SANTRAUKA I IR II TIPO CUKRINIO DIABETO LABORATORINIŲ RODIKLIŲ ANALIZĖ KLAIPĖDOS UNIVERSITETINĖJE LIGONINĖJE Epidemiologiniai tyrimai rodo, kad sergančiųjų cukriniu diabetu (CD) daugėja. Todėl ypatingas dėmesys skiriamas tyrimams, skirtiems tobulinti CD diagnozavimą ir veiksmingą ligos kontrolę. Darbo tikslas: išanalizuoti I ir II tipo CD sergančių asmenų laboratorinių rodiklių ypatumus. Tikslui pasiekti buvo išsikelti tokie uždaviniai: nustatyti ir įvertinti I ir II tipo CD priklausomybę nuo paciento amžiaus ir lyties, įvertinti gliukozės, glikozilinto hemoglobino, kreatinino, C-peptido koncentracijas sergančiųjų I ir II tipo CD tiriamųjų grupėse bei laboratorinių rodiklių reikšmę diagnozuojant I ir II tipo CD. Tyrimo medžiaga ir metodai: buvo ištirti 452 pacientai, sergantys I ir II tipo CD, hospitalizuoti Klaipėdos universitetinėje ligoninėje. Gliukozės koncentracija nustatyta fermentiniu amperometriniu metodu, glikozilinto hemoglobino (HBA1c) tyrimas atliktas chromatografijos metodu, C-peptido tyrimas imunometriniu chemiliuminescencijos metodu, kreatinino tyrimas imunofermentiniu metodu. Rezultatai: I tipo CD sergančių pacientų buvo 22,8 proc., o sergančių II tipo CD — 77,2 proc. Statistiškai reikšmingi teigiami ryšiai tarp cukrinio diabeto ir sergančiųjų amžiaus parodė, kad I tipo CD dažniau serga jaunesni, o II tipo CD — vyresni asmenys. Nustatėme statistiškai reikšmingą ryšį tarp CD ir pacientų lyties: I tipo CD statistiškai reikšmingai dažnesnis vyrams, o II tipo CD —... [toliau žr. visą tekstą] / SUMMARY THE ANALYSIS OF LABORATORY INDICES OF TYPE I AND TYPE II DIABETES MELLITUS AT KLAIPEDA UNIVERSITY HOSPITAL Epidemiological studies suggest that number of patients with diabetes mellitus (DM) is increasing. Therefore, special attention is given to studies necessary to improve the diagnosis of DM and effective control of the disease. The aim: to analyse the peculiarities of laboratory indices of patients with type 1 and 2 DM. In order to reach this, the following goals were presented: to determine and evaluate type 1 and 2 DM’s dependency on patient’s age and sex; to assess the concentrations of glucose, glycosylated heamoglobin, creatinine, C-peptide in patient groups with type 1 and 2 DM and importance of laboratory indices for diagnosis of type 1 and 2 DM. Material and methods: 452 patients with I and II type MD were examined and hospitalized at Klaipeda University Hospital. Glucose concentration was determined by amperometric method, glycosylated heamoglobin (HbA1c) test was conducted by chromatographic method, C-peptide test was conducted by chemiluminescent immunometric method, creatinine test was conducted by immunoassay method. The results: 22.8 % of patients have type 1 DM, and 77.2 % - type 2. According to the statistically significant positive relationship, type 1 is more common among younger, and type 2 DM is more common among older patients. The statistically significant positive relationship between DM and the patient’s sex was determined: statistically... [to full text]

Savitvarkės biomolekulės, gebančios formuoti beta klosčių struktūras, gali būti pritaikomos nanomedžiagų su naujomis elektrinėmis, optinėmis, katalitinėmis ir/ar mechaninėmis savybėmis, kūrimui. Šiame darbe buvo siekiama kurti nanodarinius, grįstus amiloidines fibriles formuojančiais baltymais (Abeta40 peptidas, a-sinukleinas (a-Syn), kumelės pieno lizocimas (EL)), kurie būtų pagrindas nano dydžio funkcinių sistemų gamybai. Šiam tikslui buvo sukonstruoti mutantiniai ir hibridiniai baltymai bei tiriamos jų fibrilinių struktūrų savybės. Sukurti Abeta40 ir a-Syn cisteino mutantai, kurie gali būti modifikuojami per cisteino tiolinę grupę. Pirmą kartą buvo pademonstruotas a-Syncys141 baltymo fibrilių modifikavimas biotinu ir aukso nanodalelėmis su neutravidinu. Sukonstruoti hibridiniai baltymai, kurie sudaryti iš Abeta40 ar a-Syn bei GDH, streptavidino ir hidrofobino. Buvo tikimasi, kad tokie baltymai formuos fibrilines struktūras, o funkciniai baltymai bus aktyvūs. Esant atitinkamoms sąlygoms, šie baltymai agregavo suformuodami skirtingos morfologijos beta klostines struktūras. Streptavidino ir Abeta40 hibridinis baltymas formavo fibrilinę struktūrą – tinklą, o