Přichycení leishmanií ve střevě permisivních přenašečů a role střevních proteinů flebotomů v interakci parazit-přenašeč

of PhD thesis "Leishmania attachment in permissive vectors and the role of sand fly midgut proteins in parasite-vector interaction" Anna Dostalova, MSc. The thesis focuses on the development of protozoan parasites of the genus Leishmania in their insect vectors, sand flies. It sums up results of three projects I was involved in during my PhD studies. Main emphasis was put on permissive sand fly species that support development of various species of Leishmania. Using a novel method of binding of fluorescently labeled leishmania promastigotes to the midguts in vitro, we studied the stage- and species-specificity of the binding. We demonstrated that Leishmania midgut binding is strictly stage-dependent, is a property of forms found in the middle phase of development (long and short nectomonad forms), but is absent in early forms occurring in within the blood meal, procyclics, and in final stages, metacyclics. Comparing the binding of several leishmania species, we showed the natural parasite is not necessarily the species that can always bind in vitro most efficiently to the midgut of its vector. In some cases, we even observed significant binding of Leishmania species that do not survive in the midgut of the particular sand fly species in vivo. We conclude that the specificity of in vitro binding...dizertační práce s názvem "Přichycení leishmanií ve střevě permisivních přenašečů a role střevních proteinů flebotomů v interakci parazit-přenašeč" Mgr. Anna Dostálová, PřfUK, 2.5.2011 Práce se zabývá vývojem prvoků rodu Leishmania v jejich hmyzích přenašečích, flebotomech. Shrnuje výsledky tří projektů, na kterých jsem se během svého doktorského studia podílela. Zaměřila jsem se především na permisivní druhy flebotomů, tedy takové, které umožňují vývoj mnoha druhů lieshmanií. S využitím nové metody vazby fluorescenčně značených leishmanií na střevo flebotomů in vitro jsme sledovali vazbu různých morfologických stádií a druhů leishmanií. Nutnost přichytit se ke střevnímu epitelu je považována za podmínku úspěšného vývoje leishmanií v jejich přenašečích a důležitý faktor určující specifitu vztahu parazit-přenašeč. Zjistili jsme, že na střeva se vážou takřka výhradně tzv. dlouhé nektomonády a leptomonády, tedy stádia nacházená ve střední fázi infekce. Naopak procyklická stádia, tedy formy nacházené v přenašeči krátce po infekci, nebyla schopná se vázat. Podobné zjištení platí i pro metacyklické promastigoty vyskytující se v pozdní fázi. Srovnání vazby několika druhů leishmanií ukázalo, že ne vždy se parazit přenášený daným vektorem váže nejlépe in vitro. Překvapivě jsme pozorovali také vazbu...Department of ParasitologyKatedra parazitologieFaculty of SciencePřírodovědecká fakult

Characterization of a midgut mucin-like glycoconjugate of Lutzomyia longipalpis with a potential role in Leishmania attachment

BACKGROUND: Leishmania parasites are transmitted by phlebotomine sand flies and a crucial step in their life-cycle is the binding to the sand fly midgut. Laboratory studies on sand fly competence to Leishmania parasites suggest that the sand flies fall into two groups: several species are termed "specific/restricted" vectors that support the development of one Leishmania species only, while the others belong to so-called "permissive" vectors susceptible to a wide range of Leishmania species. In a previous study we revealed a correlation between specificity vs permissivity of the vector and glycosylation of its midgut proteins. Lutzomyia longipalpis and other four permissive species tested possessed O-linked glycoproteins whereas none were detected in three specific vectors examined. RESULTS: We used a combination of biochemical, molecular and parasitological approaches to characterize biochemical and biological properties of O-linked glycoprotein of Lu. longipalpis. Lectin blotting and mass spectrometry revealed that this molecule with an apparent molecular weight about 45-50 kDa corresponds to a putative 19 kDa protein with unknown function detected in a midgut cDNA library of Lu. longipalpis. We produced a recombinant glycoprotein rLuloG with molecular weight around 45 kDa. Anti-rLuloG antibodies localize the native glycoprotein on epithelial midgut surface of Lu. longipalpis. Although we could not prove involvement of LuloG in Leishmania attachment by blocking the native protein with anti-rLuloG during sand fly infections, we demonstrated strong binding of rLuloG to whole surface of Leishmania promastigotes. CONCLUSIONS: We characterized a novel O-glycoprotein from sand fly Lutzomyia longipalpis. It has mucin-like properties and is localized on the luminal side of the midgut epithelium. Recombinant form of the protein binds to Leishmania parasites in vitro. We propose a role of this molecule in Leishmania attachment to sand fly midgut

