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Antimicrobial properties of silver (I) ion and its complexes are well recognized. However, recent studies suggest that both silver (I) ion and its complexes possess anticancer activity associated with oxidative stress-induced apoptosis of various cancer cells. In this study, we aimed to investigate whether silver nitrate and its complexes with metronidazole and 4-hydroxymethylpyridine exert anticancer action against human pancreatic cancer cell lines (PANC-1 and 1.2B4). In the study, we compared decomposition speed for silver complexes under the influence of daylight and UV-A (ultraviolet-A) rays. We employed the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazonium bromide) assay to evaluate the cytotoxicity and the alkaline comet assay to determine genotoxicity of silver nitrate and its complexes. Flow cytometry and the Annexin V-FITC/PI apoptosis detection kit were used to detect the apoptosis of human pancreatic cancer cells. We found a dose dependent decrease of both pancreatic cancer cell line viability after exposure to silver nitrate and its complexes. The flow cytometry analysis confirmed that cell death occurred mainly via apoptosis. We also documented that the studied compounds induced DNA damage. Metronidazole and 4-hydroxymethylpyridine alone did not significantly affect viability and level of DNA damage of pancreatic cancer cell lines. Complex compounds showed better stability than AgNO3, which decomposed slower than when exposed to light. UV-A significantly influences the speed of silver salt decomposition reaction. To conclude, obtained data demonstrated that silver nitrate and its complexes exerted anticancer action against human pancreatic cancer cells.

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The interaction of polyamine conjugates with DNA double helix has been studied. Binding properties were examined by ethidium bromide (EtBr) displacement and DNA unwinding/topoisomerase I/II (Topo I/II) activity assays, as well as dsDNA thermal stability studies and circular dichroism spectroscopy. Genotoxicity of the compounds was estimated by a comet assay. It has been shown that only compound 2a can interact with dsDNA via an intercalative binding mode as it displaced EtBr from the dsDNA-dye complex, with Kapp = 4.26 × 106 M-1; caused an increase in melting temperature; changed the circular dichroism spectrum of dsDNA; converted relaxed plasmid DNA into a supercoiled molecule in the presence of Topo I and reduced the amount of short oligonucleotide fragments in the comet tail. Furthermore, preliminary theoretical study has shown that interaction of the discussed compounds with dsDNA depends on molecule linker length and charge distribution over terminal aromatic chromophores.

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In this study, we compared the cellular uptake, intracellular localization and cytotoxicity of two groups of anthracycline derivatives in cultured H9c2(2-1) rat cardiomyoblasts. The first group consisted of doxorubicin (DOX) and two of its derivatives containing a formamidino group (-N = CH-N<) at the C-3' position with a morpholine (DOXM) or a hexamethyleneimine (DOXH) ring. The second group consisted of daunorubicin (DRB) and its derivatives containing a morpholine (DRBM) or a hexamethyleneimine (DRBH) ring. DOXH and DRBH were taken up by cardiomyoblasts more efficiently than estimated for other tested anthracyclines. The cellular uptakes of DOXM and DRBM were reduced compared to those of the parent compounds. Applied structural modifications of DOX and DRB influenced the subcellular localization of the tested derivatives. DOX and DOXH were localized primarily in nuclei, whereas the other anthracyclines were found in the nuclei and cytoplasm. The percentages of the compounds that accumulated in the nuclei were 80.2 and 54.2 % for DOX and DOXH, respectively. The lowest nuclear accumulation values were observed for DRBM (19.9 %), DRBH (21.9 %) and DOXM (23.7 %). The ability of anthracyclines to accumulate in the nuclei correlated with their DNA binding constants (r = 0.858, P = 0.029). A correlation was found between the accumulation of the tested anthracyclines in the nuclei of cardiomyoblasts and their cardiotoxicity in vivo, which was observed in our previous study. We suggest that cytotoxicity and the anthracycline accumulation level in the nuclei of cultured cardiomyoblasts could be used for early prediction of their cardiotoxicity.

