Objectives : The main objective of the study was to determine whether the distribution fsaand th Gastric cancer progression in correlation with distribution and density of T-lymphocytes e level of fsa: The main objective of the study was to determine whether the distribution and the level of density of tumor infiltrating CD4+ and CD8+ T-lymphocytes correlates with standard prognostic factors for gastric cancer and whether it has impact on tumor progression. Methods: The study included 60 tissue samples of operable gastric carcinomas of known regional lymph node status, stained by standard hematoxylin eosin and immunohi Objectives: The main objective of the study was to determine whether the distribution and the level of density of tumor infiltrating CD4+ and CD8+ T-lymphocytes correlates with standard prognostic factors for gastric cancer and whether it has impact on tumor progression. Methods: The study included 60 tissue samples of operable gastric carcinomas of known regional lymph node status, stained by standard hematoxylin eosin and immunohistochemical method in order to determine standard pathologic prognostic factors for gastric cancer and to evaluate distribution and density of tumor infiltrating T- lymphocytes. Results: CD8+ T lymphocytes were predominantly distributed along the margin of carcinoma infiltration, while inside the cancer tissue there were generally few. CD4+ lymphocytes were few in almost all three analyzed zones (margin of carcinoma infiltration, cancer stroma and cancer tissue). The density of CD8+ showed significant positive correlation with CD8+ T lymphocytes within the cancer’s stroma. There was statistically significant difference in density of CD4+ T lymphocytes distributed along the margin of carcinoma infiltration and histological tumor grade, as well as in tumor grade according to Goseki. Conclusion: CD8+ T lymphocytes are densely arranged along the margin of carcinoma infiltration and they correlate with histological grade of gastric carcinoma. Keywords: gastric cancer, tumor microenvironment, T- lymphocytes stochemical method in order to determine standard pathologic prognostic factors for gastric cancer and to evaluate distribution and density of tumor infiltrating T- lymphocytes. Results: CD8+ T lymphocytes were predominantly distributed along the margin of carcinoma infiltration, Gastric cancer progression in correlation with distribution and density of T-lymphocytes while inside the cancer tissue there were generally few. CD4+ lymphocytes were few in almost all three analyzed zones (margin of carcinoma infiltration, cancer stroma and cancer tissue). The density of CD8+ showed significant positive correlation with CD8+ T lymphocytes within the cancer’s stroma. There was statistically significant difference in density of CD4+ T lymphocytes distributed along the margin of carcinoma infiltration and histological tumor grade, as well as in tumor grade according to Goseki. Conclusion: CD8+ T lymphocytes are densely arranged along the margin of carcinoma infiltration and they correlate with histological grade of gastric carcinoma. Keywords: gastric cancer, tumor microenvironment, T- lymphocytes density of tumor infiltrating CD4+ and CD8+ T-lymphocytes correlates with standard prognostic factors for gastric cancer and whether it has impact on tumor progression. Methods : The study included 60 tissue samples of operable gastric carcinomas of known regional lymph node status, stained by standard hematoxylin eosin and immunohistochemical method in order to determine standard pathologic prognostic factors for gastric cancer and to evaluate distribution and density of tumor infiltrating T- lymphocytes. Results : CD8+ T lymphocytes were predominantly distributed along the margin of carcinoma infiltration, while inside the cancer tissue there were generally few. CD4+ lymphocytes were few in almost all three analyzed zones (margin of carcinoma infiltration, cancer stroma and cancer tissue). The density of CD8+ showed significant positive correlation with CD8+ T lymphocytes within the cancer’s stroma. There was statistically significant difference in density of CD4+ T lymphocytes distributed along the margin of carcinoma infiltration and histological tumor grade, as well as in tumor grade according to Goseki. Conclusion : CD8+ T lymphocytes are densely arranged along the margin of carcinoma infiltration and they correlate with histological grade of gastric carcinoma. Keywords : gastric cancer, tumor microenvironment, T- lymphocytes