The midgut transcriptome of Phlebotomus (Larroussius) perniciosus, a vector of Leishmania infantum: comparison of sugar fed and blood fed sand flies

<p>Abstract</p> <p>Background</p> <p>Parasite-vector interactions are fundamental in the transmission of vector-borne diseases such as leishmaniasis. <it>Leishmania </it>development in the vector sand fly is confined to the digestive tract, where sand fly midgut molecules interact with the parasites. In this work we sequenced and analyzed two midgut-specific cDNA libraries from sugar fed and blood fed female <it>Phlebotomus perniciosus </it>and compared the transcript expression profiles.</p> <p>Results</p> <p>A total of 4111 high quality sequences were obtained from the two libraries and assembled into 370 contigs and 1085 singletons. Molecules with putative roles in blood meal digestion, peritrophic matrix formation, immunity and response to oxidative stress were identified, including proteins that were not previously reported in sand flies. These molecules were evaluated relative to other published sand fly transcripts. Comparative analysis of the two libraries revealed transcripts differentially expressed in response to blood feeding. Molecules up regulated by blood feeding include a putative peritrophin (<it>PperPer1</it>), two chymotrypsin-like proteins (<it>PperChym1 </it>and <it>PperChym2</it>), a putative trypsin (<it>PperTryp3</it>) and four putative microvillar proteins (<it>PperMVP1</it>, <it>2</it>, <it>4 </it>and <it>5</it>). Additionally, several transcripts were more abundant in the sugar fed midgut, such as two putative trypsins (<it>PperTryp1 </it>and <it>PperTryp2</it>), a chymotrypsin (<it>PperChym3</it>) and a microvillar protein (<it>PperMVP3</it>). We performed a detailed temporal expression profile analysis of the putative trypsin transcripts using qPCR and confirmed the expression of blood-induced and blood-repressed trypsins. Trypsin expression was measured in <it>Leishmania infantum</it>-infected and uninfected sand flies, which identified the <it>L. infantum</it>-induced down regulation of <it>PperTryp3 </it>at 24 hours post-blood meal.</p> <p>Conclusion</p> <p>This midgut tissue-specific transcriptome provides insight into the molecules expressed in the midgut of <it>P. perniciosus</it>, an important vector of visceral leishmaniasis in the Old World. Through the comparative analysis of the libraries we identified molecules differentially expressed during blood meal digestion. Additionally, this study provides a detailed comparison to transcripts of other sand flies. Moreover, our analysis of putative trypsins demonstrated that <it>L. infantum </it>infection can reduce the transcript abundance of trypsin <it>PperTryp3 </it>in the midgut of <it>P. perniciosus</it>.</p

Methods of Technical Reserves Alocation of Life and Non-Life Insurance

Diplomová práce se zabývá metodami alokace technických rezerv u životního a neživotního pojištění. Konkrétně rezervou na výplatu pojistných plnění na škody, které již nastaly, ale pojišťovně nebyly dosud nahlášeny. Úvodní část pojednává o úloze pojišťovnictví, seznámení se základními pojmy. Práce podrobně popisuje tvorbu a užití rezervy. V praktické části jsou potom všechny metody přiblíženy na datech s cílem nastínit představu o tom, jak celý proces tvorby rezerv probíhá.This thesis deals with methods of allocation of technical reserves for life and non-life insurance. Specifically, for the payment of claims for damages that have already occurred but not yet reported to the insurance company. The first section describes the role of insurance, introduction to basic concepts. The work describes the creation and use of reserves. In the practical part are all the methods described on the data in order to outline an idea of how the whole process of creating reserves in progress.Ústav matematiky a kvantitativních metodStudentka předvedla velmi krátkou obhajobu, komise není spokojena. Obhajoba byla zaměřena pouze na kapitolu 11 DP, nic z dalšího v DP nebylo zmíněno. Odpovědi na otázky vedoucího i oponenta velmi krátké a mlhavé. Pacáková: Vysvětlete podrobně řetězovou-žebříkovou metodu. Odpovědi studentky nejsou uspokojivé, plete si počty a výši pojistných plnění. V obhajobě chybí levá horní část vývojového trojúhelníku, jsou pouze data pod uhlopříčkou. Studentka neumí vysvětlit koeficienty vývoje lambda. Máchová: Vliv. poj. tech. rezerv na konkurenceschopnost pojišťovny je v zad. listu, ale není v práci. Jonášová: Rozporuje fiktivní data dosazená do metod. Studentka uvádí, že reálná data z pojišťovny nedostala. Pacáková uvádí problematičnost dostupnosti dat obecně