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BACKGROUND: Type I collagen proin pro-in expression in a damaged supraspinatus tendon is thought to be dependent on the distance from the edge of the tear and the local expression of pro-inflammatory, anti-proliferative, and pro-proliferative cytokines. The study evaluates the expression of type I collagen, pro-inflammatory interleukin (IL) 1ß, anti-proliferative interferon-γ (IFN-γ), and pro-proliferative IL-4 and IL-13 cytokines along a 1-cm section taken from the edge of a torn supraspinatus tendon. Three sections were taken: 3 mm distal to the tear, 3 mm proximal to the tear, and the 4-mm middle section between them. METHODS: Nine patients (average age, 58 years) were included in the study. All fulfilled strict inclusion criteria regarding tear morphology and reconstruction technique. Samples were taken from the ruptured supraspinatus tendon at the time of arthroscopic repair. Quantitative real-time polymerase chain reaction assay was used for analysis. RESULTS: The expression of type I collagen, IL-4, and IL-13 significantly increased and that of IL-1ß and IFN-γ decreased from the distal to the proximal parts of the tendon edge (P < .05). CONCLUSIONS: The expression of type I collagen is dependent on the distance from the edge of the torn supraspinatus tendon, the balance between anti-proliferative IFN-γ and pro-proliferative IL-4 and IL-13, and the expression of pro-inflammatory IL-1ß. Hence, whereas resection of the distal 3 mm of the torn supraspinatus tendon edge eliminates its least valuable part, resection between 4 and 7 mm may enhance the healing process by reaching a reasonable compromise between the mechanical features of the tendon characterized by collagen type I expression and the technical abilities of reconstruction.

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BACKGROUND: We hypothesize that the expression of proapoptotic and antiapoptotic molecules and cytokines is dependent on the distance from the torn supraspinatus tendon edge and this expression may influence its potential for healing. The aim of this work is to evaluate the expression of proapoptotic Bax molecule and caspases 3, 8, and 9; antiapoptotic Bcl-2 molecule; and proinflammatory tumor necrosis factor (TNF) α and anti-inflammatory interleukin 10 (IL-10) in 3 sections taken from a 1-cm section of the edge of a torn supraspinatus tendon: 3 mm distal and 3 mm proximal, as well as the remaining 4-mm middle section between them. METHODS: Nine patients, with a mean age of 58 years, were included in the study. All fulfilled strict inclusion criteria regarding the morphology of the tear and reconstruction technique. Samples were taken from the ruptured supraspinatus tendon at the time of arthroscopic repair. Quantitative real-time polymerase chain reaction assay was used for analysis. RESULTS: The expression of caspases 9, 8 and 3; Bax; and TNF-α significantly decreased from the distal to the proximal parts of the tendon edge (P < .05). However, a significant increase in Bcl-2 and IL-10 expression was also found in the same direction (P < .05). CONCLUSIONS: Tenocytes can reduce the expression of proapoptotic caspases 3, 8, and 9 and Bax, as well as proinflammatory TNF-α, by increasing the expression of Bcl-2 and IL-10 within 1 cm of the supraspinatus edge in a distal to proximal direction. Resection 4 to 7 mm from the edge of the torn supraspinatus tendon may enhance the healing process by reaching a reasonable compromise between molecular homeostasis of apoptotic and inflammatory processes and mechanical aspects of rotator cuff reconstruction.

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Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that cleave protein components of extracellular matrix such as collagens, laminin, fibronectin, proteoglycans and contribute to cell migration by eliminating the surrounding extracellular matrix and basement membrane barriers. However, the extracellular matrix is not simply an extracellular scaffold because, for example, it contains sites that can bind growth factors; therefore, degradation of the extracellular matrix components by MMPs can alter cellular behavior. MMPs also cleave a variety of non-ECM proteins, including cytokines, chemokines, and growth factors, activating or inactivating them, or generating other products that have biological consequences. The immune system is also influenced by MMPs. For that reason, the function of MMPs is much more complex and subtle than simple demolition. MMPs are essential for embryonic development and morphogenesis, however, exuberant expression of these enzymes has been associated with a variety of destructive diseases, including tumor progression, cardiovascular diseases and autoimmune diseases.

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The influence of procyanidin extract from Japanese quince fruit on the activities of matrix metalloproteinases MMP-2 and MMP-9 secreted to culture medium by human peripheral blood mononuclear cells (PBMC) and by human leukemia HL-60 cells was investigated by gelatin zymography. The extract proved to be an effective inhibitor of the enzymes activities (for MMP-2 and MMP-9 secreted by PBMC IC50 = 16-19 microg extract/mL and 22-25 microg extract/mL, respectively). To identify the most effective components of the extract it was fractionated by means of column chromatography on TSKgel Toyopearl HW-40 (S) bed. The obtained fractions were analyzed by TLC, HPLC, and MALDI-TOF MS. Their antioxidant activity was measured as cation radicals ABTS(.+) scavenging efficiency. The fractions VIII-XIV containing oligomers from trimer to hexamer (and probably higher oligomers) appeared to be the most effective inhibitors of MMP-2 and MMP-9 activity (IC50 value close to 4.6 microg total polyphenols/mL). To the best of our knowledge, it is the first report on gelatinase-inhibitory activity of Japanese quince fruit polyphenol extract. We conclude that polyphenols from Japanese quince can be used in cancer chemoprevention, although further studies are needed to elucidate the mechanisms underlying their biological activities.

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