Krūties vėžys yra dažniausia Lietuvos ir viso pasaulio moterų onkologinė liga. Ši liga pasižyminti nevienoda eiga, todėl molekulinė vėžio analizė yra labai svarbi. Šiuo metu ligos eiga prognozuojama, remiantis riboto informatyvumo klinikinių žymenų sistema, o gydymui tik pavieniais atvejais skiriami atrankūs vaistai, nukreipti į ligą sukėlusį genetinį pakitimą. Naviko molekulinė analizė padeda aptikti pažaidas vėžio genuose ar jų raiškos pakitimus ir informuoja apie ligos išsivystymo priežastis, padeda prognozuoti ligos progresavimo tikimybę, atskleidžia taikinius naujos kartos gydymo priemonėms. Siekiant įvertinti krūties vėžio epigenetinių biožymenų efektyvumą mes tyrėme reguliacinių genų, dalyvaujančių ląstelės ciklo kontrolėje, signalų perdavime, apoptozėje ir DNR reparacijoje, promotoriaus sekų hipermetilinimą. Tyrimui buvo atrinktos 76 pirminės ankstyvos stadijos (pT1-2) krūties karcinomos. Metilinimo pakitimai promotoriaus sekoje buvo tiriami septyniuose naviką slopinančiuose genuose (p14, p16, RARβ, RASSF1A, DAPK, GSTP1 ir MGMT), taikant metilinimui jautrią PGR. „Tikro laiko“ PGR metodas buvo įdiegtas epigenetinių pakitimų tyrimams cirkuliuojančioje vėžio DNR, išskirtoje iš ligonių kraujo plazmos. Didžioji dalis ankstyvos stadijos krūties karcinomų (63/76) turėjo bent vieno tirto geno hipermetilinimą. Bendras epigenetinių žymenų informatyvumas – 83%. Dažniausiai hipermetilinimas krūties karcinomose nustatytas gene RASSF1A (56/76). Statistinė analizė parodė... [toliau žr. visą tekstą] / Breast cancer is the most prevalent malignancy of women in Lithuania and word-wide. The course of disease differ markedly among patients, therefore molecular characterisation of tumour is very important. However, current prognostic markers are mainly based on clinical parameters and do not enable a reliable selection of the patients with high risk of disease progression. Molecular characterisation of tumour through detection of changes in cancer-related genes can help to estimate the risk of cancer progression, detect the molecular targets for modern treatment strategies. In order to evaluate the suitability of epigenetic biomarkers for molecular characterisation of breast cancer we analysed promoter hypermethylation in a wide panel of regulatory genes involved in cell cycle control, signalling, apoptosis and DNA repair. 76 primary breast carcinomas of early stage (pT1-2) were selected for the study. Aberrant methylation in promoter regions of seven tumour suppressor genes (p16, p14, RARβ, RASSF1A, DAPK, GSTP1 and MGMT) was analysed by means of methylation-specific PCR. The “Real Time” PCR method was adapted for detection of epigenetic changes in circulating tumour DNA from plasma of cancer patients. Most of the early-stage breast tumours (63/76) exhibited hypermethylation in at least one gene involved in analysis. The overall sensitivity of the epigenetic biomarkers was 83%. Gene RASSF1A was the most frequently (56 of 76 cases) hypermethylated gene in breast tumours... [to full text]

A endometriose é uma doença caracterizada pelo crescimento de tecido endometrial ectópico histologicamente similar ao do endométrio eutópico. Dentre sua sintomatologia, encontra-se relatos de infertilidade a qual está relacionada à diminuição da receptividade endometrial e a resistência à progesterona, importante hormônio envolvido no estabelecimento desse processo e na manutenção da gestação. A etiologia da endometriose não é bem conhecida, mas estudos sugerem que essa desordem possa estar ligada a mecanismos epigenéticos de regulação da expressão genica, tais como a metilação do DNA. A receptividade endometrial à progesterona é mediada por receptores (PR-A e PR-B) a qual parece estar suprimida nessas pacientes, possivelmente por diminuição ou inatividade destes receptores, o que poderia ser um mecanismo envolvido na infertilidade associada à endometriose. Estudos de metilação no gene codificante das isoformas A e B do receptor de progesterona (PGR) mostram que o exon 1 da isoforma PR-B encontra-se hipermetilado e tem sua expressão silenciada em pacientes portadoras da doença. Entretanto ainda não foi avaliado o perfil de metilação deste gene em mulheres inférteis com a doença, o que poderia estar envolvido na infertilidade apresentada por essas pacientes. O objetivo desde estudo foi avaliar o padrão de metilação do gene PGR em endométrio eutópico de mulheres com infertilidade relacionada à endometriose e controles inférteis, durante a fase secretora. 