Mobile genetic elements – mechanism and consequences of transposition

Retrotransposons represent a significant part of the genome in eukaryote organisms. With DNA transposons, they belong to mobile genetic elements. There are two classes of transposons, DNA transposons and retrotransposons. Retrotransposons have three genes in their structure (gag, pol, env), responsible for the activation and encoding of structural proteins and envelope proteins. There is a group of retroelements without LTR (non-LTR). This group consists LINE (Long Interspersed Nuclear Elements) and SINE (Short Interspersed Nuclear Elements) sequences. LINE sequences represent 20% of human genetic material. Sequences of retrotransposons can move within the genome of a particular organism, occasionally subjected to replication as a result of enzyme activity, i.e. reverse transcriptase. The process of retro transposition is imperfect. These processes often cause mutation (most often: insertion, deletion), genetic instability, they contribute to the development of diseases on the genetic basis, including cancer. Retrotransposons are also used to analyze genetic variation as genetic markers

Synthesis and application of (18F) fluorodeoxyglucose in oncology diagnosis

[18F] Fluorodeoxyglucose ([18F] FDG) is the most commonly used radiopharmaceutical in clinical positron emission tomography (PET) in oncology. Cancer cells create their own specific microenvironment to survive and grow. Specific tumor microenvironment contributes to cancer metabolic reprogramming. Therefore, even with sufficient oxygen availability, cancer cells choose anaerobic glycolysis. Cancer cells compensate less energy efficient process by increasing the intensity of anaerobic glycolysis. Tumor cells have a high rate of metabolism and because of this, they take up more of the radioactive glucose (FDG). This makes the tumor cells appear more visible than other areas on the PET scan pictures. This paper presents nucleophilic synthesis of the [18F] FDG marker and basics of tumor development which can affect the [18F] FDG biochemical significance. Reference was made to clinical images obtained in PET technology using the [18F] FDG radiopharmaceutical

Impact of anti-inflammatory corticosteroids on changes in selected cytoimmunological parameters in selected interstitial lung diseases

Introduction. Glucocorticoids (GKS) hormones with strong immunosuppressive and anti-inflammatory activity are used, inter alia, in interstitial lung diseases (ILD). GKS inhibit excessive activity of inflammatory genes in the airways to induce apoptosis of immune cells, such as alveolar lymphocytes (AL). Objective. Assessment of cytoimmunological changes including apoptosis occurring in alveolar lymphocytes in patients with selected ILD after treatment with systemic corticosteroids. Methods. The material of the bronchoalveolar lavage (BAL) derived from patients with sarcoidosis (PS, n=66), idiopathic pulmonary fibrosis (IPF, n=27) and non-specific interstitial pneumonia (NSIP, n = 25; adequate number of patients treated with systemic GKS were 23, 8 and 6) were analyzed in cytoimmunological tests: a) percentage and the total values of BAL inflammatory cell populations; b) AL subset typing; c) CD4/CD8 index calculation; d) AL cell cycle analysis (DNA staining with propidium iodide, PI); in techniques mentioned in b) to d) items flow cytometry was used. Results. In all patient groups, treatment with GKS resulted in a decrease in the total cell number, e.g. for PS (untreated: 310 ± 80x103/ml; treated: 188 ± 43x103/ml, median ± SEM, p<0.05) and lymphocytes (untreated: 113 ± 71x103/ml; treated: 43 ± 25x103/ml median ± SEM, p<0.05). There has also been significantly lower percentage of eosinophils in all groups in GKS-treated subgroups, e.g. for IPF (untreated: 4,6 ± 3,0%, treated: 0,3 ± 0,5%), for NSIP (untreated: 1,5 ± 0,7% and treated: with 0,4 ± 0,2%; median ± SEM, p<0.05 for both). In contrast, AL apoptosis rate was significantly higher in treated patients, e.g. for PS (untreated: 0,6 ± 0,5%; treated: 4,0 ± 2,5%), NSIP (untreated: 3,4 ± 1,8%; treated: 13,1 ± 5,1%; median ± SEM, p<0.05 for both). In all GKS-treated groups corticosteroid therapy caused lower CD4/CD8 index, but only on the level of statistical tendency (e.g. for PS untreated: 4,7 ± 0,5%, treated: with 2,9 ± 0,9%) for IPF (untreated: 1,2 ± 0,5%, treated: with 1,1 ± 0,5%; median ± SEM for both). Conclusions. Systemic inflammatory glucocorticoid therapy (GKS) in all included groups of patients results in a decrease in the total number of alveolar lymphocytes, which is most likely related to the significant increase apoptosis rate of these cells. GKS medication in a similar extent caused probably both the death of helper (Th) and cytotoxic lymphocytes (Tc), since the decline in the value of CD4/CD8 index in treated patients compared with untreated ones was insignificant. A characteristic BAL cytological change in all tested ILD after GKS administration was a remarkable decrease in eosinophil percentage

