This diploma thesis deals with the evaluation of clinical manifestations in infants with allergic colitis and their overall nutritional status, anthropometry, blood count and antibodies against cow's milk proteins. Allergic colitis is one of the most common manifestations of infant allergy to cow`s milk protein with gastrointestinal involvement. The theoretical part summarizes the anthropometric evaluation of growth and development, infant nutrition, the most important laboratory indicators of nutritional status. The part is generally devoted to food allergies, mainly allergy to cow's milk protein. The practical part deals with the evaluation of a sample of examined patients in gastroenterology outpatient clinics and nutritional outpatient clinic of the Department of Pediatrics and Adolescent Medicine of the 1st Faculty of Medicine, Charles University and General Hospital in Prague. The observed group of infants with allergic colitis includes infants exclusively breastfed, breastfed by a mother following a non-dairy diet, infants on an extensive hydrolyzate or on an amino acid formula. The main goal of the diploma thesis was to trace the most common clinical manifestations and whether there are any changes in their nutritional status during the disease and its treatment. A total of 45 randomly...

Title: Cerebral projection of haptic contact via sLORETA imaging Objectives: The aim of this work is to evaluate changes in intracerebral source activity via sLORETA imaging during haptic stimulation of hands, while this contact is modified by a mirror illusion in comparison to calm state with open eyes. The work seeks to specify localization of such activity. Methods: Ten healthy volunteers aged 23-42 participated in the experiment. The electrical brain activity was detected with scalp EEG. The experiment was divided in 5 phases. First we measured the brain activity during calm state with open and with closed eyes, each for 5 minutes. Afterwards the subjects were seated at a table with a mirror occluding their right hand and reflecting their left hand. The brain activity was than recorded during 4 modifications of the experiment in duration of 2 minutes each. The first modification contained symmetrical haptic contact on both hands, modification 2 involved stimulation on the left hand only, during modification 3 the stimulus was applied on the right hand only and modification 4 had no tactile stimulus on neither side. The order of modifications for each individual was randomized. The EEG data were converted into sLORETA program, which allows to localize the source of the recorded brain activity...

IGFBP-3 expressing rekombinant vaccinia viruses used for tumor therapy Insulin-like growth factor-binding protein-3 (IGFBP-3) is a major regulator of endocrine effects of IGF and is capable to suppress the growth of variety of cancer. Several studies have shown that IGFBP-3 can induce the apoptosis of cancer cells via IGF-dependent and IGF-independent mechanisms. In our study, we have constructed recombinant vaccinia viruses (VACV) expressing IGFBP-3 under the control of the early H5 and synthetic early/late (E/L) promoter to investigate the potential effect on cancer growth in our cervical cancer model. We have shown that the expression of IGFBP-3 alone had no effect on tumor growth. On the other hand, the co-expression of IGFBP-3 enhanced the anti-cancer effect of immunization with the fusion protein SigE7LAMP, which gave rise to the anti-cancer immunity directed against HPV16 induced tumors. We have shown that the double-recombinant P13-SigE7LAMP-H5-IGFBP-3 can enhance the protective immune responses against MK16/ABC induced tumors. Furthermore, we have show that both double-recombinant viruses P13-SigE7LAMP-H5- IGFBP-3 and P13-SigE7LAMP-E/L-IGFBP-3 can increase the anti-cancer effect of SigE7LAMP expression in the therapy of TC-1 induced tumors. Key words: IGFBP-3, IGF, VACV, HPV16, E7 oncoprotein,...

Obesity negatively affects quality of life of children and its high prevalence is a global problem. Obesity is a multifactorial disease, whose formation influence genetic and exogenous factors. Exogenous factors can act directly on obese individuals and can be defined by the markers of personal history, or may act within the family and then they are define by the markers of family history. The health risks associated with obesity are high and therefore you need to focus on the treatment of obesity in childhood. In childhood it is still possible to change unhealthy habits and lifestyle. The aim of this work is to describe the relationship of severity of obesity of obese children in the selected data from their personal and family history and analyze the success of reducing treatment of children in dependence selected aspects of their personal and family history. Monitored set includes 398 girls and 216 boys aged 6 to 18 years who were examined anthropometric at the beginning and in the end of the stay in the hospital, Dr. Philip in Poděbrady, and whose parents responded to the survey questions for parents of obese children. Data where colected between years 2000 and 2010. The following parameters were monitored: body weight, body height, BMI, skinfolds thickness, circumferential and width parameters...

3 Selectively substituted cyclodextrins for analytical and pharmaceutical applications Abstract This thesis is focused on the selective modification of cyclodextrins, and its primary aim is the preparation and characterization of mono- and persubstituted derivatives of cyclodextrins in a regioselective and straightforward manner. The work is divided into two main parts describing synthetic strategies and applications of modified cyclodextrins with one or several substituents, respectively. The first section deals with the introduction of a single chromophoric moiety on the cyclodextrin scaffold such as cinnamyl, rhodaminyl, fluoresceinyl and eosinyl groups. The complete set of monocinnamyl-α-cyclodextrin regioisomers has been prepared by direct alkylation, and the self-assembling properties of the corresponding regioisomers were thoroughly investigated by dynamic light scattering and NMR experiments. These investigations revealed that the different isomers (mono-6-O-, mono-2-O- and mono-3-O- cinnamyl-α-cyclodextrin) form distinct supramolecular species through intermolecular association. A fast method for the unambiguous identification of the pure regioisomers has also been developed based on a series of 2D NMR measurements. Xanthene-modified β-cyclodextrins, other representatives of monosubstituted...

