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Objective: The primary goal of this study was to determine the difference of abundance of CD4+, CD8+ and CD56+ bronchoalveolar fluid’s lymphocytes and their subpopulations between cancerous lung and healthy lung from the same patient. Methods: Mini-bronchoalveolar lavage was taken from 55 patients from lung with cancer and healthy lung. After laboratory processing and addition of CD4, CD8, CD27, CD28 and CD56 antibody, the material was analyzed by flow cytometer. Results from lung with cancer were compared to the ones from the healthy lung. The examined patients were the test and the control group at the same time. Results: CD27+28+ forms of CD4+ and CD8+ lymphocytes are more activated in the cancerous lung compared to healthy lung, while the CD27-28- forms are less activated in diseased lung. CD4+ forms of CD56+ lymphocytes are more activated in cancerous lung compared to the health lung, while the CD8+ forms are less activated in diseased lung. Conclusion: Immature helper and cytotoxic T lymphocyte response, as well as regulatory NK and NKT cell response are more activated in cancerous lung compared to the health lung of the same patient.

Ermina Iljazović, S. Tomić, Jasminka Mustedanagić-Mujanović, Z. Karasalihović, Majda Kuljanin, Z. Fatušić, Elvira Konjić, Edin Husarić et al.

Sarcomas of the female genital tract in general are rare and ovarian sarcomas comprise less than 1% of ovarian malignancies. In the literature there are 15 reported angiosarcomas of patients 21 year old and younger with no one originated in the ovary. We report a case of ovarian angiosarcoma in an 11 year old girl, presented with left side hip pain. MRI of abdomen and pelvis confirmed expansive solid and cystic mass occupied both ovaries. Imunohistochemistry staining was performed, CD34, Factor VIII, CD31, in order to confirm the diagnosis. Final diagnosis was angiosarcoma. The patient received 6 cycles of chemotherapy, according to the CWS-2002P protocol. 8 months after the diagnosis was established, there were no signs of any tumors according to the ultrasound, CT scan, and MRI. Although, extremely rare, angiosarcoma can also affect children and this diagnosis should be considered carefully in tumor with rich vascular network, necrosis and brisk mitotic activity.

AIM To determinate the difference of abundance of CD4+, CD8+ and CD56+ bronchoalveolar fluid's lymphocytes and their subpopulations between non- and small cell lung cancer. Also, the differences of abundance of examined lymphocytes were compared between main clinical stages of lung cancer. METHODS Mini-bronchoalveolar lavate was taken from lungs of 55 patients with cancer. After laboratory processing and adding CD3, CD4, CD8, CD27, CD28 and CD56 antibody, the material was analysed by flow cytometer. Results of Mini-BAL for non- and small cell lung cancer were compared, as well as the different clinical stages of the disease. RESULTS Immature and regulatory forms of lymphocytes are more activated, while mature and activated forms are less activated in small cell lung cancer compared to non small type. With an increase of the clinical stage of disease, immunological reaction of T lymphocytes is better expressed because of increasing of abundance of immature and regulatory forms of different subpopulations of lymphocytes. CONCLUSION All components of local CD4+ and CD8+ T lymphocyte, as well as NK and NKT cells response were more activated in lungs with small cell lung cancer, and these reactions were more expressed with an increase in the clinical stage.

Azra Jahić, Ermina Iljazović, Aida Arnautović-Custović, A. Halilbasić, Vlastimir Simendić, A. Zabic