Preliminary flagellin gene transfection into tumor cells - attempts of generating anti-tumor response in experimental model

The development of new research techniques, especially molecular ones, creates hopes for improved treatment efficacy and a better prognosis in lung cancer. The starting point for very important experiments in the field of immunotherapy is the sensitization of the immune system to the tumor antigen . The aim of the study was to determine whether using flagellin can induce innate antitumor effective, and thus, increase the immunogenicity of the tumor. The test material was the established mouse cell line LLC, derived from Lewis lung cancer. Vectors were constructed by cloning FliC inserts into the pCDH-MSC-T2A Puro plasmid. The cells were transfected directly and indirectly (with pseudoviruses, produced by previously transfected packaging cells HEK 293T). The transfection efficiency was confirmed by RT-PCR. The cells thus prepared were implanted subcutaneously in mice. The control group received non-transfected LLC cells. Then, after 6 weeks, the mice were scarified. The animals were sectioned to isolate the tumor and lungs in which the presence or absence of metastases was assessed. In mice that received immunotherapeutic vaccines containing transfected LLC cells, less tumor mass was found or a complete lack of it, and the lifespan was noticeably prolonged. In addition, no metastases occurred in the group vaccinated with cells transfected with direct technique. It can be concluded that flagellin is effective as an adjuvant in the process of presenting tumor antigens to T cells Thus, in the light of recent studies and reports, it is likely that in this experiment effectively increased immunogenicity of the tumor. Activating anti-tumor cellular responses using flagellin is a promising target for lung cancer immunotherapy

The influence of Sm-153 therapy on bone marrow function

Aim of the study: Studies about possible risks connected with β-emitterradiotherapy concentrate mainly on potential myelotoxicity. Results of previously published analysis based on white blood cells (WBC) and platelet (PLT) counts – before and after radionuclide treatment – are quite varied. The aim of our study was to present the greatest possible impact of Samarium-153 on bone marrow function in clinical practice. Material and methods: The study included the blood test results of 175 patients with bone metastases treated with Sm-153 in the years 2012–2014. We compared levels of WBC, PLT, red blood cells (RBC), and haemoglobin (HGB) from two blood tests – one performed directly before the therapy and the other 2–6 weeks after isotope injection. Results and conclusions : The study showed decreased mean level of WBC in a control test performed after therapy in comparison to output results at about 27.1%. In our study 1.1% of patients developed the third-grade toxicity in CTCAE (Common Terminology Criteria for Adverse Events). Mean decrease of PLT was about 18%. Three patients (1.7% of all) result qualified as third-grade toxicity in a control test, one as fourth-grade. Analysis of RBC level showed 5.7% reduction of output values. The same calculation was seen for HGB – 5.1%. The greatest but acceptable decrease in haematological parameters was observed in WBC and PLT. Analysis of changes in WBC and PLT level showed them to be similar or smaller than was proven in previously published studies

Potencjał diagnostyczny śliny w screeningu chorób wirusowych = Diagnostic potential of saliva in the screening of viral diseases

