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Hemoglobin-mediated oxidation of marine liposomes
Škrabalová, Lada ; Mozuraityte, Revilija (referee) ; Rustad, Turid (advisor)
Cílem této práce bylo studium mechanismu oxidace lipidů katalyzované hovězím methemoglobinem a zhodnocení účinků různých experimentálních podmínek a antioxidantů (EDTA, askorbová kyselina, kávová kyselina, a-tokoferol, d-tokoferol, astaxanthin a L-askorbyl-6-palmitát) na methemoglobinem zprostředkovanou oxidaci lipidů v modelovém systému liposomů připravených z fosfolipidů. K monitorování oxidace lipidů při pH 5,5 a teplotě 30 °C bylo použito spotřeby kyslíku. Pro zhodnocení antioxidační aktivity v modelovém systému liposomů se ukázaly být důležitými faktory typ prooxidantu a koncentrace prooxidantu a antioxidantu. Dalšími důležitými faktory jsou struktura molekuly antioxidantu, jeho hydrofilita/lipofilita a umístění v systému. Všechny testované antioxidanty ve všech koncentracích (kromě koncentrace 0.1 % astaxanthinu and 0.1 % askorbyl palmitátu) inhibovaly oxidaci vyvolanou methemoglobinem. Účinnost antioxidantu stoupala s jeho zvyšující se koncentrací. Koncentrace 0.1 % astaxanthinu neměla žádný vliv na oxidaci liposomů. Koncentrace 0.1 % askorbyl palmitátu měla prooxidační efekt, který lze vysvětlit prooxidačním působením radikálu askorbylu, který může urychlit štěpení hydroperoxidů. Volné železo uvolněné z methemoglobinu se podílelo jen velmi málo na oxidaci liposomů, zatímco část prooxidační aktivity methemoglobinu byla přisouzena tvorbě singletového kyslíku (methemoglobin jako fotosenzitizátor). Antioxidační aktivita astaxanthinu, askorbyl palmitátu a tokoferolu byla z části přisouzena schopnosti zhášet singletový kyslík. Ovšem hlavním prooxidačním mechanismem methemoglobinu se ukázal být rozklad lipidových hydroperoxidů, tvorba volných radikálů a hypervalentních forem hemoglobinu. EDTA utlumila oxidaci liposomů díky chelataci přechodných kovů obsažených v liposomech a chelataci volného železa přítomného v methemoglobinovém roztoku. Velmi důležitým antioxidačním mechanismem (který vykazují askorbyl palmitát, askorbová a kávová kyselina) se ukázala být redukce hypervalentních forem hemoglobinu. Askorbová kyselina, kávová kyselina, tokoferoly a astaxanthin inhibovaly methemoglobinem zprostředkovanou oxidaci lipidů odstraňováním volných radikálů. Při použití peroxidu vodíku nebyl pozorován žádný vliv na oxidaci liposomů vyvolanou methemoglobinem. Působení vysoké teploty (tepelná denaturace) mírně utlumilo oxidaci. Významná inhibice oxidace byla pozorována u liposomů obsahujících TPP (triphenylphosphin), což značí, že je methemoglobinem vyvolaná oxidace liposomů závislá na přítomnosti již vzniklých lipidových peroxidů. Výsledky této práce přispívají k hlubšímu pochopení prooxidačních a antioxidačních mechanismů a faktorů, které ovlivňují oxidaci liposomálních roztoků, buněčných membrán a emulzí typu olej ve vodě stabilizovaných fosfolipidy.
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Electrochemical biosensors with spatially separated enzymatic and detection parts for selective analysis in flow-through arrangement
Tvorynska, Sofiia ; Barek, Jiří (advisor) ; Labuda, Ján (referee) ; Korecká, Lucie (referee)
This dissertation thesis presents the newly developed four highly reusable, stable as well as simple, and cost-effective electrochemical (bi)enzymatic biosensors for the selective and reliable determination of choline, acetylcholine, uric acid, and L-lactic acid in flow injection analysis. All biosensors are based on the concept of the spatial separation of the biorecognition part from detection one and amperometric monitoring of the enzymatically consumed oxygen via its four-electron reduction at the highly negative detection potential. In this way, the design of the biosensors includes an easily replaceable enzymatic mini-reactor(s) connected upstream to the flow cell that contains the appropriate silver amalgam-based transducer. The enzymatic mini-reactor based on choline oxidase, uricase, or lactate oxidase was used for choline, uric acid, or L-lactic acid biosensors, respectively. The acetylcholine bienzymatic biosensor includes the consequently connected choline oxidase- and acetylcholinesterase-based mini-reactors. The first part of this thesis focuses on the construction of two different silver amalgam-based electrodes. Specifically, this section discusses the fabrication of a silver solid amalgam electrode covered by mercury film operating in a wall-jet cell and also highlights the...
