Agnieszka Chwiłkowska

Biological activity and structure dependent properties of cuprous iodide complexes with phenanthrolines and water soluble tris (aminomethyl) phosphanes.

This paper presents the biological activity of copper(I) iodide complexes with 1,10-phenanthroline (phen) or 2,9-dimethyl-1,10-phenanthroline (dmp) and three tris (aminomethyl) phosphanes: P(CH2N(CH2CH2)2NCH3)3 (1), P(CH2N(CH2CH2)2O)3 (2) and P (CH2N(CH3)CH2CH2OH)3 (3). Crystallographic and DFT data indicate a significantly stronger binding ability of 3 in the complexes [CuI (phen) P (CH2N (CH3)CH2CH2OH)3] (3P) and [CuI(dmp)P(CH2N(CH3)CH2CH2OH)3] (3N) in comparison to the 1 or 2 ligands. Most probably, this is caused by the relatively small steric requirements of 3. The complexes with dmp exhibit a very high in vitro activity against the Staphylococcus aureus strain (MIC - minimal inhibitory concentration: 2.5-5 μg/mL) and Candida albicans diploid fungus (MIC: 1.25-2.5 μg/mL). All the tested complexes also show a strong in vitro antitumor activity against human ovarian carcinoma cell lines: MDAH 2774 (IC50: 7-2 μM) and cisplatin-resistant SCOV3 (IC50: 3-2 μM). Interestingly, the complexes with dmp of higher biological activity more weakly interact with bovine serum albumin (BSA) and less efficiently cleave the pBluescriptSK+ plasmid.

Oxidative modulation of marcaine and lekoptin in H9C2 rat myoblasts

AbstractAim:The cytotoxicity of marcaine was estimated in combination with a calcium channel blocker. In addition, the influence of marcaine and marcaine plus lekoptin on a model system using the H9C2 cardiac cell line was investigated.Methods:Cells were incubated for five hours with marcaine, lekoptin, or with both drugs simultaneously. Apoptotic cells were detected using the TUNEL assay and the alkaline comet assay. Mitochondrial cell function after drug uptake was examined using the MTT assay. The concentration of MDA (malondialdehyde) — the final product of fatty-acid peroxidation, was quantified spectrophotometrically. The expression of glutathione S-transferase π (GST-π) was detected by immunofluorescence (IF) and Western blotting (WB) and inducible nitric oxide synthase (iNOS) was assessed by immunocytochemical staining (ABC).Results:Incubation with marcaine resulted in the highest number of apoptotic cells. After incubation with both marcaine and lekoptin, moderate damage to cells (54.2%±1.775% of DNA destruction) was observed. The highest levels of iNOS and GST-π expression were observed in cells treated with marcaine and marcaine plus lekoptin. The characteristic nuclear GST-π expression was observed in cells treated with both drugs.Conclusion:Lekoptin stimulated cells to proliferate. Marcaine caused membrane damage and ultimately cell death.

The content of collagen type II in human arteries is correlated with the stage of atherosclerosis and calcification foci

OBJECTIVEnThe signature processes during atherosclerosis development are arterial calcification and accumulation in the arterial walls of proteins that are specific to bone and cartilage, e.g., collagen type II. The purpose of this study was to characterize localization of collagen type II and quantify its content in human arteries.nnnRESULTSnThe study was conducted on sections of thoracic and abdominal aortas (n=97) subjected to histological evaluation and classified into six grades according to the Stary scale of the atherosclerosis severity. Three types of samples were distinguished from the group of arteries: (1) without macroscopically visible calcifications, (2) with macroscopically visible calcifications dispersed within the arterial wall, and (3) calcium deposits isolated from the walls tested with respect to the segment of the artery from which they had originated. The results demonstrate that both cholesterol and collagen type II content are significantly higher in samples with calcification, whereas collagen type II is localized mainly in the tissue around the calcium deposit. A positive correlation has been shown between the levels of collagen type II and cholesterol (r=0.57, P<.05). A similar trend was observed with respect to the grade of atherosclerosis (r=0.43, P<.05).nnnCONCLUSIONSnThe amount of collagen type II is higher in the tissue around the calcium deposit. The correlation was observed between the quantityof collagen type II, the grade of atherosclerosis, and cholesterol.