10 Foram realizadas biópsias endometriais de 23 pacientes, divididas em dois grupos: 12 mulheres inférteis sem endometriose (Controle infértil) e 11 mulheres inférteis com endometriose (Endometriose). Todos os endométrios coletados foram confirmados na fase secretora do ciclo através de análise histológica clássica segundo os critérios de Noyes. O DNA genômico foi extraído com o QIAamp DNA Mini Kit e modificado utilizando o EpiTec Bissulfite Kit. O padrão de metilação das isoformas PR-A e PR-B foi avaliado pela técnica HRM (High Resolution Melting) A análise de metilação da isoforma PR-A mostrou que a região analisada encontra-se hipometilada, em que a porcentagem de metilaçao encontrada foi 0% em ambos os grupos com e sem endometriose. No entanto, no grupo com endometriose, a isoforma PR-B mostrou-se parcialmente metilada na maioria das pacientes com porcentagem de metilaçao de 50% (n=8), somente uma paciente apresentou 80% de metilacão na isoforma B. Para o grupo sem endometriose a isoforma PR-B mostrou-se hipometilada (0%). A hipometilação da isoforma PR-A sugere que este gene encontrar-se na sua forma ativa, sem alterações em sua expressão. No entanto, a hipermetilação da isoforma PR-B nas mulheres com endometriose pode estar relacionada com o silenciamento gênico ou redução da sua expressão, sugerindo que a baixa resposta à progesterona nas mulheres com endometriose pode estar diretamente ligada à redução do número de seus receptores nessas pacientes / Endometriosis is a disease characterized by ectopic growth of endometrial tissue histologically similar to the eutopic endometrium. Among this symptoms, is infertility accounts which is related to decreased endometrial receptivity and resistance to progesterone, important hormone involved in the establishment of this process and the maintenance of pregnancy. The etiology of endometriosis is not well known, but studies suggest that this disorder can be linked to epigenetic mechanisms of regulation of gene expression such as DNA methylation. The endometrial responsiveness to progesterone is mediated by receptors (PR-A and PR-B) which seem to be suppressed in these patients, possibly reducing or inactivity of these receptors, which could be a mechanism involved in the infertility associated with endometriosis. Studies in methylation of the gene encoding the A and B isoforms of progesterone receptor (PGR) showed that exon 1 of the PR-B isoform is hypermethylated and silenced have its expression in patients with the disease. However it has not been rated the methylation profile of this gene in infertile women with the disease, which could be involved in the infertility presented by these patients. The goal from study was to evaluate the methylation pattern of the PGR gene in eutopic endometrium of women with infertility related to endometriosis and infertile controls during the secretory phase. Endometrial biopsies were performed on 12 23 patients, divided into two groups: 12 infertile women without endometriosis (infertile Control) and 11 infertile women with endometriosis (Endometriosis). All endometrium collected were confirmed in the secretory phase of the cycle by classical histological analysis according to the criteria of Noyes. Genomic DNA was extracted with the QIAamp DNA Mini Kit and using the modified EpiTec Bissulfite Kit. The methylation pattern of PR-A and PR-B isoforms was assessed by HRM technique (Melting High Resolution) Methylation analysis of PR isoform The analyzed showed that the region is hipometilada, wherein the percentage of methylation was found 0% in both the groups with and without endometriosis. However, in the group with endometriosis, the PR-B isoform showed partially methylated in most patients with methylation percentage of 50% (n = 8), only one patient showed methylation in 80% of isoform B. In the group without endometriosis the PR-B isoform was shown hipometilada (0%). The hypomethylation of PR-A isoform suggests that this gene be in its active form, without change in his expression. However, hypermethylation of the PR-B isoform in women with endometriosis may be associated with gene silencing or reducing the expression, suggesting that the low response to progesterone in women with endometriosis can be directly linked to the reduction of its receptors in these patients

