Glycogen storage disease type II (GSDII) is an autosomal recessive lysosomal storage disorder caused by acid alpha-1,4-glucosidase (GAA) deficiency. This study aimed to provide an in-depth description of a late-onset GSDII (LO-GSDII) cohort (n=36) and assess potential genotype-phenotype correlation. We performed a clinical record-based study, some patients (n= 19) were also followed prospectively. Phenotypes were highly variable. We focused our clinical assessment onrespiratory failure, as it is the most frequent cause of death in LO-GSDII. In addition to standard spirometric measures, in a subgroup of patients (n = 10) we utilized a new tool, optoelectronic plethysmography (OEP), to investigate the pathophysiology of respiratory muscle impairment.The GAA gene was sequenced in every patient, and pathogenic mutations were identified inall of them. Almost all (35/36) patients carried the same mutation on one allele, IVS1-32-13T>G, which was in compound heterozygosity with a variety of other GAA mutations. To investigate genotype-phenotype correlation, we divided the patient cohort in two groups, according to the severity of the mutation on the second allele. The respiratory function study focused on diaphragmatic weakness. According to the change in forced vital capacity in supine position (ΔFVC), we defined patients with ΔFVC>25% ashaving diaphragmatic weakness (DW) and those with ΔFVC<25% as without diaphragmatic weakness (noDW). We measured pulmonary function and chest wall volumes using OEP inboth groups. We found a good correlation between the supine abdominal contribution to tidal volume (%VAB) and ΔFVC. Patients showed reduced chest wall and abdominal inspiratory capacity and low abdominal expiratory reserve volume. In terms of genotype-phenotype correlation, we counted more subjects in the group with severe second mutations (n=21) who had severe motor disability and respiratory dysfunction. However, this finding remains preliminary because differences were not significant, likely because of small sample size. Finally, in two smaller substudies, we investigated the occurrence of urinary and fecal incontinence in LO-GSDII, and reported a possibly non-fortuitous association of LO-GSDII and hydromyelia in two individuals. Overall, this work 1) provided new insight into genotype-phenotype correlation in GSDII, suggesting that it is of complex nature; 2) refined the analysis of respiratory muscle impairment and showed the utility of OEP for respiratory assessment in this neuromuscular disorder, and possibly in others as well; 3) indicated some so far little studied phenotypic features of LO-GSD-II that deserve further investigation. / Doctorat en Sciences médicales (Médecine) / info:eu-repo/semantics/nonPublished

Staigios kūdikio mirties sindromo diagnostika ir etiopatogenezė taikant šiuolaikinius metodus išlieka ribota ir net iki 0,5% mirties priežasčių nepavyksta nustatyti, atsakoma ne į visus klausimus susijusius su sindromo išsivystymu ir diagnoze. Viena iš šiuolaikinių metodų perspektyvų yra imunohistocheminių bei molekulinių metodų taikymas diagnozės patvirtinimui arba atmetimui. Minėti tyrimai yra labai svarbūs priežastinių ryšių nustatymui ir etiopatogenezės pagrindimui. Medžiagų apykaitos sutrikimai, tokie kaip glikogenozės I tipo liga – viena iš aktualiausių ir dažniausiai pasitaikančių, tačiau mažiausiai ištyrinėtų ligų. Darbo tikslas diagnozuoti glikogenozės I tipo atvejį taikant naujausias imunohistocheminių bei molekulinių tyrimų metodologijas. Tyrime naudojama naujausia parafine įlietų audinių technologija, audiniai įvertinti histologiškai, atliktos imunohistocheminės reakcijos. Iš parafine įlietų audinių išskirta DNR, kokybė įvertinta atlikus spektrofotometrinį tyrimą ir elektroforezę gelyje. Išskyrus DNR iš širdies ir kepenų buvo atlikta polimerazės grandininė reakcija naudojant tikslinius glikogenozės I tipo pradmenis. Gauti rezultatai parodė glikogeno kiekio padidėjimą kepenyse. Histologiniuose preparatuose matoma kasos, širdies miokardo, antinksčių hipertrofijos bei hiperplazijos. Imunohistocheminio tyrimo metu pastebėtas glikogeno kaupimasis kasos ląstelėse. Naudojantis gliukozės-6-fosfatazės žymenimis PGR metodu glikogenozės atvejo patvirtinti nepavyko. Iškeltos... [toliau žr. visą tekstą] / Diagnosis and etiopathogenesis of Sudden infant death syndrome are limited even nowadays, and up to 0.5 % of death the cause remains undiagnosed, not all the questions, related to the syndrome and diagnosis are answered. One of the applicable methods and perspectives for diagnosis rejection or approval are immunohistochemistry analysis and molecular methods. A very important determination of causal links and proof of etiopathogenesis are very important and possible only after the already mentioned tests. Metabolic disorders, especially glycogenosis type I disease - one of the sorest and the most common, but least-studied cases. The aim of this study is to diagnose glycogenosis type I using the latest immunohistochemistry and molecular methods. The latest technology of paraffin embedded tissue was used in the study, tissues were evaluated histologically, immunohistochemical reaction was carried-out. DNA was extracted from paraffin embedded tissues, quality was measured by spectrophotometric analysis and gel electrophoresis. Using the heart and liver - DNA polymerase chain reaction was performed using targeted glycogenosis type I primers. The results showed an increase of glycogen in the liver. Pancreas, heart, adrenal hypertrophy and hyperplasia were detected in histological samples. During immunohistochemical study an accumulation of glycogen in pancreas cells was observed. Using glucose-6-phosphatase PCR markers of glycogenosis could not be confirmed. Hypotheses for further... [to full text]