Introduction: Tumor-associated macrophages (TAM) are the most common inflammatory cells in the tumor microenvironment (TM). As a response to microenvironmental signals, they polarize into tumor resisting M1 or promoting M2 macrophages. TAM and tumor-associated dendritic cells (TADC) can either promote tumor growth and tissue invasiveness or have anti-tumor activity. The aim of the study was the examination of prognostic value in the individual cell population in TM and their correlation with clinicopathological parameters of gastric cancer. Materials and Methods: The study analyzed 60 samples of gastric cancer, known status of regional lymph nodes and without dissemination at the time of diagnosis. The control group was normal gastric tissue samples. Traditional parameters of biological aggressiveness, tumor size, histological grade, and lymphovascular invasion, are determined after standard hematoxylin-eosin staining. TAM and TADS have been evaluated using the immunohistochemical method with CD68 (TAM), TNFa (TAM-M1), CD163 (TAM-M2), and S100 (TADC) antibodies. Expression evaluation of the tissue antigen was carried out by semiquantitative methods. Results: There were statistically significant differences of TAM density (P 0.05). Conclusion: TAMs and TADC have shown potential as biomarkers for evaluating the gastric cancer staging and progression. They showed promising prediction in depth of invasion, histological grade of tumor and tumor size. Keywords: gastric cancer, tumor microenvironment, TAM, TAM-M1/M2, TADC

SUMMARY The aim of the study was to analyze correlation between morphological characteristics of intracranial meningiomas and Ki67 labeling index (Ki67 LI), and their influence on peritumoral brain edema (PTBE). There were 41 consecutive patients with intracranial meningiomas surgically treated at the Department of Neurosurgery, Zenica Cantonal Hospital, Zenica, Bosnia and Herzegovina, during the period from January 2010 to December 2015. We reviewed clinical data including patient age, gender, magnetic resonance imaging (MRI) characteristics of the tumor and peritumoral edema, tumor margins, intraoperative characteristics, histopathologic grade and Ki67 LI. In all cases, follow up MRI was obtained at about three months after resection and PTBE was analyzed. Our research showed the tumor volume, tumor margins, and intraoperative signs of arachnoidal and pial invasion to be associated with PTBE in intracranial meningiomas. Ki67 LI expression correlated with PTBE. This study showed the resolution of PTBE to depend on invasive behavior of meningioma and KI67 LI. PTBE, pial/cortical and arachnoidal invasion significantly influence the extent of surgical resection.

Introduction: to evaluate the frequency and significance of immunohistochemistry-based molecular subtypes of breast cancer and investigate their association with traditional pathological features for breast cancer among Bosnian women. Materials and methods: this study included 100 female patients with primary invasive breast cancer. Immunohistochemical analyses for estrogen receptor (ER), progesterone receptor (PR), HER-2 and Ki-67 were performed to define four biological subtypes: luminal A, luminal B, HER-2-positive and triple-negative. Results: the frequency of luminal A, luminal B, HER2-positive and triple-negative subtypes of breast cancer was 44%, 39%, 11% and 6%, respectively. Molecular subtypes of breast cancer among Bosnian women showed to be independent of traditional pathological features (p>0.05). Ki-67 showed significant di erence regarding luminal B tumor type, where high (≥14%) Ki-67 score was predominantly represented in 36 (92.3%) cases (p<0.001). Conclusion: immunohistochemistry-based molecular subtypes of breast cancer in Bosnian women somehow vary in pathological features, i.e. luminal A subtype in this sample comprised mostly ductal histological type, moderate di erentiation with the involvement of lymph nodes, known as worse prognostic factors, although with no statistical significance.

