Microvesicles from Human Immortalized Cell Lines of Endothelial Progenitor Cells and Mesenchymal Stem/Stromal Cells of Adipose Tissue Origin as Carriers of Bioactive Factors Facilitating Angiogenesis.

Endothelial progenitor cells (EPCs) and mesenchymal stem/stromal cells (MSCs) are associated with maintaining tissue homeostasis and tissue repair. Both types of cells contribute to tissue regeneration through the secretion of trophic factors (alone or in the form of microvesicles). This study investigated the isolation and biological properties of microvesicles (MVs) derived from human immortalized MSC line HATMSC1 of adipose tissue origin and EPC line. The human immortalized cell line derived from the adipose tissue of a patient with venous stasis was established in our laboratory using the hTERT and pSV402 plasmids. The human EPC line originating from cord blood (HEPC-CB.1) was established in our previous studies. Microvesicles were isolated through a sequence of centrifugations. Analysis of the protein content of both populations of microvesicles, using the Membrane-Based Antibody Array and Milliplex ELISA showed that isolated microvesicles transported growth factors and pro- and antiangiogenic factors. Analysis of the miRNA content of isolated microvesicles revealed the presence of proangiogenic miRNA (miR-126, miR-296, miR-378, and miR-210) and low expression of antiangiogenic miRNA (miR-221, miR-222, and miR-92a) using real-time RT-PCR with the TaqMan technique. The isolated microvesicles were assessed for their effect on the proliferation and proangiogenic properties of cells involved in tissue repair. It was shown that both HEPC-CB.1- and HATMSC1-derived microvesicles increased the proliferation of human endothelial cells of dermal origin and that this effect was dose-dependent. In contrast, microvesicles had a limited impact on the proliferation of fibroblasts and keratinocytes. Both types of microvesicles improved the proangiogenic properties of human dermal endothelial cells, and this effect was also dose-dependent, as shown in the Matrigel assay. These results confirm the hypothesis that microvesicles of HEPC-CB.1 and HATMSC1 origin carry proteins and miRNAs that support and facilitate angiogenic processes that are important for cutaneous tissue regeneration.

Expression and activity of multidrug resistance proteins in mature endothelial cells and their precursors: A challenging correlation.

Active cellular transporters of harmful agents-multidrug resistance (mdr) proteins-are present in tumor, stem and endothelial cells, among others. While mdr proteins are broadly studied in tumor cells, their role in non-tumor cells and the significance of their action not connected with removal of harmful xenobiotics is less extensively documented. Proper assessment of mdr proteins expression is difficult. Mdr mRNA presence is most often evaluated but that does not necessarily correlate with the protein level. The protein expression itself is difficult to determine; usually cells with mdr overexpression are studied, not cells under physiological conditions, in which a low expression level of mdr protein is often insufficient for detection in vitro. Various methods are used to identify mdr mRNA and protein expression, together with functional tests demonstrating their biological drug transporting activities. Data comparing different methods of investigating expression of mdr mRNAs and their corresponding proteins are still scarce. In this article we present the results of a study concerning mdr mRNA and protein expression. Our goal was to search for the best method to investigate the expression level and functional activity of five selected mdr proteins-MDR1, BCRP, MRP1, MRP4 and MRP5-in established in vitro cell lines of human endothelial cells (ECs) and their progenitors. Endothelial cells demonstrated mdr presence at the mRNA level, which was not always confirmed at the protein level or in functional tests. Therefore, several different assays had to be applied for evaluation of mdr proteins expression and functions in endothelial cells. Among them functional tests seemed to be the most conclusive, although not very specific.

Inhibition of proliferation, migration and invasiveness of endothelial murine cells culture induced by resveratrol.

