Prenyl Ammonium Salts – New Carriers for Gene Delivery: A B16-F10 Mouse Melanoma Model

Purpose Prenyl ammonium iodides (Amino-Prenols, APs), semi-synthetic polyprenol derivatives were studied as prospective novel gene transfer agents. Methods AP-7, -8, -11 and -15 (aminoprenols composed of 7, 8, 11 or 15 isoprene units, respectively)were examined for their capacity to form complexes with pDNA, for cytotoxicity and ability to transfect genes to cells. Results All the carriers were able to complex DNA. The highest, comparable to commercial reagents, transfection efficiency was observed for AP-15. Simultaneously, AP-15 exhibited the lowest negative impact on cell viability and proliferation—considerably lower than that of commercial agents. AP-15/DOPE complexes were also efficient to introduce pDNA to cells, without much effect on cell viability. Transfection with AP-15/DOPE complexes influenced the expression of a very few among 44 tested genes involved in cellular lipid metabolism. Furthermore, complexes containing AP-15 and therapeutic plasmid, encoding the TIMP metallopeptidase inhibitor 2 (TIMP2), introduced the TIMP2 gene with high efficiency to B16-F10 melanoma cells but not to B16-F10 melanoma tumors in C57BL/6 mice, as confirmed by TIMP2 protein level determination. Conclusion Obtained results indicate that APs have a potential as non-viral vectors for cell transfection

Glosa do biografii ks. Karola Koziołka

The priest, Karol Koziołek is a parson of the Grabin parish near to Prudnik, and is one of the head activists of the Polish national movement in Opole Silesia. In 1928 he became a member of the Provincial Assembly, he also attempted to gain a seat in the Prussian parliament and the parliament of German Reich, although was not successful. In the years 1930–1935 he performed the function of chairman of the District of the Polish Union in Germany. He worked with a huge amount of dedication to preserving the use of the Polish language in the Catholic Church. His speeches in defence of the Polish language in schools and the Catholic Church resulted in many attacks and threats directed at his name by nationalist societies in Germany. Priest Koziołek played a special role in the formation of the local unions and social life in Opole Silesia. His huge authority amongst locals, and their attachment to the Catholic faith was not left without an effect on the process of the formation of Polish national attitude.Ks. Karol Koziołek, proboszcz parafii w Grabinie koło Prudnika, to jeden z czołowych działaczy polskiego ruchu narodowego na Śląsku Opolskim. W 1928 r. został posłem do sejmiku prowincjonalnego. Ubiegał się także, chociaż bezskutecznie, o mandat do sejmu pruskiego i parlamentu Rzeszy Niemieckiej. W latach 1930–1935 sprawował funkcję prezesa Dzielnicy I Związku Polaków w Niemczech. Z ogromnym zaangażowaniem działał na rzecz zachowania języka polskiego w Kościele. Jego wystąpienia w obronie języka polskiego w szkole i Kościele stały się powodem wielu ataków i pogróżek kierowanych pod jego adresem przez środowiska nacjonalistyczne w Niemczech. Ks. Koziołek odegrał szczególną rolę w kształtowaniu życia społecznego lokalnych skupisk na Śląsku Opolskim. Jego ogromny autorytet wśród miejscowej ludności i jej przywiązanie do wiary katolickiej nie pozostało bez wpływu na proces kształtowania polskich postaw narodowych

Glosa do biografii ks. Karola Koziołka

The priest, Karol Koziołek is a parson of the Grabin parish near to Prudnik, and is one of the head activists of the Polish national movement in Opole Silesia. In 1928 he became a member of the Provincial Assembly, he also attempted to gain a seat in the Prussian parliament and the parliament of German Reich, although was not successful. In the years 1930–1935 he performed the function of chairman of the District of the Polish Union in Germany. He worked with a huge amount of dedication to preserving the use of the Polish language in the Catholic Church. His speeches in defence of the Polish language in schools and the Catholic Church resulted in many attacks and threats directed at his name by nationalist societies in Germany. Priest Koziołek played a special role in the formation of the local unions and social life in Opole Silesia. His huge authority amongst locals, and their attachment to the Catholic faith was not left without an effect on the process of the formation of Polish national attitude.Ks. Karol Koziołek, proboszcz parafii w Grabinie koło Prudnika, to jeden z czołowych działaczy polskiego ruchu narodowego na Śląsku Opolskim. W 1928 r. został posłem do sejmiku prowincjonalnego. Ubiegał się także, chociaż bezskutecznie, o mandat do sejmu pruskiego i parlamentu Rzeszy Niemieckiej. W latach 1930–1935 sprawował funkcję prezesa Dzielnicy I Związku Polaków w Niemczech. Z ogromnym zaangażowaniem działał na rzecz zachowania języka polskiego w Kościele. Jego wystąpienia w obronie języka polskiego w szkole i Kościele stały się powodem wielu ataków i pogróżek kierowanych pod jego adresem przez środowiska nacjonalistyczne w Niemczech. Ks. Koziołek odegrał szczególną rolę w kształtowaniu życia społecznego lokalnych skupisk na Śląsku Opolskim. Jego ogromny autorytet wśród miejscowej ludności i jej przywiązanie do wiary katolickiej nie pozostało bez wpływu na proces kształtowania polskich postaw narodowych

New cationic polyprenyl derivative proposed as a lipofecting agent

Cationic linear poly-cis-isoprenoid prepared from natural plant polyprenol in a mixture with dioleyl phosphatidylethanolamine was found to be an effective lipofection agent for eukaryotic cells. The transfecting activity is related to the poly-cis structure of the polyprenyl chain