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Direito, Programa de Pós-Graduação em Direito, 2015. / Submitted by Ana Cristina Barbosa da Silva (annabds@hotmail.com) on 2015-04-27T13:57:58Z No. of bitstreams: 1 2015_KeltondeOliveiraGomes_Parcial.pdf: 891096 bytes, checksum: 6bca05a01d0ffbb247afc65078e49fc1 (MD5) / Approved for entry into archive by Ruthléa Nascimento(ruthleanascimento@bce.unb.br) on 2015-04-28T18:10:30Z (GMT) No. of bitstreams: 1 2015_KeltondeOliveiraGomes_Parcial.pdf: 891096 bytes, checksum: 6bca05a01d0ffbb247afc65078e49fc1 (MD5) / Made available in DSpace on 2015-04-28T18:10:30Z (GMT). No. of bitstreams: 1 2015_KeltondeOliveiraGomes_Parcial.pdf: 891096 bytes, checksum: 6bca05a01d0ffbb247afc65078e49fc1 (MD5) / A presente dissertação apresenta um estudo empírico em torno da atuação da Procuradoria Geral da República (PGR) no âmbito do controle concentrado de constitucionalidade no Brasil, focando Ações Diretas de Inconstitucionalidade (ADI) ajuizadas pela PGR, entre 1988 e 2012, e julgadas procedentes pelo Supremo Tribunal Federal (STF), totalizando mais de 900 decisões analisadas. O trabalho parte de uma dupla contextualização histórica da pesquisa, sobre a trajetória do Ministério Público no Brasil e sobre o papel desempenhado pela PGR na história do controle concentrado de constitucionalidade no País, ressaltando o caráter historicamente contingente da vinculação do Parquet à tarefa de defesa da sociedade. Com apoio desses subsídios históricos, realiza-se uma análise quantitativa e qualitativa das ADI selecionadas, apresentando um levantamento do número de ADI que foram anualmente ajuizadas pela PGR, variações por mandato dos Procuradores, tipologia dos atos impugnados e estratégias de fundamentação recorrentes nas ações. Cruzando esses dados com os subsídios históricos anteriormente mencionados, a pesquisa conclui que a atuação da PGR no controle concentrado teve pouco impacto em casos de violação a direitos e garantias fundamentais e grande êxito em demandas envolvendo restrição da atividade política dos Estados e em questões corporativas sobre carreiras do serviço público com as da magistratura e do próprio Ministério Público. Cruzando esses achados com os subsídios históricos anteriormente colhidos, conclui-se que a atuação da PGR nessa esfera após 1988 representa certa continuidade da atuação dessa instituição antes do processo de redemocratização, sugerindo que o sistema de controle concentrado de constitucionalidade reproduz as assimetrias do pacto federativo brasileiro de forma relativamente independente do regime político no qual ele funciona. / This work presents an empirical analysis on the content of the Direct Actions of Unconstitutionality (ADI) filed by the Brazilian Federal Prosecutor’s Office’s (PGR) and adjudicated by the Brazilian Supreme Court (STF) between 1988 and 2012, totaling more than 900 decisions analyzed. These lawsuits, modeled after the constitutional complaints from European judicial review systems and with its use restricted to a very selective group of legitimated actors, are usually regarded as a more efficient and secure way to protect fundamental rights against unconstitutional federal acts. The work also contains a historical contextualization of the research, investigating the history of the Prosecution Office in Brazil. A quantitative and qualitative analysis of the selected ADI is then presented, with a survey of the number of ADI filed annually by the PGR, variations related to the mandates of the Attorneys, typology of the contested acts and recurrent reasoning strategies in these actions. The research concludes that the performance of the PGR using ADI had little impact in cases of violation of fundamental rights and great success in lawsuits involving restriction of political activity of the States and corporate issues on public service, especially concerning the Judiciary and the Prosecution Office itself. Crossing these findings with the historical elements previously presented, we conclude also that the performance of the PGR in this lawsuits after 1988 Constitution is a continuity of its behavior before the enactment of that Constitution, suggesting that this kind of judicial review in Brazil reproduces the asymmetries of the Brazilian federal system regardless of the characteristics of the political regime in which it work.

