Heat shock proteins - - their role in diagnosis and prognosis of pregnancy related complications

Proteiny tepelného šoku zvyšují svou genovou expresi po vystavení buňky, respektive organismu, určité formě stresu, kterou může být vysoká teplota, infekce, zánět, hypoxie, nedostatek živin nebo vody. Stresovou situací pro organismus jsou i těhotenské komplikace související s placentární insuficiencí - preeklampsie a intrauterinní růstová retardace, ale i další těhotenské komplikace jako fetální růstová retardace a gestační hypertenze. Proto jsem také ve své diplomové práci zjišťovala, zda má výskyt těhotenských komplikací (preeklampsie, fetální růstová retardace, gestační hypertenze) vliv na genovou expresi proteinů tepelného šoku. Detekovala jsem 5 hsp systémů, kterými byly Hsp27, Hsp60, Hsp70, Hsp90 a HspBP1. Detekci jsem prováděla na vzorcích placentární tkáně a plné periferní krve matky. K dispozici jsem měla vzorky jak od žen s fyziologickým průběhem těhotenství, tak od žen, u kterých se během gravidity vyvinula určitá komplikace (PE, FGR, GH). Ze vzorků jsem izolovala RNA. Detekce hsp exprese byla provedena pomocí RT-PCR v reálném čase s využitím komparativní Ct metody. Zjišťovala jsem změny genové exprese testovaného vzorku oproti vzorku referenčnímu. K posouzení rozdílu exprese hsp mezi fyziologickými graviditami a graviditami s vybranými těhotenskými komplikacemi jsem využila analýzy...Heat shock proteins increase their gene expression after exposure of cells or organisms to some forms of stress, which may be high temperature, infection, inflammation, hypoxia, lack of nutrients and water. Stressful situations for the body are also pregnancy-related complications associated with placental insufficiency - preeclampsia and IUGR, as well as other pregnancy-related complications such as fetal growth restriction and gestational hypertension. Therefore, I examined whether the occurrence of pregnancy-related complications (preeclampsia, fetal growth retardation, gestational hypertension) affect the gene expression of heat shock proteins. Five hsp systems was detected, namely Hsp27, Hsp60, Hsp70, Hsp90 and HspBP1 in placental tissue samples and whole maternal peripheral blood. Samples came from women with physiological pregnancy and from women with certain pregnancy-related complications (PE, FGR, GH). RNA was isolated from the samples. Detection of hsp expression was performed by using real-time RT-PCR and the comparative Ct method. Changes in gene expression between the test samples and reference sample were examined. To assess the difference between physiological pregnancies and pregnancies with selected pregnancy- related complications, an analysis of variance (ANOVA) was used....Department of Anthropology and Human GeneticsKatedra antropologie a genetiky člověkaFaculty of SciencePřírodovědecká fakult

Monitoring of CAR-T lymphocytes in the course of hemato-oncological therapy

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author Bc. Lenka Dvořáková Supervisor RNDr. Ondřej Souček, Ph.D. Title of Thesis Monitoring of CAR-T lymphocytes in the course of hemato-oncological therapy The diploma thesis deals with the therapy of T cells with a chimeric antigen receptor (CAR) in the treatment of blood malignancies. Specifically, it focuses on monitoring the population of CAR-T cells in patients over time, after the administration of a therapeutic product. Measurements were made from whole blood samples by measurement on a flow cytometer. The theoretical part is devoted to hematological malignancies: their diagnosis and treatment, as well as to CAR-T cells: history, production, clinical use, side effects and determinations. The experimental part is dedicated to processing of the measured results. The goal of the thesis was to prepare a research on the given topic and evaluate the results of monitoring the population of CAR-T cells over time. The performed technique was able to demonstrate CAR-T cells in the blood of all patients, flow cytometry using a test tube from the company Exbio and using the antiFMC63 scFv antibody from the company Acro Biosystems appears to be a suitable method. The rate of expansion of...Univerzita Karlova v Praze, Farmaceutická fakulta v Hradci Králové Katedra biologických a lékařských věd Autor Bc. Lenka Dvořáková Vedoucí RNDr. Ondřej Souček, Ph.D. Název diplomové práce Monitorování populace CAR-T lymfocytů během léčby krevních malignit Diplomová práce se zabývá terapií T lymfocytů s chimérickým antigenním receptorem (CAR) u léčby krevních malignit. Konkrétně se zaměřuje na monitorování populace CAR-T lymfocytů u pacientů v čase, po podání léčebného přípravku. Měření bylo prováděno ze vzorků plné krve měřením na průtokovém cytometru. Teoretická část je věnována hematologickým malignitám: jejich diagnostice a léčbě, dále CAR-T lymfocytům: historii, výrobě, klinickému využití, nežádoucím účinkům a stanovením. Experimentální část je potom věnována zpracování naměřených výsledků. Cílem diplomové práce bylo zpracovat odbornou rešerši o daném tématu a vyhodnotit výsledky sledování populace CAR-T lymfocytů v čase. Provedenou technikou se podařilo prokázat CAR-T lymfocyty v krvi u všech pacientů, průtoková cytometrie s využitím zkumavky od firmy Exbio a použitím protilátky antiFMC63 scFv od firmy Acro Biosystems se jeví jako vhodná metoda. Míra expanze CAR-T lymfocytů byla u jednotlivých pacientů individuální. U dvou pacientek byly...Katedra biologických a lékařských vědDepartment of Biological and Medical SciencesFaculty of Pharmacy in Hradec KrálovéFarmaceutická fakulta v Hradci Králov