Wędrowska Ewelina, Simińska Edyta, Wandtke Tomasz, Wędrowski Mateusz, Piskorska Elżbieta. Potencjał diagnostyczny śliny w screeningu chorób wirusowych = Diagnostic potential of saliva in the screening of viral diseases. Journal of Education, Health and Sport. 2016;6(1):147-156. eISSN 2391-8306. DOI http://dx.doi.org/10.5281/zenodo.45142 http://ojs.ukw.edu.pl/index.php/johs/article/view/45142 https://pbn.nauka.gov.pl/works/704038       The journal has had 7 points in Ministry of Science and Higher Education parametric evaluation. Part B item 755 (23.12.2015). 755 Journal of Education, Health and Sport eISSN 2391-8306 7 © The Author (s) 2016; This article is published with open access at Licensee Open Journal Systems of Kazimierz Wielki University in Bydgoszcz, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 15.12.2015. Revised 12.01.2016. Accepted: 25.01.2016.       Potencjał diagnostyczny śliny w screeningu chorób wirusowych Diagnostic potential of saliva in the screening of viral diseases   Ewelina Wędrowska1, Edyta Simińska2, Tomasz Wandtke1, Mateusz Wędrowski3, Elżbieta Piskorska4   1Zakład Genoterapii, Uniwersytet Mikołaja Kopernika w Toruniu, Collegium Medicum im. L. Rydygiera w Bydgoszczy 2Katedra i Zakład Toksykologii, Uniwersytet Mikołaja Kopernika w Toruniu, Collegium Medicum im. L. Rydygiera w Bydgoszczy 3Zakład Pozytonowej Tomografii Emisyjnej i Diagnostyki Molekularnej, Uniwersytet Mikołaja Kopernika w Toruniu, Collegium Medicum im. L. Rydygiera w Bydgoszczy 4Katedra Patobiochemii i Chemii Klinicznej, Uniwersytet Mikołaja Kopernika w Toruniu, Collegium Medicum im. L. Rydygiera w Bydgoszczy   1Department of Gene Therapy, Faculty of Medicine, Nicolaus Copernicus University Collegium Medicum, Bydgoszcz, Poland 2Department of Toxicology, Nicolaus Copernicus University Collegium Medicum, Bydgoszcz, Poland 3Department of Positron Emission Tomography and Molecular Diagnostics, Nicolaus Copernicus University Collegium Medicum, Bydgoszcz, Poland 4Department of Pathobiochemistry and Clinical Chemistry, Nicolaus Copernicus University Collegium Medicum, Bydgoszcz, Poland     Streszczenie   Wprowadzenie: W ostatnich latach potwierdzono, że większość substancji znajdujących się w surowicy krwi, obecnych jest również w ślinie. Wykazano także korelację pomiędzy poziomem substancji występujących w ślinie, ze zmianami zachodzącymi w surowicy krwi. Coraz bardziej podkreśla się liczne zalety śliny jako materiału do badań diagnostycznych, w porównaniu z innymi płynami ustrojowymi. W ostatnich dekadach wskaźniki zachorowalności i umieralności z powodu chorób wirusowych uległy znaczącej poprawie. Jednakże zbyt późna wykrywalność zakażeń wirusowych jest nadal poważnym problemem. Cel pracy: Celem pracy jest przedstawienie możliwości wykorzystania śliny w screeningowych badaniach chorób wirusowych. Skrócony opis stanu wiedzy: W pracy przytoczono rezultaty badań, do których włączono pacjentów z wirusem RSV, HIV, HAV, HBV, HCV. Potwierdzają one obecność markerów diagnostycznych poszczególnych zakażeń w ślinie, wskazując tym samym na możliwość wykorzystania śliny w diagnostyce chorób wirusowych. Podsumowanie: Wysoka czułość, dokładność, silna korelacja ze stopniem zaawansowania choroby, to bez wątpienia cechy, jakie wykazują badania wykonywane w surowicy krwi. Wymagają one jednak zaawansowanej aparatury, są kosztowne, dość inwazyjne, trudne w przechowywaniu. Ważne jest zatem, aby poszukiwać nowych metod, możliwych do wykorzystania w badaniach screeningowych. Badania takie powinny łączyć technologie szybkiego wykrywania z zastosowaniem łatwego do pobrania materiału. Coraz częściej podkreśla się potencjał śliny, jako materiału diagnostycznego do tego rodzaju badań.   Słowa kluczowe: ślina, wirusy, diagnostyka, infekcje.     Summary Introduction: Results in recent years have confirmed that most of substances contained in the blood serum also exist in the saliva. Significant correlations were found between salivary and serum levels. Saliva as a diagnostic test material in compare to other body fluids has many advantages and its importance is growing. During the last decades viral diseases morbidity and mortality rates improved. However late detection of viral infections is still the most Aim of the study: The aim of the study is to present the possibilities of using saliva in the  screening of viral diseases. Short description of state of knowledge: The study presents results of studies which included patients with RSV, HIV, HAV, HBV, HCV. These confirms the presence of the diagnostic markers in saliva, indicating the possibility of using saliva in the diagnosis of viral diseases. Summation: Analysis made in the blood serum display characteristics such as high sensitivity, precision and a strong correlation with stadium of disease. However, they require the advanced apparatus, are expensive, very invasive, difficult to store. It is important to looking for a new methods that can be used in screening test which should connect rapid detection technology with using easy-to-sample collection. Increasingly, emphasizes potential of saliva as a diagnostic material for this kind of research.   Key words: saliva, viruses, diagnostics, infection

