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Evolutionary aspects of Alzheimer's disease. Major role of the Apolipoprotein E
Škubica, Patrik ; Daňková, Pavlína (advisor) ; Jirák, Roman (referee)
Alzheimer's disease is one of the most discussed conditions, due to its huge social and economic consequences. Whereas molecular and genetic aspects causing early onset of the disease are relatively well known, it still remains to be clearly shown how genetic risks and environmental factors interact to ultimately cause the late onset form. Major molecular-genetic factors affecting risk of developing Alzheimer's disease are APOE gene and its product apolipoprotein E. This gene occurs in humans in three common variants that differ among each other in exon sequence by one single nucleotide polymorphism. Similar difference exists between human and mammal APOE gene that served as an origin for human forms. This thesis discusses possible evolutionary scenarios of APOE gene and links acquired information to molecular and environmental ascpects of Alzheimer's disease.
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Examination of the polymorphism of the gene ApoE
NOVOTNÁ, Tereza
Alzheimer´s disease (AD) belongs to the most frequent cause of dementia in the world. It is determined that it is about 65-75 % of all the dementias (Preiss, Přikryková Kučerová, 2006). The term Alzheimer Dementia, as well as Pre-senile Form (AD) is used with the patients at the age up to 65 years (Seidl, 2015). After the age of 65 years we use the term Senile Dementia of the Alzheimer Type. (SDAT). Up to the age of 45 this desease is rare (Seidl, Obenberger, 2004). Causal treatment of this disease does not exist, because etiopathogenesis of the disease is not entirely clarified (Seidl, 2015). The aim of this bachelor´s work was to write a professional search to the given topic. In this theoretical part there is summed up the information about neurodegenerative disease, about dementia and (AD). Most importantly there are described the symptoms, the causes, phase, treatment, diagnostics and genetic disposition of AD. The description of polymerase chain reaction (PCR) and reverse hybridization on strips is also an important part of this theoretical part. In the practical part, the main goal was to master molecular biological methods. To achieve the goals and to get the best results, I chose two methods. In addition to optimizing the PCR ARMS method, reverse hybridization was used on strips. The above-mentioned second method was performed using the commercially available ApoE StripAssay? kit from ViennaLab. In this part of the paper are described all the procedures by which I achieved the results. Molecular biological methods were performed in a laboratory at the Health and Social Faculty of the University of South Bohemia in České Budějovice.
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Significance of detection of the ApoE gene and MTHFR gene polymorphisms for osteoporosis.
ŠILHOVÁ, Markéta
Osteoporosis is the sceletal metabolic disease characterized by changes of bone tissue and disfunction of mechanic bone resistance which can lead to numerous bone fractures. Osteoporosis is frequently diagnosed in patients older than 60 years, also advanced age patients are not an exception for this disease. The most frequently affected group of patients is postmenopausal women. Prevalence of osteoporosis still rises. Nowadays more than 200 millions of people suffer from osteoporosis and one million of patients from Czech republic. Osteoporosis have became a serious complication affecting not only medical but also an economic field. Based on these findings, osteoporosis should be considered as an important topic even from the perspective of genetic risk factors. The aim of my bachelor thesis is to write a specialized research focused on introduced topic, summarize accessible information about osteoporosis, including diagnosis and genetic predisposition. I have focused especially on genes ApoE and MTHFR677. ApoE plays an important role in sceletal homeostasis. The highest risk for osteoporosis development is the presence of alel 4 gene for alipoprotein E. Gen MTHFR677 is mostly associated with bone mineral density located in femoral neck. An experimental part of bachelor thesis was based on laboratory practice in molecular biology field inluding isolation of DNA from peripheral blood, PCR technique, detection of PCR products by gel agarose electrophoresis, analysis of ApoE gel by MyTaq Red DNA Polymerase kit and 2x PCRBIO HS Taq Mix, analysis of MTHFR677 gene by MyTaq Red DNA Polymerase kit. Analysis of ApoE gene is based on ARMS PCR method, on the other hand analysis of MTHFR677 is based on RFLP PCR method. I have performed the laboratory practice within the Laboratory of genetics GENLABS in České Budějovice.
