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Trunk muscle activity and walkin
Svobodová, Jaroslava ; Opálková, Michaela (advisor) ; Davídek, Pavel (referee)
Bakalářská práce Aktivita trupového svalstva při chůzi Abstract This thesis refers to the anatomy and the kinesiology of trunk, spine and pelvis. In addition, it deals with the control of gait and gait biomechanics. Nevertheless, the main aim of this thesis is to summarize the information on the activity of trunk muscles during walking. This includes also trunk muscle activity during modified walking. This part reviews the changes of trunk muscle activity in reaction to low back pain and to the changes in walking speed. Thesis is focused on the activity of abdominal and back muscles such as m. rectus abdominis, m. obliguus externus abdominis, m. obliguus internus abdominis, m. transversus abdominis and m. erector trunci. The thesis also includes a case study of a patient who suffers from chronic low back pain. The activity of some of her trunk muscles was recorded by surface electromyography. In addition, the patient was tested using the zebris Rehawalk FDM-T system.
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Stress proteins in the cytoplasmic membrane fraction of Bacillus subtilis
Šemberová, Lenka ; Svobodová, Jaroslava (advisor) ; Krásný, Libor (referee)
Stress proteins in the cytoplasmic membrane fraction of Bacillus subtilis A primary habitat of the Gram-positive bacterium Bacillus subtilis is the upper layer of the soil. Within this ecosystem, B. subtilis experiences a wide variety of environmental challenges and nutrient limitations that induce several mechanisms to help the cells to survive. A process known as a general stress response belongs among them. Expression of about 150 genes is enhanced and their products called general stress proteins (GSP) minimize the cell damage. Although the size and structure, as well as the regulation, of many stress proteins have been fairly well elucidated the information about stress membrane proteins is very limited. We characterized the membrane proteome of Bacillus subtilis 168 trp2- exposed to acidic pH and ethanol. Cells were 1) grown under optimum conditions in complex medium (pH 7.0) at 40řC with aeration; 2) grown in complex medium at pH 5.0 for 20 hours or 3) challenged with 3% v/v ethanol for 30 minutes during exponential growth. The cultures were harvested in the mid-exponential phase by rapid filtration. Isolated membrane fractions were analysed by an optimized two-dimensional gel electrophoresis. Two alternative methods of protein detection were compared: silver staining and radioactive labeling with...
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Mechanism of surfactin self-resistance in the Bacillus subtilis cytoplasmatic membrane
Seydlová, Gabriela ; Svobodová, Jaroslava (advisor) ; Spížek, Jaroslav (referee) ; Julák, Alois (referee)
Surfactin, a lipopeptide surfactant and antibiotic produced by Bacillus subtilis, exhibits a strong membrane perturbation. One of the drawbacks hindering its commercial applications is the unknown mechanism of surfactin self-resistance in the producer. Therefore, the aim of this study was to examine the self-protective mechanisms of the cytoplasmic membrane of B. subtilis against the deleterious effect of surfactin. In order to reach this task, two isogenic pairs of strains differing only in surfactin production were constructed. It was found out that the early response of the producer is realized by increasing the amount of total lipid content in the membrane already with the onset of surfactin synthesis. This process leads to lowering the surfactin-lipid ratio in the membrane. In parallel with the growing surfactin concentration the content of anionic phospholipids with cardiolipin as the major representative rises up to 24 % of the total. Together with the fall of phosphatidylethanolamine these changes promote the membrane stabilization and protect it against the interaction with surfactin. These alterations result in higher rigidity both of the polar head and hydrophobic chain region of the membrane as the steady state anisotropy of DPH and TMA-DPH showed. After 24 h of cultivation induction of...
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Cytoplasmic membrane adaptation to surfactin in Bacillus subtilis non-producing strain
Stružinská, Olga ; Svobodová, Jaroslava (advisor) ; Fišer, Radovan (referee)
Cytoplasmic membrane adaptation to surfactin in Bacillus subtilis 168 non-producing strain Surfactin, the most potent surface active compound and antibiotic is produced by bacteria of the genus Bacillus. Surfactin interacts with membrane bilayers, that results in destabilization and permeabilization of this structure. However mechanism of surfactin self-resistance in the producer's membrane is not understood. The aim of this study was to characterize the adaptive processes occurring at the level of cytoplasmic membrane of surfactin non-producing strain B. subtilis 168, which was exposed to exogenously added surfactin during the exponential phase of growth. The cultivation protocol of B. subtilis growth on agar media plates supplemented with surfactin was developed. Two surfactin concentrations that inhibit (400 g/ml) and even stimulate (300 g/ml) the growth of B. subtilis 168 strain were assessed. Surfactin brought about the growth arrest for 3 hours and the restored growth rate decreased in the case of inhibitory concentration, whereas the stimulatory concentration increased the growth rate and resulted in higher final density of the population. TLC was performed to analyze the polar head groups of membrane phospholipids. The portion of phosphatidylserine was found to increase at both surfactin...
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Cold adaptation in stationary phase in Bacillus subtilis
Beranová, Anna ; Konopásek, Ivo (advisor) ; Svobodová, Jaroslava (referee)
Cold adaptation in stationary phase in Bacillus subtilis One of the most important abiotic factor which influences life of bacterial cells is the ambient temperature. A decrease of this temperature is usually accompanied usually with the loss of the fluidity of bacterial cytoplasmatic membrane. While the mechanisms of the responses to the cold shock during the exponential phase of growth are well known for Bacillus subtilis, the responses of stationary phase cells had not been studied yet (despite the stationary phase is the most common state of microorganism in the nature). There are two independent mechanisms which restores much needed fluidity in Bacillus subtilis - short-term adaptation and long-term adaptation. Short-term adaptation is based on the function of fatty acid desaturase coded by des gene. Long-term adaptation relies on the change in ratio of iso- and anteiso- branched fatty acids. In this work we examinated membrane adaptation during stationary phase under two different conditions, namely under cultivation at stable low temperature and after cold shock. The highest activity of Pdes was observed for cultivation at 25 řC and for the cold shock applied from cultivation in 37 řC to 25 řC. Anisotropy measurements and fatty acids analysis were also performed. Results indicated, that the...
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Study of phosphorylation of inorganic pyrophosphatase from Streptococcus pneumoniae
Štechová, Michaela ; Svobodová, Jaroslava (referee) ; Doubravová, Linda (advisor)
The human patogen Streptococcus pneumoniae encodes a single copy of eukaryotic-like Ser/Thr protein kinase StkP. StkP regulates virulence, competence, stress resistence, gene expression and plays a role in the regulation of cell division cycle. Analysis of phosphoproteome maps of the wild type and stkP mutant strain of S. pneumoniae showed that in vivo StkP phosphorylates several putative substrates including Mn-dependent inorganic pyrophosphatase PpaC. Mass spectrometry analysis identified two phosphorylation sites in an active site of the protein. Pyrophosphatases are essential enzymes that catalyze hydrolysis of inorganic pyrophosphate produced during various biosynthetic reactions that utilize ATP. Changes in pyrophosphatase activity have been described to have global effects on cell metabolism, growth and division of bacteria. The aim of this thesis was to investigate the phosphorylation of inorganic pyrophosphatase PpaC in S. pneumoniae. Gene ppaC was cloned, expressed in E. coli and protein was purified via affinity chromatography. Phosphorylation of PpaC by StkP was examined in a kinase assay but we did not confirm that PpaC is a direct substrate of StkP in vitro. Further we prepared a set of mutants in ppaC gene. We replaced two presumable phosphoaminoacids identified by mass-spectrometry with...
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