SUMMARY – The purpose of the study was to assess the level of serum malondialdehyde (MDA) concentration and its association with the stage and histopathologic sizes of colorectal cancer (CRC). One hundred and two patients having undergone surgical treatment of CRC between January 2014 and December 2015 were included in this cross-sectional study. The patients were divided into four groups (stage I-IV) according to the TNM classification. Control group included 30 subjects with no signs of malignancy and inflammatory diseases. In each patient, preoperative blood samples were obtained for determination of MDA concentration by ELISA immunoassay. Serum levels of MDA were progressively increased in patients with CRC, reaching the highest value in the fourth stage of CRC. Serum concentrations of MDA were significantly higher in pT4 group as compared with pT3 and pT2 groups of CRC patients (p<0.01). Significantly higher levels of MDA were found in the N1 and N2 groups of CRC patients as compared with N0 group, as well as in patients with metastatic disease as compared with those without metastasis (p<0.001). In conclusion, the progression of CRC is associated with a significant increase in serum MDA levels.

The Tomašica grave-site near Prijedor in the north of Bosnia is reported to be the largest primary mass grave discovered thus far relating to the 1992-95 war. A total of 275 complete bodies and 125 body parts were exhumed from it in 2013. Post mortem examinations of the victims showed that nearly all had died from gunshot injuries but an additional striking feature was the degree of preservation of many of the bodies, even 21 years on, with skin, soft tissues and internal organs still present in abundance and gross structures clearly identifiable. Histology was performed on 68 samples of soft tissue from a total 13 bodies, on both skin and internal organs, and the degree of preservation was assessed in terms of the ability to recognize microscopic structure. Further comparison was made with samples taken a month or so later (56 tissue samples from 9 bodies, all but one different from the first group), after the bodies had been covered in salt as a means of general preservation. Generally, at a microscopic level, skin and subcutaneous tissues were better preserved than internal organs, while tissues sampled at the time of autopsy were better preserved than those sampled weeks later.

Background Tumor development and growth are driven in many cases by inflammatory cells, which can produce cytokines and other factors that can stimulate the development of the malignant process. The aim of this study was to evaluate interleukin-6 (IL-6), C-reactive protein (CRP), matrix metalloproteinase-9 (MMP-9), serum levels in patients with colorectal cancer (CRC), and their association with the stage of CRC. Methods IL-6, MMP-9, and CRP serum levels were measured in 75 patients with CRC just before surgical treatment, as well as in 20 healthy individuals as controls. Surgically obtained tissue material was subjected to pathological analysis. Results Significant increase in CRP and IL-6 serum concentration is associated with increasing stage of CRC (p <0.05), where MMP-9 serum level was significantly higher in stages III and IV compared to the stage II CRC. Significant correlation was found between IL-6 and MMP-9 serum levels (rho=0.478; p <0.001) as well as between IL-6 and CRP serum levels (rho=0.720; p <0.001) and between MMP-9 and CRP serum levels (rho=0.379; p <0.001). Serum levels of MMP-9 and CRP have been shown to be independent predictors of the CRC stage. Conclusion Combined quantification of IL-6, MMP-9, and CRP serum levels seems to be a reliable index of inflammation-related processes during colorectal carcinogenesis.

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).

Aim To determine the serum levels of matrix metalloproteinase 9 (MMP-9) concentration and their association with the stage and histopathologic sizes of colorectal cancer (CRC). Methods One hundred and two patients with clinically diagnosed and histologically confirmed colorectal cancer ready for surgical treatment were included in the study. In each patient, preoperative peripheral venous blood samples were taken for determination of the concentration of MMP-9 using ELISA immunoassay test. Resected tumour specimens were studied pathologically according to the criteria of the TNM classification. All patients were divided into groups according to the TNM classification. The control group presented 30 subjects of the appropriate age and gender with no family history of cancer, clinical signs of malignancy or inflammatory bowel disease. Results The serum levels of MMP-9 were progressively increased in patients with CRC reaching the highest value in the fourth stage of CRC. It was also confirmed that the serum concentrations of MMP-9 were significantly higher in patients with pericolonic lymph nodes involvement compared to the patients with no involvement of lymph nodes, 456.4 (445.9-464.7) ng/mL vs. 438.4 (418.4-447.8) ng/mL (p<0.001). Significantly higher serum levels of MMP-9 were found in the patients with metastatic CRC, 458.5 (452.0-468.1) ng/mL compared with the CRC patients without metastasis, 445.8 (436.9-456.5) ng/mL (p<0.001). Conclusion It was confirmed that serum concentration of MMP-9 presented the significant independent risk factors for the progression of CRC.