No description available.

Clematis spp. L. is a twining vine covered in showy blooms. Typical growth of hybrids is from the main leader, producing a thin, unbranched plant with one cyme. Apical dominance is released by cutting back the vine during production. Cutting back, or pinching, of a plant is labor intensive and compromises bloom for vegetative growth at time of sales. The purpose of this project was to eliminate manual pinching by treating young plants with chemical plant growth regulators (PGRs) that enhance branching without removal of the apical meristem. The first project evaluated the use of Atrimmec (dikegulac sodium), Fascination (BA+GA4+7), Florel (ethephon), and Dropp 50 (thidiazuron) on Clematis cultivars Ernest Markham, and Hagley Hybrid, and Clematis viticella 'Polish Spirit.' Plants treated with 800 mg·L-1 Atrimmec, or 800 or 1200 mg·L-1 Fascination experienced an increase in branch numbers. The second experiment manipulated the ratio of the components of Fascination, 6-BA and GA4+7, to reduce phytotoxicity experienced in the first experiment. The optimal ratio to enhance branching was 1:1, which is the stock solution for Fascination. All ratios produced phytotoxic symptoms. A third experiment tested lower rates of thidiazuron and added CPPU (forchlorfenuron) to the list of PGRs to test. The last experiment took the most effective PGR treatments, Atrimmec at 800 mg·L-1, and Fascination at 800 or 1200 mg·L-1, and compared them to the current production practices of pinching. Large flowering cultivars of clematis were used, including 'Comotesse de Bouchard,' 'Ernest Markham,' and 'Hagley Hybrid.' Atrimmec increased branch numbers and suppressed leader lengths without a mechanical pinch. Results from Fascination varied by cultivar. / Master of Science

Dans les thylakoïdes des végétaux supérieurs, la réaction photochimique de la photosynthèse implique un transfert d’électrons linéaire ayant lieu entre les photosystèmes II et I et aboutit à la production de pouvoir réducteur (NADPH) et d’ATP. En parallèle à ce transfert linéaire, le transfert cyclique d’électrons autour du PS I court-circuite le PS II et fait intervenir distinctement les complexes PGR (PGR5/PGRL1) et NADPH déshydrogénase (NDH). Ces deux voies alternatives permettent la production d’ATP sans accumulation de NADPH dans le stroma et particulièrement en conditions de stress environnemental. Ces voies sont essentielles pour la physiologie de la plante mais leur régulation demeure mal connue. Une hypothèse a été émise quant à leur régulation redox en étudiant plusieurs thiorédoxines chloroplastiques. Des expériences menées in vivo et in vitro sur des mutants d’Arabidopsis, de tabac et de tomate ont mis en évidence le rôle très spécifique de la thiorédoxine m4 dans la régulation négative des voies dépendantes de PGR et NDH. Une stratégie originale a été mise en place pour capturer les cibles putatives de cette thiorédoxine en mutant le site actif de la protéine chez Arabidopsis. Les résultats obtenus ont montré l’existence de plus de 160 cibles potentielles et certaines d’entre elles pourraient être impliquées dans la régulation des voies dépendantes des complexes PGR et NDH. / In plants thylakoids, the photochemical reaction of photosynthesis implies a linear electron flow between photosystems II and I which produces reducing power (NADPH) and ATP. In addition of linear electron flow, the cyclic electron flow around PS I bypasses PS II and involves distinctly the PGR (PGR5/PGRL1) and the NADPH dehydrogenase (NDH) complexes. These two alternative pathways lead to the production of ATP without accumulation of NADPH in the stroma especially under environmental stress conditions. Both pathways are essential for the plant physiology but their regulation remains unclear. An assumption concerning a redox regulation has been considered by surveying several chloroplastic thioredoxins. In vivo and in vitro studies of Arabidopsis, tobacco and tomato mutants revealed that the m4 thioredoxin plays a very specific role in the down-regulation of the PGR and NDH dependant pathways. An original strategy to capture putative targets of this thioredoxin has been established by mutating the active site of the protein in Arabidopsis. The results showed the existence of more than 160 potential targets and some of them could be involved in the regulation of PGR and NDH dependant pathways.