Development and application of dedicated positron emission tomography (PET) systems in prostate cancer diagnostics

Introduction: One of the most important parameter in prostate cancer diagnostics is fast and accurate diagnosis. The important role plays finding cancer markers in blood but also advanced medical imaging systems technology, that allows for faster and more efficient objects projection. There are ongoing efforts to develop new multimodal imaging systems and find new PET radiopharmaceuticals. Aim of the study: The aim of this study is presentation of chosen information about dedicated PET systems in fusion with US and description of the most important PET radiopharmaceuticals for prostate cancer diagnosis that could confirm the meaning of new imaging technologies. Short description of state of knowledge: The most interesting recent scientific trends in prostate cancer medical imaging is presented in this work. Discussion is based on grant projects under realization. Potential clinical application problems were analyzed. The motivation of these projects was explained. Advantages and drawbacks of most recent radiopharmaceuticals for prostate cancer diagnosis was also discussed. Summation: New ideas in prostate cancer therapy are based on replacement the blind biopsy with guided biopsy in new meaning. Advanced medical imaging technology and application of new PET radiotracers in Poland (– the country with high ageing factor) may allow for cancer slowdown in early stage and minimize the treatment costs

Niewirusowy transfer genów do komórek skóry – wybrane metody = Non-viral gene transfer into skin cells – selected methods