The impact of development of atherosclerosis on delamination resistance of the thoracic aortic wall

The aim of this work is to determine the impact of development of atherosclerosis on dissection of the human thoracic aorta on the basis of an analysis of the mechanical properties of the interfaces between its layers. The research material consisted of 17 pathologically unchanged aortae and 74 blood vessels with atherosclerotic lesions, which were classified according to the histological classification by Stary. The subject of the analysis were the interfaces between the adventitia and the media-intima complex (A-MIC) and between the intima and the media-adventitia complex (I-MAC). The mechanical properties of the above interfaces were determined by the peeling test in the longitudinal and circumferential directions. The results indicate that development of atherosclerosis reduces vessel wall resistance to delamination. The greatest risk of dissection occurs at stage IV of the disease. In this case, energy values are lower by about 28% for the I-MAC interface and by 39% for the A-MIC interface compared with normal tissues. Lower values of mean force and energy were obtained for the I-MAC interface, indicating that this interface is more susceptible to delamination. The mechanical properties of the A-MIC interfaces are directional.

Correction to: Advanced glycation end products as a source of artifacts in immunoenzymatic methods

The original version of this article unfortunately contained a mistake in the author group section. Author A. Bronowicka-Szydełko’s surname was inadvertently interchanged to “Szydełko-Bronowicka”.

Advanced glycation end products as a source of artifacts in immunoenzymatic methods

The most abundant proteins in the arteries are those of extracellular matrix, ie. collagen and elastin. Due to their long half-lifes these proteins have an increased chance to undergo glycation. The aim of this study was to determine relationship between the content of the main extracellular matrix proteins and the advanced glycation end products (AGEs) in arteries. In this study 103 fragments of aorta were analyzed by ELISA and immunobloting for the content of collagens type I, III and IV and elastin and the content of advanced glycation end-products (AGE). A negative correlation between the content of collagens type III and IV and AGE (rxa0=xa0−0,258, pxa0=xa00,0122, and a weak negative correlation between collagen type III and age of the sample donor (rxa0=xa00,218, pxa0=xa00,0262) were demonstrated. This result comes as a surprise and it contradicts an intuitive assumption that with more glycation substrate, i.e. matrix proteins, more AGE products are expected. We have concluded that the results of the ELISA tests must have been influenced by the glycation. As a consequence, either modified protein molecules were not being recognized by the antibodies, or the glycation, and formation of crosslinks have blocked access of the antibodies to the antigen. It will conceal the effect of the linear dependence between the result (absorbance/densitometry) from the quantity of protein to which the antibody is directed.

Estrogen Receptors in Cell Membranes: Regulation and Signaling

Estrogens can stimulate the development, proliferation, migration, and survival of target cells. These biological effects are mediated through their action upon the plasma membrane estrogen receptors (ERs). ERs regulate transcriptional processes by nuclear translocation and binding to specific response elements, which leads to the regulation of gene expression. This effect is termed genomic or nuclear. However, estrogens may exert their biological activity also without direct binding to DNA and independently of gene transcription or protein synthesis. This action is called non-genomic or non-nuclear. Through non-genomic mechanisms, estrogens can modify regulatory cascades such as MAPK, P13K, and tyrosine cascade as well as membrane-associated molecules such as ion channels and G-protein-coupled receptors. The recent studies on the mechanisms of estrogen action provide an evidence that non-genomic and genomic effects converge. An example of such convergence is the potential possibility to modulate gene expression through these two independent pathways. The understanding of the plasma membrane estrogen receptors is crucial for the development of novel drugs and therapeutic protocols targeting specific receptor actions.