Introduction: Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are involved in the progression of several tumors, including breast cancer. Our aim was to investigate the association of immunohistochemical expression of protein MMP-2, and -9 and tissue inhibitors TIMP-1,-2,-3 by tumoral cells in the process of angiogenesis and to define their relation with clinicopathological features for breast cancer. Methods: Immunohistochemical analysis of MMP-2,-9, TIMP-1,-2,-3, endoglin/CD105, estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) status was performed on 79 tissue samples of breast cancer with axillary lymph node dissection. Results: Statistically significant difference was found between mean age of patients and tissue inhibitors of metalloproteinase (TIMP-1) expression status (p=0.008), i.e., women with TIMP-1 negative tumors were on average younger (mean age 46.5) compared to women with TIMP-1 positive tumors (mean age 58.1); TIMP-2 expression status showed association with ER status (p=0.017), while TIMP-3 negative tumors were on average more frequently ER and PR negative (p=0.016; p=0.027). Status of protein expression of MMP-9 was associated with TIMP-1 protein expression status (p=0.033), i.e., breast cancers with overexpression of protein MMP-9 were more frequently TIMP-1 protein positive. Conclusion: Only TIMPs were associated with clinicopathological features for breast cancer. TIMP-2 expression was associated with worse (TIMP-2 positive tumors were frequently ER-negative), while TIMP-3 expression in tumoral cells was associated with better clinicopathological features for breast cancer (TIMP-3 positive tumors were frequently ER and PR positive).

In the early stages of cutaneous malignant melanoma (MM), it is extremely difficult to predict adequately the risk from hematogenic and lymphatic metastasis. We investigate whether the immunohistochemical expression of Ki-67 and estrogen receptor beta (ER&bgr;) in cells of MM could predict the status of regional lymph nodes. A total of 55 tissue samples of primary cutaneous melanomas with known status of regional lymph nodes were retrospectively evaluated for Ki-67 and ER&bgr; expression by quantitative immunohistochemistry and then correlated with the status of regional lymph nodes and relevant clinicopathologic parameters. The ER&bgr;-positive expression was detected in 38 of 55 tumors (69.09%). The Clark level showed a strong correlation with ER&bgr; expression, as well as pT stage. All cases of MM showed Ki-67-positive expression and an elevated Ki-67 expression was strongly associated with increased Breslow thickness, Clark level, ulceration, lymphovascular invasion, number of mitosis, and pT stage. Logistic regression analysis showed that when ER&bgr; levels increase by 1%, the risk of positive lymph nodes decreases by 7% (odds ratio=0.930; 95% confidence interval, 0.87-0.99; P=0.036), and, when the Ki-67 expression increases by 1%, the risk of lymph nodes’ positivity increases by 10% (odds ratio=1.108; 95% confidence interval, 1.02-1.19; P=0.009). Correlation between expression of Ki-67 and ER&bgr; and the status of lymph nodes has better prognostic significance than the relationship between melanoma thickness and the status of lymph nodes. Our study showed a significant prognostic value of Ki-67 expression in predicting the behavior of MM and the potential prognostic significance of ER&bgr;.

Background : Controversy exists regarding the topography of lymph vessels in breast cancer, their usefulness as prognostic factors and whether active lymphangiogenesis occurs within the tumour. The goal was to investigate the presence of intratumoural lymphatic vessels, the existence of lymphangiogenesis and their role in tumour dissemination. Materials and methods:  Lymphatic vessel distribution was investigated in 75 specimens of invasive breast carcinoma by immunostaining using a D2-40 antibody. Intratumoural and peritumoural lymphatic vessel density (LVD) was assessed using the Chalkley counting method and correlated with clinicopathologic parameters. Endothelial cell proliferation in lymphatic vessels was analyzed by dual-color immunohistochemistry with D2-40 and Ki-67. Results:  We demonstrated the existence of intratumoural lymphatic vessels in invasive breast carcinoma. Intratumoural LVD was significantly lower (p=0, 0001) compared to peritumoural. Decrease of intra and peritumoural LVD compared to fibrocystic breast disease was observed (p=0,002). The density of intratumoural lymphatic vessels was correlated with the expression of progesterone receptors in breast cancer (p=0,036). Intratumoural endothelial cell proliferation in lymphatic vessels was minimal. Conclusion: Intratumoural lymphatic vessels are present in breast cancer. High intratumoural LVD is correlated with the expression of the progesterone receptor and thereby it may possibly serve as an indicator of a low destructive potential of the tumour, and vice versa. Whether they are tumour induced and newly formed or not is beyond the scope of this study and does not change the fact that they are tumour-associated and do not constitute a risk factor for tumour metastasis. Key words: breast carcinoma, intratumoural lymphatic density, lymphangiogenesis, progesterone