Angiogenesis is a multi-stage process of new vessel development which involves migration, proliferation and differentiation of endothelial cells. Pathological angiogenesis plays a crucial role in the pathomechanism of various ischemic, malignant and inflammatory disorders. Among eye diseases, macular degeneration (AMD) and proliferative diabetic retinopathy are a major public health issue as the most common causes of blindness. Since angiogenesis plays a crucial role in these conditions, there has been an increased interest in evaluating anti-angiogenic agents in their treatment. The polyphenol resveratrol found in the skin of red grapes, red wine, peanuts and other natural sources, controls proliferation of the cells, induces differentiation and induces apoptosis in various malignant cell lines. Modulation of angiogenesis by this compound has been considered as a very exciting topic and subject of further investigations. The aim of our study was in vitro assessment of resveratrol's influence on proliferation, migration and invasion of an immortalized murine endothelial cell line from peripheral lymph node HEC clone a10. Resveratrol was shown to inhibit the proliferation of the endothelial cells in MTT (at 1, 10 and 50 µM) and AlamarBlue (at 50 µM) assays, and at a concentration of 50 µM significantly inhibited migration of endothelial cells. A concentration-dependent decrease in invasion potential of endothelial cells incubated with resveratrol 10 µM and 50 µM was detected. These promising in vitro results might encourage investigators to test efficacy and safety of resveratrol more extensively in the clinical practice, as a natural and safe anti-angiogenic agent.

Antitumor properties of diastereomeric and geometric analogs of vitamin D3.

Analogs of 1,25-dihydroxyvitamin D3 with a reversed configuration at C-1 or C-24 and E or Z geometry of the double bond at C-22 in the side chain or at C-5 in the triene system were examined for their antiproliferative activity in vitro against a spectrum of various human cancer cell lines. The analogs coded PRI-2201 (calcipotriol), PRI-2202 and PRI-2205, such as calcitriol and tacalcitol (used as a referential agents), revealed antiproliferative activity against human HL-60, HL-60/MX2, MCF-7, T47D, SCC-25 and mouse WEHI-3 cancer cell lines. The toxicity studies in vivo showed that PRI-2202 and PRI-2205 are less toxic than referential agents. Even at total doses of 2.5-5.0 mg/kg distributed during 5 successive days, no changes in body weight were observed. Calcitriol and tacalcitol showed toxicity in the same protocol at 100 times lower doses. Calcipotriol was lethal to all mice after administration of a total dose of 5.0 mg/kg. The analog PRI-2205 appeared to be more active in mouse Levis lung cancer tumor growth inhibition than calcitriol, calcipotriol or PRI-2202. This analog did not reveal calcemic activity at doses which inhibit tumor growth in vivo nor at higher doses.

[Thermotolerant Campylobacter occurrence in slaughtering animals]. / Wystepowanie termotolerancyjnych drobnoustrojów z rodzaju Campylobacter na powierzchni tusz zwierzat rzeznych.

The aim of the study was detection and quantification of thermotolerant Campylobacter spp. in swab samples taken from carcasses of slaughter animals, i.e. poultry, cattle and pig. It has been examined 287 poultry, 176 pork and 114 beef carcass samples. Additionally, 22 poultry carcasses taken directly from slaughter line operation after evisceration were examined on the level of Campylobacter spp. contamination. Among the 287 isolates of Campylobacter spp. from poultry the most frequent was C. jejuni (59.1%), followed by C. coli (34.2%) and C. lari (6.3%). A few isolates strains from pork or beef carcasses were classified as C. coli (3.4%) or C. jejuni (0.9%).

[Gas gangrene--case diagnosis]. / Zachorowanie na zgorzel gazowa--opis diagnozowania przypadku.

Anaerobic spore-forming bacteria from species Clostridium perfringens is commonly present in soil and in the intestines of men and animals. Clostridium perfringens type A are responsible for gas gangrene men and animals. This paper describes gas gangrene case after limb amputation at 69-year-old man, treated for diabetes since several years. Used multiplex PCR generated amplification product 900 bp size (cpa) responsible for coding alpha toxin (CPA) present at Clostridium perfringens type A.