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The impact of intervention breathing programme on breathing economy during increasing load
POUL, Daniel
Breathing is essential part of our life. Breathing exercise can help acquirement of proper breathing. Thanks to this process, sports performance can be improved. The aim of our work is to determine differences in values and compare results of spiroergometric parameters from testing on a bicycle ergometer in the laboratory of functional load diagnostics. We obtained the results by an entrance test and a final test tested after eight weeks. Between testing, selected individuals performed a respiratory intervention program. Nine performance university athletes, especially focused on football, got involved in our research. Data at the rest position and load were crucial. We focused on dynamic ventilation parameters such as tidal volume, respiratory rate, minute ventilation or oxygen consumption. In the output test, we also recorded changes in values in the respiratory segments, abdominal, thoracic and subclavian areas. We verified the results from the point of view of material and statistical significances. Both of these significances were evaluated in this thesis. The materially and statistically significant differences in the results of measurement were improved in the oxygen consumption by our probands. The studied probands were able to improve in various phases of load, for example, the material significance was detected at the respiratory rate during the load of three watts per kilogram. Ascertained results were not improved in all monitored parameters. The most significant changes were discovered in the three-watt load during the respiratory rate from 32,8 +- 7,2 to 30,9 +- 5,1 breaths per minute. Probands reduced oxygen consumption by 14.8 percent.
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Physiological responses on standardized climbing task in sport climbers
Gajdošík, Jan ; Baláš, Jiří (advisor) ; Radvanský, Jiří (referee)
Title: Physiological responses on standardized climbing task in sport climbers Purpose: To determine the effect of height, wall angle, climbing speed and climbing ability on physiological responses in sport climbers. Methods: The study was divided into three parts. 75 sport climbers (36 female and 39 male) completed differing tests on climbing wall and motorized climbing ergometer. Perceived exertion was assessed on a scale suggested by Borg. Indirect calorimetry, venous blood samples and near-infrared spectroscopy were used to assess physiological response, hormonal response and muscle oxygen saturation, respectively. Results: Perceived exertions were higher when climbing to height as opposes to climbing low to the ground on the treadwall (+5,3%; P = 0,013; ηp 2 = 0,149) (Study 1A). The physiological response was higher on the climbing wall as opposed to the treadwall: V̇ O2 (+6%; P = 0,03; ηp 2 = 0,22), SF (+4%; P = 0,04; ηp 2 = 0,20), V̇ E (+9%; P = 0,01; ηp 2 = 0,30) a EC (+16%; P < 0,001; ηp 2 = 0,48). There was an interaction for climbing ability and post- climbing catecholamine concentration (P < 0,01, ηp 2 = 0,28) (Study 1B). With increasing climbing speed greater differences were found for V̇ O2 (P < 0,001, ηp 2 = 0,923) than for StO2 (P < 0,001, ηp 2 = 0,448). Between-subject effect...
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Effect of climbing ability on energy expenditure of climbing
Kodejška, Jan ; Baláš, Jiří (advisor) ; Malý, Tomáš (referee)
The aim of the study was to evaluate the relation between climbing performance and energy demands while climbing through a specific submaximal test. The sample consisted of twenty-six climbers (age 26,8 ± 3,3 years, weight 70,6 ± 6,2 kg, height 177,6 ± 6,5 cm). Climbers were deliberately chosen so that their performance covers as much scale difficulty of UIAA (Union Internationale des Associations d'Alpinisme) as possible. The performance was given by the climbers currently the highest ascent in the style of RP (Red Point). Performance of these climbers ranged between 4 to 10 degrees UIAA. Climbers were subjected to submaximal climbing test, which lasted three minutes in one slope. They moved at a constant speed of 25 movements. min-1 on climbing route, already known in advance, the slope after 3 minutes changed from 90 ř to 105 ř. VO2 (oxygen consumption), SF (heart rate) and VE (minute ventilation) significantly correlated with climbing performance of RP (90ř, VO2, r = - 0,82; SF, r = -0,66, VE, r = - 0,77; 105ř, VO2, r = - 0,84; SF, r = -0,78; VE, r = - 0,80 ). Respiratory rate during submaximal climb was about 25 breaths. min-1, which refers to a link with a climbing speed. VO2 during submaximal climbing can be used to evaluate the economy of movement. Keywords sport climbing, submaximal test,...