Background: Malignant melanoma (MM) is one of the tumors with fastest-growing incidence, also deadliest form of skin cancer. In the early stages of the disease it is extremely difficult to adequately predict risk from hematogenic and lymphatic metastasis. We assumed that the determination of Ki-67 and estrogen receptor beta (ER beta) in cells of MM could predict their biological behavior. Objectives: To examine whether the level of expression of Ki-67 and ER beta in cells of MM are in correlation with status of regional lymph nodes (LN), and compare the relationship between melanoma thickness (pT) and status of LN. Methods: Expression of Ki-67 and ER beta was examined by immunohistochemistry, and then staining positivity assessed and correlated with relevant clinical and pathological parameters. Results: Logistic regression analysis showed that when ER beta levels increase by 1% the risk of positive LN decreases by 7% (OR = 0.930, 95%CI 0.87–0.99, p = 0.036), and when the Ki67 expression increases by 1% that the risk of LN positivity increases by 10% (OR = 1.108, 95%CI 1.02 to 1.19, p = 0.009). Conclusions: Correlation between melanoma thickness and the status of LN has less prognostic significance than the relationship between expression of Ki-67 and ER beta and the status of LN. Our study showed a significant prognostic value of Ki67 expression in predicting the behavior of MM, and potential prognostic significance of ER beta.

Introduction: The lymphatic vasculature is an important route for the metastatic spread of human cancer. However, the extent to which this depends on lymphangiogenesis or on invasion of existing lymph vessels remains controversial. The goal of this study was to investigate the existence of lymphangiogenesis in invasive breast carcinoma: by measuring the lymphatic vessels density (LVD) and lymphatic endothelial cell proliferation (LECP) and their correlation with various prognostic parameters in breast cancer, including lymphovascular invasion (LVI).Methods: Lymphatic vessels density was investigated in 75 specimens of invasive breast carcinoma by immunostaining for D2-40 using the Chalkley counting method. Endothelial proliferation in lymphatic vessels was analyzed by dual-color immunohistochemistry with D2-40 and Ki-67.Results: Decrease of intra and peritumoral LVD in invasive breast carcinoma compared to fibrocystic breast disease was detected (p=0.002). Lymphatic endothelial cell proliferation was significantly higher in invasive breast cancer (p=0.008) than in the fibrocystic breast disease. LECP showed a correlation with histological grade of the tumor (p=0.05). Involvement of axillary lymph nodes with metastatic tissue was in strong correlation only with existence of lymphatic vascular invasion (p=0.0001).Conclusion: These results suggest that development of breast cancer promotes proliferation of lymphatic endothelial cells whose level correlates with histological grade of tumor, but in a scope that is insufficient to follow growth of tumor tissue that invades them and destruct them. This might explain the decrease of lymphatic vessels density.

