The pig appears to be the most promising animal donor of organs for use in human recipients. Among several types of pathogens found in pigs, one of the greatest problems is presented by porcine endogenous retroviruses (PERVs). Screening of the source pig herd for PERVs should include analysis of both PERV DNA and RNA. Therefore, the present study focuses on quantitative analysis of PERVs in different organs such as the skin, heart, muscle, and liver and blood of transgenic pigs generated for xenotransplantation. Transgenic pigs were developed to express the human α-galactosidase, the human α-1,2-fucosyltransferase gene, or both genetic modifications of the genome (Lipinski et al., Medycyna Wet 66:316-322, 2010; Lipinski et al., Ann Anim Sci 12:349-356, 2012; Wieczorek et al., Medycyna Wet 67:462-466, 2011). The copy numbers of PERV DNA and RNA were evaluated using real-time Q-PCR and QRT-PCR, respectively. Comparative analysis of all PERV subtypes revealed the following relationships: PERV A > PERV B > PERV C. PERV A and B were found in all samples, whereas PERV C was detected in 47 % of the tested animals. The lowest level of PERV DNA was shown in the muscles for PERV A and B and in blood samples for PERV C. The lowest level of PERV A RNA was found in the skin, whereas those of PERV B and C RNA were found in liver specimens. Quantitative analysis revealed differences in the copy number of PERV subtypes between various organs of transgenic pigs generated for xenotransplantation. Our data support the idea that careful pig selection for organ donation with low PERV copy number may limit the risk of retrovirus transmission to the human recipients.
It acts through membrane receptors (MT1 and MT2), nuclear receptors (ROR/RZR -retinoid orphan receptors/ retinoid Z receptors), and nonreceptor-mediated mechanisms (Slominski et al., 2012). Melatonin, through its antioxidant properties, protects against damage of cellular components including DNA, cytosolic proteins, and cell membrane lipids (Yürüker et al., 2015). Studies on the antioxidant properties of melatonin are most commonly associated with agents that induce free radicals in cells (Nazıroğlu et al., 2013). Antibiotics also induce the generation of free radicals. One of them, amphotericin B (AmB), causes lipid peroxidation through generation of reactive oxygen species (ROS). AmB belongs to the group of polyene antibiotics and is an effective antifungal drug commonly used to treat systemic mycoses. It is believed that the fungicidal activity is due to the binding of AmB with the ergosterol present in the cell membrane of fungi, resulting in membrane permeabilization and osmotic imbalance (Brajtburg et al., 1990;Paulo et al., 2013). One of the mechanisms leading to fungal cell death is leakage of potassium ions caused by formation of ion channels in the cell membrane as a consequence of AmB binding with ergosterol (Chudzik et al., 2015). This situation leads to the generation of ROS and lipid peroxidation (Mesa-Arango et al., 2012). Unfortunately, AmB, besides its affinity to ergosterol, also shows an affinity to cholesterol present in human cell membranes, causing nephrotoxic and hepatotoxic side effects (Antonowicz-Juchniewicz et al.
Our findings corroborate metabolic genes interactions, especially for CYP1A1 462Val alleles and GSTM1 homozygous deletion, in the development of head and neck cancer in the investigated population groups in Poland.
Retinal pigment epithelium (RPE) is a specialized structure essential for proper vision, which is constantly exposed to oxidative damage. With aging, this damage accumulates within the RPE cells, causing various diseases, including age-related macular degeneration (AMD). Numerous antioxidant substances are used to prevent this process in humans, including lutein. This study aims to determine the differences in the expression patterns of pyroptosis genes in senescent human retinal pigment epithelial cell line ARPE-19 exposed to lutein. Changes in the expression of pyroptosis-related genes were assessed by oligonucleotide microarrays, and the results were validated by real-time RT-qPCR. The microarray analysis showed seven transcripts were differentially expressed both in the H2O2-treated cells versus the controls and in the lutein/H2O2-treated cells compared to the H2O2-treated cells (FC > 2.0). Depending on the used lutein, H2O2, or co-treatment of ARPE-19 cells, statistically significant differences in the expression of TXNIP, CXCL8, BAX, and CASP1 genes were confirmed by the RT-qPCR (p < 0.05). A STRING database analysis showed that the proteins encoded by the analyzed genes form a strong interaction network (p < 0.001). These data indicate that lutein modulates the expression level of pyroptosis-related genes, which may be useful for the development of new methods preventing pyroptosis pathway activation in the future.
We have tested titanium (Ti) plates that are used for bone reconstruction in maxillofacial surgery, in combination with five types of novel long-resorbable biomaterials: (i) PCL0—polycaprolactone without additives, (ii) PCLMWCNT—polycaprolactone with the addition of multiwall carbon nanotubes (MWCNT), (iii) PCLOH—polycaprolactone doped with multiwall carbon nanotubes (MWCNT) containing –OH hydroxyl groups, (iv) PCLCOOH—polycaprolactone with the addition of multiwall carbon nanotubes (MWCNT) containing carboxyl groups, and (v) PCLTI—polycaprolactone with the addition of Ti nanoparticles. The structure and properties of the obtained materials have been examined with the use of Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), and/or X-ray powder diffraction (XRD). Titanium BR plates have been covered with: (i) PCL0 fibers (PCL0BR—connection plates), (ii) PCLMWCNT fibers (PCLMWCNTBR—plates), (iii) PCLOH fibers (PCLOHBR—plates), (iv) PCLCOOH (PCLCOOHBR—plates), (v) PCLTI fiber (PCLTIBR—connection plates). Such modified titanium plates were exposed to X-ray doses corresponding to those applied in head and neck tumor treatment. The potential leaching of toxic materials upon the irradiation of such modified titanium plates, and their effect on normal human dermal fibroblasts (NHDF) have been assessed by MTT assay. The presented results show variable biological responses depending on the modifications to titanium plates.
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