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Physiological response and physical activity during climbing wall in school age children
Kalábová, Monika ; Panáčková, Michaela (advisor) ; Strejcová, Barbora (referee)
Title of master thesis Physiological response and physical activity during climbing wall in school age children Work objectives Determinaton of specific oxygen uptake during climbing in the climbing wall in children. Methods The study involved 10 boys and 9 girls. Their climbing ability was in range 4 to 5+ degree of UIAA (Union Internationale des Associations d'Alpinisme). Maximal oxygen consumption was measured on climbing wall. The climbers climbed two routes, the first vertical profile (90ř) and second overhanging profile (110˚). The participants climbed the wall with a self-selected speed. They climbed the route twice to steady state of physiological response during climbing and to simulate real average wall (15 m). To make conditions equal, everybody had 30 s to get down and start climbing again. Results The girls achieved average of specific oxygen consumption VO2peak 37,1 ± 4,8 ml·kg- 1 ·min-1 in the vertical profile and in the overhanging profile was 40,6 ± 11,5 ml·kg- 1 ·min-1 . The boys achieved slightly higher average of specific oxygen consumption in both profile 39,1 ± 4,8 ml·kg-1 ·min-1 in the vertical profile and 42,1 ± 2,6 ml·kg-1 ·min-1 in the overhanging profile. The girls had average peak heart rate 179 ± 10 beats·min-1 in the vertical profile and boys 170 ± 14 beats·min-1 ....
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The energy performance at the growth rate in ski-mountaineering
Hepnar, Jan ; Vomáčko, Ladislav (advisor) ; Jindra, Matouš (referee)
Title The energy performance at the growth rate in ski-mountaineering Objectives Measure group skialpinists of different ages and performance of the ski-mountaineering simulator with a gradient 21ř and speed 1,4 km/h to 5,4 km/h with increasing 0,4 km/h in every minute. Determine their energy expenditure in different speeds as the speed increases and with constant slope. Methods Seven ski-mountaineerers of different levels were testing in laboratory of ski-mountaineering simulator. We calculate of maximum oxygen consumption and anaerobic threshold. Results The probands attained average energy expenditure 8,18 kJ per minute at the end of the first minutes and they attained average energy expenditure 29,32 kJ per minute at the end of the 10th minutes. The average energy expenditure of probands increased by 2,35 kJ per minute an average with a standard deviation 0,69 kJ per minute. Keywords ski-mountaineering, heart rate, oxygen consumption, ski-mountaineer trainer.
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Comparison of human physiological response in different one-rope ascending techiques
Doležal, Jan ; Michalička, Vladimír (advisor) ; Baláš, Jiří (referee)
Title: Comparison of the physiological response rate of the organism using different one-rope ascent techniques Objectives: The aim of this study was to compare the rate of the physiological response of the organism using 3 different one-rope ascent techniques during constant speed. Methods: It was an empirically based study of experimental character (quasi-experiment). The measured research group (n = 12) consisted of the military students (21 ± 1.1 years; 183 ± 4.9 cm; 80.6 ± 7.8 kg) of full-time study at the Faculty of Physical Education and Sport of Charles University (VO FTVS UK). The physiological response rate was measured with the Cortex Metamax 3b and Polar sporttester during one-rope ascending techniques ("using Prusik knot" = P", "using jümar = B", "using Garda knot = G") at a constant speed of 3 m·min-1 for 7 minutes. The rating of the perceived exertion was also recorded on the Borg RPE scale. The data were comparatively analyzed in the SPSS statistic program, furthermore the analysis of variance (ANOVA) was also employed. Results: The study demonstrated statistical differences (p ≤ 0.05) between techniques: while B is the easiest, the greatest differences were observed between technique B, P and G. The G technigue appeared to be the most difficult. The average HR was: 162 ± 9 bpm...
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