NM 23 protein was originally identified as a metastasis suppressor protein. The expression of NM23 has been correlated with tumour metastatic potential in various human carcinoma, mostly in ductal breast and colorectal carcinomas. Evidence for their expression in gastric cancer is rather contradictory, both for protein expression status and prognostic value. This study was done to analyze the immunohistochemical expression of NM23 in gastric carcinoma, and correlation of the degree of staining with clinicopathological parameters was investigated. In a retrospective immunohistochemical study specimens obtained from 56 gastric cancer patients who had undergone gastrectomy with perigastric lymphadenectomy were analysed, in correlation with classical clinical-pathological parameters of tumours, WHO-, Lauren-, Goseki-, and Ming- classification. NM 23 gene expression was compared in gastric adenocarcinoma and tumour-adjacent non-neoplastic gastric mucosa. A semiquantitative immunostaining evaluation (score 0-3) was used, counting the percentage of stained cells. Statistical analysis was performed using Kolmogorov-Smirnov test, and Spearman rank correlation test. The investigated group consisted of 40 males and 16 females (2.5:1) with a mean age of 63 years (range: 48-81 years). The percentage of positive expression of NM23 (score 3) were in 30 (53.5%) specimens in non-neoplastic mucosa in adjacent gastric carcinoma, and negative (score 0-2) in all 56 (100%) specimens of gastric adenocarcinoma. NM23 expression was higher in non-neoplastic mucosa than in adjacent gastric adenocarcinoma tissue (p<0.0001). NM23 protein expression did not correlate with gender (p=0.115), tumour size (p=0.844), tumour grade (p=0.172), lymphovascular invasion (p=0.606), lymph node metastases (p=0.311), Lauren classification (p=0.426), Goseki classification (p=0.458) and Ming classification (p=0.212). Our series did not show a significant correlation between NM23 expression and analysed clinico-pathological variables, but these results suggest that protein NM23 may have a role in gastric carcinoma pathogenesis.

Interdigitating dendritic cell sarcoma is extremely rare neoplasm that mainly occurs in the lymph nodes. Only 45 cases have been reported in the literature to date. We report a case of this sarcoma arising from the liver and lung, a previosly unreported site for this neoplasm. An 19-year-old girl deteriorated rapidly after artificial abortion and died 4 weeks later. Autopsy showed markedly enlarged liver and lung with numerous nodules up to 0.5 centimeters in diameter. Microscopically, nodules was composed of large pleomorphic cells that were immunohistochemically positive for proteins S-100 and vimentin, some of them expressed positivity to fascin and CD 68, with a rich small CD3 positive T lymphocytic infiltrateite around them. Based of these findings, the present case was diagnosed as interdigitating dendritic cell sarcoma, a neoplasm that remains a diagnostic and clinical challenge, because it can mimic a wide variety of other malignant tumors and tumor-like lesions.

Tissue inhibitor of metalloproteinase-1 (TIMP-1) is a natural inhibitor of matrix metalloproteinases (MMPs). Aim of this study was to assess the immunohistochemical expression of TIMP-1 in invasive breast carcinomas, and to examine its association with classical clinico-pathological parameters, oestrogen receptor, progesterone receptor and Her-2/neu protein expression. Immunohistochemistry was used to determine the expression of TIMP-1 on 38 paraffin-embedded breast tissue specimens - 18 with invasive ductal carcinoma, 10 with invasive lobular carcinoma, and 10 specimens from patients with fibrocystic breast disease. TIMP-1 protein was immunodetected in the carcinoma cells, fibroblasts and inflammatory cells of the stroma in 92,9%, 65,8%, and 65,8% of cases, respectively. TIMP-1 protein expression in carcinoma cells showed positive correlation with TIMP-1 protein expression in peritumoural fibroblasts (p=0,010). Positive peritumoural fibroblast TIMP-1 expression was associated with histological tumour type with higher frequency in ductal carcinomas (p=0,023). Negative association was found between TIMP-1 protein expression in carcinoma cells and HER-2/neu nuclear staining (p=0,005). TIMP-1 may be particularly useful as a predictive marker in breast carcinoma when evaluated along with HER-2/neu protein being a promising indicator of favourable prognosis in breast carcinoma.