Optimization of biologic therapy in children with inflammatory bowel disease (IBD) using modern biomarkers Abstract to thesis Study programme: Biochemistry and Pathobiochemistry Introduction: In adults, infliximab (IFX) levels correlate with disease activity and antibodies to IFX (ATIs) predict treatment failure. We aimed to determine the association of IFX levels and ATIs with disease activity in paediatric population. Methods: This study was performed as a prospective observational study. We prospectively collected blood, stool, and clinical data from 65 patients (age 10.5-15.1 years) with Crohn's disease (CD) before IFX administration, and measured IFX trough levels, ATIs, and faecal calprotectin levels (CPT). We used multivariate analysis to identify the predictors of IFX levels. IFX and ATIs levels were meassured using ELISA. Results: Lower levels of IFX were associated with ATIs positivity (OR [odds ratio] 0.027, CI [confidence interval] 0.009-0.077). Higher C-reactive protein (CRP) level, erythrocyte sedimentation rate (ESR) and CPT levels were found in patients with lower IFX levels. The optimal combination of specificity (50%) and sensitivity (74%) for disease activity was calculated for IFX levels ≥ 1.1 µg/ml using CRP level < 5 mg/l as a marker of laboratory remission. In a model that used CPT ≤...

Introduction: The inflammation plays the essential role in the bone loss in juvenile idiopathic arthritis (JIA). Proinflammatory cytokines and also glucocorticoids (GCs) may activate bone resorption by osteoclasts. Simultaneously, bone formation can be attenuated, especially by inhibitors of proteins, which control the osteoblast differentiation. The aim was to verify the hypothesis that in patients with highly active JIA, reduction of bone formation via Wingless (Wnt) proteins inhibitors - Dickkopf 1 (Dkk-1) and sclerostin could be found. Except the densitometry measurements of bone and lean mass, we assessed markers of disease activity, bone metabolism and remodeling in young adult patients with JIA before and during 2 years of anti TNFα (tumour necrosis factor α) treatment, which decreases disease activity. Results: In patients with JIA before antiTNFα treatment, bone mineral density (BMD, g/cmš) was significantly reduced compared to controls. Values of BMD and body composition in JIA significantly depended on disease duration and GCs treatment. Serum concentration of sclerostin was significantly elevated in JIA compared to values in healthy controls. Values of the other monitored markers did not differ between JIA and controls. In patients with JIA, Dkk-1 correlated positively with C-reactive...

The aim of the thesis was development of an innovative treatment of bone defects. Human multipotent mesenchymal stromal cells (MSC) play a crucial role in bone healing. Clinical applications of MSC require large amount of cells, which could be obtained by autologous expansion of MSC harvested from bone marrow. As a first step, the standard protocol of MSC expansion based on αMEM medium and fetal bovine serum (FBS) was used. Experiments replacing FBS by pooled human serum (HS) in the culture medium concluded in patenting of a new MSC cultivation protocol (EU 1999250, CR 301141). This one-step cultivation protocol and xenogeneic protein-free cultivation medium is based on CellGro® for Hematopoietic Cells' Medium, HS, human recombinant growth factors, dexamethasone, insulin and ascorbic acid. The preclinical in vitro and in vivo experiments with MSC from both expansion protocols were carried out. Fibrillar polylactic scaffolds were seeded with MSC, cultured, differentiated and implanted in immunodeficient mice (NOD/LtSz-Rag1-). Bone-like mineralized tissue containing vessels was observed. The MSC cultured according to patented method were classified as Advanced-therapy Medicinal Product and has to fulfil the European Medicines Agency regulations to enter the clinical trials. Nevertheless the use of MSC seems...

This diploma thesis deals with laboratory functional diagnostics of malabsorption syndromes. Especially the functional diagnosis of lactose intolerance. The golden standard for the functional diagnosis of lactose intolerance is the breath test. The aim of the study was to determine the prevalence of lactose intolerance (LI) using a lactose breath test when consuming a dose of 20 g of lactose in patients with gastrointestinal symptoms and in volunteers who do not show gastrointestinal problems. Part of the research was also to examine the symptoms that led the individual to see a doctor and undergo a lactose breath test. The method consisted in the evaluation of the results of breath tests, which were performed on patients and volunteers in the gastroenterological laboratory of the 1st Medical Faculty of Charles University and the General Hospital. A questionnaire was given to everyone to evaluate the symptoms of LI and the phenomena leading to malabsorption. Lactose intolerance was confirmed in 92 % of patients and 79 % of volunteers. The most reported symptoms in the patient sample were typical gastrointestinal LI-related problems: gastric and intestinal problems after milk consumption (85 % of patients), gastric and intestinal problems after consumption of dairy products (85 % of patients),...

Title: Modern technologies used in physiotherapy at children with cerebral palsy Objectives: The goal of this diploma thesis is evaluate the effect of robotic and virtual therapy on motor functions of lower limbs at children with cerebral palsy. Furthermore, the work is focused on summarizing current knowledge about cerebral palsy, robotic and virtual technologies. Methods: The diploma thesis is processed as a literature review from available literary resources. First part of thesis contains all theoretical bases and knowledge's about cerebral palsy, robotic and virtual technologies use in physiotherapy lower limbs at children with this diagnosis. Second part has a descriptive - analytical character and contains analysis of studies, which was used treatment with robotic and virtual technologies. The thesis is concluded with a discussion, which focused mainly on three basic research questions of the diploma thesis. Results: A total of 8 randomized controlled trials that met the criteria for inclusion in this thesis were described. When making recommendations for using robotics and virtual technologies, is necessary to take into account the type of cerebral palsy, the degree of disability according to GMFCS, age, motor deficit and individual abilities and goals of the therapy. Studies show that the...