Oral melanoma (OM) occurs from activated or genetically altered epidermal melanocytes. There is no scientific evidence that OM can be linked to physical, chemical and thermal irritation, or to other risk factors of the oral cavity. According to fi gures from various countries, OM accounts for 0.2% to 7.5% ( Japan) of all cases of melanoma of the skin and mucous membrane. The male to female ratio of OM is 2:1. About 80% of OMs are located in the mucosa of the palate and maxillary gingiva. This paper presents two cases of oral mucosal melanoma of the upper and lower lips in women aged 62 and 59 years. Diagnosis, differential diagnosis and therapy are reported.

Objectives: The association of inflammatory reactions with almost all types of cancer supports the concept that inflammation is a critical component of tumor progression. The present study aimed to evaluate the relationship of serum markers of chronic inflammation with the stage of and histopathological size of colorectal carcinoma (CRC). Methods: This cross-sectional study included 90 patients of both sexes, mean age 66.2 (range 47-78) years, with clinically and histologically confirmed CRC, who were admitted to the Clinic for abdominal surgery UCCS for surgical treatment of CRC. The patients according to the stage of disease were divided into three groups (stage II–IV). The control group consisted of 30 subjects with no signs of malignancy and acute inflammatory diseases. Staging of CRC was done according to the TNM classification. In each patient, the preoperative blood samples were taken for determination of the parameters of inflammation: the erythrocyte sedimentation rate, white blood cells, C-reactive protein (CRP), fibrinogen and alpha 2 globulins. Results: It was confirmed that increasing markers of inflammation followed increasing stages of colorectal cancer, depth of tumor invasion and the occurrence of metastatic disease. CRP is a biomarker that consistently and significantly increases from the second to the fourth stage of colorectal cancer (7.2 (2.3-14.6) mg/L vs. 21.85 (12.3-41) mg/L vs. 38.6 (21.5-79) mg/L; p<0.01) and significantly correlates positively with the stage of CRC (r= 0.783, p<0.001), and the tumor size (r=0.249, p<0.05). Conclusion: The study results point to an increase in the degree of chronic inflammation throughout the progression of colorectal cancer. The most consistent marker of chronic inflammation that accompanies the progression of colorectal carcinoma is CRP.

Nine Lactobacillus strains known for surface proteinase activity were chosen from our collection and tested for their ability to grow in pea seed protein-based medium, and to hydrolyze purified pea proteins in order to produce peptides with antioxidant (AO) activity. Two strains, Lactobacillus rhamnosus BGT10 and Lactobacillus zeae LMG17315, exhibited strong proteolytic activity against pea proteins. The AO activity of the pea hydrolysate fraction, MW <10 kDa, obtained by the fermentation of purified pea proteins with Lactobacillus rhamnosus BGT10, was tested by standard spectrophotometric assays (DPPH, ABTS, Fe3+-reducing capacity) and the recently developed direct current (DC) polarographic assay. The low molecular weight fraction of the obtained hydrolysate was separated using ion exchange chromatography, while the AO activity of eluted fractions was determined by means of a sensitive DC polarographic assay without previous concentration of samples. Results revealed that the fraction present in low abundance that contained basic peptides possessed the highest antioxidant activity. Based on the obtained results, it can be concluded that Lactobacillus rhamnosus BGT10 should be further investigated as a candidate strain for large-scale production of bioactive peptides from legume proteins. [Projekat Ministartsva nauke Republike Srbije, br. 173005 i br. 173026]

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

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.

Phenazines, secondary metabolites of fluorescent Pseudomonas, represent a group of heterocyclic nitrogen-containing compounds showing a broad spectrum of antibiotic properties. Phenazines producing fluorescent Pseudomonas species are studied extensively for their application in plant disease management. In this study, we examined the antifungal activity of different indigenous Pseudomonas isolates (Q16, B25 and PS2) against the phytopathogenic fungus Alternaria tenuissima, which had infected cardoon (Cynara cardunculus L., Asteraceae). An in vitro experiment demonstrated the antifungal activity of selected indigenous isolates. In addition, an in vivo experiment under gnotobiotic conditions showed suppression of C. cardunculus disease caused by A. tenuissima. The quantification of phenazines revealed significant amounts of phenazine-1-carboxylic acid (PCA) and 2-hydroxy-phenazine-1-carboxylic acid (2-OH-PCA). PCR analysis confirmed the presence of PCA genes in all examined indigenous Pseudomonas isolates. Based on our results, we assume that these Pseudomonas isolates have potential in controlling plant diseases caused by A. tenuissima. [Projekat Ministarstva nauke Republike Srbije, br. III46007 and br. TR31018]