streptavidinas buvo aktyvus. Šiame darbe pirmą kartą aprašoma rekombinantinio EL gamyba E. coli ląstelėse. Aktyvus EL gali formuoti panašias į natyvaus EL fibrilines struktūras. Sukonstruoti nauji hibridiniai ir mutantiniai baltymai, gebantys formuoti amiloidines fibriles, yra geras pagrindas, kuriant funkcionalizuotus... [toliau žr. visą tekstą] / Self-assembly of biomolecules into beta-sheet structures can be applied in the creation of nano-materials with novel electrical, optical, catalytical, or/and mechanical characteristics. This work was directed towards the construction of nano-derivatives based on amyloid fibrils forming proteins (Abeta40 peptide, a-Synuclein (a-Syn), equine lysozyme (EL)). Such nanostructures can be used to produce nanoscale functional systems. Herein, different mutant and hybrid proteins, which were able to form fibrillar structures, were constructed and the properties of fibrils were investigated. Designed cysteine mutants of Abeta40 and a-Syn can be modified through thiol group of cysteine. Herein, for the first time, it was demonstrated that a-Syncys141 fibrils could be modified with biotin and gold nanoparticles with neutravidin molecules. Hybrid proteins of Abeta40 or a-Syn and other non-amyloid proteins were designed on purpose to obtain fibrils with active functional non-amyloid proteins. Under appropriate conditions, these proteins aggregated into beta-sheet structures. Hybrid protein of streptavidin and Abeta40 formed a net-like fibrillar structure, and streptavidin was active. For the first time it was described the production of recombinant EL in E. coli. Moreover, active EL can form fibrils, which are similar to the fibrils formed by native EL. Constructed novel hybrids and mutants that are able to form amyloid fibrils, can be applied for the creation of functionalized nanodevices... [to full text]

Tripeptidyl-peptidase II (TPP II) is a subtilisin-like serine peptidase that forms a gigantic homooligomeric complex, and is involved in the degradation of peptides in the cytosol. In addition, TPP II has been implicated in specific cellular processes, such as apoptosis control and adipogenesis, but if this is dependent on its endo- or exopeptidase activity remains to be determined. This work is devoted to the structure and function of TPP II, and to finding connections between the two. Evolutionarily conserved regions of TPP II have been identified, and sequence signatures have been constructed as an aid in identification of TPP II homologues. The conserved regions highlight amino acid residues of potential importance to structure, function or both. In addition, the first TPP II homologue in a prokaryote has been documented, which was likely the result of a horizontal gene transfer. Substrate binding for the exopeptidase activity of TPP II has been studied through mutagenesis of Glu-331, which revealed a molecular ruler mechanism that positions substrates for cleavage at the third peptide bond from the N-terminus. Thus, the well-known tripeptidyl-releasing property of TPP II could be explained. The exopeptidase activity was also probed by pH dependence studies, which revealed that a substrate with a smaller residue in the P1 position could bind non-productively to the active site. Furthermore, a difference in the pH dependence of KM between TPP II from Drosophila and homologues from mammals indicated a difference in the configuration of the binding pockets between these species. The endopeptidase activity of TPP II has also been investigated, and was found to differ from the exopeptidase activity. The endopeptidase activity appeared to be promiscuous and the preference for basic amino acid residues in the P1 position reported earlier could not be substantiated. In conclusion, many structural and mechanistic features have been observed in this work. This might be of value to future drug discovery efforts towards TPP II, and in elucidating the physiological role of this gigantic enzyme. / Felaktigt tryckt som Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 721