Analysis of the hybrid proline-rich protein families from seven plant species suggests rapid diversification of their sequences and expression patterns

<p>Abstract</p> <p>Background</p> <p>Plant hybrid proline-rich proteins (HyPRPs) are putative cell wall proteins consisting, usually, of a repetitive proline-rich (PR) N-terminal domain and a conserved eight-cysteine motif (8 CM) C-terminal domain. Understanding the evolutionary dynamics of HyPRPs might provide not only insight into their so far elusive function, but also a model for other large protein families in plants.</p> <p>Results</p> <p>We have performed a phylogenetic analysis of HyPRPs from seven plant species, including representatives of gymnosperms and both monocot and dicot angiosperms. Every species studied possesses a large family of 14–52 HyPRPs. Angiosperm HyPRPs exhibit signs of recent major diversification involving, at least in <it>Arabidopsis </it>and rice, several independent tandem gene multiplications. A distinct subfamily of relatively well-conserved C-type HyPRPs, often with long hydrophobic PR domains, has been identified. In most of gymnosperm (pine) HyPRPs, diversity appears within the C-type group while angiosperms have only a few of well-conserved C-type representatives. Atypical (glycine-rich or extremely short) N-terminal domains apparently evolved independently in multiple lineages of the HyPRP family, possibly via inversion or loss of sequences encoding proline-rich domains. Expression profiles of potato and <it>Arabidopsis HyPRP </it>genes exhibit instances of both overlapping and complementary organ distribution. The diversified non-C-type <it>HyPRP </it>genes from recently amplified chromosomal clusters in <it>Arabidopsis </it>often share their specialized expression profiles. C-type genes have broader expression patterns in both species (potato and Arabidopsis), although orthologous genes exhibit some differences.</p> <p>Conclusion</p> <p>HyPRPs represent a dynamically evolving protein family apparently unique to seed plants. We suggest that ancestral HyPRPs with long proline-rich domains produced the current diversity through ongoing gene duplications accompanied by shortening, modification or loss of the proline-rich domains. Most of the diversity in gymnosperms and angiosperms originates from different branches of the HyPRP family. Rapid sequence diversification is consistent with only limited requirements for structure conservation and, together with high variability of gene expression patterns, limits the interpretation of any functional study focused on a single <it>HyPRP </it>gene or a couple of <it>HYPRP </it>genes in single plant species.</p

Comparison of the carbohydrate content in apples and carrots grown in organic and integrated farming systems

The aim of this study was to compare some quality parameters of apples and carrots from organic and integrated farming systems. In addition, the cultivars of carrots were grown in two plant densities (600 or 900 thousand plants per hectare). The fructose, glucose, saccharose and dry matter content of seven apple cultivars (Florina, Zvonkové, Topaz, Šampion, Ontario, Melrose and Idared) and two carrot cultivars (Afalon F1 and Cortina F1) were analysed by HPLC and gravimetric methods, respectively. Significant differences were found between organic and integrated apple samples. The interactions between cultivars and farming methods were also significant (p &lt;0.0001). The dry matter and sugar level tendencies were not the same for all apple cultivars. Conversely, more consistent data were obtained for the two carrot cultivars. The bio carrots of both cultivars showed significantly lower dry matter content (p = 0.0004) and higher carbohydrate content (pfructose = 0.0303, pglucose = 0.0003, psucrose = 0.0083) than the samples from integrated production. Other factors like cultivar and plant density also played an important role in sugar content in carrots. Different densities of plants significantly affected the glucose content (p = 0.0373). Cultivar Aftalon F1 showed higher concentration of monosaccharides compared to Cortina F1 (pfructose = 0.0001 and pglucose &lt;0.0001)

The birth prevalence of lysosomal storage disorders in the Czech Republic: comparison with data in different populations

The aim of this retrospective study was to determine the prevalence of lysosomal storage disorders (LSDs) in the Czech Republic. The data on cases diagnosed between 1975 and 2008 were collected and analyzed. The overall prevalence of LSDs in the Czech population (12.25 per 100,000) is comparable to that reported for the countries with well-established and advanced diagnostics of LSDs such as the Netherlands (14 per 100,000), Australia (12.9 per 100,000) and Italy (12.1 per 100,000). Relatively higher prevalence of LSDs was reported in the north of Portugal (25 per 100,000). Thirty-four different LSDs were diagnosed in a total of 478 individuals. Gaucher disease was the most frequent LSD with a birth prevalence of 1.13 per 100,000 births. The most frequent LSD groups were lipidoses, mucopolysaccharidoses, and neuronal ceroid lipofuscinoses, with combined prevalences of 5.0, 3.72, and 2.29 per 100,000 live births, respectively. Glycoproteinoses (0.57 per 100,000 live births), glycogenosis type II (0.37), and mucolipidoses (0.31) rarely occur in the Czech population, and a range of other LSDs have not been detected at all over the past three decades. Knowledge of the birth prevalence and carrier frequency of particular disorders is important in genetic counselling for calculation of the risk for the disorder in the other members of affected families. Earlier diagnosis of these disorders will permit timely intervention and may also result in lowering of the number of newborns with LSDs