INTRODUCTION The clinical course and outcome of B-CLL is various and so far unpredictible. Defining prognostic parameters potentiating division of patients in groups with favorable and unfavorable prognosis which could help the benefit assessment of early treatment, improve treatment effects, and potentiate treatment modification for each patient. AIM To analyze the bone-marrow (BM) pattern and immunophenotypic score at diagnosis of B-CLL and determine the correlation of BM pattern with the clinical stage of disease and immunophenotypic score. METHODS A sample of 40 untreated patients with B-CLL was divided into two groups: group with clinical stage Binet A and group with clinical stages Binet B and Binet C. BM patterns were observed as a diffuse, interstitial, nodular and mixed. BM immunophenotyping included CD5, CD23, CD22, and CD20 as an indirect indicator of FMC7. RESULTS The overall sample mean age was 62.88 years +/- 11.10, without significant difference in the age of two compared groups (63.15 +/- 10.53 years vs. 62.60 +/- 11.50 years) (t = 0.16, df= 38, p = 0.88). Proportion of men was significantly higher in stages Binet B and C (12/20) compared to stage Binet A (5/20) (Z=2.24, p=0.025). The percentage of women was higher than men in Binet A stage (75% vs. 25%). The BM patterns in Binet A stage were observed as follows: mixed 50% (10/20), interstitial 30% (6/20), nodular 15% (3/20) and diffuse 5% (1/20). The BM patterns in Binet B and C stages were observed as follows: diffuse 50% (10/20), mixed 40% (8/20), interstitial 5% (1/20) and nodular 5% (1/20). Clinical stage and the BM patterns were significantly associated (c2=8.02, p=0,005). The chance for non-diffuse patterns was 19 times higher in stage Binet A compared to stages Binet B and C, respectively, analyzing 95% CI at least 2 times higher (95% CI: 2.02-866.6). Immunophenotypic score in total sample was observed as follows: score 4: 5% (2/40), score 3: 72.5% (29/40), score 2: 20% (8/40) and score 1: 2,5% (1/40). Immunophenotypic score 3 and > 3 had 77.5% of patients (31/40), but there was no significant association between the immunophenotypic score and the BM patterns (c2=0.76, p=0.38). CONCLUSIONS Diffuse BM pattern was significantly associated with the clinical stages Binet B and C, compared to non-diffuse BM patterns which were significantly associated with the clinical stage Binet A. Diffuse BM pattern represent the parameter of progressive disease compared to the non-diffuse BM patterns which are more often represented in stable disease. Immunophenotypic score improves diagnostic accuracy of B-CLL, but should not be used as a prognostic parameter of B -CLL.

A. Latifagić, Ermina Iljazović, Belkisa Čolić, N. Mladina

UNLABELLED Lymphadenopathy is defined as an abnormality in the size or character of lymph nodes, is caused by the invasion or propagation of either inflammatory cells or neoplastic cells into the node. Numerous factors, such as age, localization, size and consistency, present and previous pathological conditions are very important in order to define the future diagnostic and therapeutic course. OBJECTIVE The aim of this study was to determine the etiological and clinical characteristics oflymphadenopathy in children in the area of the Tuzla Canton. PATIENTS AND METHODS This retrospective-prospective study analyzed the medical records of the Department of Pediatrics in Tuzla of 334 patients in age from 0 to 14 years, in which the clinical signs of palpable lymph nodes of one or more regions was diagnosed in the period from January 1st 1998 to June 30th 2003. The anamnesis data, clinical findings, diagnostic procedures results, therapeutic approach and disease outcome etiology defined lymphadenopathy were analyzed. RESULTS Out of 334 children, localized lymphadenopathy have been verified in 230, and generalized in 104. Male/female ratio was 1:1.8. Final results of our study have shown the etiologies as following: Infectious etiologies, 79.34%, neoplastic 11.34%, and non-neoplastic 9.28%. In neoplastic etiologies, lymphoblastic leukemia has been the most often verified neoplastic disease (68.4%), not related to the age or sex of patient, and equally presented as localized and generalized lymphadenopathy. In this study lymphomas were presented by generalized lymphadenopathy. CONCLUSION The regional and generalized lymphadenopathy in children depends on their etiology and has significant prognostic value for the disease.