Subclinical Hypothyroidism after Radioiodine Exposure: Ukrainian–American Cohort Study of Thyroid Cancer and Other Thyroid Diseases after the Chornobyl Accident (1998–2000)

BackgroundHypothyroidism is the most common thyroid abnormality in patients treated with high doses of iodine-131 (131I). Data on risk of hypothyroidism from low to moderate 131I thyroid doses are limited and inconsistent.ObjectiveThis study was conducted to quantify the risk of hypothyroidism prevalence in relation to 131I doses received because of the Chornobyl accident.MethodsThis is a cross-sectional (1998-2000) screening study of thyroid diseases in a cohort of 11,853 individuals < 18 years of age at the time of the accident, with individual thyroid radioactivity measurements taken within 2 months of the accident. We measured thyroid-stimulating hormone (TSH), free thyroxine, and antibodies to thyroid peroxidase (ATPO) in serum.ResultsMean age at examination of the analysis cohort was 21.6 years (range, 12.2-32.5 years), with 49% females. Mean 131I thyroid dose was 0.79 Gy (range, 0-40.7 Gy). There were 719 cases with hypothyroidism (TSH > 4 mIU/L), including 14 with overt hypothyroidism. We found a significant, small association between (131)I thyroid doses and prevalent hypothyroidism, with the excess odds ratio (EOR) per gray of 0.10 (95% confidence interval, 0.03-0.21). EOR per gray was higher in individuals with ATPO < or = 60 U/mL compared with individuals with ATPO > 60 U/mL (p < 0.001).ConclusionsThis is the first study to find a significant relationship between prevalence of hypothyroidism and individual (131)I thyroid doses due to environmental exposure. The radiation increase in hypothyroidism was small (10% per Gy) and limited largely to subclinical hypothyroidism. Prospective data are needed to evaluate the dynamics of radiation-related hypothyroidism and clarify the role of antithyroid antibodies

Efficient, non-toxic gene delivery by negatively charged polyprenyl-based lipoplexes: Application in RNA delivery and the effects on cell physiology

The development in the field of DNA and RNA delivery into cells and progress in understanding pathogenesis of many diseases resulted in nucleic acids becoming actually drugs and their delivery one of the top molecular biology techniques applicable in clinics. Still, one of the major challenges facing the development of gene therapy is lack of efficient and safe gene vectors. We have examined a new class of polyprenyl-based cationic lipids for gene transfer. Studies have shown that semisynthetic polyprenyltrimethylammonium iodides (PTAI) in formulations with co-lipids (DOPE, DC-cholesterol, DOPC) have the ability to effectively transfect plasmid DNA in a wide range of cell types in vitro both in the presence and absence of serum. Although generally it is considered that bigger lipoplexes bearing positive zeta potential are more efficient, our data clearly demonstrate that small (90 – 150 nm), negatively charged (about -30 mV) polyprenyl-based lipoplexes are efficient and have parameters making them promising candidates for in vivo gene delivery. As it was demonstrated that lipofection procedure may have several side effects on cell physiology, we tested the effects of PTAI formulation on cell motility, proliferation, viability and gap junctional intercellular coupling (GJIC). We have tested four derivatives: amino-Pren-7, amino-Pren-8, amino-Pren-11 and amino-Pren-15. Cell motility of a model DU-145 (human prostate cancer) cells was estimated by time-laps monitoring of movement of individual cells and GJIC intensity measured using donor cells labelled with calcein plated onto monolayers of acceptor cells transfected with PTAI-based lipoplexes. The dynamics of calcein transfer from donor to acceptor cells was analyzed. Antimicrobial activity was evaluated by colony reduction assay and the hemolytic activity against human red blood cells (RBCs) was tested using PBS suspension prepared from fresh blood. The results show that lipoplexes based on PTAI have no effects on cell physiology that is cell viability, proliferation and morphology. Moreover, they also occurred to have no effect on GJIC and cell motility (24 hours after transfection all the cells cover the distance of about 210-240 μm showing a displacement of 70-80 μm). Some PTAI-based vectors exhibit potent bactericidal activity against Streptococcus aureus and Escherichia coli, while showing no toxic effect on eukaryotic cells, which can be beneficial during prolonged storage of formulations. Furthermore, (as we suggest in vivo application of PTAI vectors) we have also proved their safety towards human RBSs, which membranes are not disrupted in the presence of all the examined concentrations of PTAI-based lipoplexes. Moreover, the formulations tested in plasmid DNA transfer into cells are also effective in gene silencing techniques utilizing RNA delivery. We have successfully introduced shRNA inducing GFP gene silencing into DU145, XC (rat sarcoma) and B16F10 (mouse melanoma) cells expressing pEGFP-C1 plasmid achieving GFP gene silencing. Additionally, PTAI-based formulations can be safely stored for extended periods (up to 18 months) at 4°C. In conclusion, lipoplexes based on PTAI provide ability to introduce DNA or RNA into cells with satisfying efficiency, easily and safety, as they exhibit no toxic activity and no side effects on cell proliferation, motility and GJIC. What is more, PTAI-based formulations show advantages important for convenient use (both – DNA and RNA delivery, antimicrobial activity, prolonged storage) and in vivo applications (no RBCs rupture in the presence of PTAI-based lipoplexes, effectiveness in the presence of serum)