Gimdos kaklelio vėžys pasaulyje yra antroji, o Lietuvoje ketvirtoji pagal dažnumą moterų onkologinė liga. Pagrindinė gimdos kaklelio vėžio priežastis yra žmogaus papilomos viruso (ŽPV) infekcija gimdos kaklelyje. Vakarų šalyse ikivėžiniams pokyčiams ar vėžinėms ląstelės nustatyti yra naudojamas citologija paremtas Papanicolaou testas (Pap) ir ŽPV DNR testas. Šiuo metu naudojami ŽPV diagnostiniai testai remiasi molekulinės biologijos tyrimo metodais. Tyrimui buvo pasirinktas polimerazės grandininės reakcijos (PGR) metodas, o ne plačiai naudojamas JAV bei Europos kokybės sertifikatus turintis Hybrid Capture 2 (HC2) metodas. PGR yra ypač jautrus, greitas, komerciškai prieinamas metodas, kuriam reikalingas nedidelis tiriamosios medžiagos kiekis, ir kuriuo galima tiksliai identifikuoti ŽPV tipus. HC2 metodas nenustato ŽPV genotipų, galimi klaidingai neigiami rezultatai, nes nėra vidinės kontrolės, o dėl kryžminių reakcijų galimi klaidingai teigiami rezultatai. Naujausi literatūros šaltiniai teigia, kad PGR turi daug perspektyvų ateityje ir manoma, kad ŽPV nustatymo standartas bus 14 DR-ŽPV tyrimas sujungtas su ŽPV-16 ir ŽPV-18 tipų nustatymu. Todėl siekiant ŽPV tyrimams pasirinkti optimaliausią iš Lietuvoje siūlomų komercinių rinkinių Valstybiniame patologijos centre buvo atliekamas šis tyrimas. Įvertinus tyrimų rezultatus, tolesniems tyrimams pasirinktas „Seeplex® HPV4A ACE Screening“ ŽPV nustatymo rinkinys. / Cervical cancer is the second oncological disease in women in the world, and the fourth one in Lithuania. The main cause of the cervical cancer is Human papillomavirus (HPV) infection in the cervix uteri. In order to detect precancerous changes or cancer cells, a Papanicolaou (Pap) test based on cytology and HPV DNA test are used in the Western world. Currently used HPV diagnostic tests are based on molecular biology testing methods. The polymerase chain reaction (PCR) method, not the extensively used Hybrid Capture 2 (HC2) method awarded the certificate of quality by both the USA and Europe, was chosen. PCR is a particularly sensitive, fast, and commercially available method that needs a small quantity of research material and can identify the types of HPV. HC2 method does not detect HPV genotypes, false-negative results are possible because there is no internal control, and due to cross reactions, false-positive results are possible. The newest written sources state that PCR will have much potential in the future and it is thought that HPV detection standard will become 14 HR-HPV test combined with the detection of the types of HPV-16 and HPV-18. Therefore, in order to chose the optimum one for HPV tests from the commercial kits offered in Lithuania, this investigation was carried out in the National Centre of Pathology. Four different HPV diagnostic kits were used for the research. Having evaluated the research results, „Seeplex® HPV4A ACE Screening“ HPV diagnostic kit was... [to full text]

Kiekvieno vaisto sukeltas tiek farmakologinis (veiksmingumo), tiek toksikologinis poveikis kiekvienam pacientui yra skirtingas, todėl gana dažnai vaistų skyrimas ir vartojimas tampa labai komplikuotas. Prancūzijoje 3.2% hospitalizacijos atvejų yra sąlygoti vaistų. Tai sudaro 320 milijonų eurų sumą per metus. Genetiniai faktoriai, sąlygojantys vaistų farmakokinetiką ir farmakodinamiką, dalinai paaiškina skirtingą vaistų poveikį žmogui. Tyrimo tikslas: 1. Patikrinti galimybę gausinti serume ir plazmoje esančią DNR nauju būdu ir atlikti genetinius tyrimus. 