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Preparation of food supplement containing probiotic bacteria and components with anti-inflammatory effect
Horňáková, Nikola ; Němcová, Andrea (referee) ; Skoumalová, Petra (advisor)
The presented thesis discusses the issues of chronic inflammatory diseases of the digestive system and suggests the possibility of the alternative natural remedies improving the health conditions or prolonging the remission phase of IBD. The main goal is to design a probiotic supplement enriched by natural anti-inflammatory agents. Phytochemicals, concretely phenolic compounds, flavonoids, and carotenoids suppress harmful inflammatory processes by direct targeting the function of the immune cells or by inhibiting damaging oxidative stress in general. Therefore, there were several plants potentially rich for these biologically active substances selected. Concretely, blueberry (Vaccinium myrtillus), lingonberry (Vaccinium vitis-idaea), turmeric (Curcuma longa), peppermint (Mentha piperita), chamomile (Matricaria recutita), cinnamon (Cinnamomum zeylanicum), ginger (Zingiber officinale) and flax (Linum usitatissimum). The suitable parts of these plants were extracted by using a total of three different solvents – water, ethanol, and hexane for obtaining lipidic extracts. The concentration of total polyphenols, total flavonoids, total carotenoids, and the ascertainment of specific polyphenols was determined in the prepared extracts. These characteristics were enhanced by the measurement of antioxidant activity of the aqueous and ethanolic extracts. The interaction of individual samples with probiotic cultures (Bifidobacterium breve, Lactobacillus acidophilus) and the ability of extracts to influence the viability of probiotics in the model digestion process has been examined. The most interesting samples (aqueous extracts of blueberries, mint, chamomile, and cinnamon) were selected for further experiments. The possible cytotoxicity towards human intestinal epithelial cells was tested in vitro by the MTT tests utilizing the CaCo-2 cell line. Extracts showing the highest levels of beneficial phytochemicals and antioxidant activity, supporting the growth of probiotic cultures, and showing minimal cytotoxic effects on human intestinal cells were then co-encapsulated with probiotics into alginate particles of a diameter of 1 mm. Water extracts of mint, chamomile, and cinnamon have been chosen. The encapsulation efficiency of successfully entrapped probiotics and phenolic compounds was determined in prepared particles. Furthermore, the particles were studied during the process of model digestion, when the release of the desired substances in the various parts of the digestive system was observed and assessed whether the components would reach the crucial point of action – the colon. For the use of the proposed probiotic mixture as a dietary supplement, a recommended dose of 1 g has been determined. Lastly, possible adjustments such as lyophilization or sheathing by an extra protective polymerous layer, e.g. chitosan, were suggested to prolong the shelf life of the particles and volatile substances stability.
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Management of multidisciplinary care in patients with inflammatory bowel disease.
KALMÁROVÁ, Diana
The diploma thesis deals with the management of multidisciplinary care for patients with inflammatory bowel diseases and is divided into a theoretical and a practical part. The theoretical part focuses on inflammatory bowel diseases, their characteristics, etiology and pathogenesis, diagnosis, treatment and psychosocial side of the disease. Another part of the theoretical part is the characteristics of management in nursing, nursing managers, the nursing profession itself, the multidisciplinary team in health care, the issue of competencies and the need for a multidisciplinary approach in the case of inflammatory bowel disease. The first aim of the thesis was to map the role of individual medical and non-medical staff involved in the care of patients with inflammatory bowel diseases, the second goal to find out how cooperation among these workers work and the third objective to identify dispensaries in patients with IBD outside hospital care. For the empirical part of the diploma thesis, a qualitative research survey was chosen, and semi-structured interviews were used as the method of data collection. Five variants of interviews were created, for nurses, doctors, nutrition therapists, psychologists and patients diagnosed with inflammatory bowel diseases. The research was conducted at the Clinical and Research Centre ISCARE in Prague and at the General University Hospital in Prague with the prior consent of the management of these facilities, except for patients who were contacted through patient groups and a psychologist who worked in his private outpatient facility. Based on interviews with nurses, doctors and a nutritional therapist, we were able to find out that as far as IBD centres are concerned, care is really interconnected and individual experts work as a multidisciplinary team led by doctors of various specialties, nurses, ostomy nurse, nutritional therapist. and a social worker. An exception is the representation of a psychologist, who unfortunately is not part of the team and there may be a problem with his / her availability, which was pointed out by some of the interviewees in the interviews. From the results of the research survey, we also managed to find out that patients have various options for better management of the disease, which are most often recommended to them by their attending physicians, but also by nurses. Nurses working in IBD centres are well versed in this issue and know the supportive options for patients with IBD. However, a problem in cooperation was found and appeared on the part of nurses, who in some cases take a passive attitude to education from an ostomy nurse, who is also an important member of a multidisciplinary team. Furthermore, based on research, we found that patients with IBD are often anxious and may have trouble trusting the nursing staff, so the main task of the nurse caring for such patients is to be more aware of these diseases, to know the supportive options, and to be able to educate clients. Respecting and understanding cooperation with other professionals are is important.