Tyrimo tikslas – įvertinti mokymo įtaka pacientų gyvenimo kokybei, funkciniam pajėgumui bei klinikinei būklei, sergant lėtiniu širdies nepakankamumu. Tyrimo uždaviniai. Parengti pacientams mokymo apie ŠN programą. Įvertinti mokymo įtaką pacientų, sergančių lėtiniu ŠN, gyvenimo kokybės, funkcinio pajėgumo, klinikinės būklės rodiklio - B-tipo natriuretinio peptido koncentracijos dinamikai. Tyrimo populiacija. Vilniaus universiteto Santariškių klinikų Ambulatorinės kardiologijos skyriuje konsultuojami pacientai, sergantys lėtiniu širdies nepakankamumu, stabilios būklės, priklausantys II-III NYHA funkcinei klasei, optimalaus medikamentinio gydymo fone. Tyrimo metodai: Prieš mokymą ir 2 mėn. po mokymo buvo vertinami: 1. Gyvenimo kokybė (GK) pagal standartizuotą Minesotos “Gyvenimas su ŠN” klausimyną (bendras GKVI, fizinį, emocinį aspektą atspindintis GKVI). . 2. Funkcinis pajėgumas atliekant spiroergometrinį tyrimą (VO2piko ml/kg/min, VO2piko l/min, AT VO2 ml/kg/min, AT VO2 ml/kg/min, AT VO2 l/min ir procentinės šių parametrų reikšmės, apkrova (W), krūvio trukmė (min), MET, NYHA, AT ŠSD, ŠSD piko, VE/VCO2, VE/VO2. Krūvio intensyvumas išreikštas MET, kurie panaudoti NYHA funkcinės klasės objektyvizavimui. 3. Klinikinė būklė pagal B-tipo natriuretinio peptido (BNP) koncentraciją kraujyje. Kiekybiniams kintamiesiems aprašomoji statistika pateikiama vidurkių – standartinių nuokrypių pavidalu. Kokybiniams - pateikiami dažniai. Lyginant rezultatus prieš ir po mokymo kiekybinių kintamųjų... [toliau žr. visą tekstą] / Aim of the work: to evaluate influence of education on quality of life, exercise capacity and clinical status in patients suffering from chronic heart failure. Tasks of the work: to create educational heart failure programme for the patients. To evaluate the influence of education on changes of quality of life in patients suffering from chronic heart failure. To evaluate the influence of education on changes of B-type natriuretic peptide levels (functional status) in patients suffering from chronic heart failure. Study population: Patients suffering from chronic heart failure in stabile condition (functional NYHA class II – III), consulted at Vilnius University Santariškių Clinic Out – patient Department of Cardiology who had been receiving optimal treatment. Methods: The following data were evaluated before tuition and 2 months thereafter: 1. Quality of life (QoL) in accordance with standardized Minnesota questionnaire “Life With Heart Failure” (generali [QoLEI], physical, reflecting emotional QoLEI aspects). 2. Exercise capacity by means of spiroergometry: VO2peak ml/kg/min, VO2peak l/min, AT VO2 ml/kg/min, AT VO2 ml/kg/min, AT VO2 l/min and percentage value of this parameters workload (W), exercise duration in minutes, MET, NYHA, heart rate during anaerobic threshold, peak heart rate, VE/CO2, VE/VO2. Workload intensity was expressed in MET in order to determine NYHA functional class more objectively. 3. Clinical condition in accordance with B-type natriuretic peptide blood... [to full text]

PDI ("potato cathepsin D inhibitor ") and NID ("novel inhibitor of cathepsin D ") from potato (Solanum tuberosum) belong to the protein family of Kunitz inhibitors (I3 family, Merops database). These 20 kDa isoinhibitors with the typical β-trefoil architecture inhibit aspartic and serine peptidases. In this thesis, the constructs for recombinant expression of PDI and NID in the yeast Pichia pastoris system were prepared and high-producing colonies were selected. Both proteins were identified in the cultivation media by mass spectrometry and N-terminal sequencing. A purification protocol for PDI with three chromatographic steps was designed. Analogous functional properties were demonstrated for the purified recombinant PDI and the native PDI isolated from a natural source. Analysis of the inhibitory specificity showed that PDI is a potent inhibitor of selected aspartic peptidases from the A1 family and serine peptidases from the S1 family, including a relevant enzyme of insect origin. This finding supports the hypothesis that Kunitz inhibitors are involved in plant defense against herbivorous insects. The inhibitors prepared within the project will be used for analysis of the reactive centers against target peptidases by protein crystallography. (In Czech) Key words: proteolytic enzymes, activity...