Tissue inhibitor of metalloproteinase- (TIMP-) is a natural inhibitor of matrix metalloproteinases (MMPs). Aim of this study was to assess the immunohistochemical expression of TIMP- in invasive breast carcinomas, and to examine its association with classical clinico-pathological parameters, oestrogen receptor, progesterone receptor and Her-/neu protein expression. Immunohistochemistry was used to determine the expression of TIMP- on  paraffi n-embedded breast tissue specimens  with invasive ductal carcinoma,  with invasive lobular carcinoma, and  specimens from patients with fi brocystic breast disease. TIMP- protein was immunodetected in the carcinoma cells, fi broblasts and infl ammatory cells of the stroma in ,, ,, and , of cases, respectively. TIMP- protein expression in carcinoma cells showed positive correlation with TIMP- protein expression in peritumoural fi broblasts (p=,). Positive peritumoural fi broblast TIMP- expression was associated with histological tumour type with higher frequency in ductal carcinomas (p=,). Negative association was found between TIMP- protein expression in carcinoma cells and HER-/neu nuclear staining (p=,). TIMP- may be particularly useful as a predictive marker in breast carcinoma when evaluated along with HER-/neu protein being a promising indicator of favourable prognosis in breast carcinoma.

We report a case of exceedingly rare cutaneous neoplasm with histological features of malignancy and uncertain biological potential. The nodular, darkly pigmented facial tumor with central exulceration, size 12 x 10 x 7 mm, of the skin 61-year-old man preauricular left was completely exised. Histologically tumor consists of atypical squamous cells, which express signs of moderate to significant pleomorphism, mitotically active, with foci forming of parakeratotic horn cysts ("pearls"). Characteristically tumor also consists of large number of atypical melanocytes with multifocal pattern, inserted between atypical squamous cells, and which contain large amount of dark brown pigment melanin. Immunohistochemically, squamous cells stain positively with keratin (CK116), melanocytes were stained with S -100 protein, HMB 45, and vimentin, but failed to stain with CK 116. To our knowledge this is the sixth reported case in world literature. The follow-up time of four years no evidence of recurrence or metastasis, similar all reported cases, but it is too short period in estimation to guarantee a benign course. However, it appears that this group of neoplasm may have different prognosis from pure squamous carcinoma or malignant melanoma.

The aim of this study was to investigate expression of cyclin D1, bcl-2, p53, Ki-67 and HER-2 proteins in 14 cases of non-small cell lung cancer and to establish their correlation to classical clinico-pathological findings, and alleged prognostic value to estimate biological potential of tumor. Retrospective pilot study of the surgically treated non-small cell lung cancer biopsy specimen, paraffin embedded, used immunohistochemical method to demonstrate expression of cyclin D1, bcl-2, p53, Ki-67 and HER-2. Protein quantification was performed by the semi-quantitative method. Achieved results were correlated with classical clinico-pathological parameters, like tumor size, histological type, differentiation level, presence of vascular invasion and metastasis in regional lymph nodes. Out of 14 cases of non-small cell lung cancer, squamous cell carcinoma was found in 7 patients, giant cell carcinoma in 3, adenocarcinoma in 2, and 1 case of pleomorphic and mucoepidermoid carcinoma. Expression of cyclin D1 was not found, while expression of HER-2 and bcl-2 protein was established in one cases each. p53 expression was noted in 8 cases (57,1%). Statistically positive significant correlation (p<0,05) was found among: presence of lymphovascular invasion to tumor tissue and appearance of nodal metastasis; proliferation Ki-67 index and level of tumor differentiation, i.e. size of tumor. Other investigated parameters showed no significant statistically dependence. p53 expression was not correlated to any of the investigated parameters what might imply the possibility that there is an independent pathway of this protein expression. Negative expression of bcl-2 protein points out to possibility that it is not included into process of tumor apoptosis, as well as that proteins cyclin D1 and HER-2 are not included into processes of the tumor genesis. Since the proliferative activity of the tumor, measured by the expression of Ki-67, is correlated to the gradus and size of the tumor mass, Ki-67 protein can be of a prognostic value to determine biological potential of non-small cell lung cancer.