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Molekulární diagnostika fungálních patogenů z klinických vzorků
CIMICKÁ, Jana
This study is focused on optimization and use of molecular methods in mycological diagnostics. The patient's life often depends on early diagnosis, which current culture methods cannot provide. The solution is based on pan-fungal primers of ITS region and next generation sequencing (NGS). These methods were used for fungal detection from FFPE tissue samples from patients with ulcerative colitis (UC), Crohn disease (CD) and 10 controls without chronic inflammatory changes in the intestine. Results of this study have three levels. Firstly, the process of optimization highlights the importance of sterile DNA isolation in a laminar box which can prevent contamination by Malassezia spp. Secondly, raw data showed balanced rate of Ascomycota and Basidiomycota in CD samples similar to negative controls, while UC samples indicated higher representation of Ascomycota than other groups. Thirdly, detailed data showed fungal genera Malassezia, Cladosporium and Toninia in all groups, while genus Candida was found only in UC samples and genera Engyodontium and Ramularia were found in CD samples. In conclusion, mycobiome plays a role in inflammatory bowel diseases; however, its compartments are still questionable. The optimization of pan-fungal nested PCR and NGS helped with introduction of mycological diagnostics from FFPE samples to Bioptická laboratoř s.r.o. and is used for differential diagnostics.
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Preparation and characterization of human cellular cofactors of retroviral integration.
Čermáková, Kateřina ; Maloy Řezáčová, Pavlína (advisor) ; Obšil, Tomáš (referee)
Lens epithelium-derived growth factor/p75 (LEDGF/p75) is a prominent cellular binding partner of Human Immunodeficiency Virus type 1 (HIV-1) integrase. It is a human nuclear protein, which has been implicated in transcriptional regulation and cell survival. The role of LEDGF/p75 in HIV integration is well characterized, the HIV integrase binding domain (IBD) was identified and structural studies, which provide detail information about this interaction, were done. However, very little is known about its physiological function. As a transcriptional co-activator, LEDGF/p75 is implicated not only in HIV replication, but also in human cancer and autoimmunity. Key feature for both, the viral and cellular role of this protein, is its ability to act as a molecular adaptor tethering proteins to the chromatin fiber. Recently, PogZ (Pogo transposable element derived protein with zinc finger domain) was identified and validated as a new cellular interaction partner of LEDGF/p75. It was shown, that their interaction is mediated by IBD of LEDGF/p75 and the C-terminal domain of PogZ. To gain more insight in this interaction, we have initiated structural studies of their complex. Structural information is crucial for understanding the LEDGF/p75 biological role and might help in design of inhibitors selectively blocking...
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Effect of intestinal microflora on the activity of higher brain centers
Arnold, Tomáš ; Hock, Miroslav (advisor) ; Melkes, Barbora (referee)
Abstract Intestinal microflora consists of large bacterial community that resides in the intestines, especially the colon, and lives in symbiosis with the host. It consists mainly representatives of the four bacterial strains (Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria). It has already been shown that the proportional representation of the bacterial strains in time does not change much, but the research of the last decade have shown irrefutable effect between the change of diet and composition of the intestinal microflora. In addition, there is evidence of changes in the representation of bacterial strains in the course of development of certain intestinal pathologies such as inflammatory bowel disease. More and more are starting to emerge papers describing the influence of altered intestinal microflora respectively its representatives, on the activity and function of the CNS. This interaction of the digestive system and CNS seems to be bidirectional and mediated by several different pathways. Finally, it shows that dysbiosis of the intestinal microflora promotes anxiety and depression that can lead to the urge to prematurely terminate life. The aim of this work is to provide a comprehensive view of this relatively young, but also very interesting...
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Preparation of food supplement containing probiotic bacteria and components with anti-inflammatory effect
Horňáková, Nikola ; Němcová, Andrea (referee) ; Skoumalová, Petra (advisor)
The presented thesis discusses the issues of chronic inflammatory diseases of the digestive system and suggests the possibility of the alternative natural remedies improving the health conditions or prolonging the remission phase of IBD. The main goal is to design a probiotic supplement enriched by natural anti-inflammatory agents. Phytochemicals, concretely phenolic compounds, flavonoids, and carotenoids suppress harmful inflammatory processes by direct targeting the function of the immune cells or by inhibiting damaging oxidative stress in general. Therefore, there were several plants potentially rich for these biologically active substances selected. Concretely, blueberry (Vaccinium myrtillus), lingonberry (Vaccinium vitis-idaea), turmeric (Curcuma longa), peppermint (Mentha piperita), chamomile (Matricaria recutita), cinnamon (Cinnamomum zeylanicum), ginger (Zingiber officinale) and flax (Linum usitatissimum). The suitable parts of these plants were extracted by using a total of three different solvents – water, ethanol, and hexane for obtaining lipidic extracts. The concentration of total polyphenols, total flavonoids, total carotenoids, and the ascertainment of specific polyphenols was determined in the prepared extracts. These characteristics were enhanced by the measurement of antioxidant activity of the aqueous and ethanolic extracts. The interaction of individual samples with probiotic cultures (Bifidobacterium breve, Lactobacillus acidophilus) and the ability of extracts to influence the viability of probiotics in the model digestion process has been examined. The most interesting samples (aqueous extracts of blueberries, mint, chamomile, and cinnamon) were selected for further experiments. The possible cytotoxicity towards human intestinal epithelial cells was tested in vitro by the MTT tests utilizing the CaCo-2 cell line. Extracts showing the highest levels of beneficial phytochemicals and antioxidant activity, supporting the growth of probiotic cultures, and showing minimal cytotoxic effects on human intestinal cells were then co-encapsulated with probiotics into alginate particles of a diameter of 1 mm. Water extracts of mint, chamomile, and cinnamon have been chosen. The encapsulation efficiency of successfully entrapped probiotics and phenolic compounds was determined in prepared particles. Furthermore, the particles were studied during the process of model digestion, when the release of the desired substances in the various parts of the digestive system was observed and assessed whether the components would reach the crucial point of action – the colon. For the use of the proposed probiotic mixture as a dietary supplement, a recommended dose of 1 g has been determined. Lastly, possible adjustments such as lyophilization or sheathing by an extra protective polymerous layer, e.g. chitosan, were suggested to prolong the shelf life of the particles and volatile substances stability.