Leishmania attachment in permissive vectors and the role of sand fly midgut proteins in parasite-vector interaction

of PhD thesis "Leishmania attachment in permissive vectors and the role of sand fly midgut proteins in parasite-vector interaction" Anna Dostalova, MSc. The thesis focuses on the development of protozoan parasites of the genus Leishmania in their insect vectors, sand flies. It sums up results of three projects I was involved in during my PhD studies. Main emphasis was put on permissive sand fly species that support development of various species of Leishmania. Using a novel method of binding of fluorescently labeled leishmania promastigotes to the midguts in vitro, we studied the stage- and species-specificity of the binding. We demonstrated that Leishmania midgut binding is strictly stage-dependent, is a property of forms found in the middle phase of development (long and short nectomonad forms), but is absent in early forms occurring in within the blood meal, procyclics, and in final stages, metacyclics. Comparing the binding of several leishmania species, we showed the natural parasite is not necessarily the species that can always bind in vitro most efficiently to the midgut of its vector. In some cases, we even observed significant binding of Leishmania species that do not survive in the midgut of the particular sand fly species in vivo. We conclude that the specificity of in vitro binding..

Leishmania attachment in permissive vectors and the role of sand fly midgut proteins in parasite-vector interaction

of PhD. thesis named "Leishmania attachment in permissive vectors and the role of sand fly midgut proteins in parasite-vector interaction", Anna Dostálová, 2011 This thesis focuses on the development of protozoan parasites of the genus Leishmania in their insect vectors, sand flies. It sums up results of three projects I was involved in during my PhD studies. Main emphasis was put on permissive sand fly species that support development of various species of Leishmania. Using a novel method of binding of fluorescently labeled leishmania promastigotes to the midguts in vitro, we studied the stage- and species-specificity of the binding. We demonstrated that Leishmania midgut binding is strictly stage-dependent, is a property of forms found in the middle phase of development (long and short nectomonad forms), but is absent in early forms occurring in within the blood meal, procyclics, and in final stages, metacyclics. Comparing the binding of several leishmania species, we showed the natural parasite is not necessarily the species that can always bind in vitro most efficiently to the midgut of its vector. In some cases, we even observed significant binding of Leishmania species that do not survive in the midgut of the particular sand fly species in vivo. We conclude that the specificity of in vitro..

Acute effect of static and dynamic stretching on vertical jump

Title Acute effect of static and dynamic stretching on vertical jump. Objectives The aim of this thesis is to assess whether it is better to warm up by static or dynamic stretching in order to improve performance in the vertical jump. Methods The first part of this thesis is theoretical and is the basis for measurement, which is described in the second part. The tested set consists of ten women volleyball players. Each player underwent three vertical jump measurements on power plates Kistler, which recorded flight time of the player over the plates. All measurements were evaluated afterwards. Five attempts were recorded in each measurement. Measurements were carried out without stretching, after dynamic stretching and after static stretching. The thesis includes a survey that monitors the perception of the players` performance. Results Results of the total tested set measurement show that inclusion of dynamic stretching during the warm up leads to a slight decline in the physical performance. This decrease presents about 0.92%. Another conclusion of this research is that the inclusion of static stretching to the warm up also results in a slight decrease in the athletic performance. In this case the decrease is about 1.72%. In a direct comparison of these two types of stretching, the inclusion of..

Collective investment

Tato bakalářská práce se zabývá kolektivním investováním. Cílem je čtenáři přiblížit podstatu kolektivního investování a vyhodnotit výnosnost investice do vybraných dluhopisových podílových fondů. Jsou zde popsány výhody a nevýhody kolektivního investování, nejčastější subjekty kolektivního investování, základní dělení podílových fondů a vhled do investování v rámci dluhopisů. V poslední části bakalářské práce jsou představeny vybrané dluhopisové otevřené podílové fondy. Tato kapitola také obsahuje názorné ukázky modelací výnosností investic v těchto vybraných podílových fondech a zhodnocení jejich vývoje za poslední roky.This bachelor thesis deals with collective investment. The aim is to introduce the reader to the nature of collective investment and to evaluate the return on investment in selected bond mutual funds. The advantages and disadvantages of collective investment, the most common collective investment vehicles, the basic divisions of mutual funds and insights into bond investing are described. In the last part of the bachelor thesis, selected bond open-end mutual funds are introduced. This chapter also includes illustrative examples of modelling the investment returns of these selected mutual funds and an assessment of their evolution over the past years