Wędrowska Ewelina, Gawroński Maciej, Wandtke Tomasz, Goede Arkadiusz, Wędrowski Mateusz, Piskorska Elżbieta. Niewirusowy transfer genów do komórek skóry – wybrane metody = Non-viral gene transfer into skin cells – selected methods. Journal of Education, Health and Sport. 2016;6(1):157-170. eISSN 2391-8306. DOI http://dx.doi.org/10.5281/zenodo.45147 http://ojs.ukw.edu.pl/index.php/johs/article/view/45147 https://pbn.nauka.gov.pl/works/704486       The journal has had 7 points in Ministry of Science and Higher Education parametric evaluation. Part B item 755 (23.12.2015). 755 Journal of Education, Health and Sport eISSN 2391-8306 7 © The Author (s) 2016; This article is published with open access at Licensee Open Journal Systems of Kazimierz Wielki University in Bydgoszcz, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 15.12.2015. Revised 12.01.2016. Accepted: 25.01.2016.       Niewirusowy transfer genów do komórek skóry – wybrane metody Non-viral gene transfer into skin cells – selected methods     Ewelina Wędrowska1, Maciej Gawroński1, Tomasz Wandtke1, Arkadiusz Goede1, Mateusz Wędrowski2, Elżbieta Piskorska3   1Zakład Genoterapii, Uniwersytet Mikołaja Kopernika w Toruniu, Collegium Medicum im. L. Rydygiera w Bydgoszczy 2Zakład Pozytonowej Tomografii Emisyjnej i Diagnostyki Molekularnej, Uniwersytet Mikołaja Kopernika w Toruniu, Collegium Medicum im. L. Rydygiera w Bydgoszczy 3Katedra Patobiochemii i Chemii Klinicznej, Uniwersytet Mikołaja Kopernika w Toruniu, Collegium Medicum im. L. Rydygiera w Bydgoszczy   1Department of Gene Therapy, Faculty of Medicine,  Nicolaus Copernicus University Collegium Medicum, Bydgoszcz, Poland 2Department of Positron Emission Tomography and Molecular Diagnostics, Nicolaus Copernicus University Collegium Medicum, Bydgoszcz, Poland 3Department of Pathobiochemistry and Clinical Chemistry, Nicolaus Copernicus University Collegium Medicum, Bydgoszcz, Poland       Streszczenie   Wprowadzenie: Skóra jako największy i najłatwiej dostępny narząd stanowi atrakcyjny cel dla terapii genowej, która od wielu lat budzi ogromne nadzieje środowiska naukowego. Jednakże, próby terapii przeprowadzone z wykorzystaniem wektorów wirusowych wykazały szereg wad i ograniczeń m.in. obserwowano indukcję odpowiedzi immunologicznej, losową integrację transgenu z genomem gospodarza i/lub niską wydajność jego ekspresji. Dlatego, wciąż poszukuje się alternatywnych, skuteczniejszych i jednocześnie bezpieczniejszych metod transferu genów. Atrakcyjnej alternatywy upatruje się w metodach niewirusowych. Cel pracy: Przedstawienie wybranych metod niewirusowego transferu genów wykorzystywanych w terapii genowej chorób skóry. Skrócony opis stanu wiedzy: Terapia genowa chorób skóry obejmuje wykorzystanie wektorów plazmidowych jako nośnika genów terapeutycznych, a także metod ich dostarczania do komórek takich jak: elektroporacja, mikroiniekcja, sonikacja, wykorzystanie nośników lipidowych i polimerów kationowych. Podsumowanie: Niewirusowe metody transferu genów oferują pewne zalety włączając niską toksyczność, brak infekcyjności oraz łatwość i niskie koszty produkcji w porównaniu z technikami wirusowymi. Niewirusowe metody wydają się być obiecującym narzędziem terapii genowej chorób skóry w szczególności nowotworów tego narządu.   Słowa kluczowe: plazmid, transfer genów, skóra, wirus, terapia genowa.     Summary Introduction: Skin, the largest and most accessible organ of the human body is considered as an ideal gene therapy target. However, various types of viral vectors used in classical gene therapy have a number of disadvantages, such as possibility of immune response induction, random integration of inserts into the host genome or low expression efficiency. Therefore, there is an urgent need for alternative, non-viral methods of gene transfer. Aim of the study: To present methods for non-viral gene transfer used in gene therapy of skin diseases. Short description of knowledge state: Gene therapy for skin diseases include the usage of plasmid vectors as a carrier for therapeutic genes and different methods for their delivery into cells such as: electroporation, microinjection, sonication, lipid carriers and cationic polymers. Summary: Non-viral gene transfer methods offer some advantages including lower toxicity, non-infectious properties, ease of production and low costs as compared to viral techniques. Non-viral approaches are the promising tool in gene therapy of skin diseases, in particular in skin cancer ceases.   Key words: plasmids, gene transfer, skin, viruses, gene therapy

Automatic Production of [¹⁸F]F-DOPA Using the Raytest SynChrom R&D Module

[18F]F-DOPA is widely used in PET diagnostics. Diseases diagnosed with this tracer are schizophrenia, Parkinson’s disease, gliomas, neuroendocrine tumors, pheochromocytomas, and pancreatic adenocarcinoma. It should be noted that the [18F]F-DOPA tracer has been known for over 30 years. However, the methods of radiosynthesis applied in the past did not allow its clinical use due to low efficiency and purity. Currently, in the market, one encounters different types of radiosynthesis using the fluorine 18F isotope and variants of the same method. The synthesis and its modifications were carried out using a Raytest Synchrom R&D module. The synthesis consists of the following steps: (a) binding of the fluoride anion 18F− on an anion exchange column; (b) elution with TBAHCO3−; (c) nucleophilic fluorination to the ABX 1336 precursor; (d) purification of the intermediate product on the C18ec column; (e) Baeyer–Villiger oxidation; (f) hydrolysis; and (gfinal purification of the crude product on a semipreparative column. The nucleophilic synthesis of [18F]F-DOPA was successfully performed in 120 min, using the ABX 1336 precursor on the Raytest SynChrom R&D module, with a radiochemical yield (RCY) of 15%, radiochemical purity (RCP) ≥ 97%, and enantiomeric purity (ee) ≥ 96%