2. Patvirtinti realaus laiko polimerazės grandininės reakcijos (PGR) metodiką aromatazės genui, panaudojus ląsteles, kurių šio geno raiška yra pakankama. Rezultatai: Patikrinta galimybė gausinti serume ir plazmoje esančią DNR, sekvenavus gautą DNR ir sekas palyginus su esančiomis duomenų bazėje : rezultatai teigiami. Nustatyta, kad, ekstrahuojant druskiniu metodu, gaunama daugiau DNR, bet ji blogesnės kokybės, nei naudojant QIAGEN kolonėles. Patvirtinta realaus laiko PGR metodika aromatazės genui, naudojant krūties vėžio ląsteles. / There is much variability in the manner individuals respond to drugs, such that the management of some drugs is problematic. In France, the incidence of hospital admissions related to adverse drug reactions is estimated to be 3.2 %, at an annual cost of over 300 millions euros. Genetic factors affecting the pharmacokinetics and pharmacodynamics of drugs partly explain interindividual variability in drug response. Aim of experiment: 1. verify, if it is possible to amplify serum and plasma DNA using new method, and make a genetic research. 2. verify real time polymerase chain reaction to aromatase gene. Find cell line, whish have a sufficient expression of this gene. Results : We verified the possibility to amplify serum and plasma DNA using new method. We made the sequencing of DNA extracts and we compared results in data base : results are positives. It`s important, that QIAGEN extracts are cleanner than salt extracts, but there are few of DNA. We confirmed real time PCR method to aromatase gene, using breast cancer cells.

Due to climate change, marked developments are tracked in various links of pathogen-host distribution. Markedly increased number of ectoparasites is observed in Central and Eastern Europe. Ticks are blood feeding wide group of arthropods of utmost medical, epidemiological and veterinary significance throughout the world. The ecology of ticks, the outcome of their interactions with their natural environment, is fundamental to the spatial and temporal variation in the risk of infection by tick-borne pathogens. The aim of the study was to investigate: the distribution of Ixodes ricinus ticks and their transmitted parasites prevalence in different biotopes in Lithuania; and to find out factors influencing the activity dynamics of ticks and prevalence of endoparasites. The scientific researches on ectoparasite-endoparasite system (I. ricinus-B.burgdorferi s.l., B.divergens and Ehrlichia sp.) in different biotopes in Lithuania have been carried out for the first time. Also, it was the first investigation of prevalence of ticks (according to development phases and different sexes) in different biotopes and ticks infectious by microparasites. Influence of the main factors (air temperature, precipitations) on seasonal and daily activity changes of ticks have been described. Experiments on ectoparasites and endoparasites have been employed by the procedures on molecular level, including PCR method. As the result of the study the specificity of ticks’ species has been identified... [to full text] / Vykstant klimato atšilimui yra stebimi ryškūs pokyčiai įvairiose parazitas-šeimininkas sistemos grandyse. Pastebimas ektoparazitų skaitlingumo padidėjimas Vidurio ir Rytų Europoje. Erkės yra įvairių zoonozių ir antropozių, kurias sukelia įvairūs virusai, bakterijos ir protistai, pernešėjai. Erkių platinamų ligų sukėlėjų patogeninių bakterijų (Borrelia sp., Ehrlichia sp. ir kt.) ir protistų (Babesia sp. ir kt.) tyrimus turi apimti visus ekologinius lygmenis: patogeninio organizmo, vektoriaus ir rezervuarinio šeimininko. Šio darbo tikslas – nustatyti Ixodes ricinus L. erkių ir jų pernešamų endoparazitų paplitimą skirtinguose biotopuose Lietuvoje bei išaiškinti veiksnius, turinčius įtakos erkių aktyvumui ir endoparazitų paplitimui. Lietuvoje pirmą kartą buvo atlikti ektoparazito - endoparazito sistemos (I. ricinus–B.burgdorferi