S. de Sanjosé, Wim Quint, L. Alemany, D. Geraets, J. Klaustermeier, B. Lloveras, S. Tous, A. Félix et al.

The aim of this study was to evaluate the incidence, clinical data and patterns of mediastinal lymph node metastasis (pN2) in non-small-cell lung cancer patients who underwent systematic mediastinal lymph node dissection (SMLND). We retrospectively studied 140 consecutive patients [125 male and 15 female, mean ages 54.61 +/- 9.23 years (range, 21-75)], underwent SMLND and major lung resections due to non-small lung cancer (NSCLC), from January 2005 till December 2009. Preoperative clinical staging for mediastinal lymph node metastasis was negative (cN0) in all patients. SMLND was defined as a complete removal of mediastinal lymph nodes. Clinicalpathological data were compared according to the pN stage. Lymph node metastasis to the mediastinum was confirmed in 13 (9.28%) patients. In squamous cell cancer pN2 were in 8 (5.71%) cases out of 82 cases with cN0. On the other side in the adenocarcinomas pN2 were in 5 (3.57%) cases out of 48 with cN0. Unvaried analysis revealed central tumor site as predictive factor for mediastinal lymph node involvement. The upper mediastinal compartment was infiltrated in 12 (8.57%) cases, middle in 8 (5.71%) and lower in 3 (2.14%) cases. Pneumonectomy was the most performed surgical procedure in pN2 patients. We concluded that SMLND improves pTNM staging in lung cancer patients who underwent major lung resections with central location of the tumour.

Introduction: Reed-Sternberg cells are typical for Hodgkin’s lymphoma. They are transformed post-germinative B cells that not achieve successful immunoglobulin gene rearrangement, and therefore predetermined to apoptosis. Several mechanisms, including latent Epstein Barr virus infection (EBV), enable survival of Hodgkin’s lymphoma neoplastic cells. Goal of this study is correlation of Hodgkin’s lymphoma EBV status with expression of apoptosis associated proteins in neoplastic cells. Association of Hodgkin’s lymphoma with EBV was determined by chromogenic hybridization in situ detection of EBER1 and EBER2. Material and Methods: Presence of apoptosis associated proteins: bcl-2, bcl-X, bax and p53, were determined immunohistochemicaly in neoplastic cells of 39 EBV positive, and 42 EBV negative Hodgkin’s lymphomas. Results: Bcl-2 protein was detected in neoplastic cells in 22.2%, Bcl-X in 87.7%, Bax in 80.2% and p53 in 67.8% of cases. In 44.4% cases simultaneous presence of Bcl-X, Bax and p53 protein was detected. Bcl-X is significantly more produced in mixed cellularity, and Bcl-X and Bax in nodular sclerosis. No significant difference was found in production of all four proteins between EBV positive and negative Hodgkin’s lymphoma. Median intensity of p53, Bcl-2, Bcl-X and Bax staining of complete examined group, EBV positive and EBV negative group show significant difference, except between Bcl-X and Bax. Discussion: Survival capacity of Hodgkin’s lymphoma neoplastic cells depends on activity of numerous regulators of programmed cell death. Expression of bcl-2, bcl-X, bax and p53 proteins is not different in neoplastic cells of EBV positive and EBV negative Hodgkin’s lymphoma.

Ermina Iljazović, D. Ljuca, A. Sahimpasić, S. Avdić

Cervical dysplasia, a premalignant lesion that can progress to cervical cancer, is caused primarily by a sexually transmitted infection with an oncogenic strain of the human papillomavirus (HPV). The HPV infections are treated through destroying the clinical lesions: laser, cryotherapy, podophyllin... The hope is that by causing local tissue inflammation that the body will be stimulated to mount an antibody response and thereby prevent recurrence. In contrast to other prevention approaches, vaccines can reduce susceptibility in uninfected partners by stimulating the immune system. Aloe vera has also been reported to retard tumour growth and stimulate the immune response to viruses. A list of possible actions of propolis includes: antibacterial, antifungal, antiviral, antioxidant, anticarcinogenic, antithrombotic and immunomodulatory. Research on the possible role of some B vitamins in preventing cancer began in the last few decades, but however this complex have an influence on immune status. The aim of our study is to try to treat the HPV infection as confirmed cause of neoplastic transformation with some herbal therapy and interferon and to try define the guidelines in the management of the HPV positive patients. Goal of this paper is to search for evidence of efficacy of any treatment for HPV infection of the cervix mostly in woman with no concomitant CIN. Fifty five woman affected by HPV genital infection were enrolled in the study from September 2005 to April 2006. Patients were classified according to the results of the HPV testing prior and after the therapy. Patients were randomized into two groups: the first group was HPV positive woman treated with other than recommended therapy (n=20), (control group); the second group was pharmacologically treated with intravaginal administration of an interferon and aloe vera-propolis in recommended scheme (n=35) with treatment of the possible fungal or bacterial genital infection prior to the specific therapy. The almost same therapy was recommended to the male partner. Patients from the second group used B complex during the therapy. Patients were retested for the HPV presence after three or six month from therapy depend of the presence bacterial or fungal genital coinfection. Three months after applied therapy HPV infection was still present in more than 90% of the patients in the first group. In the second group treated according to the recommended therapy scheme HPV infection disappeared in 71.42% of the patients after three months and in 100% of patients after six months. Samples of the cervical smear for the HPV analysis were being taken during routine gynecological examinations, by using sticks with cotton, taken from the Digene Specimen Collection Kit, from the whole surface of a portion, and by mild rotating moves from the outer cervical entrance. Our results suggest that the combination of interferon and herbal therapy with B complex is effective, atraumatic and simple non-surgical treatment of HPV infection. Since prospective efficacy trials will take several years to complete, considering alternative approaches is also worthwhile.