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Microbiota and inflammatory bowel diseases
Gajdárová, Zuzana ; Kostovčíková, Klára (advisor) ; Grobárová, Valéria (referee)
Inflammatory bowel diseases (IBD) are an autoimmune illnesses affecting gastrointestinal tract. The main types include ulcerative colitis and Crohn's disease. Recently, primary sclerosing cholangitis (PSC) has also been associated with IBD. PSC is a chronic liver disease associated with bile duct stenosis. The exact pathogenesis and etiology of these diseases is not clear, despite the great efforts of the scientific community. They are multifactorial diseases that are associated with dysbiosis of intestinal microbiota. Their diagnosis is based on for patients unpleasant endoscopic examinations and therefore the search for new serum biomarkers is needed and appreciated target of scientific interest. In the first part of diploma thesis, we focused on the reactivity of peripheral blood cells of IBD patients to 10 selected representatives of typical intestinal microbiota: Lactobacillus plantarum, Bifidobacterium adolescentis, Blautia coccoides, Roseburia intestinalis, Eubacterium rectale, Faecalibacterium prausnitzii, Ruminococcus flavefaciens, Bacteroides thetaiotaomicron, Prevotella ruminicola and Escherichia coli. Reactivity of CD, UC and PSC- IBD patients was increased after stimulation with Faecalibacterium, Lactobacillus and Prevotella. However, we got low percentage of cytokine-producing cells,...
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Effect of intestinal microflora on the activity of higher brain centers
Arnold, Tomáš ; Hock, Miroslav (advisor) ; Melkes, Barbora (referee)
Abstract Intestinal microflora consists of large bacterial community that resides in the intestines, especially the colon, and lives in symbiosis with the host. It consists mainly representatives of the four bacterial strains (Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria). It has already been shown that the proportional representation of the bacterial strains in time does not change much, but the research of the last decade have shown irrefutable effect between the change of diet and composition of the intestinal microflora. In addition, there is evidence of changes in the representation of bacterial strains in the course of development of certain intestinal pathologies such as inflammatory bowel disease. More and more are starting to emerge papers describing the influence of altered intestinal microflora respectively its representatives, on the activity and function of the CNS. This interaction of the digestive system and CNS seems to be bidirectional and mediated by several different pathways. Finally, it shows that dysbiosis of the intestinal microflora promotes anxiety and depression that can lead to the urge to prematurely terminate life. The aim of this work is to provide a comprehensive view of this relatively young, but also very interesting...
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Quality of life of patients with inflammatory bowel disease (IBD)
HAVELKOVÁ, Kristýna
Inflammatory bowel disease is often discussed topic among the medics in the field of diagnostics, treatment and prognosis, however what is usually ignored is quality of life. The bachelor thesis aims at revealing the quality of life of patients with idiopathic bowel disease. The theoretical part introduces inflammatory bowel disease with its principal types of Crohn´s disease and ulcerative colitis. The empiric part is focused on IBD patients´ quality of life. The research part of the thesis is based on standardized questionnaire Short Form 36, aimed at quality of life of patients in South Bohemia. For reaching the goal, three main hypotheses were selected: H1: Patients with inflammatory bowel disease (IBD) have mental problems. H2: Patients with inflammatory bowel disease (IBD) have physical problems. H3: Patients with inflammatory bowel disease (IBD) have social problems. The questionnaire construction was conducted in Word Excel programme using graphs and basic statistics. A positive output from the respondents was very surprising because the most of the respondents answered the questions concerning their quality of life positively. It is possible to state that this situation may be caused by the latest available treatment, timely diagnosis and also by a certain effort within our society to deal with chronically ill patients with kindness and respect. This survey convinced me that also people with chronical diseases can have good quality of life. The topic of this bachelor thesis was highly interesting for me; my original presumptions were different, therefore the results were surprisingly pleasing. Creating this bachelor thesis was very helpful for me. This bachelor thesis can be used as informative material for nurses in hospitals or for students of non-medical health care branches. Furthermore, this thesis can be used for further study.
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