s.l., B.divergens ir Ehrlichia sp.) tyrimai skirtinguose Lietuvos biotopuose. Buvo įvertintas bendras erkių (pagal vystymosi stadiją ir lytį) paplitimas skirtinguose biotopuose, jų užsikrėtimas mikroparazitais ir išanalizuoti veiksniai, lemiantys erkių aktyvumo sezoninius ir paros kitimus, gausumą ir užsikrėtimą. Ektoparazitų ir endoparazitų tyrimams buvo įdiegtas polimerazės grandininės reakcijos metodas ir identifikuotas erkių rūšinis specifiškumas, nustatyti mikroparazitai bei jų genotipai. Buvo nustatytas mikroparazitų paplitimas skirtinguose Lietuvos biotopuose ir ištirti veiksniai, galintys sąlygoti šį paplitimą. Pirmą kartą Lietuvoje buvo... [toliau žr. visą tekstą]

Paukščių maliariniai parazitai sukelia pavojingas naminių ir laukinių paukščių ligas. Šie parazitai dažnai sutinkami įvairiose paukščių grupėse ir yra plačiai paplitę Europoje, tame tarpe ir Baltijos regione. Dabartinių maliarinių parazitų studijų ypatumas yra tas, kad informacija apie šių parazitų ekologiją, paplitimą, įvairovę ir kitus biologijos aspektus yra surenkama iš laisvai gyvenančių paukščių. Vertinant maliarinių parazitų įtaką paukščių fizinei būklei, elgsenai, lytinei atrankai ir parazitų-šeimininkų ko-evoliucijai yra būtina surinkti informaciją apie maliarinių parazitų specifiškumą, virulentiškumą ir parazitemijos vystymąsi skirtinguose paukščiuose. Tokie duomenys gali būti gaunami atliekant eksperimentinius tyrimus. Deja, tokių studijų kol kas yra nedaug. Disertacijos tikslas – lauko ir eksperimentinių tyrimų pagalba surinkti naują medžiagą apie paukščių maliarinių parazitų biologiją bei susieti PGR paremtais metodais gautą informaciją su tradicinės parazitologijos duomenimis. Šių tyrimų metu įrodyta, kad PGR paremtų ir tradicinės parazitologijos (mikroskopijos) metodų, naudojamų tiriant paukščių maliarinius parazitus ir kitas hemosporidijas, tikslumas nesiskiria. Abu metodai turi savų privalumų ir trūkumų, todėl rekomenduojame naudoti mikroskopijos ir PGR paremtų metodų kombinaciją. Taip pat išvystytas Plasmodium relictum (linija SGS1) ir P. circumflexum (TURDUS1) molekulinis identifikavimas, bei sukurti nauji paukščių maliarinių ir susijusių kraujo parazitų... [toliau žr. visą tekstą] / Avian malaria parasites are responsible for severe diseases in some domestic and wild birds. These parasites are cosmopolitan in distribution; they are widespread in Europe, including the Baltic region. A peculiarity of current studies of avian Plasmodium species is that information about ecology, distribution, prevalence and other aspects of their biology has been accumulated using free-living birds. To elucidate the significance of malaria infections and their impact on host fitness, behaviour, sexual selection and parasite-host co-evolution, experimental information about Plasmodium spp. virulence, specificity and dynamics of parasitemia in different avian hosts is crucial. Unfortunately such studies remain uncommon. The objective of this study was to obtain new field and laboratory experimental data about the biology of avian malaria parasites and to link PCR-based information with data from traditional parasitology. It was demonstrated that prevalence of avian malaria and other haemosporidian parasites is estimated equally well by microscopy and currently used nested PCR-based methods. Both methods have advantages and disadvantages. So we encourage using both these tools in parallel during studies of haemosporidians. Lineages for molecular identification of P. relictum (lineage SGS1) and P. circumflexum (TURDUS1) were determined, also new methods of single cell dissection, DNA extraction and PCR-based analysis of avian malaria and closely related blood parasites were... [to full text]