Z. Karasalihović, Ermina Iljazović, D. Ferluga, E. Čičkušić, Jasminka Mustedanagić Mujanović, J. Stahov, Ina Skaljic

The needle biopsies from 60 transplanted and native kidneys have been processed and a prospective analysis of pattern, intensity and distribution of immunoglobulin deposits (IgA, IgG and IgM) and complement components (C3c and C1q) identified in these lesions has been carried out by immunohistochemistry with three step immunoperoxidase, in the period from 2000 to 2004. Those deposits were previously detected and analyzed by immunofluorescence. The samples consisted of 30 renal biopsies, previously diagnosed with glomerulonephritis and positive immunofluorescence and 30 renal biopsies without morphologic changes and deposits on immunofluorescence. 78,7% of the analyzed samples showed the identical results of the deposits of immunoglobulin and components of the complement with both, immunohistochemistry and immunofluorescence method. Sensitivity of the immunohistochemistry method with three step immunoperoxidase for all analyzed immunoglobulin and complement components is high (0,93), while specificity for the same method is 0,79. Standardized method of the three step immunoperoxidase on the paraffin embedded, formalin fixed needle renal biopsies could successfully replace the immunofluorescence method in diagnostic of GN, with the emphasis on a follow up and control of each single step in the procedure of the method.

In about 70% of cases, ovarian carcinoma has been diagnosed at an advanced stage. Invasion and metastasis of solid tumors request protease activity resulting in basal membrane destruction and surrounding matrix. In that process, urokinase plasminogen activator (uPA) and its receptor, urokinase plasminogen activator receptor (suPAR) play a key role, that via plasmin activation lead to basal membrane and matrix degradation in surrounding of the tumor, enable to its invasion and metastasis. Determination of serum concentration of those tumor markers can be useful in preoperative as well as in postoperative period. Their serum concentrations in ovarian cancer patients may help in good monitoring of remission or progression during chemotherapy treatment. In late 1950s and ear1y 1960s, when it was found out that malignant ovarian tumors were chemosensitive, their chemotherapy treatment has begun. In the beginning it was used only mono-therapy, and by discovering new cytostatics it was replaced by poly-chemotherapy. Now days, in the therapy of advanced stages of ovarian carcinoma combination of cisplatine or carboplatine with paclitaxel is considering as standard treatment. Aim of this study was to determine serum uPA, suPAR and CEA in FIGO II and III patients with different histological type (serous, mucinous, clear cell tumor) before and after PT chemotherapy protocol during following three cycles. In this prospective study we have analyzed 17 patients with ovarian carcinoma, those have been after surgery treated by chemotherapy. Serum levels of uPA and suPAR have been determined by ELISA-test (Imubind uPA, Imubind uPAR, American Diagnostica), and CEA by OPUS Imunoassay method. Results of this study have shown that uPA, suPAR and CEA met criteria for prognostic markers for monitoring of successfulness of platina/taxol chemotherapy protocol for serous, mucinous and clear cell tumor FIGO II and III stage of ovarian carcinoma. In case of PT chemotherapy protocol suPAR was better prognostic marker for monitoring of chemotherapy successfulness (Pearson coefficient 0,9 do 1,0; p<0,00l) than uPA (Pearson coefficient between 0,86 and 0,92; p<0,02) and CEA (Pearson coefficient 0,5 do 0,89; p<0,04).

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