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Determination of spontaneus abortions - the role of Apo E gene polymorphism, importance of selected congenital thrombophilias and thyroid function during the pregnancy
Kašparová, Dita ; Fait, Tomáš (advisor) ; Procházka, Martin (referee) ; Límanová, Zdeňka (referee)
Introduction: Spontaneous abortion (SA) is the most common complication in pregnancy. The aim of the study was to investigate the causality of selected genetic factors - Apolipoprotein E (Apo E) gene polymorphisms, factor V Leiden (FVL), Prothrombin (PT G20210A) and nongenetics factors - Thyroid stimulating hormone (TSH), free thyroxine (fT4), antibodies against thyroid peroxidase (a-TPO) in the role of early SA. Materials and methods: For genotyping of APO E polymorphism was used PCR-RFLP. The detection of mutations in genes FV and FII was performed using by HRM. Laboratory markers of thyroid (TSH, a-TPO and fT4) were determined by an automated analyzer using chemiluminescent immunoassay. Results: APOE genotypes of investigated group of 410 samples abortioned embryonic/ fetal tissues were not significantly different from 2 606 adult controls (P = 0.653). In observed infertile group of 75 women with isolated SA was FVL detected in heterozygous constitution with a prevalence of 12 %. The prevalence of FVL in a group of women with early insulated SA was significantly higher than 76 controls (12 % vs. 2.6 %, P = 0.031). The difference of PTG20210A prevalence between women with isolated SA and controls was not significant (4 % vs. 5.3 %, P = 1). The prevalence of elevated TSH levels (higher than 2.5...
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Evolutionary aspects of Alzheimer's disease. Major role of the Apolipoprotein E
Škubica, Patrik ; Daňková, Pavlína (advisor) ; Jirák, Roman (referee)
Alzheimer's disease is one of the most discussed conditions, due to its huge social and economic consequences. Whereas molecular and genetic aspects causing early onset of the disease are relatively well known, it still remains to be clearly shown how genetic risks and environmental factors interact to ultimately cause the late onset form. Major molecular-genetic factors affecting risk of developing Alzheimer's disease are APOE gene and its product apolipoprotein E. This gene occurs in humans in three common variants that differ among each other in exon sequence by one single nucleotide polymorphism. Similar difference exists between human and mammal APOE gene that served as an origin for human forms. This thesis discusses possible evolutionary scenarios of APOE gene and links acquired information to molecular and environmental ascpects of Alzheimer's disease.
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The importance of predisposition examination of late Alzheimer´s disease using methods of molecular biology
KOPAČKOVÁ, Michaela
Alzheimer's disease is the most common type of dementia. It affects up to 70% of cases from approximately 38 million people worldwide suffering from some type of dementia. Alzheimer's disease usually affects patients over 65 years of age, however, there are exceptions. It is presumed that the number of people with Alzheimer's disease will grow constantly in the future too. Its etiopathogenesis is not yet fully understood, that is why it is important to investigate dementia further, whether in terms of specific environmental and genetic risk factors, or in view of the economic burden for the society. The presented bachelor's thesis summarises findings about Alzheimer's disease, its neurobiology, etiology, diagnostics and genetic predispositions, in particular the ApoE, APP, PSEN1 and PSEN2 genes. The presence of epsilon 4 gene allele for apolipoprotein E provably contributes to the increased risk of the late form of Alzheimer's disease. On the contrary, mutation in APP, PSEN1 and PSEN2 genes are connected with Alzheimer's disease with the early onset of the disease, before 65 years of age. The objective in the experimental part was to manage the practical isolation of DNA from peripheral blood and buccal swab, preparation and execution of PCR reaction, detection of PCR products by gel electrophoresis and independent examination of ApoE gene by a certified CVD StripAssay Kit from ViennaLab company (PentaGen). The work also includes the optimisation of another molecular genetic method for the detection of variant alleles of ApoE gene. This method was carried out by a certified DiaPlexQTM ApoE Kit from SolGent Co., Ltd. The principle of detection in this case is based on real-time PCR.
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