conditions for simultaneous separation of all three analytes such as mobile phase composition, proportion and pH. A satisfactory separation of all three drugs was achieved with a mobile phase of 0.01 M KH?PO? buffer (pH adjusted to 2.6 with phosphoric acid):acetonitrile:methanol 20:40:40 v/v/v at flow rate of 0.8 ml/min. The proposed method was validated for linearity, precision and accuracy. The validated method can be successfully applied for simultaneous quantification of clopidogrel, acetylsalycilic acid and atorvastatin either in combination or in single dosage form in quality control analysis. The analytical conditions can also be performed for human plasma analysis.

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14. Correlation between Tgf-?1 and Fsp-1 Expression in Chronic Viral Hepatitis - an Immunohistochemical Study

DANIELA ELISE TACHE, S.G. FIRU, ANA MARINA ANDREI, ANCA PREDESCU, STEFANA OANA PURCARU, AMELIA MIHAELA DOBRESCU, ANICA DRICU, CATALINA PISOSCHI, ELENA CAMELIA STANCIULESCU

Infection with hepatitis C virus (HCV) is the most important stimulus for chronic hepatitis and subsequent progression to cirrhosis and hepatocellular carcinoma. Fibrosis that follows inflammation represents the main complication. One of the mechanisms that could be associated with development of liver fibrosis is epithelial-mesenchymal transition (EMT). Transforming Growth Factor ?1 (TGF-?1) is an important mediator of fibrosis and also able to trigger phenotypic changes in EMT. Fibroblast-specific protein 1 (FSP-1), a marker of fibroblasts in organs undergoing tissue remodeling, is used to identify cells that derive from EMT. In this study, we assessed the expression of TGF-?1 and FSP-1 in liver biopsies obtained from HCV-infected patients using immunohistochemistry and correlated them in order to evaluate the relation between fibrosis and EMT in liver disease progression. Staining of liver sections revealed increased amount of type III collagen and clusters of inflammatory cells invading portal spaces. The number of TGF-?1-positive cells was directly proportional to the incidence of liver injury. In cases of mild fibrosis, FSP-1 positive cells were observed in cells lining sinusoids. As fibrosis progressed, increased number of FSP-1 positive fibroblasts, isolated cholangiocytes and hepatocytes was observed. Even EMT via the activation of TGF-? signaling pathway is recognized as a pathogenic mechanism of HCV-induced liver disease, FSP-1 alone couldn’t be used as a valuable marker for cells that undergo EMT.

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15. Dermatoscopic and Histopatological Aspect of Preneoplasia and Skin Cancers – Study on 74 Patients

LOREDANA ELENA STOICA, MADALINA VOICULESCU, CLAUDIA CIRSTEA

Dermoscopy is an important in-vivo, non-invasive diagnostic technique that allows visualization of morphological features not macroscopic visible. It has a major contribution in enhancing the diagnostic accuracy for pigmented skin lesions. Recent studies have shown that it also aids in the diagnosis of non-pigmented keratinizing skin lesions, including actinic keratosis and Bowen’s disease. We performed a retrospective study in Dermamed Clinic Craiova, between January and June 2014, with the aim to correlate the dermoscopic and histopathological aspect of skin tumors. This study included a total of 74 patients, aged between 16 and 76 years. The dermatoscopic examination revealed 12 skin cancers, 51 precancerous lesions and 11 benign tumors. Those 12 subjects diagnosed with malignant tumors by dermatoscopic examination, had also histopathological examination and confirmation of the diagnosis, also 11 patients identified with benign tumors with dermatoscopic examination were subsequently confirmed by histopathological assesment. Out of the 51 patients with premalignant tumors, 48 patients had histopatological confirmation of it, and for 3 patients the result was squamous cell carcinoma. Thus dermatoscopic examination identified the malignant lesions with a specificity of 100% and a sensitivity of 80% with a positive predictive value of 100% and a negative predictive value of 94.12%, p<0.000.

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